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Title: DEVELOPMENT AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO SUBGROUP J AVIAN LEUKOSIS VIRUS

Author
item QIN, AIJIAN - YANGZHOU UNIVERSITY PRC
item Lee, Lucy
item Fadly, Aly
item Hunt, Henry
item CUI, ZHIZHONG - YANGZHOU UNIVERSITY

Submitted to: Avian Diseases
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 5/16/2001
Publication Date: 12/31/2001
Citation: QIN, A., LEE, L.F., FADLY, A.M., HUNT, H.D., CUI, Z. DEVELOPMENT AND CHARACTERIZATION OF MONOCLONAL ANTIBODIES TO SUBGROUP J AVIAN LEUKOSIS VIRUS. AVIAN DISEASES. 2001. V. 45. P. 938-945.

Interpretive Summary: Subgroup J avian leukosis virus (ALV-J) is an emerging, economically important virus infection that can cause cancer-like disease and other production problems in meat-type chickens. The virus was first reported in 1991 in the United Kingdom and in 1993 in the United States. In an attempt to develop specific diagnostics and effective control programs, we developed four immunological reagents (monoclonal antibodies) specific for ALV-J. Using these reagents, we detected ALV-J in cells infected with various isolates of the virus from United States. The information obtained from this research is of great interest to scientists in industry and academia as these reagents will aid in developing diagnostic kits for detection of ALV-J infection in the poultry farm.

Technical Abstract: In an attempt to develop a specific diagnostic test for ALV-J strain Hcl, four monoclonal antibodies (Mabs JE9, G2, 145 and J47) were generated which are specific for ALV-J envelope (env) glycoprotein, gp85. Polymerase chain reaction (PCR) was used to amplify genomic pro-viral DNA of ADOL-Hcl and ADOL-4817 envelope (env) genes. Both open reading frames(ORF) encoding glycoproteins gp85 and gp37 were cloned into baculoviruses. Abundant expression of gp85 and gp37 was detected in the recombinant viruses using specific antibody to Hc1 strain of the ALV-J. The expressed proteins were used for immunization of mice to produce hybridoma cell lines secreting monoclonal antibodies (Mab) specific to ALV-J env protein. A panel of Mabs was generated by fusing NS1 myeloma cells and spleen cells from mice immunized with the recombinant baculoviruses. Using an immunofluorescence assay (IFA), three Mabs (JE9, G2, 145) reacted with ALV-J, but not with subgroups A, B, C, D or E of ALV. Mab J47 reacted with all exogenous subgroups of ALV including A, B, C, D, and J but not with endogenous subgroup E viruses. Western Blot analysis was performed using all four Mabs against recombinant baculovirus and Hc1 infected chicken embryo fibroblasts (CEF) lysates. A major band with a molecular weight about 90 kDa corresponding to the size of ALV-J envelope was consistently obtained. Using these Mabs we detected the Hc1 antigen in CEF infected with several ALV-J viruses isolated from the United States and also tissue sections from infected chickens.