Chuntharapai A, Lee J, Hébert C A, Kim K J
Department of Bioanalytical Technology, Genentech, Inc., South San Francisco, CA 94080.
J Immunol. 1994 Dec 15;153(12):5682-8.
mAbs previously reported to be specific for IL-8R type A (IL-8R-A) and mAbs specific for IL-8R type B (IL-8R-B), which are described in this paper, were used to investigate the expression of each receptor on various types of cells. We generated mAbs specific for IL-8R-B, 4D1, and 10H2 by immunizing mice with 293 cells that expressed IL-8R-B and by selecting hybridoma cell lines that secreted mAbs that bind to human neutrophils. Flow cytometry showed that mAbs 4D1 and 10H2 were specific for IL-8R-B, as determined by their exclusive binding to 293-27 cells that expressed IL-8R-B, but not to 293-71 cells that expressed IL-8R-A. Epitopes recognized by these IL-8R-B-specific mAbs were shown to be within the N-terminal residues 1-18 of the IL-8R-B on the basis of their binding to various N-terminal peptides, as measured by ELISA. These IL-8R-B-specific mAbs were able to inhibit up to 90 and 50% of the 125I-labeled IL-8 binding to 293-27 cells and human neutrophils, respectively. The combination of mAb 9H1 (anti-IL-8R-A) and mAb 10H2 (anti-IL-8R-B) inhibited approximately 70% of 125I-labeled IL-8 binding to human neutrophils. Flow cytometry showed a wide range of donor variation in the expression levels of IL-8R-A and IL-8R-B on various human peripheral blood leukocytes. All neutrophils, all monocytes, and 5 to 25% of total lymphocytes (CD8+ T cells and CD56+ NK cells) expressed IL-8R. Neutrophils expressed the highest level of both IL-8R-A and IL-8R-B, at an approximately equal ratio, whereas monocytes and IL-8R+ lymphocytes expressed higher levels of IL-8R-B than IL-8R-A. Double-color flow cytometric analysis showed that 7 to 42% of CD8+ T cells and 39 to 76% of CD56+ NK cells, but no CD 20+ B cells or CD4+ T cells, expressed IL-8R.
先前报道的对A型白细胞介素8受体（IL-8R-A）具有特异性的单克隆抗体（mAbs）以及本文中描述的对B型白细胞介素8受体（IL-8R-B）具有特异性的单克隆抗体，被用于研究每种受体在各种类型细胞上的表达。我们通过用表达IL-8R-B的293细胞免疫小鼠，并筛选分泌能与人中性粒细胞结合的单克隆抗体的杂交瘤细胞系，制备了对IL-8R-B具有特异性的单克隆抗体4D1和10H2。流式细胞术显示，单克隆抗体4D1和10H2对IL-8R-B具有特异性，这是通过它们仅与表达IL-8R-B的293-27细胞结合，而不与表达IL-8R-A的293-71细胞结合来确定的。基于酶联免疫吸附测定（ELISA）所测结果，这些对IL-8R-B具有特异性的单克隆抗体识别的表位显示在IL-8R-B的N端1-18位残基内。这些对IL-8R-B具有特异性的单克隆抗体分别能够抑制高达9（此处原文可能有误，推测应为90）%和50%的125I标记的IL-8与293-27细胞和人中性粒细胞的结合。单克隆抗体9H1（抗IL-8R-A）和单克隆抗体10H2（抗IL-8R-B）的组合抑制了大约70%的125I标记的IL-8与人中性粒细胞的结合。流式细胞术显示，在各种人类外周血白细胞上，IL-8R-A和IL-8R-B的表达水平存在广泛的个体差异。所有中性粒细胞、所有单核细胞以及5%至25%的总淋巴细胞（CD8 + T细胞和CD56 + 自然杀伤细胞）表达IL-8R。中性粒细胞表达的IL-8R-A和IL-8R-B水平最高，且比例大致相等，而单核细胞和IL-8R + 淋巴细胞表达的IL-8R-B水平高于IL-8R-A。双色流式细胞术分析显示，7%至42%的CD8 + T细胞和39%至76% 的CD56 + 自然杀伤细胞表达IL-8R，但CD20 + B细胞或CD4 + T细胞不表达。
