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Restoration of the Lipopolysaccharide‐Responsive Phenotype in C3H/HeJ Peritoneal Macrophage‐P388D1 Cell Hybrids
Author(s) -
Zuckerman Steven H.,
Surprenant Yvonne M.,
Carlson Donald G.,
Beutler Bruce
Publication year - 1988
Publication title -
journal of leukocyte biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.819
H-Index - 191
eISSN - 1938-3673
pISSN - 0741-5400
DOI - 10.1002/jlb.43.2.133
Subject(s) - biology , macrophage , lipopolysaccharide , tumor necrosis factor alpha , microbiology and biotechnology , phenotype , antigen , hypoxanthine guanine phosphoribosyltransferase , immunology , genetics , in vitro , gene , mutant
The fusion of thioglycollate‐elicited peritoneal macrophages from the lipopolysaccharide (LPS)‐nonresponsive C3H/HeJ mouse strain to a hypoxanthine phosphoribosyltransferase (HPRT)‐negative variant of the murine macrophage cell line P388D1 has resulted in the derivation of eight hybrid clones following HAT selection. Propidiumiodide staining followed by flow cytometry has demonstrated that the DNA content of the hybrids represents the sum of the parents. Codominant expression of class I antigens from both parental haplotypes is observed in the hybrids. While class II antigens are inducible following a 72‐hr induction with gamma interferon‐containing supernatants, the amount of each haplotype varies between clones. These hybrids demonstrate Fc‐mediated crythrophagocytosis in contrast to P388D1. In distinction to the C3H/HeJ primary peritoneal‐macrophage parent, LPS treatment of the hybrids results in the Increased release of both interieukin‐1 (IL‐1) and cachectin/tumor necrosis factor (TNF) into culture supernatants. Therefore, cell fusion has resulted in the stable restoration of the LPS‐responsive phenotype in C3H/HeJ macrophage hybrids. These macrophage hybrids should serve as useful models In understanding the regulation of macrophage effector functions in response to environmental stimuli.