Pike B L, Boyd A W, Nossal G J
Proc Natl Acad Sci U S A. 1982 Mar;79(6):2013-7. doi: 10.1073/pnas.79.6.2013.
The clonal anergy theory of induction of immunological tolerance states that differentiating B lymphocytes that encounter multivalent antigen at the pre-B to B cell transition stage can receive and store a negative signal, which renders them anergic to later triggering stimuli. The theory was tested by using an anti-mu chain monoclonal antibody, E4, as a model tolerogen. The fluorescence-activated cell sorter was used to select B cell-free cell populations from adult murine bone marrow or newborn spleen, and later, to analyze B cell neogenesis in vitro. The presence of E4 at greater than or equal to 1 microgram/ml was required to impede the development of normal numbers of B cells with full receptor status. The subsequent capacity of these B cells to respond in vitro to mitogens was assessed in a filter-cell free microculture system that allows single B cells to proliferate and differentiate. Concentrations of E4 far below those required to affect B cell neogenesis had profound inhibitory effects on the subsequent functional capacity of the B cells. In fact, 10(-3) micrograms/ml of E4 markedly impaired both proliferation and antibody formation, and 10(-1) micrograms/ml, which had no effect on Ig receptor development, abrogated functional capacity. Thus B cells formed in the presence of E4 at 10(-1) micrograms/ml, though possessing the receptor status typical of B cells, were functionally entirely anergic. Exposure to E4 appeared to accelerate the spontaneous death rate of newly formed B cells in vitro. Whether the anergic cell would also have a shortened life span in vivo is not known.
免疫耐受诱导的克隆失能理论指出，在从前B细胞到B细胞转变阶段遇到多价抗原的分化B淋巴细胞能够接收并储存一个负信号，这使它们对后续的触发刺激产生失能。该理论通过使用抗μ链单克隆抗体E4作为模型耐受原进行了验证。利用荧光激活细胞分选仪从成年鼠骨髓或新生鼠脾脏中选择无B细胞的细胞群体，随后在体外分析B细胞的新生情况。需要大于或等于1微克/毫升的E⁴才能阻碍具有完整受体状态的正常数量B细胞的发育。在一个无滤过细胞的微培养系统中评估这些B细胞随后在体外对有丝分裂原作出反应的能力，该系统允许单个B细胞增殖和分化。远低于影响B细胞新生所需浓度的E⁴对B细胞随后的功能能力有深远的抑制作用。事实上，10⁻³微克/毫升 的E⁴显著损害增殖和抗体形成，而对Ig受体发育无影响的10⁻¹微克/毫升则消除了功能能力。因此，在10⁻¹微克/毫升的E⁴存在下形成的B细胞，尽管具有典型的B细胞受体状态，但在功能上完全失能。在体外，暴露于E⁴似乎加速了新形成B细胞的自发死亡率。尚不清楚这种失能细胞在体内的寿命是否也会缩短。
