Bich-Thuy L T, Samarut C, Brochier J, Revillard J P
J Immunol. 1981 Oct;127(4):1299-303.
During short incubation in serum-free medium, polymorphonuclear neutrophils (PMN) release soluble material that can be characterized as receptors for Fc IgG (Fc gamma R) on the following evidence: it agglutinates erythrocyte-IgG antibody (EAG) complexes, it prevents the binding of EAG to EAG-rosette-forming cells, and it binds to EAG-rosette-forming cells after modulation of their Fc gamma R, allowing the formation of 'passive' rosettes. These Fc gamma R were isolated by affinity chromatography on sepharose 4B-IgG. This material was shown to interfere with the differentiation of peripheral blood B cells into Ig-secreting cells in cultures stimulated by pokeweed (PWM) or Nocardia opaca (NOC) extracts. The number of Ig-secreting cells determined by a reverse hemolytic plaque assay using protein A-coated sheep erythrocytes was decreased by addition of Fc gamma R over a wide range of dilutions. The number of Ig-containing cells was diminished in PWM-stimulated cultures, but not in cultures stimulated with NOC. Fc gamma R did not affect cell viability, nor did they interfere with plaque-forming cell assay. Fc gamma R was not suppressive when added before the 3rd day or after the 6th day of culture. The suppressor factor was shown to remain associated with Fc gamma R after elution at acidic pH; it was removed by absorption on Sepharose 4B-IgG, but not on pepsin-digested F(ab')2 fragments. The suppressive factor as well as the capacity to restore EAG rosette formation by modulated lymphocytes were destroyed by heating (56 degrees C, 30 min) or by freezing and thawing. Properties of Fc gamma R released from PMN in this system are similar to those of Fc gamma R released from unstimulated human peripheral blood lymphocytes (PBL) and to those of mouse Ig-binding factor produced by alloactivated T cells or T cell lines.
在无血清培养基中短期孵育期间,多形核中性粒细胞(PMN)释放出一种可溶性物质,基于以下证据可将其鉴定为Fc IgG受体(FcγR):它能凝集红细胞-IgG抗体(EAG)复合物,阻止EAG与EAG花环形成细胞结合,并且在其FcγR被调节后能与EAG花环形成细胞结合,从而形成“被动”花环。这些FcγR通过在琼脂糖4B-IgG上进行亲和层析分离得到。已证明该物质会干扰在商陆(PWM)或 opaque诺卡氏菌(NOC)提取物刺激的培养物中,外周血B细胞分化为Ig分泌细胞的过程。在广泛的稀释范围内添加FcγR后,使用蛋白A包被的绵羊红细胞通过反向溶血空斑试验测定的Ig分泌细胞数量减少。在PWM刺激的培养物中,含Ig细胞的数量减少,但在NOC刺激的培养物中则没有减少。FcγR不影响细胞活力,也不干扰空斑形成细胞试验。在培养的第3天之前或第6天之后添加FcγR时没有抑制作用。已证明在酸性pH下洗脱后,抑制因子仍与FcγR相关联;它可通过在琼脂糖4B-IgG上吸附而去除,但不能在胃蛋白酶消化的F(ab')2片段上吸附。抑制因子以及通过调节淋巴细胞恢复EAG花环形成的能力会因加热(56℃,30分钟)或冻融而被破坏。在该系统中从PMN释放的FcγR的特性与从未刺激的人外周血淋巴细胞(PBL)释放的FcγR以及同种异体激活的T细胞或T细胞系产生的小鼠Ig结合因子的特性相似。