Malley A, Bradley L M, Shiigi S M
J Immunol. 1987 Aug 15;139(4):1046-53.
Cultures of normal spleen cells with anti-idiotypic antibody (anti-Id) or antigen B (AgB)-specific T suppressor factor (Tsf1) in mini-Marbrook chambers for 4 days at 37 degrees C lead to the in vitro induction of AgB-specific T suppressor (TS) cells. These TS cells significantly suppress a secondary AgB-specific IgE response, but they do not affect a secondary AgB-specific IgG response. Depletion of both B cells and macrophages from normal spleen cells by panning on anti-Ig-coated petri dishes provides an enriched T cell population. These enriched T cells when cultured with anti-Id or Tsf1 in mini-Marbrook chambers do not produce AgB-specific TS cells, and mice treated with cells harvested from the mini-Marbrook chambers have normal secondary AgB-specific IgG and IgE responses. The addition of as few as 1000 bone marrow-derived macrophages (BMDM) to cultures of the enriched T cells with anti-Id, or Tsf1 restores the ability of these cultures to produce significant levels of AgB-specific TS cells. Further studies reveal that the macrophage population must be histocompatible and express a cell surface I-J antigen. Attempts to pulse BMDM with anti-Id or Tsf1 at 4 degrees C and to culture in mini-Marbrook chambers 10(3) pulsed BMDM with enriched T cells were unsuccessful in producing AgB-specific TS cells. However, pulsing BMDM with anti-Id or Tsf1 at 37 degrees C, and adding 10(3) of these pulsed BMDM to enriched T cells in culture led to the formation of significant levels of AgB-specific TS cells.
在微型马尔布鲁克培养室中,将正常脾细胞与抗独特型抗体(抗Id)或抗原B(AgB)特异性T抑制因子(Tsf1)于37℃培养4天,可在体外诱导出AgB特异性T抑制(TS)细胞。这些TS细胞可显著抑制二次AgB特异性IgE反应,但不影响二次AgB特异性IgG反应。通过在抗Ig包被的培养皿上淘选,从正常脾细胞中去除B细胞和巨噬细胞,可得到富集的T细胞群体。这些富集的T细胞在微型马尔布鲁克培养室中与抗Id或Tsf1一起培养时,不会产生AgB特异性TS细胞,并且用从微型马尔布鲁克培养室收获的细胞处理的小鼠具有正常的二次AgB特异性IgG和IgE反应。向富集的T细胞与抗Id或Tsf1的培养物中添加低至1000个骨髓来源的巨噬细胞(BMDM),可恢复这些培养物产生显著水平AgB特异性TS细胞的能力。进一步研究表明,巨噬细胞群体必须是组织相容性的,并表达细胞表面I-J抗原。尝试在4℃用抗Id或Tsf1脉冲BMDM,并在微型马尔布鲁克培养室中用富集的T细胞培养10³个脉冲的BMDM,未能产生AgB特异性TS细胞。然而,在37℃用抗Id或Tsf1脉冲BMDM,并将10³个这些脉冲的BMDM添加到培养的富集T细胞中,导致形成显著水平的AgB特异性TS细胞。