Stukart M J, Vos A, Melitef C J
Eur J Immunol. 1981 Mar;11(3):251-7. doi: 10.1002/eji.1830110316.
A method for infection of lymphocytes with Moloney(Abelson) murine leukemia virus [M(A)-MuLV] is described. Only lymphoblasts obtained after stimulation of normal spleen cells by the B cell mitogen lipopolysaccharide (LPS) were satisfactory targets for virus-specific, secondary cytotoxic T lymphocytes (CTL), whereas spleen cells stimulated by the T cell mitogen concanavalin A were not. The secondary CTL response against M(A)-MuLV could be efficiently measured using M(A)-MulV-infected LPS blasts as stimulating cells for secondary in vitro restimulation and as target cells for virus-specific destruction. Cold target inhibition demonstrated virus specificity of CTL. The T cell character of the cytotoxic cells was demonstrated by their sensitivity to anti-Thy-1.2 treatment. Using syngeneic virus-infected LPS blasts as target and stimulator, CTL responses were measured with effector cells from C57BL mice of the H-2b haplotype and of recombinant haplotypes sharing either K or D alleles with H-2b. In analogy with previous studies on Moloney virus-specific CTL, it was observed that C57BL/6 (H-2b) effector cells predominantly lysed Db-compatible, virus-infected target cells; B10.A(5R), (KbDd) effector cells showed a poor CTL response against syngeneic, virus-infected target cells. The combined findings indicate the existence of an Ir gene in the H-2D region regulating the CTL response against Moloney leukemia virus.
描述了一种用莫洛尼(阿贝尔森)鼠白血病病毒 [M(A)-MuLV] 感染淋巴细胞的方法。只有在用B细胞有丝分裂原脂多糖(LPS)刺激正常脾细胞后获得的淋巴母细胞才是病毒特异性二级细胞毒性T淋巴细胞(CTL)的满意靶细胞,而用T细胞有丝分裂原伴刀豆球蛋白A刺激的脾细胞则不是。使用感染了M(A)-MuLV的LPS母细胞作为二级体外再刺激的刺激细胞和病毒特异性破坏的靶细胞,可以有效地检测针对M(A)-MuLV的二级CTL反应。冷靶抑制证明了CTL的病毒特异性。细胞毒性细胞的T细胞特性通过它们对抗Thy-1.2处理的敏感性得以证明。使用同基因病毒感染的LPS母细胞作为靶细胞和刺激细胞,用来自H-2b单倍型的C57BL小鼠以及与H-2b共享K或D等位基因的重组单倍型的效应细胞测量CTL反应。与先前关于莫洛尼病毒特异性CTL的研究类似,观察到C57BL/6(H-2b)效应细胞主要裂解与Db相容的病毒感染靶细胞;B10.A(5R),(KbDd)效应细胞对同基因病毒感染靶细胞的CTL反应较差。综合研究结果表明,在H-2D区域存在一个Ir基因,其调节针对莫洛尼白血病病毒的CTL反应。