Inducing the loss of immunoglobulin lambda light chain production and the rearrangement of a previously excluded allele in human plasma B cell lines with concanavalin A.
We investigated the expression of differential lambda light chains in human B cell lines secreting immunoglobulin (Ig). When these cell lines were cultured with concanavalin A for a long period of time, a subpopulation of some but not all of these cell lines was induced to express new lambda light chains replacing the original lambda chain (light chain shifting). Production of the new lambda chain, which replaces the original lambda chain, results from a VJ rearrangement at a previously excluded allele and a dramatic reduction of the original lambda chain transcript, although no difference was found in the level of heavy chain transcript. Recombination activating genes RAG-1 and RAG-2, which are normally expressed during specific early stages of lymphocyte development, were expressed in not only the light chain shifting-inducible lines but also in the non-inducible cells. Treatment of these Ig secreting cell lines with dibutyryl cAMP, which is known to enhance expression of the RAG genes, could not induce the creation of new lambda light chain-producing cells from the inducible lines, suggesting that the expression of the two RAG genes is not sufficient for inducing new lambda light chain production. Concanavalin A induced a gradual but significant production lost of the original lambda chain in a subpopulation of the light chain shifting-inducible cells but not in the non-inducible cells. Association of new lambda light chain production with loss of original lambda chain raises the possibility that, when the RAG genes are expressed, concanavalin A may act on a novel intracellular mechanism controlling lambda light chain allelic exclusion in these plasma cell lines.
Tachibana
H
H
Graduate School of Genetic Resources Technology, Kyushu University, 6-10-1, Hakozaki, Fukuoka, Japan.
Ushio
Y
Y
Krungkasem
C
C
Shirahata
S
S
D000230:Adenocarcinoma; D000483:Alleles; D001402:B-Lymphocytes; D001483:Base Sequence; D002051:Burkitt Lymphoma; D002460:Cell Line; D003208:Concanavalin A / Q000494:pharmacology*; D017931:DNA Primers; D004268:DNA-Binding Proteins; D005434:Flow Cytometry; D015321:Gene Rearrangement / Q000187:drug effects*; D005803:Genes, Immunoglobulin / Q000187:drug effects*; D018398:Homeodomain Proteins; D006801:Humans; D006825:Hybridomas; D007146:Immunoglobulin lambda-Chains / Q000096:biosynthesis* / Q000235:genetics*; D008175:Lung Neoplasms; D008969:Molecular Sequence Data; D009687:Nuclear Proteins; D016133:Polymerase Chain Reaction; D014176:Protein Biosynthesis; D014158:Transcription, Genetic / Q000187:drug effects*
D017931:DNA Primers; D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; D007146:Immunoglobulin lambda-Chains; D009687:Nuclear Proteins; C499311:RAG2 protein, human; C064655:V(D)J recombination activating protein 2; D003208:Concanavalin A; C064658:RAG-1 protein