Time course of arrest of immunoglobulin expression in heterokaryons and early hybrids of human lymphoma cells and mouse fibroblasts. A study of transcriptional and translational events.
Early events in arrest of immunoglobulin expression were investigated at the levels of both translation and transcription in heterokaryons and early hybrids between human Daudi lymphoma cells and mouse cl. 1D cells. Large populations of 1s: 1s hybrids, isolated by fluorescence-activated cell sorting (FACS) a few hours after fusion, were grown for up to 5 days. A survey at the light-microscopical level of peroxidase-antiperoxidase-immunostained cell populations showed that arrest of expression of IgM heavy chain (mu) occurred in up to 98% of the cells. Furthermore, quantitation of mu chain contents, by using an ELISA technique, suggested that synthesis of IgM was blocked shortly after fusion. The levels of cytoplasmic mRNA specific for mu and kappa chains, respectively, decreased at rates similar to those induced in unfused Daudi cells by treatment with actinomycin D. It is concluded that arrest of immunoglobulin expression in these hybrids occurs immediately or very shortly after fusion by mechanisms that affect the levels of their cytoplasmic mRNAs.
D000818:Animals; D001483:Base Sequence; D002469:Cell Separation; D005347:Fibroblasts / Q000378:metabolism; D005434:Flow Cytometry; D016650:Fluorescein-5-isothiocyanate; D005452:Fluoresceins; D005786:Gene Expression Regulation; D006801:Humans; D006822:Hybrid Cells / Q000378:metabolism*; D007136:Immunoglobulins / Q000235:genetics*; D007700:Kinetics; D008223:Lymphoma / Q000378:metabolism*; D051379:Mice; D014176:Protein Biosynthesis; D012333:RNA, Messenger / Q000032:analysis; D013861:Thiocyanates; D013937:Thymidine Kinase / Q000378:metabolism; D014158:Transcription, Genetic
D005452:Fluoresceins; D007136:Immunoglobulins; D012333:RNA, Messenger; D013861:Thiocyanates; D013937:Thymidine Kinase; D016650:Fluorescein-5-isothiocyanate