Identification and characterization of a monoclonal antibody to an antigen expressed on activated macrophages

T. P. Koestler, D. Rieman, K. Muirhead, R. G. Greig, George Poste

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

A hybridoma clone secreting a monoclonal antibody, designated MA158.2, that reacts with an antigen expressed on lymphokine-treated marcrophages was produced by fusion of mouse myeloma cells with rat spleen cells immunized against C57BL/6 peritoneal macrophages rendered tumoricidal in vitro by incubation with the lymphokine macrophage-activating factor. The specificity of the antibody for activated macrophages and lack of reactivity with histologically diverse cell types was determined by radioimmune indirect binding and flow cytometry. MA158.2 antibody binds to mouse peritoneal macrophages elicited by nonspecific inflammatory agents and to tumoricidal macrophages elicited with Corynebacterium parvum. Resident peritoneal, splenic, and alveolar macrophages were only weakly positive. Several macrophage cell lines (P388D1, WEH1-231, J774, RAW 264.7), murine fibroblasts, and neutrophils did not bind detectable amounts of MA158.2. Radioimmune indirect binding analysis demonstrated that cell suspensions prepared from C57BL/6 mouse spleen, thymus, and lymph node as well as polymorphonuclear leukocytes, lymphocytes, and T- and B-cell murine lymphomas were MA158.2 negative. Expression of the reactive antigen on the macrophage cell surface was enhanced 3-fold following in vitro activation of elicited macrophages with macrophage-activating factor and the kinetics of activation to the tumoricidal state paralleled the increased expression of the antigen recognized by MA158.2. MA158.2 is a rat IgG2a antibody containing a single specific heavy and light chain that does not detect a polymorphic determinant. This monoclonal antibody will be a useful tool for monitoring the efficacy of agents in activating murine macrophages to the tumoricidal state and in analyzing the sequence of biochemical events that culminate in macrophage activation.

Original languageEnglish (US)
Pages (from-to)4505-4509
Number of pages5
JournalProceedings of the National Academy of Sciences of the United States of America
Volume81
Issue number14 I
StatePublished - 1984
Externally publishedYes

Fingerprint

Macrophages
Monoclonal Antibodies
Antigens
Peritoneal Macrophages
Macrophage-Activating Factors
Macrophage Activation
Lymphokines
Neutrophils
Spleen
Propionibacterium acnes
Antibody Specificity
Antibodies
Hybridomas
Alveolar Macrophages
B-Cell Lymphoma
Inbred C57BL Mouse
Thymus Gland
Suspensions
Flow Cytometry
Clone Cells

ASJC Scopus subject areas

  • Genetics
  • General

Cite this

Identification and characterization of a monoclonal antibody to an antigen expressed on activated macrophages. / Koestler, T. P.; Rieman, D.; Muirhead, K.; Greig, R. G.; Poste, George.

In: Proceedings of the National Academy of Sciences of the United States of America, Vol. 81, No. 14 I, 1984, p. 4505-4509.

Research output: Contribution to journalArticle

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