Poncet P, Huetz F, Marcos M A, Andrade L
Laboratory of Immunobiology, Pasteur Institute, Paris, France.
Eur J Immunol. 1990 Jul;20(7):1583-9. doi: 10.1002/eji.1830200726.
We have studied the relationship between B cell reactivity to bromelain-treated autologous mouse erythrocytes (BrMRBC) and expression of the VH11 gene family in splenic, peritoneal and pleuropericardial cell populations from normal C57BL/6 mice. B lymphocytes producing antibodies to BrMRBC were selectively enriched or depleted from normal populations by rosette formation with BrMRBC, followed by centrifugation over density gradients. This selection method, based on the presence of functional receptors (membrane IgM), is harmless for the cells and allowed subsequent cloning in agar (colony-forming unit-B). The utilization of the 10 VH gene families was then scored in mRNA colony blot assays. The analysis of greater than 650 anti-BrMRBC clones and greater than 350 VH11-expressing colonies indicates that about half of those antibody reactivities are encoded by VH11 genes. Furthermore, it appears that all VH11-expressing B cells in the peritoneal cavity produce anti-BrMRBC antibodies.
我们研究了正常C57BL/6小鼠脾细胞、腹腔细胞和胸膜心包细胞群体中，B细胞对菠萝蛋白酶处理的自体小鼠红细胞（BrMRBC）的反应性与VH11基因家族表达之间的关系。通过与BrMRBC形成玫瑰花结，然后在密度梯度上离心，从正常群体中选择性富集或去除产生抗BrMRBC抗体的B淋巴细胞。这种基于功能性受体（膜IgM）存在的选择方法对细胞无害，并允许随后在琼脂中克隆（集落形成单位-B）。然后在mRNA集落印迹分析中对10个VH基因家族的利用情况进行评分。对超过650个抗BrMRBC克隆和超过350个表达VH11的集落的分析表明，这些抗体反应性中约有一半由VH11基因编码。此外，似乎腹腔中所有表达VH11的B细胞都产生抗BrMRBC抗体。
