Inhibition of the T-cell receptor-mediated signal transduction by microinjection of anti-Lck monoclonal antibody into T-cells

Kazuhiko Nakamura, Yasuhiro Koga, Hiroki Yoshida, Kazuo Tanaka, Masafumi Sasaki, Genki Kimura, Kikuo Nomoto

研究成果: ジャーナルへの寄稿記事

9 引用 (Scopus)

抄録

Engagement of T-cell receptor (TcR)/CD3 complexes on T-cells rapidly provokes tyrosine phosphorylation of cellular proteins, which is thought to be an essential step to the following events of T-cell activation. p56lck, a member of src-related, non-receptor type protein tyrosine kinases, is expressed predominantly in lymphocytes. Accumulating data suggest that p56lck is one of the kinases responsible for TcR-mediated protein tyrosine phosphorylation. To investigate the role of p56lck in TcR-signaling in detail, we injected anti-Lck monoclonal antibody (mAb), MOL171 or MOL294, both specfically suppress Lck kinase activity in vitro, into Jurkat T-cells by the erythrocyte-ghost procedure in order to block the activity of p56lck. In Jurkat cells injected with anti-Lck mAb, intracellular Ca2+ mobilization induced by TcR-stimulation was markedly reduced in comparison with control mouse IgG-injected samples. This block of Ca2+ influx seems to be specific for TcR-signaling because anti-Lck mAb-injection did not cause significant suppression of phytohaemagglutinin-induced Ca2+ increase. Furthermore, injection of anti-Lck mAb inhibited TcR-mediated protein tyrosine phosphorylation of 100 kDa protein and phospholipase Cγ1. These results confirm that p56lck is an indispensable element of TcR-signaling and p100 and phospholipase Cγ1 are strongly presumed to be candidates for substrates for p56lck.

元の言語英語
ページ(範囲)495-505
ページ数11
ジャーナルBBA - Molecular Cell Research
1224
発行部数3
DOI
出版物ステータス出版済み - 12 30 1994

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Microinjections
T-Cell Antigen Receptor
Signal Transduction
Monoclonal Antibodies
T-Lymphocytes
Tyrosine
Jurkat Cells
Phosphorylation
Proteins
Lymphocyte Specific Protein Tyrosine Kinase p56(lck)
CD3 Antigens
Injections
Erythrocyte Membrane
Phytohemagglutinins
Protein-Tyrosine Kinases
Phosphotransferases
Immunoglobulin G
Lymphocytes

All Science Journal Classification (ASJC) codes

  • Molecular Biology
  • Cell Biology

これを引用

Inhibition of the T-cell receptor-mediated signal transduction by microinjection of anti-Lck monoclonal antibody into T-cells. / Nakamura, Kazuhiko; Koga, Yasuhiro; Yoshida, Hiroki; Tanaka, Kazuo; Sasaki, Masafumi; Kimura, Genki; Nomoto, Kikuo.

:: BBA - Molecular Cell Research, 巻 1224, 番号 3, 30.12.1994, p. 495-505.

研究成果: ジャーナルへの寄稿記事

Nakamura, Kazuhiko ; Koga, Yasuhiro ; Yoshida, Hiroki ; Tanaka, Kazuo ; Sasaki, Masafumi ; Kimura, Genki ; Nomoto, Kikuo. / Inhibition of the T-cell receptor-mediated signal transduction by microinjection of anti-Lck monoclonal antibody into T-cells. :: BBA - Molecular Cell Research. 1994 ; 巻 1224, 番号 3. pp. 495-505.
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abstract = "Engagement of T-cell receptor (TcR)/CD3 complexes on T-cells rapidly provokes tyrosine phosphorylation of cellular proteins, which is thought to be an essential step to the following events of T-cell activation. p56lck, a member of src-related, non-receptor type protein tyrosine kinases, is expressed predominantly in lymphocytes. Accumulating data suggest that p56lck is one of the kinases responsible for TcR-mediated protein tyrosine phosphorylation. To investigate the role of p56lck in TcR-signaling in detail, we injected anti-Lck monoclonal antibody (mAb), MOL171 or MOL294, both specfically suppress Lck kinase activity in vitro, into Jurkat T-cells by the erythrocyte-ghost procedure in order to block the activity of p56lck. In Jurkat cells injected with anti-Lck mAb, intracellular Ca2+ mobilization induced by TcR-stimulation was markedly reduced in comparison with control mouse IgG-injected samples. This block of Ca2+ influx seems to be specific for TcR-signaling because anti-Lck mAb-injection did not cause significant suppression of phytohaemagglutinin-induced Ca2+ increase. Furthermore, injection of anti-Lck mAb inhibited TcR-mediated protein tyrosine phosphorylation of 100 kDa protein and phospholipase Cγ1. These results confirm that p56lck is an indispensable element of TcR-signaling and p100 and phospholipase Cγ1 are strongly presumed to be candidates for substrates for p56lck.",
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AU - Tanaka, Kazuo

AU - Sasaki, Masafumi

AU - Kimura, Genki

AU - Nomoto, Kikuo

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