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人淋巴细胞功能相关抗原3(hLFA3)是一种重要的T细胞黏附分子,其与CD2结合后可促进T细胞活化。恒河猴被广泛用作人类免疫疾病动物模型。因免疫系统存在严格的种属特异性,有必要研究恒河猴LFA3的功能。本文通过PCR扩增获得mmLFA3Sh基因,将其与人IgG1Fc片段融合,插入毕赤酵母表达载体构建表达质粒pPIC9K-mmLFA3Sh-Ig。诱导表达后获得融合蛋白mmLFA3Sh-Ig,产量为3~4mg/L。mmLFA3Sh-Ig能够特异性与CD2阳性细胞结合,能够抑制Con A和同种抗原刺激的猴和人淋巴细胞增殖。这些实验结果表明,mmLFA3Sh-Ig可能作为一种新的生物制剂用于研究恒河猴免疫疾病发病机制和实验治疗。
Human lymphocyte function-associated antigen 3 (hLFA3) is an important T cell adhesion molecule that, when combined with CD2, promotes T cell activation. Rhesus macaques are widely used as animal models of human immunodeficiency. Due to the strict species-specific immune system, it is necessary to study the function of rhesus LFA3. In this paper, we amplified the mmLFA3Sh gene by PCR and fused it with the human IgG1 Fc fragment. The recombinant plasmid pPIC9K-mmLFA3Sh-Ig was constructed by inserting into Pichia pastoris expression vector. After induction of expression, the fusion protein mmLFA3Sh-Ig was obtained and the yield was 3 ~ 4mg / L. The ability of mmLFA3Sh-Ig to specifically bind CD2-positive cells inhibits both Con A and alloantigen-stimulated monkey and human lymphocyte proliferation. These experimental results show that the mmLFA3Sh-Ig may be used as a new biological agent for studying the pathogenesis and experimental treatment of immune disorders in rhesus monkeys.