Chen Y W, Word C J, Jones S, Uhr J W, Tucker P W, Vitetta E S
J Exp Med. 1986 Aug 1;164(2):548-61. doi: 10.1084/jem.164.2.548.
We have subcloned the in vitro-adapted murine B cell leukemia, BCL1.B1, to obtain a variant that expresses both IgM and IgG1. By fluorescence analysis, radioiodination, and immunoprecipitation of cell surface Ig, and by RIA of medium from limiting dilution cultures, we have shown that: (a) all the cells express and secrete both isotypes. The heavy chains of both IgG1 and IgM have the apparent molecular weights of membrane mu and gamma 1 chains; (b) both isotypes bear the same idiotype as determined by immunoprecipitation with antiidiotypic antibody, and both use the same VDJ rearrangement as shown by Southern blotting; and (c) the cells express the membrane and secreted forms of mRNA for both mu and gamma 1 but not gamma 2b or gamma 3. Taken together, the data suggest that all the cells are synthesizing, expressing on their surface, and secreting two isotypes that use the same VDJ rearrangement in the DNA and express the same serologically-defined idiotype. The molecular basis responsible for the production of the two isotypes in a single cell is the subject of the accompanying paper.
我们对体外适应的小鼠B细胞白血病BCL1.B1进行了亚克隆,以获得一种同时表达IgM和IgG1的变体。通过对细胞表面Ig进行荧光分析、放射性碘化和免疫沉淀,以及对有限稀释培养物的培养基进行放射免疫分析,我们发现:(a)所有细胞均表达并分泌这两种同种型。IgG1和IgM的重链具有膜μ链和γ1链的表观分子量;(b)通过抗独特型抗体免疫沉淀确定,这两种同种型具有相同的独特型,并且通过Southern印迹法显示两者使用相同的VDJ重排;(c)细胞表达μ链和γ1链的膜形式和分泌形式的mRNA,但不表达γ2b或γ3。综合来看,这些数据表明所有细胞都在合成、在其表面表达并分泌两种同种型,它们在DNA中使用相同的VDJ重排并表达相同的血清学定义的独特型。单个细胞中产生两种同种型的分子基础是随附论文的主题。