Huff T F, Uede T, Ishizaka K
J Immunol. 1982 Aug;129(2):509-14.
Mesenteric lymph node (MLN) cells of rats infected with Nippostrongylus brasiliensis (Nb) were obtained 8 days after infection and were incubated overnight with rat IgE for the formation of IgE-binding factors. These cells were then fused with BW 5147 mouse T lymphoma cells. Two hybrid clones (A4 and B6) obtained by the fusion formed IgE-binding factors upon incubation with IgE. It was found that 1-hr incubation with 3 micrograms/ml of rat IgE was sufficient to induce hybridoma cells to form IgE-binding factors. Both hybrid clones express rat T cell markers; the A4 clone bears Fc epsilon R, whereas the B6 clone bears Fc gamma R on their surface. The IgE-binding factors formed by both clones bound to rat IgE-coated Sepharose and could be eluted from the beads at pH 3.0. The factors also have affinity for mouse IgE but not human IgE nor rat IgG. IgE-binding factors formed by the A4 clone had a m.w. between 26,000 and 30,000; the B6 clone formed IgE-binding factors of 26,000 and 13,000 daltons. The factors of both high and low m.w. failed to bind to lentil lectin and Con A, but had affinity for peanut agglutinin. Purified IgE-binding factors of 13,000 daltons selectively suppressed the IgE-forming cell response of DNP-ovalbumin-primed MLN cells to homologous antigen, whereas the factor of 26,000 to 30,000 daltons neither suppressed nor enhanced the IgE response.
在感染巴西日圆线虫(Nb)8天后获取大鼠的肠系膜淋巴结(MLN)细胞,将其与大鼠IgE孵育过夜以形成IgE结合因子。然后将这些细胞与BW 5147小鼠T淋巴瘤细胞融合。通过融合获得的两个杂交克隆(A4和B6)在与IgE孵育后形成IgE结合因子。发现用3微克/毫升的大鼠IgE孵育1小时足以诱导杂交瘤细胞形成IgE结合因子。两个杂交克隆均表达大鼠T细胞标志物;A4克隆表面带有FcεR,而B6克隆表面带有FcγR。两个克隆形成的IgE结合因子与大鼠IgE包被的琼脂糖结合,并可在pH 3.0时从珠子上洗脱下来。这些因子对小鼠IgE也有亲和力,但对人IgE和大鼠IgG没有亲和力。A4克隆形成的IgE结合因子的分子量在26,000至30,000之间;B6克隆形成分子量为26,000和13,000道尔顿的IgE结合因子。高分子量和低分子量的因子均不与扁豆凝集素和刀豆蛋白A结合,但对花生凝集素有亲和力。纯化的13,000道尔顿的IgE结合因子选择性地抑制了用二硝基苯卵清蛋白致敏的MLN细胞对同源抗原的IgE形成细胞反应,而26,000至30,000道尔顿的因子既不抑制也不增强IgE反应。