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Genet Mol Res ; 15(3)2016 Aug 29.
Artigo em Inglês | MEDLINE | ID: mdl-27706644

RESUMO

The envelope protein (Env) of the Jaagsiekte sheep retrovirus (JSRV) is known to be a unique oncoprotein responsible for inducing ovine pulmonary adenocarcinoma (OPA). The objective of this study was to prepare a specific monoclonal antibody (mAb) against the JSRV Env protein using bioinformatic analysis. According to the structure and epitope prediction results of JSRV Env, the JSRV-Env572-615 antigen was prepared via peptide synthesis (amino acid sequence 572-615, denoted as JSRV-Env572-615). BALB/c mice were immunized to prepare the anti-JSRV-Env572-615 mAb. Spleen cells were fused with SP2/0 myeloma cells after being screened by indirect ELISA and cloned by limiting dilution. The specificity of mAb was evaluated by western blot analysis and immunohistochemistry assays. Western blot results showed that the JSRV Env protein was able to bind to mAb with high specificity. Immunohistochemistry assays demonstrated that the mAb was able to recognize JSRV Env in adenomatous hyperplasia of the lung. Furthermore, JSRV was detected in peripheral blood leukocytes during the pre-clinical period of OPA in 2 of the 25 sheep using this newly synthesized mAb. Therefore, this mAb may be a useful tool for the detection of JSRV in sheep.


Assuntos
Adenocarcinoma/diagnóstico , Adenocarcinoma/veterinária , Anticorpos Monoclonais/biossíntese , Anticorpos Antivirais/biossíntese , Retrovirus Jaagsiekte de Ovinos/imunologia , Neoplasias Pulmonares/diagnóstico , Neoplasias Pulmonares/veterinária , Adenomatose Pulmonar Ovina/diagnóstico , Adenocarcinoma/imunologia , Adenocarcinoma/virologia , Adenocarcinoma de Pulmão , Sequência de Aminoácidos , Animais , Anticorpos Monoclonais/química , Anticorpos Monoclonais/isolamento & purificação , Anticorpos Antivirais/química , Anticorpos Antivirais/isolamento & purificação , Especificidade de Anticorpos , Biologia Computacional , Diagnóstico Precoce , Epitopos/química , Epitopos/imunologia , Produtos do Gene env/química , Produtos do Gene env/imunologia , Retrovirus Jaagsiekte de Ovinos/isolamento & purificação , Leucócitos Mononucleares/imunologia , Leucócitos Mononucleares/virologia , Pulmão/imunologia , Pulmão/virologia , Neoplasias Pulmonares/imunologia , Neoplasias Pulmonares/virologia , Camundongos , Camundongos Endogâmicos BALB C , Peptídeos/administração & dosagem , Peptídeos/síntese química , Peptídeos/imunologia , Adenomatose Pulmonar Ovina/imunologia , Adenomatose Pulmonar Ovina/virologia , Ovinos , Carneiro Doméstico , Baço/citologia , Baço/imunologia
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