Detection of Jaagsiekte sheep retrovirus in the peripheral blood during the pre-clinical period of ovine pulmonary adenomatosis

Abstract

ABSTRACT. 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-Env 572-615 antigen was prepared via peptide synthesis (amino acid sequence 572-615, denoted as JSRV-Env 572-615 ). BALB/c mice were immunized to prepare the anti-JSRV-Env 572-615 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.

Clinical-histopathological and molecular study of ovine pulmonary adenocarcinoma in Awassi sheep in Al-Qadisiyah Province, Iraq

“…PCR analysis of the all the samples obtained revealed the presence of OPA in the sheep in the studied regions to be 25%, indicating the reliability of PCR for early screening of the disease, even in subclinical cases [17,18]. This result is in agreement with those of the previous studies in which the virus in the sheep was detected using PCR for the 1 st time in Ireland [19-21]. The finding is also supported by those of many studies in which OPA was diagnosed using PCR, such as the ones conducted in Iraq (5.55%) [10], India (8%) [12], Iran (13.75%) [22], and Northwest Iran (18%) [23].…”

Exogenous Jaagsiekte Sheep Retrovirus type 2 (exJSRV2) related to ovine pulmonary adenocarcinoma (OPA) in Romania: prevalence, anatomical forms, pathological description, immunophenotyping and virus identification

“…Currently, this method is not extensively used because the sample collection requires sedation. Furthermore, intravitam diagnosis of exJSRV infection by PCR from blood offers many false negative results probably because the numbers of infected white blood cells is very low [26,27]. Additionally, the viral genetic material or proteins can also be detected by PCR and immunohistochemistry (IHC) respectively, in pulmonary and lymphoid tissues [16,17,28].…”

Exogenous Jaagsiekte Sheep Retrovirus type 2 (exJSRV2) related to ovine pulmonary adenocarcinoma (OPA) in Romania: prevalence, anatomical forms, pathological description, immunophenotyping and virus identification

“…Currently, this method is not extensively used because the sample collection requires sedation. Furthermore, intravitam diagnosis of exJSRV infection by PCR from blood offers many false negative results probably because the numbers of infected white blood cells is very low [26,27]. Additionally, the viral genetic material or proteins can also be detected by PCR and immunohistochemistry (IHC) respectively, in pulmonary and lymphoid tissues [16,17,28].…”