Delespesse G, Sarfati M, Rubio-Trujillo M, Wolowiec T
Eur J Immunol. 1986 Jul;16(7):815-21. doi: 10.1002/eji.1830160716.
A monoclonal antibody (mAb) specific for lymphocyte IgE receptors (ER) was employed in a rosette assay for the detection of cells bearing IgE receptors (Fc epsilon R). The specificity of the assay was documented by inhibition studies with soluble immunoglobulins (Ig) and anti-Ig antibodies. Moreover, similar results were obtained by employing the F(ab')2 fragment of mAbER instead of intact molecule. Circulating mononuclear cells isolated from normal or allergic adults and from umbilical cord blood contained approximately 8% of Fc epsilon R-bearing cells with values ranging from 0.3 to 17%. Tonsillar lymphocytes contained about 30% of Fc epsilon R+ cells. After the removal of adherent cells, there was a small but significant reduction of the proportion of Fc epsilon R+ cells. When mononuclear cells were separated into T and B cell fractions by two-cycle rosetting with 2-aminoethylisothiouronium bromide hydrobromide-treated sheep red blood cells, most of the Fc epsilon R+ cells were in the B cell fraction; however, a small proportion of Fc epsilon R+ was also found in the enriched T cells and double-labeling experiments confirmed that these cells were indeed T lymphocytes. Fc epsilon R+ cells were purified by rosetting with mAbER-coated erythrocytes and their phenotype was compared to that of Fc epsilon R- cells; Fc epsilon R+ cells contained about 90% of B cells (B1+) together with a small proportion of OKT3+, Leu 7+ and Mo2+ cells. The bulk of T cells, macrophages and natural killer (NK) cells was found in the Fc epsilon R- cells which contained fewer B cells than the fraction of Fc epsilon R+ cells. These data thus indicated that the great majority of Fc epsilon R-bearing cells are B cells but that a small proportion of NK cells, macrophages and T lymphocytes also express Fc epsilon R. Upon incubation at 37 degrees C, B cells lost their Fc epsilon R and this phenomenon was selectively inhibited by IgE; however, purified T cells seemed to express more Fc epsilon R after overnight incubation at 37 degrees C and this was not influenced by IgE. It is finally shown that the expression of Fc epsilon R is cyclic and that Fc epsilon R-bearing B cells do not represent a functionally distinct subpopulation of B lymphocytes.
一种针对淋巴细胞IgE受体(ER)的单克隆抗体(mAb)被用于玫瑰花环试验,以检测带有IgE受体(FcεR)的细胞。通过用可溶性免疫球蛋白(Ig)和抗Ig抗体进行抑制研究,证明了该试验的特异性。此外,使用mAbER的F(ab')2片段而非完整分子也获得了类似结果。从正常或过敏的成年人以及脐带血中分离出的循环单核细胞中,约8%的细胞带有FcεR,其值范围为0.3%至17%。扁桃体淋巴细胞中约30%的细胞为FcεR+细胞。去除贴壁细胞后,FcεR+细胞的比例有小幅但显著的下降。当通过用氢溴酸2-氨基乙基异硫脲处理的绵羊红细胞进行两轮玫瑰花环试验将单核细胞分离为T细胞和B细胞组分时,大多数FcεR+细胞存在于B细胞组分中;然而,在富集的T细胞中也发现了一小部分FcεR+细胞,双标记实验证实这些细胞确实是T淋巴细胞。通过用mAbER包被的红细胞进行玫瑰花环试验纯化FcεR+细胞,并将其表型与FcεR-细胞的表型进行比较;FcεR+细胞中约90%为B细胞(B1+),还有一小部分OKT3+、Leu 7+和Mo2+细胞。在FcεR-细胞中发现了大量的T细胞、巨噬细胞和自然杀伤(NK)细胞,其B细胞数量比FcεR+细胞组分中的少。因此,这些数据表明,绝大多数带有FcεR的细胞是B细胞,但一小部分NK细胞、巨噬细胞和T淋巴细胞也表达FcεR。在37℃孵育后,B细胞失去其FcεR,这种现象被IgE选择性抑制;然而,纯化的T细胞在37℃过夜孵育后似乎表达更多的FcεR,且这不受IgE的影响。最终表明,FcεR的表达是周期性的,且带有FcεR的B细胞并不代表B淋巴细胞中一个功能上不同的亚群。