Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera

Takuya Tamatani, Fujiko Kitamura, Keisuke Kuida, Makoto Shirao, Manabu Mochizuki, Makoto Suematsu, Geert W. Schmid-Schönbein, Kazuyoshi Watanabe, Sasumu Tsurufuji, Masayuki Miyasaka

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Abstract

We have characterized the rat LECAM-1 (L-selectin) by the use of newly generated hamster anti-rat LECAM-1 monoclonal antibodies (mAb) (HRL1, HRL2, HRL3, HRL4), with respect to the biochemistry, cellular distribution and function, and developed an ELISA system to detect the soluble form of rat LECAM-1. In the rat, lymphocyte and neutrophil LECAM-1 have apparent molecular masses of 65 and 62 kDa, respectively, and differential glycosylation may account for the molecular heterogeneity. Readily detectable levels of LECAM-1 are expressed on peripheral blood lymphocytes and neutrophils, but not on thymocytes. Lymphocyte LECAM-1 is rapidly shed from the cell surface upon cell activation with PMA, but not with interleukin (IL)-8. In contrast, neutrophil LECAM-1 showed rapid shedding upon stimulation with phorbol 12-myristate 13-acetate (PMA) or IL-8. Concomitantly there is up-regulated expression of Mac-1 in PMA- and IL-8-stimulated neutrophils. Neutrophil rolling in mesenteric venules was significantly inhibited by administration of functionblocking anti-rat LECAM-1 mAb HRL3, but not by non-blocking HRL4, indicating that LECAM-I plays a significant role in leukocyte rolling. Given that LECAM-1 is rapidly shed from the cell surface, we attempted to develop an ELISA system for detecting LECAM-1 is soluble form, and measured the levels in experimental autoimmune uveitis. The circulating levels of LECAM-1 increased from day 4, which preceded the appearance of clinical signs of uveitis and remained high until uveitis subsided, suggesting that soluble LECAM-1 is potentially a useful parameter to monitor certain types of inflammatory or immune disorders.

Original languageEnglish
Pages (from-to)2181-2188
Number of pages8
JournalEuropean Journal of Immunology
Volume23
Issue number9
Publication statusPublished - 1993 Sep

Fingerprint

L-Selectin
Monoclonal Antibodies
Serum
Neutrophils
Uveitis
Interleukin-8
Acetates
Lymphocytes
Enzyme-Linked Immunosorbent Assay
Leukocyte Rolling
Venules
Immune System Diseases
Thymocytes
Glycosylation
Cricetinae
Biochemistry

Keywords

  • L-Selectin
  • LECAM-1
  • Lymphocyte homing
  • Rat

ASJC Scopus subject areas

  • Immunology

Cite this

Tamatani, T., Kitamura, F., Kuida, K., Shirao, M., Mochizuki, M., Suematsu, M., ... Miyasaka, M. (1993). Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera. European Journal of Immunology, 23(9), 2181-2188.

Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera. / Tamatani, Takuya; Kitamura, Fujiko; Kuida, Keisuke; Shirao, Makoto; Mochizuki, Manabu; Suematsu, Makoto; Schmid-Schönbein, Geert W.; Watanabe, Kazuyoshi; Tsurufuji, Sasumu; Miyasaka, Masayuki.

In: European Journal of Immunology, Vol. 23, No. 9, 09.1993, p. 2181-2188.

Research output: Contribution to journalArticle

Tamatani, T, Kitamura, F, Kuida, K, Shirao, M, Mochizuki, M, Suematsu, M, Schmid-Schönbein, GW, Watanabe, K, Tsurufuji, S & Miyasaka, M 1993, 'Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera', European Journal of Immunology, vol. 23, no. 9, pp. 2181-2188.
Tamatani, Takuya ; Kitamura, Fujiko ; Kuida, Keisuke ; Shirao, Makoto ; Mochizuki, Manabu ; Suematsu, Makoto ; Schmid-Schönbein, Geert W. ; Watanabe, Kazuyoshi ; Tsurufuji, Sasumu ; Miyasaka, Masayuki. / Characterization of rat LECAM-1 (L-selectin) by the use of monoclonal antibodies and evidence for the presence of soluble LECAM-1 in rat sera. In: European Journal of Immunology. 1993 ; Vol. 23, No. 9. pp. 2181-2188.
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