Madridge Journal of Vaccines

ISSN: 2638-1966

2nd International Conference and Expo on Vaccines and Vaccination

November 20-21, 2017, Dubai, UAE
Scientific Session Abstracts
DOI: 10.18689/2638-1966.a1.006

Development of a Recombinant Fusion Protein Vaccine towards Middle Eastern Respiratory Syndrome (MERS)

Lee Smith1*, Ian Jones2 and Alexander Edwards2

1Anglo Biopharma Ltd, United Kingdom
2University of Reading, United Kingdom

Anglo Biopharma Ltd. is developing a Rapid Epitope & Antigen Discovery Platform (READ) technology in collaboration with the University of Reading, UK. This technology is being used to generate candidates for use in a safe and efficacious vaccine towards Middle Eastern Respiratory Syndrome (MERS). MERS is an emerging infectious disease related to SARS that was described by the director of the World Health Organisation (WHO) as a “a threat to the entire world”. The landscape of this disease and the potential for a vaccine will be discussed. Furthermore, the construction and expression of candidate vaccine constructs from the READ platform will be described, highlighting the expression of good yields of MERS fusion proteins as structurally intact and conformationally correct protective antigens. In addition, data will be discussed that has been obtained from serum analysis following candidate protein immunizations during preclinical studies. This is to understand the affinity of serum antibodies against MERS CoVneutralising epitopes known to be important for protection by end-point titer (EPT) ELISA. The utility and validity of using serum to neutralise the infectious ability of a “pseudotyped” MERS virus will also be discussed and their ability todemonstrate useful vaccine efficacy data compared to the classic downstream challenge models.

Dr. Lee Smithʼs experience spans over 20 years in biopharmaceutical CMC, process, analytical, formulation pre-clinical and clinical assay development as well as experience in product characterization and regulatory submissions and interactions. He is regularly involved in applying QbD and advising on the use of DoE and data analysis for processes, formulation and assays, with a particular expertise in bioassays. He has worked on a wide range of vaccines including Dengue, influenza, meningitis B, Zika and MERS. This includes their development, optimization and validation of related processes and methods.

Oral Salmonella-Based Vaccine for Type 1 Diabetes

Mohamed I. Husseiny*, Fouad Kandeel and Kevin Ferreri

Beckman Research Institute of City of Hope, USA

Type 1 diabetes (T1D) is a metabolic disease caused by the autoimmune destruction of pancreatic insulin-producing beta-cells due to loss of tolerance to specific self-antigens (autoantigens). A promising approach to restore immune balance is the oral administration of diabetic autoantigens which diminishes the islet-specific destructive responses and induces regulatory responses. We recently reported the development of an oral vaccine for T1D based on live attenuated Salmonella expressing preproinsulin (PPI) as the autoantigen combined with host cell expression of TGFβ. We showed that oral vaccination with the combined PPI+TGFβ prevented the onset of diabetes in non-obese diabetic (NOD) mice. In this study we extended this approach by evaluating another autoantigen, GAD65, as well as the addition of a complementary treatment, partial T-cell ablation with anti-CD3. Our results showed that GAD65+TGFβ and the combination of PPI with TGFβ+IL10 and a sub-therapeutic dose of anti-CD3 prevented diabetes in the NOD mice and restored normal glucose tolerance. We also found the oral combination therapy of PPI+TGFβ+IL10 combined with anti-CD3 effectively reversed diabetes in the majority of diabetic NOD mice after replacement of arginine with glutamic acid (R22E) and glutamic acid to glycine (E21G) in Insulin B9-23. Initially we have shown that combined vaccine therapy increases the Tregs in splenocytes, and local Tregs in PLN and pancreas of vaccinated NOD mice. Additionally, the combination therapy significantly increased regulatory cytokines (IL10 and IL2) and inhibited the inflammatory IFNγ. Together these results indicated that the vaccine suppressed the autoimmunity and increased regulatory mechanisms leading to a conclusion that a Salmonella-based oral vaccine expressing autoantigens in combination with tolerogenic cytokines is a promising therapy for the prevention and treatment of T1D.

Dr. Mohamed I. Husseiny, completed his master of microbiology and immunology from Faculty of Pharmacy, Zagazig University, Egypt. He received his Ph.D from Institute for Clinical Microbiology, Immunology and Hygiene, Friedrich-Alexander University, Germany and postdoctoral studies from LA Biomed at Harbor-UCLA Medical Center, California, and Childrens Hospital Los Angeles at USC California. Currently he is an Assistant Research Professor at Beckman Research institute of City of Hope, California, USA he is working with Salmonella-based vaccine for over 16 years and I published this methodology in reputed journals.

Development of Novel Vaccines Utilizing a Recombinant Nanoparticle Technology Platform

Atin Tomar

CPL Biologicals Private Limited, India

A novel technology platform for recombinant vaccines with significant advantages over conventional vaccines:carefully designed genetic constructs enable production of immunogenic protein-based nanoparticles against key antigenic components of pathogens, which enable functional immunity and protection against infection and disease.The platform is based on genetically engineered baculovirus constructs that in Spodopterafrugiperda (Sf9) [Lepidopteran] cells (BV/Sf9) efficiently produce correctly folded, complex antigens as nanoparticles that induce durable, neutralizing and protective immunity (demonstrated in pre-clinical and clinical studies).Our innovation introduces a significant immunologic advantage, embodied by the presentation of antigens as highly immunogenic particles in their native configuration, and sound principles of development, as a scalable, efficient recombinant vaccine production system. Unlike traditional vaccine manufacturing, infectious agents are not required for manufacturing based on this platform. This platform technology is completely recombinant, enabling efficient and safe manufacture of vaccines against highly pathogenic targets. The platform manufacturing process uses no animal source material, there is no requirements for higher BSL facilities and, therefore, no safety threats.This recombinant technology platform is the basis for a portfolio of novel vaccines that represent important advances and address current unmet medical needs. H1N1 (Swine Flu) and Seasonal Influenza vaccines have already been licensed for commercialization in India. Other products currently under development include vaccines against Rabies Virus, Varicella Zoster Virus, Zika Virus, MERS Virus, Ebola Virus etc. The technology also enables Halal certification for the vaccines.H1N1 (Swine Flu) vaccine and Seasonal Influenza vaccines have already been Halal certified.

Atin Tomar is President & CEO of CPL Biologicals, a joint venture of Cadila Pharmaceuticals, India and NovavaxInc, USA. Atin brings with him comprehensive experience in the Biotechnology industry, globally. Atinʼs experience and expertise spans from Process Development, Technology Transfer and Manufacturing to Business Development, Licensing and Commercialization. Atin completed his B.Tech (Hons) in Biotechnology and Biochemical Engineering from the Indian Institute of Technology (IIT) Kharagpur, India, MS in Biological Engineering from the University of Georgia, Athens, USA and Post-Graduate Programme in Management (MBA) in Finance and Strategy & Leadership from the Indian School of Business (ISB), Hyderabad, India.

Immune Response Enhanced by DNA Priming followed by Proteinic Form of Vaccine

Kripa N. Nand1,3*, Jagdish C. Gupta1, Amulya K. Panda2, S. K. Jain3 and G. P. Talwar1

1The Talwar Research Foundation, New Delhi, India
2National Institute of Immunology, New Delhi, India
3JamiaHamdard, New Delhi, India

Introduction: Immunization against human chorionic gonadotropin (hCG) prevents pregnancy in sexually active women of proven fertility, as shown by previous Phase II efficacy trials. In order to make the vaccine consistent in its linkage to the carrier, we developed a recombinant vaccine linking hCGβ to LTB, B subunit of heat labile enterotoxin of E.coli which isa potent mucosal adjuvant. The hCGβ-LTB vaccine was fairly immunogenic in mice of different genetic strains.Since a vaccine for control of fertility should ideally be effective in every recipient and be potent enough to generate above protective threshold antibody titres to prevent pregnancy, it was decided to investigate if prime-boost approach employing a combination of anti-hCG DNA and protein vaccines, can enhance the immune response.

Methodology: hCGβ-LTB protein vaccine was made and purified using yeast Pichia pastoris pPIC9k/GS115 host-vector system.DNA version of the vaccine was prepared by incorporating the gene encodinghCGβ-LTB in eukaryotic plasmid VR1020(DJ).Mycobacterium indicuspranii (MIP)was used as an immuno-modulator. Female inbred Balb C mice received 100 μg of DNA vaccine in saline along with 5x106 cells of MIP/animal/dose route twice fortnightly followed by 2 μg of alum adsorbed hCGβ-LTB along with 5x106 cells of MIP by intramuscular route. Second group of mice was immunized by only protein version of the vaccine along with MIP.

Results and Conclusions: Immunization with the DNA form of the recombinant hCGβ-LTB vaccine twice at fortnightly interval followed by the proteinic form of the vaccine induced distinctly higher antibody response than the proteinic vaccine alone.DNA is not only cheaper to make, it is thermostable and does not require cold chain. Hence the employment of DNA for primary immunization is expected to reduce the cost besides the benefitof enhancing antibody response

Dr. Kripa N. Nand is currently working at Talwar Research Foundation, India. His research interest is based on Rec. protein expression in prokaryotes & eukaryotes, purification of recombinant protein, Molecular Biological Techniques, Cell Culture, Animal handling & immunization, Bioreactor, Immunology Techniques, Prostate Cancer. He has published many articles in reputed journals.

Leishmania Antioxidants: Peroxidoxin 1 as a Promising Target for Vaccine Development

Nada Daifalla1,2, AbebeGenetu Bayih1,3 and Lashitew Gedamu1

1University of Calgary, Canada
2Imam Abdulrahman Bin Faisal University, Saudi Arabia
3Gondar University, Ethiopia

Leishmania antioxidants, like peroxidoxins, represent potential vaccine candidates against leishmaniasis. We assessed the immunogenicity and protective efficacy of recombinant Leishmaniadonovani peroxidoxin 1-GST fusion protein (rPxn1-GST) in BALB/c mouse model. Mice were immunized with three s. c injections of rPxn1-GST with or without CPG-ODN in three-week intervals. Four weeks after the last immunization the mice were challenged with Leihsmania major and the level of protection was evaluated as the reduction in lesion size and parasite burden. Recombinant Pxn1-GST protein with CpG demonstrated significant level of protection as compared to rPxn1-GST without adjuvant and all other control groups. The rPxn1-GST/CpG ODN regimen showed marked reduction in the parasite load and footpad swelling of BALB/c mice. Recombinant Pxn1-GST alone and GST with or without adjuvants gave similar results as controls. There was higher level of IgG2a and IFN-γ in rPxn1-GST/CpG vaccinated mice, as compared to protein alone or adjuvant alone control groups, which indicates that the protection in this group is associated with a biased Th1-type immune response. Despite the enhanced Th1 immune response in mice immunized with GST/CpG, these mice showed progression of the infection similar to other control groups. This result be speak that the partial protection seen in mice receiving rPxn1-GST/ CpG is antigen dependent.

Dr. Nada Daifalla obtained her master degree in Immunology from the Institute of Endemic Diseases (Dr. H. Ghalibʼs lab), University of Khartoum, Sudan and her Ph.D in Molecular Immunology from the University of Calgary (Dr. L. Gedamuʼs lab). Nada has several years of expertise in the area of immunology and molecular biology focusing on infectious diseases including leishmaniasis, tuberculosis, and lyme disease. She worked in different projects including antigen discovery, diagnostic and vaccine development, biomarker identification, and stem cell research. Nada worked in research and teaching in renowned institutions including The Institute of Endemic Diseases, Sudan; Infectious Disease Research Institute, Seattle, USA; University of Calgary, Canada; The Forsyth Institute (Harvard University), Massachusetts, USA. Currently she is an Assistant Professor at Imam Abdulrahman Bin Faisal University, KSA.

Vaccine for Psychiatric Disorders

Srinivasa K. Rao, Sourabh Khanna, Vasundhara D. Kalasapudi and Srinivas V. Kaveri

North East Biotech LLC, USA

Mental health is a major public health problem. Mental disorders include: depression, bipolar affective disorder, schizophrenia and other psychoses, dementia, intellectual disabilities and developmental disorders including autism. They are generally characterized by a combination of abnormal thoughts, perceptions, emotions, behavior and relationships with others. Globally, an estimated 300 million people are affected by depression alone. Although there are various strategies to treat psychiatric disorders, there is a need for new ones.

We examined the advances in immunology to find the suitable immunological approaches for psychiatric disorders. Recent advances in research show that inflammation as one of the triggers for psychiatric disorders. Treating inflammation using immune therapy approach is gaining momentum. Inflammatory markers can act as detectable secondary biological markers for psychiatric disorders. Treating inflammation can help mitigate the psychiatric disorders to the extent that they are causing it.

Some studies have shown that there is an increased proinflammatory response in patients diagnosed with depression, indexed by an elevation in C-reactive protein, and acute phase cytokines such as IL-6 and TNF-alpha. Antidepressants have been shown to suppress the inflammatory response, whereas electroconvulsive therapy results in acute elevations in proinflammatory markers. This demonstrates the nuanced nature between inflammation and depression that warrants further investigation.

Intravenous Immunoglobulins(IvIg) are well known as a broad-spectrum treatment option for many diseases involving inflammation, with minimal side effects. We present possible use of IvIgsinitially, and proposeother prophylactic and therapeutic vaccines for psychiatric illness.

Dr. Srinivasa K. Rao is a biotechnology, life sciences, biomedical scientist with experience in teaching, basic research, R&D, training, liaison and business development. Currently he is popularizing, farming and processing quinoa in India with the support of farmers in India and Bolivia. He edited a book ‘Quinoa: Nutritional & Health Benefitsʼ. He launched a brand, “Dr. Quinoa” in the Indian market to make the grain more affordable.
Dr. Rao has teaching experience in cell & molecular biology, developmental biology and biotechnology at City University of New York (CUNY). His Ph.D. thesis from the University of Paris, France in 1988 was on the molecular genetics of hemoglobinpathies. His post-doctoral work at Columbia University & the Albert Einstein College of Medicine in New York was on gene expression and regulation. He was a PI at the LIJMC (Albert Einstein College of Medicine) worked in matrix biology and molecular diseases created an enzyme detection kit. Dr. Rao published over 30 peer reviewed scientific papers. He participated in, presented at, organized, co-chaired and chaired several scientific meetings over the last 25 years in France, India and the USA.Dr. Rao led business development for Shantha Biotech in India & the United States. His efforts led to the addition of new vaccines (dengue, rubella, pertussis, human papilloma virus (HPV) to the companyʼs pipeline.Currently, Dr. Rao is the founder and President of North East Biotech, LLC. In addition to quinoa, the companyʼs other projects in the area of malnutrition include the development of a growth meter and Poshak, a software for improving nutrition and food security using Neglected and Under Utilized Species (NUS). The company also created value for small oranges in India. Dr. Rao is frequently invited to lecture and motivate students in pharma, life sciences and biotechnology. His vision is to make the next generation part of the biotech revolution to solve problems such as malnutrition. At present working on book – Vaccines in Psychiatry.

Barriers to Adult Immunizations and Strategies to Improve Coverage

Litty Varghese

SEHA/Ambulatory Healthcare Services, UAE

Vaccination against childhood communicable diseases through the Expanded Program on Immunization (EPI) is one of the most cost-effective public health interventions available. Most causes of vaccine preventable illness among children are relatively uncommon due to high rates of vaccination and very effective vaccines.

Adult vaccination is one of the most important tool for reducing morbidity and mortality not just in the elderly but in other members of the society by increasing herd immunity, reducing severe clinical outcome and helping in eradication of the disease.

The immunization program overall has been very successful with kids. So is there is a problem? Yes, there is. Weʼre not doing so well with adults.

Adults are recommended to receive vaccinations based on their age, underlying medical conditions, lifestyle, prior vaccinations, and other considerations.

Although provider recommendation is a key predictor of vaccination, more often clients report not receiving vaccine recommendations at healthcare provider visits. Other barriers to vaccinating adults, including the cost of providing vaccination services, inadequate or inconsistent payment for vaccines and vaccine administration, and acute medical care taking precedence over preventive services. Despite these challenges, we introduced a number of strategies which have been demonstrated to substantially improve adult vaccine coverage. Providers are encouraged to incorporate routine assessment of their adult patientsʼ vaccination needs during all clinical encounters to ensure patients receive recommendations for needed vaccines and are either offered needed vaccines or referred for vaccination.

Litty Varghese RN, RM, MSN is working as Senior Charge Nurse, Nursing Education, AHS/SEHA, UAE. She has 34 years of experience in the field of PHC and Ambulatory Health care Services, UAE and has expertise as JCIA chapter lead and tracer team member, quality improvement coordinator, diabetic clinic project coordinator, community program organizer, nurse educator, newsletter editor and researcher. She holds Masterʼs Degree in Community Nursing, Post Graduate Diploma in Nursing Administration and obtained certification in Principles of Epidemiology and Biostatistics from CDC, Atlanta. She taught Vaccination in the residency program in Faculty of Medicine UAE for 3 years. Her awards and recognitions with her continuance in excellent professionalism, she won the first seha best employee award 2012(one among top 12), the 3rd best abstract award at the pan arab hypertension conference in Abu Dhabi Feb. 5-9, 2000, and actively participated in winning the following awards: ARAB HEALTH AWARD FOR PHC Chronic Disease program; 2009 Arab health award for Nursing HR development: 2010. Her Philosophy is “Passionately leading and caring, the best teacher should be the best lifelong student, and we generate value for ourselves by adding value to others”

Conjugation Technology – Current Trends and Impact in Vaccine Industry

B.M.V. Srikanth Babu1*, Yuan Jun1, Charles Sia1, Michel Klein1 and Wu Ke1

Formulation and Conjugation Research Unit, Pilot Research Department, BravoVax Co. Ltd, China

Bacterial diseases are the leading cause for children deaths accounting more than 1.5 million per annum, which could be prevented by vaccines. The introduction of conjugate vaccines against bacterial pathogens such as Haemophilus influenzae type b (Hib), Streptococcus pneumonia and Neisseria meningitidis lead to a substantial reduction in mortality rate and incidence of diseases. The impact of conjugate vaccines is considerable and covers vaccination with either monovalent conjugate or in combination with other vaccines.

The preparation of conjugate vaccines is an art of science involving preparation of suitable candidate antigen (polysaccharides) which are covalently bound to a carrier protein that ultimately induces long-term immunity by evoking T-cell responses. The preparations of polysaccharides, selection of carrier protein and conjugation techniques are the major imperative factors for the development of conjugate vaccines. The range of conjugation technology is ever expanding using different carriers/ antigens and conjugation methodologies in manufacturing high-quality conjugate vaccines which ultimately provides a significant protection against infectious diseases.

The World Health Organization (WHO) has recommended the widespread immunization of Hib, pneumococcal and meningococcal vaccines in epidemic disease areas/countries. Such initiatives along with continuous development of quality conjugate vaccines will significantly contribute in reducing childhood mortality. Knowing the importance of conjugate vaccines improving global health, BravoVax Co. Ltd. (China) is focusing on the development of conjugate vaccines for various diseases as per international regulations at affordable prices.

Dr. Srikanth, a proficient with fifteen years of industrial experience in downstream processing, conjugation and formulation research of vaccines and currently working as a Research Scientist, Pilot Research department, BravoVax Co. Ltd., Wuhan, China. Did Masterʼs in Biotechnology, Doctorate degree (PhD) in Human Genetics and MBA (project management) from Indiaʼs reputed universities (Perriyar, Osmania and Manipal Universities) and also authored for ten manuscripts in various journals. During last fifteen years involved in process development, technology transfer and manufacturing of various vaccines (HPV, rHBsAg, Pneumococcal, Haemophillus, Typhoid, Cholera Vaccines etc) in Biotech vaccine industries viz. Shantha A SANOFI company, Indian Immunological and Unique Biotechnics Limited.

The Role of Bacteria in Cancer Therapy

Esmael Besufikad

Mizan-Tepi University, Ethopia

Currently, bacteria have shown promising and significant potency in preclinical mouse tumor models because of their tumor-targeting capability and ability to deliver therapeutic genes. However the successful translation of these pre-clinical strategies into clinical practice will depend on the outcome of clinical trials. Amongst all these, bacteria like S. typhimurium vector-mediated cancer therapy and immunotherapy are very promising. The use of bacterial toxins and spores has also shown great promise for cancer treatment. The exact mechanism of tumor clearance by bacteria needs to be elucidated. Focusing on the therapeutic effect of bacteria alone could be insufficient, and combination therapy is necessary to pave the way for routine application of bacteria for cancer treatment in clinics. Challenges like bacterial toxicity, stability and efficiency could be addressed by using synthetic biology.

Immunizing Pregnant Women, Benefits and Challenges

Litty Varghese

SEHA/Ambulatory Healthcare Services, UAE

Introduction: Vaccine-preventable infectious diseases are responsible for significant maternal, neonatal, and young infant morbidity and mortality. Changes in the immune response in pregnant women – which are thought to occur in order to allow the woman to tolerate the semi-allogeneic foetus – may interfere with the development of the specific immune response to pathogens. These immunological changes may alter the susceptibility of the woman and the foetus to certain infectious diseases and increase the risk of more serious outcomes. This paper discuss the benefits and challenges of immunization of pregnant women.

Immunization of pregnant women can protect them directly against vaccine-preventable infections, and in so doing potentially protect the foetus. It can also directly protect the foetus and infant via specific antibodies transferred from the mother during the pregnancy.

Pregnant women and their babies are at increased risk for influenza-related complications, including premature labour and preterm birth. Additionally, pertussis outbreaks continue to occur with infants at highest risk of severe illness and death. However, immunization coverage among pregnant women is suboptimal. Clients report concerns for the safety of vaccines and not receiving vaccine recommendations at healthcare provider visits.

Department of Health and Human services recommendation includes Assess the immunization status of each patient, Recommend the indicated vaccines to each patient, Administer any necessary vaccines or, if you do not stock the vaccine, refer the patient to a provider or location that can vaccinate the patient, Document the vaccinations that your patient is given, ideally in your state or local immunization registry.

Conclusion: Health care providers play a crucial role in helping keep pregnant women and their new born healthy. Assuring our patients are protected by recommended vaccines is key.

Litty Varghese RN, RM, MSN is working as Senior Charge Nurse, Nursing Education, AHS/SEHA, UAE. She has 34 years of experience in the field of PHC and Ambulatory Health care Services, UAE and has expertise as JCIA chapter lead and tracer team member, quality improvement coordinator, diabetic clinic project coordinator, community program organizer, nurse educator, newsletter editor and researcher. She holds Masterʼs Degree in Community Nursing, Post Graduate Diploma in Nursing Administration and obtained certification in Principles of Epidemiology and Biostatistics from CDC, Atlanta. She taught Vaccination in the residency program in Faculty of Medicine UAE for 3 years. Her awards and recognitions with her continuance in excellent professionalism, she won the first seha best employee award 2012(one among top 12), the 3rd best abstract award at the pan arab hypertension conference in Abu Dhabi Feb. 5-9, 2000, and actively participated in winning the following awards: ARAB HEALTH AWARD FOR PHC Chronic Disease program; 2009 ARAB HEALTH AWARD FOR Nursing HR development; 2010. Her Philosophy is “Passionately leading and caring, the best teacher should be the best lifelong student, and we generate value for ourselves by adding value to others”

Sub-Domain Frap of Cell Surface Molecules to Monitor Cell Surface Topography in Living Cells

Maha Al-Jumaa1* and Maurice B Hallett2

1,2Neutrophil Signalling Group, IMEM, Cardiff University School of Medicine, UK

It is becoming increasingly clear that many cells have a reservoir of excess plasma membrane in the form of cell surface wrinkles and micro-ridges. The released of these surface structures allow cells to change shape rapidly by permitting an apparent increase in cell surface area eg during phagocytosis or cell spreading. Although indirect methods point to this conclusion, it is only possible to visualise the cell surface topography under scanning electron microscopy and therefore cannot be used to follow surface topography changes to be monitored in living cells during changes in cell morphology. In this abstract, we describe a novel methodological approach which allows this using the measurement of fluorescence recovery after photo bleaching in sub domain at defined 2 dimension distances from the bleach front. Assuming that the diffusion of the fluorescent molecules is constant to rate of recovery at defined points reflects the diffusion distance and hence indicates the surface topography between the bleach front and the measurement domain.

Human neutrophils were isolated from blood and there plasma membrane labelled with DiI. The dye can be excited using a 543nm laser line and effectively photobleached using the 488nm line transiently elevated. A photo-bleaching zone across the region of the cell of interest was chosen and the image of the cell recording during bleaching and recovery. The bleach/recovery cycle could be repeated.

This approach was showed to reflect the cell surface topography by experimentally increasing the wrinkledness of the cell surface using hyperosmotic media. The measured apparent diffusion length increased in the increased wrinkled state and was returned to close to its pre-shrunk value by restoration of isotonicity. The methodology showed that whereas the body of neutrophils has significant ridge-like topographical features, the phagosome and the extending pseudopodia are smooth. This method also showed that elevation of cytosolic Ca2+ (by uncaging IP3) influenced the cell surface topography.

This methodology opens a novel way of monitoring an important cell characteristic which has not be possible to study previously.

Fluorescent Isothiocyanate Dextran Evaluates the Permeability of Blood-Brain Barrier in Rabies Infected Mice Model

Waqas Ahmad1,2*, Liu Xinyue1, Li Yingying1, Xinyu Wang1, Ming Duan1, Zhenhong Guan1 and Zhang Maolin1

1Key Laboratory of Zoonosis, Ministry of Education, Institute of Zoonosis, Jilin University Peopleʼs Republic of China
2Section of Epidemiology and Public Health, College of Veterinary and Animal Sciences, Jhang, Pakistan

Rabies is primarily a horrifying viral zoonosis that annually accounts for 55,000 deaths worldwide. Acute encephalitis develops as the rabies virus (RABV) enters to the central nervous system by crossing the blood brain barrier (BBB) which is a tight junction of endothelial cells. In this study, three different molecular weights (70 kDa, 150 kDa and 200 kDa) of fluorescent isothiocyanate dextrans (FITC-Dextrans) were used to measure the extent of BBB damage and subsequent leakage patterns in brain tissues of rabies infected mice which were post-immunized with neutralizing antibodies to observe whether it has positive effect on infected mice by decreasing the death ratio. The brains were processed for immunofluorescence to observe the neutralizing antibodies and its relevant compatibility with the leakage of FITC- Dextrans. Results showed that 70 kDa and 150 kDa FITC-Dextrans efficiently crossed BBB, and produced fluorescent illumination mainly in the cerebral cortex of brain. The enhancement of BBB permeability was significant at 5th day of post-immunization, while the neutralizing antibody neutralized some rabies virus particles by crossing BBB, but it did not present enough treatment effect to the dying mice. These findings suggest that FITC-Dextran is an important fluorescent marker to investigate the integrity of BBB permeability.

Keywords: Rabies virus, blood-brain barrier, Evans blue, FITC-Dextrans, neutralizing antibody.

Mr. Waqas Ahmad, Completed matriculation in 2006 and F.Sc Pre-Medical in 2008, both from Pakistan Urdu School & Higher Secondary Education - Bahrain via F.B.I.S.E - Islamabad. Completed D.V.M with a Gold Medal in 2013 form University of Veterinary Sciences-Lahore. Joined Small Animal Practice as Teaching Assistant at Pet Centre-UVAS after graduation in 2013. Got promoted to the status of Lecturer at Pet Centre in 2015. His hobbies include collecting coins and postal stamps, gardening, video gaming, fish keeping, etc. Area of interest include soft tissue surgery, ultrasonography, exotic animal medicine, human-pet bonding, mobility and pain management, injury and surgery rehabilitation

The Trans-Boundary Disease and the Problem of Continues Vaccination

Samia Ahmed Kamal

Animal Health Research Institute, Egypt

How to address the continued presence of rift valley fever virus in neighboring countries that do not take any measures to resist the virus and import live animals. This problem is ongoing and cannot be tolerated or ignored. Because the risk of weather fluctuations exists and the possibility of a suitable environment for those viruses exists. The effect of these conditions has been evident in the foot-and-mouth disease, which has always taken place in the Egyptian environment. The various infectious diseases, which do not exist in Egypt, enter through the port of Sudan to represent a direct outlet for other African countries that do not have a border. The continued importation of camels and cattle from Sudan is considered a permanent threat. The continued vaccination of animals with Rift Valley Fever vaccine for decades is also inappropriate behavior to address this problem. As the possibility of cheating in the vaccine, especially in a country like Egypt, puts the country at risk of a pandemic through the vaccine, which has already happened before. The application of vaccine safety tests is not enough, because it does not include the experiment on the target animals so fully that we observe the pathological effect of the tested vaccine, which include strict and rigorous pathological examination. Ignoring the source of natural infection and continuing to import these animals is the real problem that must be faced. The control of disease-carrying insects must be effective and severe. Both in countries exporting disease and receiving countries.

Dr. Samia Ahmed Kamal is (Associate professor) first researcher in virology department in AHRI of Egypt. She graduated from Faculty of Veterinary medicine, ZagazigUniversity in 1989. She worked for Animal Health Research Institute in 1991 till now. Her postgraduate studies are; Master degree in Veterinary Pathology, Zagazig University (1996), Ph.D. in Veterinary Pathology, Cairo University (2005) and Diploma of public health (microbiology) from Alexandria University (2009). In the present time, she focused on her researches in the field of virology, vaccines and public health (microbiology).

Development and Evaluation of Eimeria Tenella (Indigenous/Local Strain) Vaccine(S) against Coccidiosis in Chickens

Mian Abdul Hafeez

University of Veterinary & Animal Sciences, Pakistan

Sporozoites of E.tenella(local isolate) were adapted on the chorioallantoicmembrane(CAM) of chicken embryos. Gametocytes were harvested from CAM and chorioallantoic fluid. Egg adapted gametocytes (E. tenella) were used to prepare adjuvanted(Amphigen) vaccine and compared with the non-adjuvanted by immunizing chicks through oral and subcutaneous route in comparison with controls. Egg adapted gametocytes (EAG)of E. tenella and wall fragments from sporulated and un-sporulated oocysts were subjected to SDSPAGE to fractionate the proteins. Five random samples taken from different aliquots of EAG were analyzed. All the samples of gametocyte showed similar protein bands having molecular weights of 48.43, 27.20, 25.75, 22.57 and 12.92 kDa. Sporozoites-specific ELISA with purified sporozoites of E. tenella revealed that antibodies produced due to EAG vaccine are sporozoite specific. Humoral response in vaccinated and control chicks was detected by IHA test and ELISA. Results of all groups with IHA geomean titer and ELISA OD values were partially comparable. A significant difference (P>0.01) in geomean titer and OD values among vaccinated and control groups at day 5th and 14th post vaccination was recorded. Cell mediated immunity (CMI) of the vaccinated and control groups were detected by Modified splenic cell migration inhibition assay. There was significant difference (P>0.01) in migration index of vaccinated groups with control group either with or without antigen and non-significant difference (P>0.01) was observed in vaccinated groups with or without adjuvanted vaccine. On 10th days post challenge, organ body weight ratio of Lymphoid was calculated. The ratio of the vaccinated chicks remains normal while increased in non-vaccinated chicks (control groups).Sporulated oocysts of mixed species of genus Eimeria were used to challenge the vaccinated and control chicks. Both vaccines check the mortality but maximum protection (71.42 %) against heavy doses of infection was recorded by adjuvanted vaccine given orally. Maximum reduction (86%) in oocyst count was also observed in group A given adjuvanted vaccine orally.

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