Differential expression of c-myc protein in B and T lymphocytes.
Overexpression of c-myc may play a role in the multistep pathogenesis of B- and T-cell malignancies. To determine whether this expression is inappropriate requires information on the normal cellular counterparts. There is no agreement in the literature on the levels of expression of c-myc mRNA and protein in normal peripheral blood lymphocytes and there are no reports on the differential expression in different lymphocyte populations. The aim of this study was to assess the state of c-myc expression in normal peripheral blood lymphocytes at the single cell level by immunocytochemistry and flow cytometry. Two monoclonal antibodies against c-myc and specific peptide inhibition controls were tested in mononuclear cells from nine healthy volunteers and the HL60 cell line. The expression of c-myc in B- and T-lymphocyte subsets was studied by two-colour immunocytochemistry and flow cytometry. Using calibrated reference standards, we quantified the c-myc protein and results were referred as molecules of equivalent soluble fluorochrome. Almost all lymphocytes express c-myc by both techniques. Two patterns of nuclear staining (weak and strong) were found by immunocytochemistry and this was confirmed by two peaks of fluorescence intensity by flow cytometry. Double immunostaining showed that the stronger pattern of c-myc staining corresponds to B lymphocytes and the weak one to T cells. Quantification confirmed these results which demonstrated a statistically significant difference in the expression of c-myc in these two lymphocyte populations (p < 0.005). Our results demonstrate for the first time that normal circulating B cells express higher levels of c-myc protein than T lymphocytes.
Lens
D
D
Academic Department of Haematology and Cytogenetics, Royal Marsden Hospital, London, UK.
Matutes
E
E
Farahat
N
N
Morilla
R
R
Catovsky
D
D
D000911:Antibodies, Monoclonal; D016175:B-Lymphocyte Subsets / Q000378:metabolism; D001402:B-Lymphocytes / Q000378:metabolism*; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D015473:Leukemia, Promyelocytic, Acute / Q000097:blood; D016271:Proto-Oncogene Proteins c-myc / Q000097:blood*; D016176:T-Lymphocyte Subsets / Q000378:metabolism; D013601:T-Lymphocytes / Q000378:metabolism*
D000911:Antibodies, Monoclonal; D016271:Proto-Oncogene Proteins c-myc