Anti-C5a receptor antibodies

Characterization of neutralizing antibodies specific for a peptide, C5aR-(9-29), derived from the predicted amino-terminal sequence of the human C5a receptor

Edward L. Morgan, Julia A. Ember, Sam Sanderson, Wolfgang Scholz, Robert Buchner, Richard D. Ye, Tony E. Hugli

Research output: Contribution to journalArticle

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Abstract

Results obtained indicate that a site-directed polyclonal antibody specific for a synthetic peptide based on the predicted ami no-terminal sequence of human C5aR (anti-C5aR(9-29)) is capable of binding to both normal human and transfected cells bearing C5aR. Flow cytometric analysis of stable murine L cell transfectants (C5aR-neo), human neutrophils, and human monocytes indicated that these cells bound anti-C5aR(9-29) in a specific manner. Moreover, F(ab′)2 fragments of anti-C5aR(9-29) specifically neutralized proinflammatory and immunoregulatory activities induced by natural human C5a. This antiserum was found to block, in a dose-dependent manner, 1) zymosan-induced neutrophil chemotaxis, 2) C5a-induced enzyme release from neutrophils, and 3) C5a-induced cytokine production (IL-6 and IL-8) from human monocytes in vitro. These results suggest that this site-directed polyclonal antiserum specifically interacts with the human C5aR molecule. To the best of our knowledge, none of the existing reports in the literature provided evidence for a site-directed antiserum to C5aR that was capable of specifically blocking C5a-mediated inflammatory/immunoregulatory activities in vitro. Studies conducted with anti-C5aR(9-29) indicated that this antiserum effectively blocked C5a-mediated cell activation but by itself did not activate either neutrophils or monocytes. Combined, these data suggest that this antiserum does not interact with the C5a "effector" site but sterically interferes with C5a binding to its receptor by interacting with the extracellular amino-terminal region of the receptor.

Original languageEnglish (US)
Pages (from-to)377-388
Number of pages12
JournalJournal of Immunology
Volume151
Issue number1
StatePublished - Jul 1 1993

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Anaphylatoxin C5a Receptor
Neutralizing Antibodies
Peptides
Immune Sera
Antibodies
Neutrophils
Monocytes
Zymosan
Chemotaxis
human C5AR1 protein
Interleukin-8
Interleukin-6
Cytokines

ASJC Scopus subject areas

  • Immunology

Cite this

Anti-C5a receptor antibodies : Characterization of neutralizing antibodies specific for a peptide, C5aR-(9-29), derived from the predicted amino-terminal sequence of the human C5a receptor. / Morgan, Edward L.; Ember, Julia A.; Sanderson, Sam; Scholz, Wolfgang; Buchner, Robert; Ye, Richard D.; Hugli, Tony E.

In: Journal of Immunology, Vol. 151, No. 1, 01.07.1993, p. 377-388.

Research output: Contribution to journalArticle

Morgan, Edward L. ; Ember, Julia A. ; Sanderson, Sam ; Scholz, Wolfgang ; Buchner, Robert ; Ye, Richard D. ; Hugli, Tony E. / Anti-C5a receptor antibodies : Characterization of neutralizing antibodies specific for a peptide, C5aR-(9-29), derived from the predicted amino-terminal sequence of the human C5a receptor. In: Journal of Immunology. 1993 ; Vol. 151, No. 1. pp. 377-388.
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abstract = "Results obtained indicate that a site-directed polyclonal antibody specific for a synthetic peptide based on the predicted ami no-terminal sequence of human C5aR (anti-C5aR(9-29)) is capable of binding to both normal human and transfected cells bearing C5aR. Flow cytometric analysis of stable murine L cell transfectants (C5aR-neo), human neutrophils, and human monocytes indicated that these cells bound anti-C5aR(9-29) in a specific manner. Moreover, F(ab′)2 fragments of anti-C5aR(9-29) specifically neutralized proinflammatory and immunoregulatory activities induced by natural human C5a. This antiserum was found to block, in a dose-dependent manner, 1) zymosan-induced neutrophil chemotaxis, 2) C5a-induced enzyme release from neutrophils, and 3) C5a-induced cytokine production (IL-6 and IL-8) from human monocytes in vitro. These results suggest that this site-directed polyclonal antiserum specifically interacts with the human C5aR molecule. To the best of our knowledge, none of the existing reports in the literature provided evidence for a site-directed antiserum to C5aR that was capable of specifically blocking C5a-mediated inflammatory/immunoregulatory activities in vitro. Studies conducted with anti-C5aR(9-29) indicated that this antiserum effectively blocked C5a-mediated cell activation but by itself did not activate either neutrophils or monocytes. Combined, these data suggest that this antiserum does not interact with the C5a {"}effector{"} site but sterically interferes with C5a binding to its receptor by interacting with the extracellular amino-terminal region of the receptor.",
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AB - Results obtained indicate that a site-directed polyclonal antibody specific for a synthetic peptide based on the predicted ami no-terminal sequence of human C5aR (anti-C5aR(9-29)) is capable of binding to both normal human and transfected cells bearing C5aR. Flow cytometric analysis of stable murine L cell transfectants (C5aR-neo), human neutrophils, and human monocytes indicated that these cells bound anti-C5aR(9-29) in a specific manner. Moreover, F(ab′)2 fragments of anti-C5aR(9-29) specifically neutralized proinflammatory and immunoregulatory activities induced by natural human C5a. This antiserum was found to block, in a dose-dependent manner, 1) zymosan-induced neutrophil chemotaxis, 2) C5a-induced enzyme release from neutrophils, and 3) C5a-induced cytokine production (IL-6 and IL-8) from human monocytes in vitro. These results suggest that this site-directed polyclonal antiserum specifically interacts with the human C5aR molecule. To the best of our knowledge, none of the existing reports in the literature provided evidence for a site-directed antiserum to C5aR that was capable of specifically blocking C5a-mediated inflammatory/immunoregulatory activities in vitro. Studies conducted with anti-C5aR(9-29) indicated that this antiserum effectively blocked C5a-mediated cell activation but by itself did not activate either neutrophils or monocytes. Combined, these data suggest that this antiserum does not interact with the C5a "effector" site but sterically interferes with C5a binding to its receptor by interacting with the extracellular amino-terminal region of the receptor.

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