Chen C, Martin T M, Stevens S, Rittenberg M B
Department of Microbiology and Immunology, Oregon Health Sciences University, Portland 97201.
J Exp Med. 1994 Aug 1;180(2):577-86. doi: 10.1084/jem.180.2.577.
We have investigated four secretion-deficient antibodies (Abs) derived from a panel of 46 mutant T15 anti-phosphocholine Abs, all of which have point mutations in the heavy chain (H) complementarity determining region 2 (CDR2). The level of secretion for these four Abs was < 10% of wild type when expressed together with the T15 light chain (L) in either SP2/0 or P3X63Ag8.653 myeloma cells although normal levels of H and L chain mRNA were produced. Moreover, abundant intracellular H and L chain proteins were detected. Three of the four mutants had little or no assembled H and L complexes intracellularly whereas one had a significant amount of intracellular immunoglobulin (Ig) which was shown to be capable of binding Ag. Thus, we demonstrate for the first time that point mutations confined to CDR2 of the H chain variable (V) region can impede Ab assembly and secretion. We then introduced the same CDR2 mutations into a related H chain which is encoded by the same T15 VH gene but different diversity (D) and joining (J) genes. When these H chains were expressed with a non-T15 L chain, the resulting Abs were secreted normally. The results thus suggest that the effects of the CDR2 mutations on Ab secretion are dependent on their interactions with L and/or H chain D-J sequences. These results also reveal a novel mechanism that could contribute to B cell wastage.
我们研究了从一组46种突变型T15抗磷酸胆碱抗体(Abs)中获得的四种分泌缺陷型抗体,所有这些抗体在重链(H)互补决定区2(CDR2)中都有单点突变。当在SP2/0或P3X63Ag8.653骨髓瘤细胞中与T15轻链(L)一起表达时,这四种抗体的分泌水平不到野生型的10%,尽管产生了正常水平的H链和L链mRNA。此外,还检测到大量细胞内H链和L链蛋白。四个突变体中的三个在细胞内几乎没有或没有组装的H链和L链复合物,而另一个有大量细胞内免疫球蛋白(Ig),显示其能够结合抗原。因此,我们首次证明,局限于重链可变(V)区CDR2的单点突变可阻碍抗体组装和分泌。然后,我们将相同的CDR2突变引入到一个相关的重链中,该重链由相同的T15 VH基因编码,但具有不同的多样性(D)和连接(J)基因。当这些重链与非T15轻链一起表达时,产生正常分泌的抗体。因此,结果表明CDR2突变对抗体分泌的影响取决于它们与轻链和/或重链D-J序列的相互作用。这些结果还揭示了一种可能导致B细胞损耗的新机制。