الفهرس 
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Abstract
Benisueif University
Faculty of veterinary medicine
Dept. of virology
Name : Wesam Hassan Mohamed Mady Mohamed
Nationality : Egyptian
Date of birth :18/2/1983
Place of birth: Cairo
Specification :Virology
Thesis title : Preparation of DNA H5 Avian Influenza vaccine
Supervisors : Prof. Dr. Hanafy Mahmoud Madbouly, Dr. Ahmed Saad Hussein, Prof. Dr. Mona MehrezAly and Dr. Abdel SatarArafa Mohamed
Abstract
Since the DNA vaccine seemed to be a promising novel approach for vaccination against influenza A virus, the goal of this study was to prepare HA1 based DNA vaccine against the H5N1 avian influenza viruses circulating in Egypt.The HA1 gene from Egyptian virus A/chicken/Egypt/1055/2010(H5N1) was extracted and cloned into PCIneo mammalian expression vector. The invitro expression of recombinant H5-plasmid DNA was confirmed in 293T human embryonic kidney (HEK) cell line and in Vero cell line by SDS-PAGE and by detection of mRNA using RT-PCR. The immunological response was investigated by intramuscular immunization of SPF chickens with PCIneo-H5 lipofectamine and nigella sativa adjuvant plasmid DNA vaccine. The immunological analysis showed that PCIneo-HA1 vaccine induced both humoral and cell mediated immune response in chickens. The protection of H5-DNA vaccine was evaluated by challenge of 3 weeks old SPF chickens vaccinated with PCIneo-H5 DNA vaccine using one dose of conc. 5µg/dose by direct intramuscular injection in thigh muscle and compared with protection afforded by commercial volvacH5N2 AI vaccine. The obtained results showed that the H5 DNA vaccine protected the chickens from the Egyptian field H5N1 virus better than the H5N2 inactivated vaccine (60% versus 40% respectively). This is the first paper describing the development and primary evaluation for DNA vaccine prepared from Egyptian H5N1 virus. Further adjustments of the newly developed H5-DNA vaccine dose and vaccination regimen are required to better protect chickens against the virulent field viruses.