A monoclonal antibody that detects a specific human neutrophil antigen involved in phorbol myristate acetate- and formyl-methionyl-leucyl-phenylalanine-triggered respiratory burst

Pedro M. Lacal, Isabel Barasoaín, Ana Sánchez, Javier García-Sancho, Ignacio Flores, Faustino Mollinedo

Research output: Contribution to journalArticle

Abstract

A mouse IgM mAb termed P1E3 was raised against resting human peripheral blood neutrophils and has been shown to recognize a cell-surface Ag with an apparent molecular mass of 155 kDa, as assessed by immunoprecipitation analysis. In addition to the main 155-kDa protein, an additional band of about 210 kDa was also recognized by P1E3 in Western blot analysis. Sequential immunoprecipitation assays showed that the Ag recognized by P1E3 differed from the CD29 and CD45 Ag. However, sequential immunoprecipitation assays carried out with two distinct anti-CD15 mAb and P1E3 showed that P1E3 reacted with CD15 or with a CD15-like Ag. P1E3 stained strongly resting human peripheral blood neutrophils, hardly reacted with peripheral blood monocytes and did not react with PBL and platelets, as assessed by immunofluorescence flow cytometry. P1E3 inhibited the respiratory burst induced by PMA or FMLP, but not the oxidative response induced by Con A or the calcium ionophores A23187 or ionomycin. Furthermore, P1E3 inhibited the activation of the Na+/H+ antiporter in response to PMA or FMLP and the phosphorylation of a protein of about 50 kDa in response to PMA. However, preincubation of neutrophils with P1E3 did not affect the increase in cytosolic free calcium concentration induced by FMLP. These data suggest that the Ag recognized by P1E3 may play a role in modulating the activation of the respiratory burst induced by PMA or FMLP, and that P1E3 seems to affect protein kinase C-mediated signal transduction mechanisms coupled to the induction of the respiratory burst.

Original languageEnglish
Pages (from-to)161-168
Number of pages8
JournalJournal of Immunology
Volume148
Issue number1
Publication statusPublished - Jan 1 1992

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methionyl-leucyl-phenylalanine
Respiratory Burst
Tetradecanoylphorbol Acetate
Immunoprecipitation
Neutrophils
Monoclonal Antibodies
Antigens
Sodium-Hydrogen Antiporter
Ionomycin
Calcium Ionophores
Calcimycin
Protein Kinase C
Fluorescent Antibody Technique
Immunoglobulin M
Monocytes
Signal Transduction
Flow Cytometry
Proteins
Blood Platelets
Western Blotting

ASJC Scopus subject areas

  • Immunology

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A monoclonal antibody that detects a specific human neutrophil antigen involved in phorbol myristate acetate- and formyl-methionyl-leucyl-phenylalanine-triggered respiratory burst. / Lacal, Pedro M.; Barasoaín, Isabel; Sánchez, Ana; García-Sancho, Javier; Flores, Ignacio; Mollinedo, Faustino.

In: Journal of Immunology, Vol. 148, No. 1, 01.01.1992, p. 161-168.

Research output: Contribution to journalArticle

Lacal, Pedro M. ; Barasoaín, Isabel ; Sánchez, Ana ; García-Sancho, Javier ; Flores, Ignacio ; Mollinedo, Faustino. / A monoclonal antibody that detects a specific human neutrophil antigen involved in phorbol myristate acetate- and formyl-methionyl-leucyl-phenylalanine-triggered respiratory burst. In: Journal of Immunology. 1992 ; Vol. 148, No. 1. pp. 161-168.
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