Pennell C A, Maynard E, Arnold L W, Haughton G, Clarke S H
Department of Microbiology and Immunology, University of North Carolina, Chapel Hill 27599.
J Immunol. 1990 Sep 1;145(5):1592-7.
Our previous analyses of peritoneal Ly-1 B cells indicate that a high percentage express VH genes of the VH11 and VH12 families, and that this bias is due to clonal selection. The antibodies encoded by these genes bind the same hapten, phosphatidyl choline (PtC). Twenty-one of 73 hybridomas generated from fusions with peritoneal Ly-1 and Ly-1 sister population B cells of B10.H-2aH-4bp/Wts mice produce anti-PtC specific antibodies. We show here that 19 of these express VH11 and VH12 family genes and two express VH36-60 family genes. To assess whether there is a bias in VH gene use among non-PtC-specific hybridomas we analyzed the remaining 52 hybridomas for VH family expression by using VH family-specific probes in an RNA dot blot assay and by Ig mRNA sequencing. We find a seven-fold increase in the expression of the VHS107 family genes, and only slight differences in the expression of VH genes of other families relative to splenic B cells. We attribute the increase in VHS107 gene expression to clonal selection inasmuch as five of the seven VHS107+ hybridomas express the same VH gene (V11) and VL association is nonrandom. The bias in VH gene use among the entire panel of 73 peritoneal hybridomas is to the extent that approximately one-third express one of three genes: the V11 gene of the S107 family, the CH34 gene of the VH11 family, and the VH12 family gene.
我们之前对腹膜Ly-1 B细胞的分析表明,很大比例的细胞表达VH11和VH12家族的VH基因,这种偏向性是由克隆选择导致的。这些基因编码的抗体结合相同的半抗原,磷脂酰胆碱(PtC)。用B10.H-2aH-4bp/Wts小鼠的腹膜Ly-1和Ly-1姐妹群体B细胞融合产生的73个杂交瘤中,有21个产生抗PtC特异性抗体。我们在此表明,其中19个表达VH11和VH12家族基因,2个表达VH36-60家族基因。为了评估非PtC特异性杂交瘤中VH基因使用是否存在偏向性,我们通过RNA斑点印迹分析和Ig mRNA测序,使用VH家族特异性探针分析了其余52个杂交瘤的VH家族表达情况。我们发现VHS107家族基因的表达增加了7倍,相对于脾B细胞,其他家族VH基因的表达只有轻微差异。我们将VHS107基因表达的增加归因于克隆选择,因为7个VHS107+杂交瘤中有5个表达相同的VH基因(V11),且VL关联是非随机的。在全部73个腹膜杂交瘤中,VH基因使用的偏向性在于大约三分之一的细胞表达三个基因之一:S107家族的V11基因、VH11家族的CH34基因和VH12家族基因。