Monoclonal Antibody Studies on the Expression of Cellular src-gene Product (pp60c-src) in Human Leukemia-Lymphoma Cell Lines Correlated with Induced Differentiation
Project/Area Number |
61570582
|
Research Category |
Grant-in-Aid for General Scientific Research (C)
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Allocation Type | Single-year Grants |
Research Field |
Hematology
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Research Institution | Kagawa Medical Shool |
Principal Investigator |
TANAKA Terukazu Kagawa Medical School, Assistant, 医学部, 助手 (20155146)
|
Co-Investigator(Kenkyū-buntansha) |
KUBOTA Yoshitsugu Kagawa Medical School, Assistant, 医学部附属病院, 助手 (90178054)
|
Project Period (FY) |
1986 – 1987
|
Project Status |
Completed (Fiscal Year 1987)
|
Budget Amount *help |
¥2,100,000 (Direct Cost: ¥2,100,000)
Fiscal Year 1987: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 1986: ¥1,400,000 (Direct Cost: ¥1,400,000)
|
Keywords | Cellular src-gene product / Monoclonal antibody / Human leukemia-lymphoma cell lines / Induced differentiation / Cell cycle / flow cytometry / セル・ソーティング |
Research Abstract |
Expression of cellular src-gene product (pp60^<c-src>) in human leukemia-lymphoma cell lines was analysed by flow cytometry using indirect immunofluorescence with a monoclonal antibody (McAb), H2B4, which recognizes pp60^<c-src> protein in human cells. In several human leukemia-lymphoma cell lines (K562,HL60, U937, Namalva),pp60^<c-src> expression was higher than peripheral mononuclear cells from healthy volunteers. Some non-lymphoid cells can be induced to differentiate into monocyte-macrophages or granulocytes by several agents including TPA, DMSO and retinoic acid (RA). The scanning electron microscopy revealed that TPA treatment induced K 562, HL60 and U937 cells to represent prominent ruffles and to abhere on substratum with pseudopods. The TPA treatment also induced K562 cells to phagocytize Enterococcus faecalis actively. After induction of differentiation of HL60 by TPA or DMSO and RA into monocyte-macrophages or granulocytes, respectively, the bactericidal sctivity of different
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iated HL60 cells was enhanced in the presence of sub-MIC <beta>-lactam antibiotics . These results indicate that these cells were induced to differentiate not only morphologically but also functionally by the agents. Flow cytometric analyses using the McAb H2B4 revealed that the amount of pp60^<c-srs> in K562 cells markedly decreased during TPA induced differentiation. The induction of differentiation altered cell distribution in each phase of cell cycle. The cell population of K562 in S phase markedly decreased after the differentiation. On the other hand, the cell population in Gl and G2+M phases increased after the differentiation. K562 cells in G1 and G2+M phases were sorted before and after the differentiation. The amount of pp60_<c-src> in K562 cells at the same phases of cell cycle were compared by flow cytometry using the McAb H2B4 before and after the differentiation. A decrease of the amount of pp60^<c-src> was observed in G1 phase but not in G2+M phase after the differentiation. These results suggest that the pp60^<c-src> expression plays a crucial role in some human leukemia-lymphoma cells especially during the induced differentiation into the monocyte-macrophage lineage. Less
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Report
(2 results)
Research Products
(11 results)