T淋巴细胞中的应用

Goose CD4 extracellular region resisting monoclonal antibody and application of goose CD4 extracellular region resisting monoclonal antibody to detecting CD4+T lymphocytes in peripheral blood of poultry

Abstract

本发明公开了抗鹅CD4胞外区单克隆抗体及其在检测禽类外周血中CD4+T淋巴细胞中的应用。本发明利用携带鹅CD4胞外区的原核表达载体pET-30a-CD4ex,采用大肠杆菌RosettaTM(DE3)pLysS表达鹅CD4胞外区基因,得到重组蛋白rGopET-30a-CD4ex。以重组蛋白rGopET-30a-CD4ex为免疫原,免疫三次BALB/c小鼠,加强免疫后,经重组蛋白rGopET-30a-CD4ex筛选获得1株稳定分泌抗鹅CD4胞外区单克隆抗体的杂交瘤细胞株,命名为5A9,保藏号是:CGMCC?NO.11192。研究表明,由该杂交瘤细胞株分泌的单克隆抗体能与纯化的重组鹅CD4胞外区蛋白rGopET-30a-CD4ex反应,同时也能够与分离的鹅、鸭以及鸡外周血淋巴细胞发生特异性反应。因此,本发明的提出为检测禽类外周血中CD4+T淋巴细胞提供了一种新的、有效的技术手段。 CGMCC NO.11192 20150914
The invention discloses a goose CD4 extracellular region resisting monoclonal antibody and application of the goose CD4 extracellular region resisting monoclonal antibody to detecting CD4+T lymphocytes in peripheral blood of poultry. Genes of goose CD4 extracellular regions are expressed by Escherichia coli RosettaTM(DE3) pLysS by the aid of prokaryotic expression vectors pET-30a-CD4ex which carry the goose CD4 extracellular regions, so that recombinant proteins rGopET-30a-CD4ex can be obtained. The recombinant proteins rGopET-30a-CD4ex are used as immunogens, BALB/c mice are immunized thrice, the recombinant proteins rGopET-30a-CD4ex are screened after the immunity of the BALB/c mice is strengthened to obtain a hybridomas cell strain capable of stably secreting thegoose CD4 extracellular region resisting monoclonal antibody, the hybridomas cell strain is named as 5A9, and a preservation number of the hybridomas cell strain is CGMCC NO.11192. The goose CD4 extracellular region resisting monoclonal antibody and the application have the advantages that as shown by research, the goose CD4 extracellular region resisting monoclonal antibody secreted by the hybridomas cell strain can react to purified recombinant proteins rGopET-30a-CD4ex of the goose CG4 extracellular regions and also can specifically react to separated lymphocytes in the peripheral blood of geese, ducks and chicken, and a novel and effective technical means is provided for detecting the CD4+T lymphocytes in the peripheral blood of the poultry.

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