Quantitative tracing of mRNAs for T- and B-lymphocyte receptor genes in individual cells by in situ hybridization with fluorochrome-labeled gene probes. I. Expression in malignancies carrying B-lineage associated antigens.
Acute and chronic lymphatic leukemias were investigated on the single-cell level for the activity of genes coding for the IgM heavy chain and the alpha and beta chains of the T-cell antigen receptor (TCR). We used a new method for preparing highly fluorochrome-labeled gene probes for in situ hybridization, which allowed rapid and quantitative detection of mRNA at the individual cell level. Leukemic cell populations classified as belonging to the B lineage according to their surface antigenic patterns revealed increasing expression of mRNA for the IgM heavy chain (mu mRNA) in a maturation-dependent fashion, which was not correlated to rearrangement of the immunoglobulin mu chain gene--only 66% of the leukemias with rearranged mu gene also transcribed it. TCR mRNA was detected in B-antigen positive leukemic cells. High levels of both TCR and mu mRNA expression in all cells of some of these leukemias allowed the conclusion that these cells simultaneously transcribed the genes for T and B cell antigen receptors. TCR mRNA was also found in what are considered relatively mature B leukemias, lineage cross-over on the mRNA level being observed at a frequency of 23% (five of 22 cases), comparable with that of "inappropriate" receptor gene rearrangement. The quantitation of mRNA with fluorochrome-labeled gene probes in situ may allow determining the degree of gene activation in individual antigenically defined cells and may thus contribute a new tool for characterization of normal and malignant cells.
Mar
P
P
GSF Institut für Immunologie München, West Germany.
Pachmann
K
K
Reinecke
K
K
Emmerich
B
B
Thiel
E
E
D000944:Antigens, Differentiation, B-Lymphocyte / Q000235:genetics*; D000951:Antigens, Neoplasm / Q000235:genetics*; D001402:B-Lymphocytes / Q000145:classification / Q000473:pathology; D014408:Biomarkers, Tumor; D002454:Cell Differentiation; D015342:DNA Probes; D005456:Fluorescent Dyes; D015322:Gene Rearrangement, B-Lymphocyte; D006801:Humans; D007148:Immunoglobulin mu-Chains / Q000235:genetics; D009693:Nucleic Acid Hybridization; D010641:Phenotype; D054198:Precursor Cell Lymphoblastic Leukemia-Lymphoma / Q000235:genetics / Q000276:immunology / Q000473:pathology; D012333:RNA, Messenger / Q000302:isolation & purification*; D011947:Receptors, Antigen, B-Cell / Q000235:genetics*; D011948:Receptors, Antigen, T-Cell / Q000235:genetics*
D000944:Antigens, Differentiation, B-Lymphocyte; D000951:Antigens, Neoplasm; D014408:Biomarkers, Tumor; D015342:DNA Probes; D005456:Fluorescent Dyes; D007148:Immunoglobulin mu-Chains; D012333:RNA, Messenger; D011947:Receptors, Antigen, B-Cell; D011948:Receptors, Antigen, T-Cell