Cytoplasmic and released μ-chain of the human cultured B-cell lines determined by enzyme immunoassay (EIA)

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Recently various types of B-lymphoid cell lines have been established from normal healthy subjects, patients with acute lymphoblastic leukemia (ALL) and with Burkitt's lymphoma leukemia. These cell lines can be divided into 3 categories by conventional cell surface markers; 1) normal B-cell lines with surface membrane immunoglobulins (including EBV-induced cell lines), 2) Burkitt's lymphoma cell lines and B-ALL cell lines, and 3) cell lines with nature of pre-B cell. Previously all of non-T, common ALL antigen (CALLA) positive cell lines were given a general term "null cell lines". Nowadays among them, pre-B cell lines have been discriminated in terms of the existence of cytoplasmic μ-chain. However, there is still a technical difficulty to prove the small amount of cytoplasmic μ-chain in the leukemic lymphoblasts.The objectives of this present study are to determine the cytoplasmic μ-chain and the μ-chain released into culture supernatants of lymphoid cell lines by means of enzyme immunoassay (EIA). To determine the cytoplasmic μ-chain, the cells were destructed by freezing and thawing, and after the centrifugation (10, 000g, 30min), the supernatants were examined for μ-chain by EIA. In addition, the culture supernatants of cells in RPMI 1640 supplemented with 10% fetal bovine serum were sequentially tested.The results indicate that quantity of cytoplasmic μ-chain and released μ-chain are definitely different among the above three B-cell groups. In 4 T-cell lines (MOLT-4F, KOPT-K1, JM, KOS-20) and 3 of 7 non-T, CALLA positive cell lines (NALM-16, Reh, NALL-1), μ-chain could not be detected by EIA in both cytoplasms and supernatants. In 4 of 7 non-T, CALLA positive, pre-B cell lines (KOPN-8, KOPN-K, NALM-1, NALM-6), cytoplasmic μ-chain (4-30ng/108 cells) and released μ-chain (3-18ng/ml) were detected. In 4 malignant B-cell lines (KOBK-96, KOBK-101, KOPB-Y1, Daudi) derived from Burkitt's lymphoma leukemia and B-ALL, cytoplasmic μ-chain and released μ-chain were 150-1, 500ng/108 cells and 35-270ng/ml, respectively. In 8 normal B-cell lines, cytoplasmic μ-chain and released μ-chain were 1, 000-15, 000 ng/108 cells and 600-39, 000ng/ml, respectively.Although there exists the slight overlapping among these 3 groups of B cell lines, they demonstrate definite difference as to either cytoplasmic or released μ-chains. This difference may be dependent upon the maturation of B lymphoid cells.
日本臨床免疫学会の論文

Cytoplasmic and released μ-chain of the human cultured B-cell lines determined by enzyme immunoassay (EIA)

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