Vijayarani, K.Kumanan, K.Roy, ParimalThangavelu, A.Gowralkar, MaheswarappaTANUVAS2016-07-252016-07-252012http://krishikosh.egranth.ac.in/handle/1/69816Chicken are the most abundant birds in the world with a population of more than 24 billion. They are mainly kept for egg and meat production. With increasing industrialization and intensification of poultry rearing systems, the disease pattern is also changing. In developing countries, the infectious diseases still interfere with economic returns from the poultry industry. Tropical countries have more problems with infectious diseases due to the climatic circumstances. Important economic lossess for the industry can occur, due to viral diseases such as Avian influenza and Newcastle disease. Vaccination is the only available tool to curb the outbreak of epidemics in poultry. Till today only few good vaccines are available to prevent the epidemic diseases. Recombinant DNA technology led to the development of recombinant vaccines and subunit vaccines but immune potency of recombinant vaccines and subunit vaccines are still not well understood in Newcastle disease vaccines. Hence, in this study, we cloned and expressed the fusion and haemagglutinin neuraminidase genes of NDV in insect cells. Immunological study was carried out to assess the immunopotency of rec-F and rec-HN antigens in commercial broilers. Humoral and cell mediated immune response of recombinant proteins were evaluated. The immunogenic potentials of recombinant antigens along with commercial ND live and inactivated ND vaccine in stimulating humoral antibody response was recorded by haemagglutination inhibition (HI) test and ELISA. Cell mediated immune responses in stimulating the peripheral blood mononuclear cells by recombinant antigens were measured using lympoproliferation assay (MTT assay). Further, expression of cytokines such as interferon alpha and interferon gamma stimulated by recombinant F and HN antigens and commercial ND vaccines were studied by reverse transcriptase real-time polymerase chain reaction. Results of this study revealed that recombinant antigens had similar immuneresponse as commercial inactivated ND vaccines and mixed antigens vaccine has significant effect in cell mediated immune response with high peak expression of INF-gamma compared to commercial live vaccines. Booster dose of fusion protein has shown higher protection percentage than booster dose of HN protein in challenged broilers and combination of both antigens showed significant protection percentage than individual antigen. However, commercial ND vaccine has shown highly significant protection percentage than recombinant proteins but we could still believe that recombinant protein vaccines were more efficient. Since, immunodominant region of virus was used instead of whole virus. Recombinant vaccines were safer and affordable in comparison to other type of commercial vaccines. Many viruses are studied in details for their apoptotic and oncolytic properties among them Newcastle disease virus (NDV) is one of the important virus in consideration. Reverse genetics approach has allowed in engineering and constructing more potent Newcastle disease virus with a better result in treatment of malignant cancers in humen and aminals. In this study, the recombinant fusion and recombinant haemagglutininneuraminidase (HN) genes of NDV9 were cloned and expressed in insect cells. The apoptotic and oncolytic properties of recombinant antigens were studied in chicken embryo fibroblast (CEF),Vero and six different cancerous cell lines (HeLa, Hep2, HepG2, A72, A549 and MCF7). Apoptotic and antineoplastic properties of recombinant fusion and recombinant-HN proteins were confirmed using MTT assay, flowcytometry, in vitro comet assay and TUNEL assay. Results indicated that recombinant fusion protein causes more apoptotic changes than recombinant HN protein at the concentration of 2 μg/ml at 48 hrs post-treatment. Similarly, antineoplastic activity of rec-F and rec-HN proteins in HeLa, Hep2, HepG2, A72, A549 and MCF7 cells revealed that recombinant fusion protein has significant anticancerous property than recombinant HN protein at the concentartion of 2 μg/ml at 48 hrs post-treatment. The results of this study strongly support the hypothesis that surface glycoproteins such as fusion and HN of NDV are responsible of apoptosis and oncolysis in the oncolytic virotherapy.enLive and inactivated vaccinesOncolyticNewcastle disease virusTUNEL assayThesis