Fidler J M
J Exp Med. 1979 Sep 19;150(3):491-506. doi: 10.1084/jem.150.3.491.
The maturation level of the B-lymphocyte subpopulations involved in trinitrophenyl (TNP)-specific immunological tolerance in adult mice induced by the injection of trinitrobenzenesulfonic acid (TNBS) was investigated using in vitro antigen-specific and nonspecific polyclonal stimulation. The maturity of the B-cell subsets being studied was defined by the antigen or polyclonal activator which evoked a response. Thus, the thymic independent (TI-1) antigen TNP-lipopolysaccharide (TNP-LPS) and the polyclonal stimulant LPS were used to activate immature, neonatal-type B lymphocytes, whereas mature, adult-type B cells were responsive to the TI-2 antigen, TNP-Ficoll, and the nonspecific activator, purified protein derivative (PPD). Whereas unresponsiveness in TNP-LPS-reactive (immature) B cells 4 d after TNBS treatment was previously shown to be the result of functional deletion, partially reversible receptor blockade was detected in this study early after tolerogen treatment. By the 24-h point, tolerance was irreversible, as assessed by 24-h of antigen-free incubation and cocultivation of tolerant cells with control splenocytes. Tolerance was induced more rapidly in immature, TI-1 B cells than in mature TI-2 B lymphocytes. B lymphocytes reactive to TNP-Ficoll were also less susceptible to receptor blockade. Using LPS as a nonspecific probe for immature B cells, 60% tolerance in high affinity TNP-specific cells was induced within 12 h of TNBS treatment, and complete unresponsiveness by 24 h. In contrast, no significant decrease in response to the mature B-cell activator, PPD, occurred until day 2. Furthermore, the 50% tolerance level was achieved in TNP-specific LPS-reactive B cells by 100 times less tolerogen than required for PPD-responsive cells. Thus, TNBS-induced unresponsiveness in cells reactive to TNP-LPS is initially a result of reversible receptor blockade which leads within 4 d to functional deletion. Immature, TI-1 B lymphocytes, which give polyclonal responses to LPS and antigen-specific responses to TNP-LPS, are rendered tolerant to TNBS more rapidly and at lower tolerogen does than mature, TI-2 mouse B cells which react polyclonally to PPD and specifically to TNP-Ficoll. Moreover, these data show that both the immature and the mature B lymphocyts with these characteristic tolerance susceptibilities and specific and nonspecific immune response patterns are present in the adult mouse spleen.
采用体外抗原特异性和非特异性多克隆刺激的方法，研究了注射三硝基苯磺酸（TNBS）诱导成年小鼠对三硝基苯基（TNP）特异性免疫耐受过程中涉及的B淋巴细胞亚群的成熟水平。所研究的B细胞亚群的成熟度由引发反应的抗原或多克隆激活剂来定义。因此，胸腺非依赖性（TI-1）抗原TNP-脂多糖（TNP-LPS）和多克隆刺激剂LPS用于激活未成熟的新生型B淋巴细胞，而成熟的成年型B细胞对TI-2抗原TNP-菲可和非特异性激活剂纯化蛋白衍生物（PPD）有反应。此前已表明，TNBS处理4天后，TNP-LPS反应性（未成熟）B细胞的无反应性是功能缺失的结果，而在本研究中，在耐受原处理后早期检测到了部分可逆的受体阻断。到24小时时，通过无抗原孵育24小时以及将耐受细胞与对照脾细胞共培养评估，耐受性是不可逆的。未成熟的TI-1 B细胞比成熟的TI-2 B淋巴细胞更快诱导出耐受性。对TNP-菲可反应的B淋巴细胞也较不易受到受体阻断的影响。使用LPS作为未成熟B细胞的非特异性探针，TNBS处理12小时内，高亲和力TNP特异性细胞诱导出60%的耐受性，24小时时完全无反应。相比之下，直到第2天，对成熟B细胞激活剂PPD的反应才出现显著下降。此外，TNP特异性LPS反应性B细胞达到50%耐受水平所需的耐受原比PPD反应性细胞所需的少100倍。因此，TNBS诱导的对TNP-LPS反应细胞的无反应性最初是可逆受体阻断的结果，这种阻断在4天内导致功能缺失。未成熟的TI-1 B淋巴细胞对LPS产生多克隆反应，对TNP-LPS产生抗原特异性反应，与对PPD产生多克隆反应、对TNP-菲可产生特异性反应成熟的TI-2小鼠B细胞相比，其对TNBS的耐受性诱导更快，所需耐受原剂量更低。此外，这些数据表明，具有这些特征性耐受易感性以及特异性和非特异性免疫反应模式的未成熟和成熟B淋巴细胞都存在于成年小鼠脾脏中。
