Good M F, Nossal G J
J Immunol. 1983 Dec;131(6):2662-9.
Immunologic tolerance to the hapten TNP was induced in adult mice through the i.v. injection of reactive TNBS. To probe the cellular basis of the tolerant state, splenic cytotoxic T lymphocyte precursors (CTL-P) were stimulated in vitro with haptenated, x-irradiated syngeneic spleen cells in the presence or absence of exogenously added growth factors derived from Concanavalin A-stimulated spleen cell conditioned medium (CAS). The cultures were either conventional bulk cultures or limit dilution cloning cultures. For the latter, cytotoxicity was assessed through a semi-automated, radioautographic 111In-release assay. Suppressive potential was assessed by mixing spleen cells from tolerant mice with normal spleen cells before culture. In the absence of CAS, bulk cultures showed profound tolerance, and suppressive capacity was clearly evident. Suppression was dependent on the presence of TNP-self during culture and affected the generation of CTL from CTL-P and not the effector function of CTL. Cyclophosphamide treatment did not prevent tolerance induction. In the presence of CAS, bulk cultures still showed marked tolerance, but mixing experiments now yielded no evidence of suppression. As documented previously, limit dilution cultures of tolerant spleen cells in the presence of CAS showed a functional clonal deletion of hapten-specific CTL-P. In the absence of CAS, limit dilution cultures became dependent on helper T cells as the limiting element. Tolerant populations showed a diminution of activatable helper T lymphocyte precursors (HTL-P), which may have been due to a functional clonal deletion of HTL-P and/or a concomitant activation of suppressor T cells. Adoptive transfer studies showed that cells from tolerant mice did not detectably influence the number of hapten-specific CTL-P in the spleens of host animals. Taken together, the results suggest that both functional clonal deletion of CTL-P and suppression of HTL-P contribute to the tolerant state induced.
通过静脉注射反应性三硝基苯磺酸（TNBS），在成年小鼠中诱导对半抗原三硝基苯（TNP）的免疫耐受。为了探究耐受状态的细胞基础，在有或没有外源性添加的源自刀豆球蛋白A刺激的脾细胞条件培养基（CAS）的生长因子存在的情况下，用半抗原化的、经X射线照射的同基因脾细胞在体外刺激脾细胞毒性T淋巴细胞前体（CTL-P）。培养物为常规批量培养或有限稀释克隆培养。对于后者，通过半自动放射自显影铟-111释放试验评估细胞毒性。通过在培养前将耐受小鼠的脾细胞与正常脾细胞混合来评估抑制潜力。在没有CAS的情况下，批量培养显示出深度耐受，并且抑制能力明显可见。抑制依赖于培养期间TNP-自身的存在，并且影响从CTL-P产生CTL，而不是CTL的效应功能。环磷酰胺处理不能阻止耐受诱导。在有CAS的情况下，批量培养仍然显示出明显的耐受，但混合实验现在没有产生抑制的证据。如先前所述，在有CAS的情况下，耐受脾细胞的有限稀释培养显示出半抗原特异性CTL-P的功能性克隆缺失。在没有CAS的情况下，有限稀释培养变得依赖于辅助性T细胞作为限制因素。耐受群体显示可激活的辅助性T淋巴细胞前体（HTL-P）减少，这可能是由于HTL-P的功能性克隆缺失和/或抑制性T细胞的伴随激活。过继转移研究表明，来自耐受小鼠的细胞没有可检测地影响宿主动物脾脏中半抗原特异性CTL-P的数量。综上所述，结果表明CTL-P的功能性克隆缺失和HTL-P的抑制都有助于诱导的耐受状态。
