Rodewald H R, Koszinowski U H, Eichmann K, Melchers I
Max-Planck-Institute for Immunobiology, Freiburg, FRG.
J Immunol. 1989 Dec 15;143(12):4238-43.
Cytotoxic T cell responses to the murine Cytomegalovirus (MCMV) were elicited in BALB/c mice (H-2d) by infectious virus. Eight days after infection, MCMV-primed local lymph node T cells were either depleted for T cells expressing a V beta 8+ TCR or separated into V beta 8+ and V beta 8- subpopulations by a cell sorter using the mAb F23.1. T cells were then expanded in vitro under limiting dilution conditions in the presence of IL-2 and in the absence of viral Ag to avoid selection by Ag in vitro. Frequencies of CTL precursors specific for the Immediate-Early-Ag 1 of MCMV and restricted to H-2Ld were determined. L cells of the endogenous haplotype H-2k cotransfected with the genes for MCMV-IE 1 and H-2Ld were used as target cells. Detection of a CTL response required previous priming of the animals by infection in vivo (less than 1/10(6) for nonimmunized animals). In primed animals CTL precursors of this specificity and restriction were three to fivefold more frequent in the V beta 8+ population (1/9.900 to 1/22.300) than in the V beta 8- population (1/57.000 to 1/87.200). Control experiments showed that frequencies were not influenced by the treatment with the anti-V beta 8-antibody and the fluorescein-labeled anti-Ig itself. V beta 8+ and V beta 8- T cells did not reveal any frequency differences when several other responses were determined (TNP-specific self-restricted CTL precursor; Th cells specific for keyhole limpet hemocyanin or Listeria monocytogenes).
通过感染性病毒在BALB/c小鼠(H-2d)中引发对鼠巨细胞病毒(MCMV)的细胞毒性T细胞反应。感染后8天,用表达Vβ8+ TCR的T细胞耗尽MCMV致敏的局部淋巴结T细胞,或者使用单克隆抗体F23.1通过细胞分选仪将其分离为Vβ8+和Vβ8-亚群。然后在有限稀释条件下,在存在IL-2且不存在病毒抗原的情况下体外扩增T细胞,以避免体外抗原选择。测定了对MCMV的立即早期抗原1特异且受H-2Ld限制的CTL前体细胞频率。将内源性单倍型H-2k的L细胞与MCMV-IE 1和H-2Ld基因共转染用作靶细胞。检测CTL反应需要事先通过体内感染使动物致敏(未免疫动物的频率小于1/10(6))。在致敏动物中,这种特异性和限制性的CTL前体细胞在Vβ8+群体(1/9,900至1/22,300)中的频率比在Vβ8-群体(1/57,000至1/87,200)中高3至5倍。对照实验表明,频率不受抗Vβ8抗体和荧光素标记的抗Ig自身处理的影响。当测定其他几种反应时(TNP特异性自身限制性CTL前体细胞;对钥孔戚血蓝蛋白或单核细胞增生李斯特菌特异的Th细胞),Vβ8+和Vβ8- T细胞未显示任何频率差异。