Bishop D K, Ferguson R M, Orosz C G
Department of Surgery, Ohio State University College of Medicine, Columbus 43210.
J Immunol. 1990 Feb 15;144(4):1153-60.
We have developed modified limiting dilution analysis (LDA) techniques that distinguish in vivo Ag-stimulated murine helper T lymphocytes (HTL) and CTL from unstimulated precursor T cells, even those with the same Ag specificity. We refer to these cells that are detectable in the modified LDA as "Ag-conditioned" T cells (cHTL and cCTL). We have used the modified LDA techniques in conjunction with conventional LDA techniques (which enumerate all Ag-specific T cells) to evaluate the in vivo distribution of Ag-conditioned cHTL and cCTL following in vivo sensitization to alloantigens via sponge matrix or skin allografts. In general, we observed the following regarding the distribution of cHTL and cCTL: 1) Ag-conditioned HTL and CTL were detectable only after in vivo sensitization with alloantigen: 2) not all Ag-reactive T cells became conditioned T cells after in vivo Ag deposition; 3) the percentage of Ag-reactive T cells that converted to conditioned T cells after Ag deposition varied among different lymphoid compartments; 4) a high percentage of cHTL, but a low percentage of cCTL, accumulated in regional lymph nodes and spleen; 5) cHTL accumulated in peripheral blood, whereas cCTL did not; 6) Ag-conditioned cHTL were detectable in various lymphoid tissues for greater than 60 days following Ag deposition, whereas cCTL were detectable for only 14 to 20 days; and 7) unlike the other lymphoid sites, the site of Ag deposition accumulated a high percentage of both Ag-stimulated cHTL and cCTL. Furthermore, cHTL and cCTL appeared to reside in phenotypically distinct T cell subsets in that in vivo treatment with anti-L3T4 mAb abrogated the accumulation of HTL, but not CTL, at the site of Ag deposition. These data demonstrate differential compartmentalization of Ag-conditioned cHTL and cCTL subsequent to in vivo Ag deposition. The implications of these findings regarding the monitoring of in vivo immune responses are discussed.
我们已经开发出改良的有限稀释分析（LDA）技术，该技术能够区分体内抗原刺激的小鼠辅助性T淋巴细胞（HTL）和细胞毒性T淋巴细胞（CTL）与未受刺激的前体T细胞，即使是那些具有相同抗原特异性的细胞。我们将在改良LDA中可检测到的这些细胞称为“抗原预处理”T细胞（cHTL和cCTL）。我们已将改良的LDA技术与传统LDA技术（用于计数所有抗原特异性T细胞）结合使用，以评估通过海绵基质或皮肤同种异体移植在体内致敏同种异体抗原后，抗原预处理的cHTL和cCTL在体内的分布情况。一般来说，关于cHTL和cCTL的分布，我们观察到以下几点：1）只有在用同种异体抗原进行体内致敏后才能检测到抗原预处理的HTL和CTL；2）并非所有抗原反应性T细胞在体内抗原沉积后都会变成预处理T细胞；3）抗原沉积后转化为预处理T细胞的抗原反应性T细胞百分比在不同淋巴区室中有所不同；4）高比例的cHTL，但低比例的cCTL，积聚在局部淋巴结和脾脏中；5）cHTL积聚在外周血中，而cCTL则不然；6）抗原沉积后60多天，在各种淋巴组织中均可检测到抗原预处理的cHTL，而cCTL仅在14至20天内可检测到；7）与其他淋巴部位不同，抗原沉积部位积聚了高比例的抗原刺激的cHTL和cCTL。此外，cHTL和cCTL似乎存在于表型不同的T细胞亚群中，因为用抗L3T4单克隆抗体进行体内治疗消除了抗原沉积部位HTL的积聚，但没有消除CTL的积聚。这些数据表明，体内抗原沉积后，抗原预处理的cHTL和cCTL存在不同的区室化。讨论了这些发现对体内免疫反应监测的意义。
