Kisaki T, Huff T F, Conrad D H, Yodoi J, Ishizaka K
J Immunol. 1987 May 15;138(10):3345-51.
A B cell hybridoma secreting monoclonal antibody against human IgE binding factors was obtained by immunization of BALB/c mice with partially purified IgE binding factors, and fusion of their spleen cells with SP-2/0-AG14 cells. The monoclonal antibody bound all of the 60,000, 30,000, and 15,000 dalton IgE binding factors from two T cell hybridomas and those from activated T cells of a normal individual. The antibody bound both IgE-potentiating factors, which had affinity for lentil lectin, and IgE-suppressive factors, which had affinity for peanut agglutinin. However, the monoclonal anti-IgE-binding factor bound neither Fc epsilon R on RPMI 8866 cells nor IgE binding factors from the B lymphoblastoid cells. A monoclonal antibody against Fc epsilon R on B cells (H107) bound the 60,000 and 30,000 dalton IgE binding factors from both T cell hybridomas and RPMI 8866 cells but did not bind the 15,000 dalton IgE binding factors from either T cells or B cells. The results indicate that T cell-derived IgE binding factors have a unique antigenic determinant that is lacking in both Fc epsilon R on B cells and B cell-derived IgE binding factors. The anti-IgE binding factor and anti-Fc epsilon R monoclonal antibodies both failed to stain cell surface components of IgE binding factor-producing T cell hybridomas. However, both antibodies induced the T cell hybridoma to form IgE binding factors. The results suggest that the T cell hybridomas bear low numbers of Fc epsilon R that share antigenic determinants with IgE binding factors secreted from the cells.
通过用部分纯化的IgE结合因子免疫BALB/c小鼠,并将其脾细胞与SP-2/0-AG14细胞融合,获得了一种分泌抗人IgE结合因子单克隆抗体的B细胞杂交瘤。该单克隆抗体与来自两种T细胞杂交瘤以及正常个体活化T细胞的所有60,000、30,000和15,000道尔顿的IgE结合因子结合。该抗体既结合了对扁豆凝集素有亲和力的IgE增强因子,也结合了对花生凝集素有亲和力的IgE抑制因子。然而,抗IgE结合因子单克隆抗体既不与RPMI 8866细胞上的FcεR结合,也不与B淋巴母细胞的IgE结合因子结合。一种针对B细胞上FcεR的单克隆抗体(H107)与来自T细胞杂交瘤和RPMI 8866细胞的60,000和30,000道尔顿的IgE结合因子结合,但不与来自T细胞或B细胞的15,000道尔顿的IgE结合因子结合。结果表明,T细胞来源的IgE结合因子具有独特的抗原决定簇,而B细胞上的FcεR和B细胞来源的IgE结合因子中都没有这种抗原决定簇。抗IgE结合因子和抗FcεR单克隆抗体都未能对产生IgE结合因子的T细胞杂交瘤的细胞表面成分进行染色。然而,两种抗体都诱导T细胞杂交瘤形成IgE结合因子。结果表明,T细胞杂交瘤上携带少量与细胞分泌的IgE结合因子具有共同抗原决定簇的FcεR。