Claudia Facciotti, biologist,PhD in Science and worked at Novartis Vaccines in Siena (Italy) since 2005 in the Protein Biochemistry Unit at the Research Center. Specialized in purification of recombinant proteins from E.coli and their characterization. Analytical studies on purified proteins as SE_UPLC/HPLC and RP_HPLC for protein integrity/purity and SCK for the protein/antibodies interactions by BiacoreT200.

The Neisserial Heparin Binding Antigen (NHBA) is a protective antigen of N. meningitidis and one of the components of the recently licensed vaccine Bexsero. NHBA was demonstrated to be an important virulence factor and to be able to elicit a robust immune response in humans against meningococcal strains expressing homologous and heterologous NHBA peptides. In order to better understand the cross protective nature of NHBA we performed a screening with the purpose to obtain monoclonal antibodies (mAbs) to be used as tools in the process of characterization. Mice were immunized with different forms of recombinant NHBA peptide 2, the same peptide present in the Bexsero formulation. When tested in the SBA assay, two of the new mAbs displayed bactericidal activity and other two mAbs were bacteriostatic against meningococcal strains expressing the homologous protein. Antibody binding and antibody affinity were measured on the recombinant proteinby enzyme-linked immunosorbent assay (ELISA) and Surface Plasmon Resonance (SPR) experiments, respectively, whereas Fluorescence-activated cell sorter (FACS) analysis was used to detect the native protein on bacterial surface. Cross-bactericidal activity was assayed by testing the selected mAbs on a panel of MenB strains expressing different NHBA peptides.Finally, a plethora of epitope mapping techniques, including protein chip analysis on NHBA fragments and Hydrogen-Deuterium Exchange (HDX-MS) were adopted to precisely map the position of the recognized epitopes. By this approach we have mapped protective epitopes on NHBA and studied their possible role on NHBA cross bactericidal activity.

Heterologous vaccination based on priming with a plasmid DNA vector and boosting with an attenuated Vaccinia virus expressing Leishmania infantum LACK antigen has shown efficacy conferring protection in murine and canine models against cutaneous and visceral leishmaniasis. Until now, MVA, M65 and M101 are the only attenuated Vaccinia virus strains described as vaccine vectors against this disease. Here, we analyzed in mouse model the immune response elicited and the protection induced by NYVAC strains of Vaccinia virus expressing LACK antigen. NYVAC-LACK and a replication competent NYVAC expressing C7L host range gene (NYVAC-LACK-C7L), induced high quality CD4+ and CD8+ adaptive and effector memory T cell responses against LACK antigen. These CD8+ T cell populations also showed an excellent proliferative capacity when stimulated with LACK antigen. Differences between the NYVAC viruses and with the MVA strain were restricted to magnitude of the elicited response. After subcutaneous L. major challenge, mice vaccinated with NYVAC-LACK-C7L showed similar reduction in lesion size as those vaccinated with MVA-LACK strain. Our findings suggest that both NYVAC-LACK and NYVAC-LACK-C7L meet the criteria to be potential vaccine candidates against leishmaniasis.

Mauro Di Pilato is a Ph.D student in the Spanish National Centre for Biotechnology (CNB), Madrid, Spain. He is working in Poxvirus and Vaccine laboratory, headed by Prof. Mariano Esteban at the Cellular and Molecular Biology Department. He published two papers in reputed journals.

Vaccinia virus (VACV) promoter early-sequences had been modified to enhance antigen expression and to improve antigen-specific T cell responses; how the spacer length between the early sequence and the gene can affect its expression, and if there is a correlation between timing of antigen expression and antigen-specific CD4 T cell response had not been demonstrated. We have generated several recombinant VACV based on an attenuated Modified Vaccinia Ankara (MVA) strain that express GFP and Leishmania infantum LACK antigen under transcriptional control of promoters with different spacer lengths. The spacer length augmented the GFP expression in vitro within the first hour of infection, and correlated with an enhancement in the LACK-specific memory CD4 T cell response in mice. These results illustrate the important role of promoter spacer length in the design of future poxviruses vaccine vectors.

Rong Xu has completed her master degree of sciences in China at the age of 27 years from Peking Union Medical College and currently is carrying on her research fellowship study in Dental School of Cardiff University. Her research project is under Dr. Xiao-Qing Wei’s supervising on a project of study cytokine biology in human health and diseases.

Candia albicans is a commensal fungal microorganism, which does not normally trigger inflammatory responses by resident macrophages such as Langerhans cells in skin. An immune tolerance of skin Langerhans cells to C. albicans challenges has been suggested, however, the mechanism(s) of such tolerance has not been elucidated. IL-34 is a recently discovered cytokine, which is constitutively expressed by keratinocytes in epidermal skin. In skin, the key function of IL-34 is to maintain Langerhans cell expansion. Resident macrophages exhibit plasticity and can be transformed into inflammatory M1 macrophages for immunity and anti-inflammatory M2 macrophages for tissue repair. The aim of this study was to investigate the role of IL-34 in regulating macrophage response following C. albicans challenge. We have previously demonstrated that inflammatory M1 macrophages produce higher levels of the inflammatory cytokine, TNFα, in response to C. albicans stimulation; this is not evident with anti-inflammatory M2 macrophages.
Method: Mouse bone marrow macrophages were cultured with 10 ng/ml GM-CSF for 7 days to drive M1 macrophage maturation. Increasing concentrations of recombinant mouse IL-34 (R&D Systems) were added in the presence of GM-CSF for different periods of culture. The production of TNFα was then determined for M1 macrophages stimulated with heat killed Candida (HKC). Expression of Toll-like receptor 2 (TLR2) and C-type lectin receptor (Dectin-1), which are key pattern recognition receptors (PPRs) for β-glucan in the yeast wall, was also determined.
Results: 1. IL-34 was found to inhibit HKC induced TNFα production in M1 macrophage in a dose dependent manner. 2. Both expression of Dectin-1 and TLR2 was significant reduced in M1 macrophage following treatment with IL-34.
Conclusion: IL-34 suppressed HKC induced TNFα production in inflammatory M1 macrophage by down regulation of Dectin-1 and TLR2 expression. This could indicate that immune tolerance of Langerhans in skin might be maintained by constitutive expression of IL-34 by keratinocytes. From a clinical perspective, neutralisation of IL-34 function in skin may have therapeutic benefit in the treatment of Candida mucosal infection; it may also have value for promoting immune responses following vaccination.

D. Vanrompay is Director of the Laboratory for Immunology and Animal Biotechnology at Ghent University and Director of the National Diagnostic Reference Laboratory for C. psittaci infections in humans. She is a member PROVAXS (www.provaxs.com), the UGhent Center for Strategic Prophylaxis and Vaccine Development.
Here research involves: i) the development of new molecular diagnostic methods for identifying bacterial pathogens in both humans and animals, ii) unravelling the cellular and molecular pathogenesis of (zoonotic) bacterial infections, and iii) gaining insight in the induction of protective mucosal immunity. Our laboratory focuses on Chlamydia infections in humans (Chlamydia trachomatis, Chlamydia psittaci) and animals (Chlamydia psittaci, Chlamydia suis and Chlamydia abortus), Escherichia coli O157:H7 infections in ruminants and Vibrio spp infections in aquatic animals. In vitro and in vivo models (e.g. C. psittaci in poultry, C. trachomatis in pigs) are used to study bacterium host interactions in order to develop innovative prophylactic tools such as virulence blockers and genetic vaccines.

Recombinant major outer membrane protein (MOMP) of a highly virulent Chlamydia psittaci genotype D strain (92/1293) was tested for its ability to induce protective immunity against challenge with the homologous C. psittaci genotype. The vaccine was prepared in transiently transfected COS-7 cells. Specific pathogen free chickens (Lohman, Germany) were divided into 2 groups of five animals, reared in negative pressure isolators. Group 1 received 500 µg recombinant MOMP per animal, administered by aerosol at the age of 7 days. Group 2 served as non-immunized control. All chickens were challenged by aerosol infection at the age of 4 weeks. The challenge infection consisted of 106 TCID50 of C. psittaci strain 92/1293. Animals were euthanized at the age of 42 days. Severe clinical signs, macroscopic and histopathological lesions were only observed in control group 2. Animals of group 1 showed a significant protective immune response compared to group 2. Details will be presented. The use of recombinant MOMP as a means of preventing severe clinical signs and lesions in a chicken model of C. psittaci infection was demonstrated.

Infection with Mycobacterium avium subspecies paratuberculosis (MAP) causes Johne’s disease (JD), anincurable enteritis in ruminants. Enzyme-linked immmunosorbent assays (ELISA) of milk and blood samples are routinely used to screen herds for exposure to MAP. Such serological investigations, however, have been shown in a single controlled study to be impacted by administration of avian and bovine purified protein derivative (aPPD and bPPD). PPD is administered to cattle during the single intradermal cervical comparative test (SICCT) for the purposes of diagnosing bovine tuberculosis (bTB). A compulsory bTB eradication scheme has been operating in Ireland since 1962. All bovines over six weeks of age are injected intradermally with aPPD and bPPD at least once annually. The aim of the current study was to examine the impact of SICCT on MAP ELISA results in an Irish dairy herd.Blood and milk samples were taken from 139 cows pre- and post-administration of the routine annual SICCT. All samples were tested using the ID Screen Paratuberculosis Indirect Screening Test (ID Vet, France) which detects anti-MAP IgG.Prior to SICCT, 6% and 8% of the herd tested serologically positive using milk and blood ELISA, respectively. Within 14 days of PPD administration a significant increase in the ELISA S/P% ratios and prevalence of positive cows was recorded. This increase peaked at a prevalence of 39% for both sample matrices and declined subsequently. S/P% ratios remained significantly higher until day 43 post-SICCT in milk (P=0.850), and day 71 in blood (P=0.602).These results may indicate of a period of reduced ELISA specificity post-SICCT with antibodies generated in response to PPD cross-reacting with the capture antigen. It should be noted that Ireland records relatively few clinical cases of JD annually, and a relatively low prevalence of MAP ELISA positive herds compared to prevalences reported internationally. It is our hypothesis that the intensive bTB testing regime implemented in Ireland may act as an annual mycobacterial antigenic stimulant (similar to a vaccine) providing immune cross-protection against MAP. Thorough investigation of this hypothesis is required as antibodies are generally regarded as ineffective in controlling MAP, but their increase post-SICCT may indicate the presence of a wider immune response that may be protective against MAP.

Christine Rueckert studied Biophysics at the Humboldt University, Berlin, Germany, and the Imperial Colllege, London, UK. In 2000, she graduated in Experimental Biophysics at the Humboldt University, Berlin, and specialized in Molecular Cell Biology over her postdoctoral period at the University of Virginia, Charlottesville, USA.Since 2011Christine Rueckerthas beena project leader in the Vaccinology Department with Carlos Guzmánat the Helmholtz Centre for Infection Research in Braunschweig, Germany. Her research focuses on elucidating the molecular mechanisms of vaccine adjuvants at the cellular level. Her objective is to understand the contributions of specific properties of adjuvant molecules to the finetuning of immune responses as the basis for rational vaccine design.

The recently discovered metazoan enzyme cyclic GMP-AMP synthase (cGAS) produces cyclic GMP-AMP (cGAMP) upon detection of pathogen-derived double-stranded DNA. The cyclic di-nucleotide (CDN) cGAMP was shown to bind to STING (stimulator of interferon genes) thereby activating an IFN-β producing pathway. Protozoan CDNs such as c-di-AMP or c-di-GMP are known to have immune modulatory activity when used as vaccine adjuvants in mouse immunization experiments. C-di-GMP was also shown to bind to and activate STING. Here we report the activity of cGAMP as a mucosal adjuvant in mouse immunization experiments. We demonstrate its potential to promote antigenspecific humoral and cellular immune responses in vivo. We further show that cGAMP can directly activate murine as well as human innate immune cells in vitro. Taken together our findings suggest cGAMP as a candidate adjuvant for mucosal vaccine development with potential also for human vaccines.

Filippo de Nicolellis graduated at Rome University “La Sapienza” in 1986, where he specialized in infectious diseases in 1990. From 1995 to 2000 he worked as a family doctor in Doberdò del Lago/ Doberdob (Gorizia) and since 2000 in Fiumicello, Udine, in Friuli Venezia Giulia, in the north-east of Italy. Since 2000 he has been the president of the medical association “Croce Medica”, which deals with the organization of Health Services and Permanent Training and particularly with Primary Health Care and Emergency.

The poster is an early report on a cultural project, focused on developing e-communication and other tools in a rural area of Friuli Venezia Giulia, Italy. The Medical Association “CROCE MEDICA” is making all efforts in Primary Health Care to increase the rate of people vaccinated against influenza. The Association is promoting the use of posters in family doctor’s offices as well as of e-mails to communicate the patients dates and procedures for the vaccination against influenza. Furthermore the Association e-mails its registered doctors a dispatch about influenza as well as a request for information about the illness seasonal evolution in the various areas of Friuli Venezia Giulia. Unfortunately the feedback has been scarce up to now. It is difficult to state if the better information provided about free vaccination for some categories of patients and its importance as well as about the dates of vaccination sessions is increasing the rate of people involved in the vaccination campaign. As a matter of fact there has been a decrease in the percentage of vaccinated people in the investigated area in the last year, a decrease due to several factors. Nevertheless I believe all efforts aiming at improving the communication with patients and explaining how to obtain the vaccination against influenza are useful in any case.

Miroslav Toman is the professor of Veterinary microbiology and immunology at the University of Veterinary and Pharmaceutical Sciences Brno, Czech Republic. He has been working at the Veterinary Research Institute Brno, Czech republic as the senior researcher at the dept. of Immunology; in 2001 – 2012 he acted also as the director of the institute. His research interest is host pathogen interaction and possibility of nonspecific and specific immunoprophylaxis, especially in pig infection models. He is also interested in clinical immunology of companion animals.

In spite of many similarities with other systems of mucous associated lymphatic tissues (MALT) the immune response in respiratory tract has its own properties and is not so much recognized as the immune response in gut. In our experiment we compared immune response of piglets against PRRS virus after immunization with four different (inactivated and modified live - MLV) vaccines. We also monitored the changes in leukocyte population and lymphocyte subpopulation in blood and lavages after immunization and infection. The first specific antibodies were detected in the serum 7 days after the second dose of inactivated vaccines. Antibodies were also detected in bronchoalveolar lavages but not in saliva. Vaccination with inactivated vaccine with oil adjuvant cause strong cellular reaction in vivo, therefore it was difficult to estimate the specific cell mediated immune response in vitro. The virus in piglets vaccinated with killed vaccine was detected in all samples including faeces 3 days after infection and the shedding was massive at the end of the experiment (21 days post infection). In piglets vaccinated with MLV, the first specific antibodies were detected 14 days after the administration of vaccine and were detected both in serum and saliva and in lavages. The cell mediated immunity tested in vitro with blood lymphocytes appeared 7 days after immunization but a strong response was detected only after experimental infection. The antigen specific interferon production was also detected in leukocytes derived from bronchoalveolar lavages. The virus in piglets vaccinated with MLV was detected in blood and saliva 3 days, and in faeces 14 days after immunization. The virus was also detected after infection but the shedding decreased with time and the virus was not detected in some of the animals 14 – 21 days post infection. The results of the project LO1218 were obtained with a financial support from the MEYS of the CR under the NPU I program and the project QJ1210120 of the Ministry of Agriculture.

Background: Nanoparticles (NPs) have been demonstrated to function as carriers to manipulate immune responses as well as specific drug delivery system. In the present study, we examined whether ovalbumin conjugated on NPs (OVA-NPs) modulate anti-tumor immune responses as well as humoral antibody responses in vivo.
Results: When C57BL/6 mice were immunized with OVA-NPs, anti-OVA IgG1 responses were potentiated with a poor IgE synthesis. Pretreatment with OVA-NPs delayed the growth of E.G7 thymic lymphoma cells expressing a model tumor antigen OVA (E.G7-OVA) that were inoculated subcutaneously. OVA-NPs encapsulating IL-7 completely blocked the growth of E.G7-OVA tumor cells, although NPs-IL-7 had a meager effect. However, the pretreatment with OVA-NPs-IL-7 failed to reduce the growth of parental thymic tumor cells, suggesting that the anti-tumor effect is antigen-specific. When tumor-free mice inoculated with OVA-NPs-IL-7 plus EG.7 cells were re-challenged with E.G7-OVA cells, they demonstrated the reduced growth compared with control mice. Moreover, vaccination with OVA-NPs-IL-7 induced the generation of cytotoxic T cells (CTLs) specific for OVA, as revealed by tetramer assay. Thus, a single subcutaneous injection of mice with OVA-NPs entrapping IL-7 induced tumor-specific cytotoxic immune response, resulting in regression of tumor cells.
Conclusion: Antigen on NPs entrapping IL-7 would be a promising carrier for developing and enhancing immune responses including humoral and cellular immunity as well as drug delivery to a specific target of interest.

Yee completed his honours degree in Biotechnology at Universiti Sains Malaysia in 2002. In 2009, he was offered a prestigious scholarship from Malaysian government to pursue his doctorate degree study in the field of parasite vaccinology at University of Queensland under the supervision of A/Prof. Malcolm Jones. His research focus was on investigating the potential vaccines targeting against schistosomes. In February 2011, he was conferred an Edward Jenner award by Australian Centre for Vaccine Development through his PhD research findings. Upon obtaining a PhD, he was appointed as a Research Officer in Molecular Immunology Laboratory at QIMR Berghofer Medical Research Institute, Brisbane led by renowned immunologist, Dr. Michelle Wykes. In August 2014, he joined Institute for Research in Molecular Medicine (INFORMM) at Universiti Sains Malaysia as a lecturer, focusing on vaccinology and immunotherapeutics.

Schistosomes are parasitic blood flukes that infect approximately 200 million people worldwide. The disease known as schistosomiasis, is the second most significant tropical parasitic disease after malaria. Only one drug currently exists for schistosomiasis and there are concerns about resistance to the drug. There has been much effort to develop vaccines against schistosomiasis to produce long-term protection in endemic regions. Surface-associated proteins, and in particular, those expressed in the body wall, or tegument, have been proposed as potential vaccine targets. Annexins are thought to be of integral importance for the stability of this apical membrane system. Here, we present the first structural and immunobiochemical characterization of an annexin from S. mansoni. The crystal structure of annexin confirms the presence of the previously predicted a-helical segment in the II/III linker. Results from ultrastructural localization confirm the occurrence of annexins in the tegument of S. mansoni. A recombinant chimeric peptide (chimeric-AT) harbouring the linker region of Anx(Sm)3 and Sm-TSP-2-EC2 was constructed for its vaccine efficacy evaluation. Immunoinformatics prediction showed that Anx(Sm)3 linker and Chimeric-AT were capable to bind with 21.6% alleles and 49% alleles, respectively. Schistosome infected mice sera reacted strongly with Anx(Sm)3 linker peptide and chimeric-AT, showing that they are immunogenic. Anx(Sm)3 and chimeric-AT delivered in alum induced high IgG1/IgG2a ratio, implying a Th2 type predominant immune response. Mice vaccinated with Anx(Sm)3 showed low protective effect while chimeric-AT showed highest reductions in worm burden (20%), liver egg burden (17%) and intestinal egg burden (40.4%). The results showed that chimeric-AT triggered a high level of antibody production, suggesting this multi-antigen construct could be a potential vaccine candidate against schistosomiasis.

Zakaria Benmansour doctor in University of Medical Sciences " Oran Algeria On parasitology and mycology medical "Laboratory of The University Hospital , Oran he is a researcher in the laboratory of infectious diseases and biologically active substances in the faculty of medicine of or an Algeria . His research work focuses on molecular fungal and virology and vaccinology Ph.d. In parasitology and mycology department of medicine of or an Algeria.His research included work on mycology and parasitology infections.

Objective: The presence of molds in a hospital environment became a subject of concern both for the healthcare professionals and for the users. Indeed, in spite of the absence of indicators allowing to measure their roles in the arisen of the fungal infections, it is established that bio contamination at the hospital is a major risk for the weakened patients, also for the certain places where are practiced the care or invasive acts. Their gravity is a real problem of Public health an in first row of morbidity, mortality. The causes are often multiple: air, water, renovation work without taking precautions standards, the cases of fungal contamination and the cases of fungal contamination we declared follow in serology realized.
Materials and methods : In our series from september 2011 to March 2013, a study was undertaken to investigate the environmental fungal flora, and systhematique serology in some units of CHU The samples sent to Laboratory for Parasitology and Mycology mycological analysis. A questionnaire was conducted in which there is information for each sample patients answering war the usual protocols of treatments Antibiotics and a fever it was asked to realize takings in aimed fungal at these patients: blood test on tube dry,. The takings treated in direct examination and in culture on specific Common agreement with the doctors (Heads of the services an environmental study is realized (harnessing of the environmental fungal flora in the respective services as well as a follow-up of the serology of the patients Galactomannane and Antibody
Results: The patients in question benefited from a serology Aspergillosiselisa and from an antifungal treatment with one followed by the serology.
Conclusion: the fungal infection is always relegated to the last rows, by misunderstanding or by absence of service competent in clinics. Nevertheless must be considered in front of a therapeutic failure in antibiotics in such departments at risks.

He is PhD in occupational Toxicology since 1993. He was nominated professor of Medicine in 2002. He runs for the last 20 years the Department of Occupational Diseases of the University hospital of Reims (Champagne County). He manages the Regional Institute of Occupational Health since 1995. He belongs to the French National University College of Occupational Researches and Practionners. He is focusing his work on occupational stress, infectious diseases and long-term exposure effects on health due to low doses toxics.

Background : infection with hepatitis B virus (HBV) can be prevented by vaccination with HB surface antigen, which induces HBS specific antibodies and T cells. But the immunization status of workers and its relationship with occupational factors are not well documented. The goal is to examine the factors of risk and the immunization status against HBV among a population of workers.
Methods : an assessment of the vaccination and the immunization status against HBV was conducted among a miscellaneous population of French workers, recruited from a medical occupational center, during a cross-sectional study. A representative sample of a population of 3000 workers enrolled, was selected.
Results : the population studied included many housemaids (18%), police officers (15%), technicians (15%), administrative agents (15%), electricians (12%) and health care workers (6%). The overall vaccination coverage (against HBV) was 39%, with an average of 4.2 doses of vaccine per worker. But the immune status was known for 18% of the studied population of which 2/3 (12% of the whole population) where immunized against HBV. Elevated risk factor to be infected by HBS concerns 6% of the studied workers (half of them where immunized). Medium risk concerns 30% of the whole population (1/3 was immunized) and low risk enrolled 64% (nobody was immunized).
Conclusion : it is known that the high risk of infection among health care workers is greater than the general working population ; but most of workers with medium occupation risk exposure were not immunized. Vaccination programs against HBV should been forced for this last population.

The vast majority of pathogens invade through or cause disease at mucosal surfaces. Development of novel immunization strategies suitable for mucosal vaccines is widely desired to protect against infectious diseases. However, very few mucosal vaccines are available for human use, none of which are recombinant proteins or subunits of pathogens, owing to the lack of potent and safe mucosal adjuvants. Therefore, novel approaches to elicit mucosal immune responses are highly needed. Given the crucial role of Vitamin A metabolites, such as retinoic acid (RA) in imprinting a mucosal homing capacity on T and B cells, as well as its potential to promote the differentiation of IgA-producing plasma cells, we evaluated the capacity of RA to improve mucosal vaccinations.
We found that mice treated with RA as compared with untreated ones, showed enhanced systemic (IgG) and mucosal (IgA) tetanus toxoid (TT)-specific antibody responses after intranasal administration of TT as antigen and cholera toxin (CT) as adjuvant. Of note, in these set of experiments, we observed an appreciable TT-specific antibodies responses after intranasal priming with Ag alone in the absence of CT in RA treated as compared with untreated intranasally primed mice. Therefore, we asked whether we could amplify this response by giving a systemic boost with Ag in the presence of systemic adjuvant alum. We found that mice treated with RA and primed intranasally with Ag alone showed a higher titer of both systemic TT-specific IgG and mucosal IgA as compared with untreated mice after systemic boost with TT and alum. Furthermore, we detected a higher frequencies of TT-specific IgG and IgA secreting cells in the bone marrow of mice treated with RA after intranasal priming with Ag alone followed by systemic boost with Ag and alum as compared to untreated mice. The persistence of the antigen-specific responses was evaluated and after 8 months we found still higher IgG TT-specific antibodies in the serum and, even if at lesser extent, higher IgA TT-specific antibodies in the vaginal compartment in RA treated as compared with untreated mice.
This approach can contribute to progress beyond the state of the art in adjuvant research by achieving mucosal immunity in the absence of mucosal adjuvants or improve the effectiveness of mucosal delivered vaccine.
The research leading to these results has received funding from the European Union’s Seventh Framework Programme (FP7/2007-2013) under Grant Agreement 280873 ADITEC.

Beatriz Basso has completed her PhD from National University of Córdoba, Argentina. She was Visitant Research at Michigan State University and Universite Libre de Bruxelles. Is Proffessor at School of Medical Sciences, Natl. Univ. of Córdoba, Director of Reserarch Projects about Immunology of Chagas` disease and Vaccination, subject in which has published more than 80 papers. She has 10 awards in national and international conferences. She is Reviewer and Editorial Board member of international journals.

Chagas’ disease affects near 16 million people in Latinamerica and is being important in non endemic countries, through congenital, transfusional and transplantation transmission. In our laboratory we have developed an experimental model of vaccination with non pathogenic Trypanosoma rangeli. The aim of this work was to compare the humoral response and the efficacy of preventive and therapeutic vaccination. BALB/c mice were vaccinated with epimastigotes of T. rangeli, at different times before (BI) or after (AI) infection with Trypanosoma cruzi. The course of infection and the pattern of antibody response (IgM, IgG, IgG1, IgG2 and IgG3) were analyzed. The results showed that both BI and AI groups had a better outcome of infection than only infected mice, with significantly lower parasitemias and less mortality. The immunological response was also similar, showing both schemes significantly increase of total IgG and IgG1. The difference was that BI induces increase of IgG2 whereas AI showed elevation of IgG3 compared to control animals. In conclusion, both vaccination schedules with T. rangeli triggers a high production of antibodies reactive with T. cruzi, and modulates the course of infection. Although both strategies are protective, the pattern of isotypes was different, suggesting that they induce distinct mechanisms of response, both of them important for the early clearance of parasite. As this vaccine also protects guinea pigs and dogs, domestic reservoirs of T. cruzi, it can be an important tool as veterinary vaccine, by disruption of epidemiological chain of Chagas’ disease, in areas with active vectorial transmission.

Beatriz Basso has completed her PhD from National University of Córdoba, Argentina. She was Visitant Research at Michigan State University and Universite Libre de Bruxelles. Is Proffessor at School of Medical Sciences, Natl. Univ. of Córdoba, Director of Reserarch Projects about Immunology of Chagas` disease and Vaccination, subject in which has published more than 80 papers. She has 10 awards in national and international conferences and serving as Referee and Editorial Board member of international journals.

Trypanosoma cruzi is a real challenge to the host immune system, because required of strong humoral and cellular immune response, for removal trypomastigotes circulating forms, and amastigotes replication forms in tissues, involving many regulators and effectors components. This protozoa is responsible for Chagas disease, a major public health problem in Latinamerica We have developed a model of vaccination with Trypanosoma rangeli, a parasite closely related to Trypanosoma cruzi, but nonpathogenic to humans, which reduces the infectiousness of mice against challenge with T. cruzi. In previous work, we demonstrated that mice vaccinated with T. rangeli showed an important phagocytic activity versus only infected groups. The aim of this work was to study in peritoneal fluid different population cells and some soluble mediators (cytokines) in mice vaccinated - infected with T. cruzi. The results showed in vaccined mice, in the first hours of challenge, increase of NK, Granulocites, Dendritic cells, regulation of IL6, IFNγ, TNFα and IL10, with marked increase of IL12, with respect to only infected controls. Furthermore it was observed an increase of Li T, Li B responsible for adaptive response.
Finally the findings showed that the innate immune response play an important role in vaccinated mice, for the early elimination of the parasites, complementary with adaptative immune response, suggesting that vaccination with T. rangeli modulate the innate response, which develops some kind of immunological memory, recognizing shared antigens with T. cruzi. These results could contribute to the knowledge of new mechanisms of the innate immune response in Chagas Disease

Dr. ChetanJawale is basically a Veterinarian, currently pursuing Ph.D in Veterinary Medicine at college of Veterinary Medicine, Chonbuk National University, South Korea. He completed his B.V.Sc. and A. H. (2002-2007) from Bombay Veterinary College, Mumbai, India, and M.V.Sc in Animal biotechnology (2007-2009) from Anand Agricultural University, Anand, India. During 2009-2010 he worked as junior scientist at Xcelris Genomics Center, Ahmedabad, India. His research primarily focuses on the development of the recombinant and genetically inactivated vaccines against the Salmonella and E. coli infections in the domestic animals.

Fowl typhoid (FT), a septicemic disease of poultry, causes acute mortality and induces severe inflammation of internal organs such as liver and spleen, caecum and yolk sac which results in significant economic losses to the poultry industry worldwide. FT is caused by facultative intracellular Gram-negative bacterium,Salmonellaenterica serovarGallinarum (SG). Bacterial ghost (BG) technology has been a new and progressive approach to construct the safe and immunogenic inactivated vaccines against wide variety of infectious diseases. In order to develop a novel, safe and immunogenic fowl typhoid (FT) vaccine candidate, aSalmonella Gallinarum ghost with controlled expression of the bacteriophage PhiX174 lysis gene Ewas constructed. The formation of the Salmonella Gallinarum ghost with tunnel formation and loss of cytoplasmic contents was observed by scanning electron microscopy and transmission electron microscopy. No viable cells were detectable 24 h after the induction of gene E expression by an increase in temperature from 37 °C to 42 °C. The safety and protective efficacy of the Salmonella Gallinarum ghost vaccine was tested in chickens. None of the immunized animals showed any adverse reactions such as abnormal behavior, mortality, or signs of FT such as anorexia, depression, or diarrhea. immunization with the Salmonella Gallinarum ghosts induced significantly high systemic IgG response in all immunized groups. Among the groups, orally-vaccinated group B showed significantly higher levels of secreted IgA. A potent antigen-specific lymphocyte activation response along with significantly increased percentages of CD4+ and CD8+T lymphocytes found in all immunized groups clearly indicate the induction of cellular immune responses. Birds were subsequently challenged with a virulent Salmonella Gallinarum strain at 3 weeks post-immunization (wpi). Significant protection against the virulent challenge was observed in all immunized groups based on mortality and post-mortem lesions compared to the non-immunized control group.Overall, these findings suggest that the newly constructed Salmonella Gallinarum ghost appears to be a safe, highly immunogenic, and efficient non-living bacterial vaccine candidate that protects against FT.

Traditionally, the quality control for Japanese Encephalitis Virus JEV) vaccine has performed in vivo potency assay using animals. The Ministry of Food and Drug Safety MFDS) established alternative in vitro assay (ELISA) replacing the in vivo assay requiring animals and many times as an official quality control method of potency test for the JEV vaccine. The in vitro potency assay showed it's faster and easy to perform without pre-treatment such as a mouse immunization. Also it had better precision and reproducibility comparing to the conventional in vivo assay. The reference material is essential in order to evaluate potency test for the JEV vaccine. The 1st and 2nd national standard for in vivo potency assay of JEV vaccine had manufactured, each established in 2001 and 2007, and have been using for the manufacturer's quality control and national lot release since then. As the need of the national standard for in vitro potency assay, this study was initiated by MFDS in 2013 to manufacture and establish the 3rd national standard for in vitro and in vivo potency assay of JEV vaccine. The in vitro and in vivo potency results of the candidate material for 3rd national standard, each were measured 1.077 and 2.761. In the study hereafter, the collaborative study of the MFDS and manufacturers will be conducted to estimate of the reliable virus content with the candidate material.

Dr. Chetan V. Jawale is basically a Veterinarian, currently pursuing Ph.D in Veterinary Medicine at college of Veterinary Medicine, Chonbuk National University, South Korea. He completed his B.V.Sc. and A. H. (2002-2007) from Bombay Veterinary College, Mumbai, India, and M.V.Sc in Animal biotechnology (2007-2009) from Anand Agricultural University, Anand, India. During 2009-2010 he worked as junior scientist at Xcelris Genomics Center, Ahmedabad, India. His research primarily focuses on the development of the recombinant and genetically inactivated vaccines against the Salmonella and E. coli infections in the domestic animals.

One of the most widely studied bacterial killing factors is the integral membrane host-toxic protein E expressed by DNA phage PhiX174, which can be utilized for genetic inactivation of Gram-negative bacterium, resulting in lysis of host bacterial cells. Protein E causes lysis in growing Gram-negative cells by blocking cell wall synthesis and this blockage is effected by specific inhibition of MraY, an enzyme involved in the pathway for murein biosynthesis.The application of genetic engineering tools is necessary for proper development of tightly regulated prokaryotic expression systems with stable maintenance of genes whose expression is detrimental to the growth of the host bacteria. The use of a regulatory promoter system that tightly represses expression of gene E during normal culture growth is essential because of its lethality to the bacterial host. Such tight regulation of gene E expression at normal growth temperatures may facilitate optimal and stable growth of host bacterial cells, ultimately resulting in higher production of ghost cell mass. In order to avoid leaky expression of the bacterial host-toxic PhiX174 lysis gene E from the λpR promoter, a convergent promoter construct was made in which gene E was placed between a sense λpR promoter and an anti-sense ParaBAD promoter. In the presence of L-arabinose, leaky transcription of lysis gene E at 28 °C from the sense λpR promoter was repressed by an anti-sense RNA simultaneously expressed from the ParaBADpromoter. The stringent repression of lysis gene E in the absence of induction temperature resulted into higher concentration of bacteria in culture suspension, and consequently higher and stable production of a Salmonella Enteritidis (S. Enteritidis) ghost.

Plague is the infection â„– 1 in the world. The natural foci of plague occupy about 40 % of the territory of Kazakhstan. In the CIS, including Kazakhstan and Russia, alive plague vaccine from Yersinia pestis EV strains (APV) is used for vaccinations against plague. However, its application requires annual revaccination. Being active enoughfor primary vaccination, it is much less effective at revaccinations. We developed two approaches to increase of given vaccine efficiency.
The first is connected with increased vaccine’s immunogenicity within the process of its production. For this purpose stock culture suspension was administrated to a rabbit intravenously at stage of vaccine operational seed lot production and further extracted from this rabbit inner organs culture was used for further stages of vaccine production. Such procedure named animalization resulted not only in confident (p<0,05) increase (up to 1.8-fold) of alive cells quantity of finished balk-vaccine, but also increased immunogenetic (protective) activity: significant (p<0,01) increased survival rates of quinea pigs (2.2 –fold) and reduced ED50 up to 9.5-fold with confidence (p<0.05)
The second approach to increaseeffectivness of APVis connected with immunomodulation. In modeling studies with infected quinea pigs it is shown that co- administration of APV and an immunomodulator (Polyoxidonium with recombinant interleukin-1 β) increases protective activity of the vaccine (p<0.01) and significantly reduces mortality rates among animal models. In addition, effect of immunomodulation on antigen - specific immune response indicators was studied in the studies where rabbits were immunized with APV with and without immunomodulators. It is shown that these preparations accelerate development of specific antigenetic response to APV at both early, and effector phase. Calculation of used immunization schemes efficiency integrated rate demonstrated better result for recombinant interleukin -1 β.
Results of our studies demonstrated effectiveness of thedescribed approaches to increase APV efficiency and the need for further research of immunomodulation, including during vaccinations of humans.

Objective: To compare varicella zoster virus-specific cell-mediated immunity and humoral immunogenicity against the herpes zoster vaccine and placebo.
Methods: A double-blind, placebo-controlled, randomized trial of herpes zoster vaccine effects in elderly people with diabetes mellitus was conducted during May 2012 – November 2013 at Kitano Hospital, Osaka, Japan. People aged 60–75 years with diabetes with 6–9.5% HbA1c levels were eligible for enrollment; immunocompromised individuals were excluded. Participants received either the herpes zoster vaccine ( Live Varicella Vaccine; BIKEN, The research foundation for Microbial Diseases of Osaka University, Japan) or placebo (0.5-mL dose) subcutaneously. They simultaneously received one dose of 0.5 ml of the 23-valent pneumococcal polysaccharide vaccine subcutaneously to promote participation. A varicella skin test, intererferon-gamma enzyme-linked immunospot assay (ELISPOT), and immunoadherence hemagglutination (IAHA) test were performed before and 3 months after vaccination. ELISPOT counts are shown as spot-forming cell (SFC) per 106 PBMC. Vaccine safety was assessed using questionnaires for 42 days along with data of development of zoster obtained during the one year observational period.
Results: Participants were 29 (56%) male and 23 female patients with respective mean ages of 65.8 and 66.8 years. Mean skin test score differences from before to 3 months after immunization in placebo and vaccine group were, respectively, 0.1+-0.2 S.E. and 0.4 +-0.2 S.E. (no significant difference). Ratios of SFC after 3 months to before vaccination in placebo and vaccine group were, respectively, 1.0 ±0.0 and 1.1 ±0.0 The mean antibody titer in the IAHA increased approximately two-fold after vaccination in each group (no significant difference). No one developed herpes zoster during the one-year observational period. No systemic adverse reaction was found.
Conclusion: The herpes zoster vaccine boosted virus-specific, cell-mediated, and humoral immunity of elderly people with diabetes as well as placebo. The herpes zoster vaccine was used safely.

Barbara Kalenik has completed her B.Sc. and M.Sc. at University of Warsaw. During studies she worked in the International Institute of Molecular and Cell Biology, Warsaw, Poland and made 3.5 month Erasmus practice at University of London, School of Pharmacy, London, GB (now University College London). She is a Ph.D. student in IBB PAS group headed by Prof. Agnieszka Sirko. She published one paper in peer-reviewed journal (as Barbara Broniatowska). She was awarded with several scholarships for academic performance by University of Warsaw and Capital City Warsaw and for oral presentation on the 7th Warsaw International Medical Congress (05.2011).

In-depth characterization of antibodies raised by vaccination is connected with cloning and selecting best binders by hybridoma or display technology. One of gold standards in this approach is antibody phage display technology which can be started from mRNA of immunized animals/people or from hybridoma. Naturally, the abilities of cloned immunoglobulin recombinant fragments (like scFv) to bind antigen of interest, need to be confirmed. Many protocols recommend ELISA with antibody-displaying-phages preparation for such assessment (here reported as standard Phage ELISA), however a quicker solution, named RISE detection, has been also proposed [1]. The cDNA sequence encoding scFv fragment from monoclonal anti-H5 HA antibody (mAb) was cloned into phagemid pSEX81 vector and analysed by Phage ELISA and RISE. In all methods, phages exposing scFv on gpIII capsid protein were incubated with BSA or one of the following H5 HA: Vietnam (A/Vietnam/1203/2004(H5N1)); Qinghai (A/Bar headed goose/Qinghai/12/05 (H5N1)) or self-made (HA_17-340) containing H1 region of HA from A/swan/Poland/305-135V08/2006 (H5N1). RISE detection method gave the best results. The mean±SD value of OD450 for HA_17-340 was 3.34±0.01, for Qinghai it was 4.02±0.03, while the background OD450 (for BSA) was 0.31±0.04. Different concentrations of phage preparations and two time periods of incubation of phages with protein (2h and 18 h) were tested, confirming that both, amount of phages exposing scFv and the time of incubation have impact on experiment results. This poster describes Pros and Cons of the used methods. [1] Vanhercke et al. 2005, J Biomol Screen. 10: 108

Jimmy Chandia graduated as a Medical Doctor at Makerere University in Uganda in 1979.He specialised in Family Medicine at the Medical University of South Africa in 1990.He is currently the Principal Investigator for Walter Sisulu University HIV Vaccine research Unit and an Associate Professor of Family Medicine in South Africa. His interest in HIV dates back to 1985 when he was involved in establishing the prevalence of HIV in South Africa. He has published in reputed journals and presented papers at national and international conferences. He is on the Editorial board of the South African Family Practice Journal.

South Africa with about 6.4 million people living with HIV infection to date has the highest number of people living with HIV in the world. Various strategies have been adopted by the country in response to the pandemic. The search for an effective, relevant and affordable vaccine against HIV is one. Indeed South Africa is at the forefront of HIV Vaccine research. It has produced the first candidate HIV vaccine in the third world. Various institutions are involved in HIV Vaccine research in the country. The latest HIV Vaccine research unit to be established is Walter Sisulu HIV Vaccine research Unit, located in the city of Mthatha in the Eastern Cape Province . It is the only research unit of its kind in the Province. The research unit was inaugurated by the Medical Research Council in 2006 . While the idea of an HIV Vaccine research unit was well received by the stakeholders i.e. Walter Sisulu University, the Department of Health and the local community, acquiring the necessary resources was a huge challenge. It was only when The Italian Institute of Health through a program of corporation between the governments of Italy and South Africa that the dream of establishing the research unit was accomplished in 2011.The research Unit has a Laboratory, pharmacy, offices, consulting rooms, data storage room housed within the Clinical Research Unit, the relevant staff and a well sensitised population. It is open to the world for HIV Vaccine and related research.

AthinaKilpeläinen is a master student at the Biomedicine programme at Karolinska Institute. She completed her bachelor’s degree in biomedical laboratory science in 2012 at the age of 21, where she performed her thesis on gene immunogen optimizations of HIV-1 Reverse Transcriptase.

By a combination of existing databases, we predicted new HLA class I binding epitopes important for HIV immune responses and discuss mutations to preserve immunogenicity of the Nef protein, modified not to confer HLA or CD4+ down-regulating activities. By Nef epitope-to-allele binding predictions, we identified previously not described epitopes for the common African HLA alleles HLA-A*02:01, A*30:01, A*30:02, B*58:01 and C*07:01 and compared them to already reported epitopes from the Los Alamos database. The smallNef gene/protein may contribute effectively to both stronger and broader cellular immunogenicity of an HIV-1 vaccine.

Pinghua Li completed her Ph.D of preventive veterinary science at Gansu Agricultural University. she is research assistant of National Foot and Mouth Disease Reference Laboratory of Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences and is now working for the development of novel and effective marker vaccine of FMDV of type A, O and Asia1 using FMDV infectious cDNA clone.

Foot and mouth disease (FMD) is a highly contagious and economically devastating disease of domestic and wild cloven-hoofed animals. The disease is distributed worldwide and has great negative economic impact not only on livestock health and production but also on international trade. Conventional FMD vaccines consisting of chemically inactivated viruses have been used for many years and proved quite effective in controlling clinical disease. However, vaccinated animals cannot be distinguished serologically from ones that have recovered from a natural infection. The availability of an antigenic marker vaccine allowing discrimination between infected and vaccinated animals (DIVA) is of great value for the control and eradication of endemic infectious diseases. Here we report construction of a recombinant FMDV containing 93-143aa deletion (this region contains a relatively conserved B-cell epitope) in the NSP 3A using a recently developed FMDV infectious cDNA clone. The recombinant marker virus, r-HN/3A93-143, had growth kinetics similar to the wild type (WT) virus in culture cell and caused a symptomatic infection in pigs. Pigs immunized with chemically inactivated r-HN/3A93-143 vaccine were fully protected from WT FMDV challenge. Furthermore, a test using the 50% pig protective dose (PD50) showed that this marker vaccine could achieve 10.05 PD50 per dose. Serum analysis demonstrated that this recombinant marker virus, in conjunction with a blocking ELISA, enabled serological differentiation between the marker virus-infected and WT virus-infected animals. Our study indicated that a DIVA FMDV vaccine can be developed by deleting an immunodominant epitope in NSP 3A.

Objective: Gastric cancer is one of the most common malignant tumors in clinical work. With the traditional treatments, such as radical operation, chemotherapy and radiotherapy, patients still have to face on a poor prognosis. Dendritic cell is the new focus of anti-tumor biotherapy these years. Many researchers have been attracted by its strong ability to present antigen and inspire more effective anti-tumor immune reactions. Gastric cancer cells are lack of differential tumor antigens, with the former methods we only can get few tumor antigens and their ability to irritate dendritc cells or succedent anti-tumor immune reactions is very tiny. In this experiment, we used cell fusion technology to get the gastric cancer cell-dendritic cell fusion vaccine. We hoped that the fusion cells could get more tumor antigen information to increase their phenotypes and have more competence to irritate the anti-tumor immune reactions. By this experiment, we hoped to find out whether the fusion vaccine has satisfying safety and to verify its ability of inducing anti-tumor immune reactions. By researching its biological effects, we hoped to get a new kind of anti-tumor vaccine for gastric cancer’s biological therapy.
Methods: Part 1. Peripheral blood mononuclear cells were separated from gastric cancer patients and co-cultured with granulocyte-macrophage colony stimulating factors, interleukin-4 and tumor necrosis factor-αto generate mature dendritic cells. The frozen stored human gastric cancer SGC7901 cell lines were resuscitated and cultured at the same time. The dendritic cells and SGC7901 cells were fusioned by using of polyethylene glycol, and the pure fusion cells were screened out by cultured with HAT and HT selective culture systems. The phenotypes of dendritic cells and fusion cells were detected by flow cytometry.
Part 2. The biological characteristics of fusion cells (such as their morphological specialities, the expression of cell surface phenotypes (CD80\CD83\CD1a\HLA-DR), their growth curves, whether they can proliferate to be a new carcinoma in nude mice, the ability to irritate cytotoxic T lymphocytes and anti-tumor immune reactions) were tested to verify their safety when be used as anti-tumor vaccines.
Part 3. The ability of fusion cells-activated T lymphocytes to kill SGC7901 gastric cancer cells was investigated by MTT method in vitro. SGC7901 cells were injected subcutaneously in nude mice to make the planted gastric cancer models. The anti-tumor effects were evalutated by detecting the growth of the new planted cancers, the apoptosis and proliferation of the planted cancer cells, the pathologic changes of the tumor tissues and the prohibitive rate for planted tumors pre and after the fusion cell vaccine’s application and the cell divide cycles.
Part 4. The expression of B7-1, B7-2 mRNA of fusion cells were detected by RT-PCR to detect the gene changes of these cells.
Results: 1.We obtained mature dendritic cells from gastric cancer patients’ peripheral blood mononuclear cells by co-cultured with granulocyte-macrophage colony stimulating factors, interleukin-4 and tumor necrosis factor-α. These mature dendritic cells had typical morphological characteristics and their phenotypes’ expression were CD83 75.54±0.73%、CD1a 64.94±0.52%、CD80 61.56±0.27%、HLA-DR 62.50±0.66%.
2. Dendritic cells and SGC7901 cells could be fusioned by PEG and we could get pure fusion cells by cultured with HAT\HT selective culture systems.
3. Fusion cells were much larger than dendritic cells or SGC7901 cells. They exhibited typical characteristics by light and electron microscope testing. They lived in suspension in the culture medium and could not proliferate to be new carcinomas in nude mice. The cell phenotypes were CD83 80.16±1.12%、CD1a 72.86±2.48%、CD80 81.24±2.76%、HLA-DR 79.54±1.56% respectively. Fusion Cells also could strongly irritate the proliferation of T lymphocytes.
4. Fusion vaccine could induce strongly anti-tumor biological effects in vivo and in vitro: (1) Tumor growth was remarkably inhibited and the tumor size in treat group was 1.298±0.021cm3, which was extensively smaller than the size of control group 2.429±0.033cm3 (P<0.01) . The prohibitive rate for planted tumors was 57.8%. (2) Detected by Annexin-V/PI double labeling flow-cytometry, tumor cells’ apoptosis rate of treat group was 54.9±1.38%, which was promoted extensively than that of control group 20.02±0.39% (P<0.01) . (3) The tumor cells’ proliferation were found to be greatly restrained by detecting the cell divide cycles. (4) In treat group, the tumor tissues had more infiltrated lymphocytes and apoptotic tumor cells than control group. The treat group had more hemorrhage and necrotic tissues than the control group.
. The mRNA expressions of B7-1, B7-2 in fusion cells were highly increased and became more higher than normal dendritic cells.
Conclusions: 1. Mature dendritic cells could be obtained from gastric cancer patients’ peripheral blood mononuclear cells by co-cultured with GM-CSF, IL-4 and TNF-α. These dendritic cells had their unique modality and phenotypes. Gastric cancer cells and dendritic cells could be fusioned by PEG, we could obtain pure fusion cells by cultured with HAT/HT selective culture systems.
2. The fusion cells had their typical morphologies and kept the suspending living characteristic like dendritic cells. They had weak proliferation abilities and could not proliferate to be new carcinomas in immune deficiency animals, but the cell surface phenotypes were remarkably increased.
3. By irritating the proliferation of T lymphocytes, fusion vaccine could induce more strong anti-tumor effects.
4. The fusion vaccine could inhibit the tumor growth and accelerate the apoptosis of tumor cells.

The continued development of new cell culture technology is essential for the future growth and application of insect cell and baculovirus biotechnology. The use of cell lines for academic research and for commercial applications is currently dominated by two cell lines; The Spodoptera frugiperda line, SF21 (and its clonal isolate, SF9), and the Trichoplusia ni line, BTI-5B1-4, commercially known as High Five cells. High Five cells has been sufficiently expressed high recombinant protein. However, a contaminated TNCL virus was reported from High Five cells which sufficiently expressed high level of recombinant proteins recently. Herein, to erase the TNCL contamination and improve ability of higher expression, a novel cell clone with higher level of recombinant proteins, QB-Tn9-CL-F(QB-CL-F) , from Trichoplusia ni QB-Tn9-4S cell line has been established. It has been adapted to SF-900 III SFM, commercial serum-free medium. The cell morphology, cell growth kinetics, and virus production of the serum-free cultures were indistinguishable in compared with that High Five cells of serum-containing cultures (TNM-FH). RAPD analysis of the genomic DNA of this clone confirmed the genetic identify as T. ni cells. To the most, The QB-Tn9-CL-F cell clones was free of the TNCL virus examined by RT-PCR.

Currently tested multi-gene DNA vaccines towards HIV fail to induce a strong immune response against reverse transcriptase (RT) which is a key enzyme in the viral escape from drugs. Design of DNA-vaccine against HIV, specifically against drug resistant virus variants, requests a considerable enhancement of RT-specific immune response. MHC class II targeting of RT by fusion with signals of lysosome targeting of lysosome-associated membrane protein I, fragment of invariant chain and the minimal Gly-Ala repeat of EBNA1 tested previously somewhat improved the cellular, but not the humoral immune response to RT. To further strengthen its immunogenic performance, we N-terminally fused RT to the leader sequence of nonstructural protein 1 (NS1) of TBE. The NS1-RT chimera was detected both on the surface of transfected cells and in the cell culture medium. BALB/c mice were immunized with RT gene chimera by intradermal injection followed by electroporation, and immune response was assessed by serology, IFN-g/IL-2 Fluorospot, ICCS for IFN-g, IL-2; and ELISA for perforin and granzyme B. NS1-RT chimeric gene induced a potent Th2-tilted immune response manifested by strong dual IFN-gamma/IL-2 production and secretion of perforin and granzyme B by murine splenocytes stimulated with RT-peptides. Moreover, mice responsed by high anti-RT antibody production reaching 80000 for IgG and 1000 for systemic IgA. Thus retargeting of RT processing and presentation by turning it into secretable protein resulted in strong potentiation of RT-specific immune response.

Amira SOUII is a Doctor in "Biological Sciences and Biotechnology" and an Associate University Assistant of Microbiology in the Higher Institute of Applied Biological Sciences of Tunis, Tunisia. She is a researcher in the laboratory of infectious diseases and biologically active substances in the Faculty of Farmacy of Monastir Tunisia. Her research work focuses on Molecular Virology and Vaccinology. She studied the RNA-protein and RNA-RNA inetractions during the initiation of translation of two Coxsackievirus B3 strains: a cardiovirulent wild type and an attenuated mutant strain. Interestingly, the mutant strain has shown very interesting results and so, it constitutes a promising vaccine candidate against Coxsackievirus B3 infections.

Coxsackievirus B3 (CVB3) is an Enterovirus of the family of Picornaviridae. The Group B coxsackieviruses includes six serotypes (B1 to B6) that cause a variety of human diseases including myocarditis, meningitis and diabetes. Among group B, the B3 strain is mostly studied for its cardiovirulence and its ability to cause acute and persistent infections, yet no effective therapeutic against CVB3 is available. Translation initiation of CVB3 RNA is directed by an internal ribosome entry site (IRES) within the 5’ untranslated region. It is a complex process in which initiator tRNA, 40S and 60S ribosomal subunits are assembled by eukaryotic initiation factors into an 80S ribosome. Host cell factors involved in this process include some canonical translation factors and additional RNA-binding proteins. We have, previously, described that the Sabin3-like mutation (U475 --> C) introduced in CVB3 genome led to a defective mutant with a serious reduction in translation efficiency. In this regard, we analyzed the efficiency of formation of ribosomal initiation complexes 48S and 80S through sucrose gradients using Rabbit reticulocyte lysate and stage-specific translation inhibitors. We demonstrated that formation of 48S and 80S ribosomal complexes within the mutant CVB3 RNA was abolished compared to the wild-type RNA. With the aim to identify proteins interacting with CVB3 wild-type and Sabin3-like IRESes and to study interactions between either HeLa cell or BHK-21 protein extracts and CVB3 RNAs, UV cross-linking assays were performed. We have observed a number of proteins that specifically interact with both RNAs. In particular, molecular weights of five of these proteins resemble to those of eukaryotic translation initiation factors 4G, 3b, 4B and PTB. According to cross-linking patterns obtained, we have demonstrated a better affinity of CVB3 RNA binding to BHK-21 proteins and a reduced interaction of the mutant RNA with almost cellular polypeptides compared to the wild-type IRES. On the basis of phylogeny of some initiation factors and on the knowledge of the translation initiation process, we focused on the interaction of both IRESes within eIF3, p100 (eIF4G) and 40S ribosomal subunit by Filter binding assays. We have demonstrated a better affinity of binding to the wild-type CVB3 IRES. Taken together, we can conclude that the reduction efficiency of Sabin3-like RNA to bind to cellular proteins involved in the translation initiation could be the reason behind inefficient IRES function.

Rudolf Toman received his Ph.D in Organic Chemistry and later D.Sc in Microbiology from Slovak Academy of Sciences, Bratislava, Slovakia. He has published more than 130 papers in reputed journals, edited several books and served as an editorial board member of international journals. He is engaged mainly in glycomic/proteomic and immunological studies of the intracellular pathogen Coxiella burnetii that is classified as a category B biological warfare agent.

Coxiella burnetii is etiological agent of Q fever. The disease is a worldwide zoonosis affecting mammals, birds and anthropods. In most cases, the human infection follows inhalation of aerosols derived from the excretions and secretions of infected animals such as cattle, sheep and goats. The recent large Q fever outbreak in the Netherlands stressed a need for a new and effective vaccine against Q fever.There were several attempts to produce safe and effective vaccines against the disease in the past. Although these vaccines showed high efficacy in establishing protection against infection, the application of them has been problematic. Vaccination often causes adverse local, or occasionally, systemic reactions in people previously sensitized to the pathogen. Therefore, skin tests, serological tests, and/or in vitro lymphocyte proliferation assays were prerequisites prior to vaccination. This makes vaccination time-consuming and costly for large-scale applications. Lipopolysaccharide (LPS) has been considered to be a major antigen determinant of virulence expression and infection of C. burnetii. The LPS is highly immunogenic and contains a noticeable amount of two sugars virenose (Vir) and dihydrohydroxystreptose (Strep), which have not been found in other LPSs and are considered as unique biomarkers of the bacterium. We have shown that both sugars are located in the O-polysaccharide chain of LPS, mostly in terminal positions. Our studies on modification/modelling of the O-specific chain of C. burnetii have indicated that Vir and Strep could be indispensable components for the resulting protective qualities of the vaccine. It appears that LPS of C. burnetii is a promising candidate molecule for the development of a new sub unit vaccine against Q fever.

Schistosoma mansoni is a blood fluke parasite responsible for schistosomiasis. The best long-term strategy to control schistosomiasis is through immunization combined with drug treatment. In this study, we cloned, expressed and purified Sm-TSP-2 fused to the N- and C-terminal halves of Sm29 and tested these chimeras as vaccine candidates.The results demonstrated that the association of Sm-TSP-2 with N- or C- terminus of Sm29 elicited a protective effect when compared to the immunization with Sm29 alone regarding the reduction of the worm burden, however these association are not effective in diminish the liver pathology. Additionally, we detected high levels of mouse specific IgG, IgG1 and IgG2a against chimeras A and B and significant amounts of IFN-γ and TNF-α and no IL-4. Finally, studies with sera from patients resistant to infection and living in schistosomiasis endemic areas revealed IgG recognition of chimeras A and B. In conclusion, the chimeric proteins tested here induced superior protection against infection compared to Sm29 alone and higher levels of IgG in serum of patients from endemic areas with different status of resistance and susceptibility, therefore they should be considered as potential vaccine candidates instead of Sm29 alone.

Engy M. El-Ghitany is an Egyptian public health specialist. She is working for Tropical Health department, High Institute of Public Health, Alexandria University, Egypt. She was graduated from the faculty of medicine, Alexandria University. She has earned her doctorate degree in 2005 from her institution and in the same year she attended the six weeks course on “ Immunology and Vaccinology applied to infectious diseases” organized by WHO in Lausanne. She has also got “Diploma of Tropical Medicine and Hygiene” from London school of hygiene and tropical medicine in 2006. In 2012, she earned a diploma in Vaccinology from Pasteur Institute, Paris. She is interested in travel medicine and is a member of international society of travel medicine. Her main research interest is viral hepatitis.

Background: Despite being highly efficacious vaccine, no response to HBV vaccination is properly evaluated among diabetics whoretainpost-vaccination seroprotective antibodies level for shorter time than healthy ones.The necessity of providing booster doses following a successful course of primary immunization is an emerging question. The present study was conducted to determine how long immunity persists in Egyptian diabetic school students previously vaccinated against hepatitis B, whether boosters are needed, and, if so, when and in whom they should be administered.
Methods:The study included two phases, acomparative-case-control(screening) phase followed by a quasi-experimental (boosting) phase. A baseline serologic screening for anti-HBs titre was carried out among 260 school students (130 diabetics and 130 healthy non-diabetics, matched for age and sex) who were in the age group 10-17 years and had received the full three-dose regimen of HBV vaccine under the EPI in Egypt. Ninetyparticipants(45 diabetic and 45healthy ones) with anti-HBs < 10 mIU/mlwere included in the quasi-experimental phase after being consented where up to three vaccine booster doses were given and anti-HBs level (mIU/ml) was determined.
Results:The median value of anti-HBs titre was significantly lower among diabetics (3.0 m IU/mL) as compared to non diabetics (6.8 m IU/mL) (p=0.002 ). The frequency of poor response (anti-HBs titre<10 m IU/ml) was higher among diabetic group as compared to non diabetics (70.8% vs.60%) respectively. Those with anti-HBs titre>100 m IU/ml were 10(7.7%) among diabetics versus 14(10.8%) among non diabetics.Independent factors shown by the stepwise logistic regression analysis showedthat beingdiabetic has 60% more risk for having a poor response.The only significant risk factor associated with poor response was the age. One year increase in age was associated with about 30% higher risk for being poor responder. One month after giving both the first and second booster doses of HBV vaccine, the diabetic students had a minimum level of anti-HBs lower than that of non-diabetics (0.0 versus 21mIU/ml and 11.0 versus 146.0 respectively). Moreover, the diabetic students showed a lower mean titre of anti-HBs (124.3±60.4 versus 149.3±32.0 and 104.2±63.0 versus 156.5±7.0 respectively) than that of the non-diabetic students (P=0.016) after the first booster dose only. In contrast to non-daiabetics, the mean titre of anti-HBs among diabetics after first, second and third booster doses revealed statistically significant difference (P=0.018*). To reach full protection, we found that only one booster dose was needed by 80.0% of diabetics compared to 91.1% of non-diabetics while two booster doses were needed by11.1% of diabetics in comparison to 8.9% of non-diabetic students. Four (8.9%) diabetic students needed a third booster dose in comparison to none of the normal students. Incorporating a booster dose of HBV vaccine would be best timed at age of 12 years old for immunocompromised diabetic students while it remains an optional one at the age of 13.5 years old for immunocompetent healthy students. We concluded that,type 1 DM adolescents express hyporesponsivness to HBV vaccination and more rapid decline of protective anti-HBs compared to healthy ones. Although such decline of anti-HBs, the presence of specific immune memory can be inferred by demonstrating anamnestic response to HBV vaccine booster doses.Adequate protection against HBV infection among students with unprotective anti-HBs level is achieved after one booster dose for healthy students and two booster doses for diabetic ones.

Molalegne Bitew has completed his Ph.D at the age of 33 years in 2014 from Indian Veterinary Research Institute. He is the assistant professor of Virology at Jimma University, college of agriculture and veterinary medicine; we have research activities in collaboration with different research institutes. He has published 20 and submitted 3 manuscripts in reputed journals and serving as an editorial board member of International journal of virology.

Recent invasion of multiple bluetongue virus serotypes (BTV) in different regions of the world necessitates urgent development of efficient vaccine that aims numerous serotypes. In this experimental study, cell mediated immune response and protective efficacy of binary ethylenimine (BEI) inactivated montanide adjuvanted pentavalent (BTV-1, 2, 10, 16 and 23) vaccine was evaluated in sheep against challenge with homologous serotypes in their respective group. Upon vaccination and challenge, it was found that both unvaccinated and vaccinated sheep peripheral blood mononuclear cells (PBMCs) exhibited expression of IFN-α, IL-2, IL-6, IL-12, IFN-γ and TNF-α transcripts. However, compared to unvaccinated ones, PBMCs of vaccinated sheep showed significant (P < 0.05) up regulation of these cytokines at certain point of time. On the other hand, there was a significant increase in Mean ± SD percentage of CD8+ T cells after 7 days post challenge (DPC) but, the Mean ± SD percentage of CD4+ T-cell population slightly declined at 7 DPC and enhanced after 14DPC. There was also significant difference (P < 0.05) of CD8+ and CD4+ T cells population between vaccinated and unvaccinated animals. The vaccine also significantly (P<0.05) reduced BTV RNA load in PBMCs of vaccinated animals than unvaccinated animals following the challenge. There were no significant difference (P > 0.05) in cytokine induction, and BTV RNA load CD8+ and CD4+ cell count among BTV-1, 2, 10, 16 and 23 serotype challenges except significant increase Mean ± SD percentage of CD8+ by BTV-2. These findings put forwarded that binary ethylenimine inactivated montanide adjuvanted pentavalent bluetongue vaccine has stimulated cell mediated immune response and most importantly reduced the severity of BTV-1, 2, 10, 16 and 23 infections following challenge in its respective group.

Background: The advanced stage of HIV-1 disease may manifest into a variety of central nervous system disorders. TheNeurons may get damaged because of direct infection with the virus as well as indirectly due to the immune response. CD8+ Cytotoxic T-lymphocytes are proposed tobe responsible for HIV associated dementia, but the exact mechanisms have yet not been elucidated.
Methodology: HIV+ femaleswith dementia score either <7.5 and non-demented HIV+ score>7.5 were recruited along with age matched healthy controls. Peripheral blood mononuclear cells were phenotyped for CTL markers along withCD127, CD45RA and CD45RO. CTLs expressing intracellular perforin and granzyme B werealso enumerated. Among cytokines IFNγ and IL-7 levels estimated. The results were analysed using appropriate statistical tools.
Results: Among HAD group the mean CD4 count was 269 cells/μL, and viral load 19095 copies/ml, while the non-demented individuals had normal CD4 count of 485 cells/μLwith lower quantity of viral RNA(7527 copies/ml).The HAD group had significantly lower number(9.8%) of CD4+CD127+ cells compared to 17.04% in ND group and 32.18% in HCs. The frequency of CTLs expressing CD127 was 14.34% among demented individuals as compared to 18.67% in non-demented patients.The significantly higher proportion of CTLs from HAD patients expressed Granzyme B and Perforin(59.4+20.9 and 34.05+19.09respectively) as compared to non-demented patients (47.5+19.55 and 26.94+16.17).. The dementia score was significantly correlated with the frequency of CD8+ cells expressing Granzyme B indicating a direct association.
Conclusions: The findings suggest that the proliferation of terminally differentiated and activated CD8+ cells in the HIV patients with prolonged disease, may be associated with enhanced killing of neuronal cells leading to dementia.

Dr. Virendra Gomase [B.Pharma, M.Sc, M.Phil, Ph.D] did his Ph.D. in Computer Science from Department of Computer Science and IT, Dr. B.A.M. University, Aurangabad, India. He has authored more than 15 Books. He published more than 300 International and National peer reviewed Journals research papers having Impact Factor. He presented more than 200 articles in International and National conferences, seminars. His research included work on Bioinformatics, Machine Learning, Computational Intelligence, Immunoproteomics, Toxicoproteomics. He is serving as member of more than 60 scientific societies. He is serving as an editorial member of several reputed journals.

The prediction of peptides which are presented by the restricting major histocompatibility complex (MHC) molecules is a crucial problem in immunology. In silico methods for the prediction of antigenic peptides binding to MHC class molecules play an increasingly important role in the identification of T-cell epitopes. Statistical and machine learning methods in particular are widely used to score candidate binders based on their similarity with known binders and non-binders. The genes coding for the MHC molecules, however are highly polymorphic, and statistical methods have difficulties building models for alleles with few known binders. In this context, recent work has demonstrated the utility of leveraging information across alleles to improve the performance of the prediction. The aim of our method is to build each and every known MHC class allele, a model to predict whether or not candidate peptides can bind to it. Our method must then learn the binding predictive models from the training set. The most relevant measure of a prediction model’s accuracy is how well it performs on novel peptide sequences that are dissimilar to those used to fit the model. A model with a large number of parameters, such as most peptide-MHC binding prediction models, can potentially overfit the data so that its accuracy is good on the training set but poor on unrelated peptide sequences. Cross-validation is one procedure that can be used to estimate the accuracy of the prediction model for novel data

Objectives: Hepatitis C virus (HCV) and human immunodeficiency virus (HIV) infections are a global health concern. Therefore, they are considered for the huge number of studies looking for effective vaccines. In a previous study, we introduced a single cycle replicable (SCR) HIV system that, completely maintained the antigenic structures of HIV-1, through its one cycle replicating properties represented a good implication as a potential vaccine candidate, Herein, we constructed a novel HIV-1 virion, capable of expressing non structural 3 (NS3) protein of HCV as potential bivalent candidate vaccine that provides a more immunogenicity, while preventing any pathologic effects with further evaluated its biological properties.
Methods: The pIPNL4-3/NS3 containing HIV genome of NL4-3 strain with a 2-kb deletion in reverse transcriptase (RT) and integrase (IN) genes and replacement of the deleted fragment with NS3 was constructed, confirmed by sequencing reactions and transfected into HEK 293T cell line. By further co-transfection of psPAX2 and pMD2.G plasmids, which encoded HIV Gag-pro-pol and vesicular stomatitis virus surface glycoprotein, into the same pIPNL4-3/NS3-harboring cells, pseudotyped virions were produced, evaluated by electron microscopy, quantified using P24 end-point ELISA assay and western blotting. Infectivity of recombinant virions and their efficiency towards the syncytium formation was evaluated on HIV-sensitive MT-2 cells.
Results: Production of HIV virions was indicated by the level of P24 protein in culture supernatant of transfected cells and was further confirmed by electron microscopy. Also, expression of NS3 protein was confirmed using western blotting. Formation of syncytia in MT-2 cells also evidenced for the functionality of the surface glycoproteins in produced pseudotyped virions. Interestingly, infectivity analysis verified that the second generation virions were completely non-replicative.
Conclusion: The results were shown that a new recombinant virion with capable to express NS3 protein completely maintained the antigenic structures of HIV-1, by its one cycle replicating properties , and represented a good implication as a potential bivalent vaccine candidate. Moreover, this guarantees further investigations toward the assessment of its immunogenicity, which are currently under process. It may also present another interesting approach towards the improvement of its application in bivalent HIV and HCV vaccine researches.

While Crimean-Congo hemorrhagic fever (CCHF) has a high mortality rate in humans, the associated virus (CCHFV) does not induce clinical symptoms in animals, but animals play an important role in disease transmission to humans. Our aim in this study was to examine the immunogenicity of the CCHFV glycoprotein when expressed in the root and leaf of transgenic plants via hairy roots and stable transformation of tobacco plants, respectively. After confirmatory analyses of transgenic plant lines and quantification of the expressed glycoprotein, mice were either fed with the transgenic leaves or roots, fed the transgenic plant material and injected subcutaneously with the plant-made CCHFV glycoprotein (fed/boosted), vaccinated with an attenuated CCHF vaccine (positive control), or received no treatment (negative control). All immunized groups had a consistent rise in antiglycoprotein IgG and IgA antibodies in their serum and feces, respectively. The mice in the fed/boosted group showed a significant rise in specific IgG antibodies after a single boost. Our results imply that oral immunization of animals with edible materials from transgenic plants is feasible, and further assessments are under way. In addition, while the study of CCHF is challenging, our protocol should be further used to study CCHFV infection in the knockout mouse model and virus neutralization assays in biosafety level 4 laboratories.

He is 23-year-old senior of M.Sc at Tehran Azad University of Medical Sciences . He had some papers in the field of New Biology and Cardiovascular Diseases ,and now he is working on HPV VLPs (Virus Like Particles) to develop the next generation of vaccine for cervical cancer at Pasteur Institute of Iran.

Background and Aim: High-risk type of human papilloma viruses (HPVs) can be considered as the etiologic agents of cervical cancer. At present, two prophylactic vaccines have been designed to prevent HPV infections. Both vaccines (Cervarix and Gardasil) are included VLP structure derived from L1 protein of HPV16 and 18. Both of these vaccines are highly immunogenic and elicit high titers of neutralizing antibody responses. Type-specific anitbody responses of L1 VLPs are one limitation of these vaccines. So, current vaccine strategy provides protection against HPV types associated with cervical cancer and it won't be able to induce immunity against other important HPV types. Alternatively, L2 minor capsid protein is a suitable candidate for next-generation of HPV vaccine. L2 has important functions in both papilloma virus assembly and the infectious process. Sequence analysis indicates L2 proteins as highly conserved with no changes in amino acid sequences during development. Indeed, L2 vaccine targeting can provide much more comprehensive protection against infection by various HPV types. However, L2 VLP typically elicits much lower neutralizing antibody titers than L1 VLP. The aim of this study is to evaluate the L1-L2 protein expression in mammalian cell lines.
Methods: Two plasmid DNA fusion constructs harboring HPV16 L1-L2 (pEGFP-L1-L2 and/or pcDNA-L1-L2) were generated by sequential cloning processes. The constructs were confirmed by sequence analysis. Then, gene expression was evaluated and compared in different cell lines (COS-7 and HEK293) using fluorescent microscopy, flow cytometry and western blot analysis.
Results: The results showed that vector modification and cell type have an effect on L1-L2 protein expression, in vitro. These parameters can influence the protein expression in vivo resulting to various immunological responses.
Conclusion: Regarding to the studies, the use of different linkers (IRES or proteasomal residues) can involve the levels of expression and subsequently stimulation of immune responses. We are going to investigate the effects of these motifs on the expression of L1-L2 protein in near future.

Mumuni Abdul Malik is 27 years old and is a currently a second year student of the University of Ghana, Accra, Ghana. He is the president of Nutrition Students of Ghana, Tamale Branch. He has been carrying research work on Obesity with his fellow students of the Faculty of Nutrition. He has written about 20 abstracts and proposals in the University Journals and Publications and is currently carrying out his Research work on Obesity at the Tamale Teaching Hospital. He has attendedconferences in other parts of West Africa namely; Cote d’ivoire, Burkina Faso and Nigeria.

Background: Diet-induced obesity is one of the leading causes of cardio vascular morbidity and mortality that correlates with the duration and intensity of adiposity. This study tested the hypothesis that the increased time of exposure to obesity intensifies the cardiac dys-function. In addition, also examined whether the functional impairment is due to increased levels of myocardial collagen.
Methods: Thirty-day-old male Wister rats were distributed in control (C) and obese (Ob) groups. The C was fed a standard diet and Ob was alternately submitted to four palatable high-fat diets for 15, 30 and 45 weeks. The obesity was determined by adiposity index. The cardiac remodeling process was assessed by structural and functional analysis.
Results: Adiposity index was higher in Ob than C overtime. Obesity promoted several co morbidities after15, 30 and 45weeks. LV mass and inter ventricular septum and posterior wall thickness in dias to le after 15 , 30 and 45 weeks were increased in Ob. However, obesity has led to an increase in LV relative wall thickness in the 30th and 45th weeks. Obesity did not promote changes in LV interstitial collagen in all periods. Obesity improved the systolic function after 30 weeks and myocardial dys-function after 15weeks, however, the duration of obesity did not intensify the cardiac dys-function in vitro.
Conclusions: Long time exposure to obesity does not intensify the cardiac dys-function and does not promote alterations on myocardial collagen.

Dr. Akusa Yuma Darlington is a 1996 graduate of Makerere University in Uganda, with MBCh.B degree. I served as the zonal coordinator for National malaria control programme from 1999 to 2002. I have published in reputable journals and presented papers at national conferences. I am currently leading a team of Health workers investigating an outbreak of Black Water Fever Malaria in North Western Uganda, based in Arua Regional Referral Hospital, in Uganda.

Introduction: Black Water Fever Malaria, an acute haemolytic disease syndrome, associated with Plasmodium falciparum infection, occurring only in non – immune children and adults could, be a disorder of the Zinc finger gene and tumour necrosis factor alfa. It is characterized by haemoglobinuria, fever, jaundice and anaemia. We now report that their immunity can be boosted with a combination of antihistamine and Zinc Sulphate to the effect of preventing further malaria attack for over a year.
Case History: 120 childen aged three months to twelve years, were followed for haemoglobinuria without a known haemoglobinopathy, with symptoms of fever, vomiting, abdominal pain, passage of dark red urine, and loose stools, epistaxis and body weakness, after treatment with chlorpheniramine and zinc sulphate in addition to anti malarials. There was a positive family history of leprosy one case and congenital malformations, ranging from cervico-facial-ano genital sinuses and tags, in 96 cases, polydactyly, in one case, to Einhoms disease, in one case and or dactylitis, in one case. Physical examination revealed fever, pallor, jaundice, dehydration, renal angle tenderness, hepatosplenomegaly and congestive cardiac failure in all of them.
Method: (i) Blood and urine samples were taken for examination. (ii) Abdominal ultrasonography was requested.
Result and Treatment:Full Haemogramme showed low haemoglobin, suggestive of severe anaemia, monocytosis, high total white and low red blood cell counts; positive rapid test for plasmodium falciparum and unspecified mixed species of plasmodium, and random blood glucose of varying degrees of hypoglycaemia. Urinalysis report revealed a positive Haem -test without the presence of Red blood cells. Renal parenchymal disease was detected on Ultrasonography in all of them.Black water fever malaria with severe anaemia and congenital pre-auricular sinus with renal disorder was diagnosed. In addition to general and specific care, • Oral chlorpheniramine, 0.35mg/kg/day in three divided doses for five days, and • Oral Zinkid (zincsulphate), 0.4mg/kg twice daily for 14 days, were administered. They were discharged between November 2012 and April 2013, and 114 of them have not had recurrence of the disease to date with exception of six, four of whom turned up at 8 months of follow up, nine at 9 months, and the other, at 10 months, with Black water Fever Malaria syndrome.
Conclusion: Black water fever Malaria syndrome patients developed ample immunity to the disease to the extent of protecting 95% of them for more than a year against Malaria.

Introduction: Although, some studies on the airway show IL-22/IL-25 play a critical role in the pathogenesis of asthma but there are little documents about the systemic production of these cytokines. Therefore, the aim of this study was IL-22/IL-25 assay in serum, in mitogen activated of whole blood (WB) and in mitogen activated of peripheral blood mononuclear cells (PBMCs) cultures of patients with sever asthma.
Materials & methods: In this cross sectional study, to determine the severity of the asthma, a questionnaire was prepared. The questionnaire asked information including clinical signs, clinical symptoms, and past medical history so all active or ex-smoker patients were excluded. Then a trained observer assessed airway reversibility, peak flowmetry and spirometry in the patients. Twenty one patients with severe asthma and simultaneously, twenty age-sexes matched healthy controls were selected. Ten ml sterile blood was taken from each person. The sera were isolated and anticoagulant bloods used to WB and PBMCs cultures and hematological tests. Phytohemagglutinin (PHA) and Lipopolysaccharide (LPS) used to activate WB and PBMCs. Data of two groups were compared with Student’s t-test and nonparametrical statistic test.
Results: Except total white blood cells count that was increased in asthmatic group, other hematological indices and IL-22/IL-25 levels in two groups were not significantly (p<0.05) different.
Conclusion: The levels of IL-22/IL-25 in patients with severe asthma are not higher than healthy people and their roles in asthma can related to local immunological process.

Background: Immunization averts an estimated 2 to 3 million deaths every year globally. In Ethiopia only quarter of children are fully immunized; the rest are remained at risk for vaccine-preventable mortality. To increase the immunization, its coverage and predictors has to be identified. This study has measured immunization coverage and identified the predictors.
Methodology: Cross-sectional community based study has been conducted within 630 age 12–23 months children in 15 districts of Arba Minch town and Arba Minch Zuria district, Southern Ethiopia in March 2013. Census was done to identify eligible children. The 2005 world health organization expanded program of immunization cluster sampling method has been used. Data were collected using semi-structured pretested Amharic version questionnaire by interviewing index children’s mothers/caretakers, copying from vaccine card and observing BCG vaccine scar. Data were processed using SPSS version 16. Associations between dependent and independent variables has been assessed and presented using three consecutive logistic regression models.
Result: Nearly three fourth (73.2%) of children in Arba Minch Town and Arba Minch Zuria district were fully immunized. The rest 20.3% were partially immunized and 6.5% received no vaccine. Mother education, mothers’ perception to accessibility of vaccines, mothers’ knowledge to vaccine schedule of their site, place of delivery and living altitude were independent predictors of children immunization status.
Conclusion: Expanded program of immunization (EPI) coverage at Arba Minch town and Arba Minch Zuria district is better than the national immunization coverage but still below the goal. Educating mother, promoting institution delivery could help to maintain and enhance current immunization coverage. More emphasis should be given to the highland areas of the area.