Modulation of Fc receptors for IgG on bovine polymorphonuclear neutrophils by interferon-gamma through de novo RNA transcription and protein synthesis.
Polymorphonuclear neutrophils (PMN) from 4 cows were preincubated (30 minutes, 37 C) in either actinomycin D (100 micrograms/ml) or puromycin (10 micrograms/ml), inhibitors of mRNA transcription and protein translation, or in medium 199. The PMN were incubated for a further 4.5 hours in medium containing 100 U of recombinant bovine interferon-gamma (rboIfn-gamma). The PMN were then incubated with bovine IgG1, IgG2, IgM, or aggregated IgG (aIgG; 4 C, 12 hours) for flow cytometric analysis, using fluoresceinated isotype-specific antibody. The percentage of PMN binding the ligand and the logarithmic mean fluorescent channel (LMFC), an indicator of the amount of receptor (R) expression, were recorded. Competitive inhibition of ligand binding was measured by incubating PMN with fluoresceinated IgG2 in the presence or absence of 100-fold excess of IgG1, IgG2, and aIgG. Activation with rboIfn-gamma induced a 4.5-fold increase in binding of IgG1 and a fivefold increase in LMFC for IgG2. These increases were inhibited by actinomycin D and puromycin. Percentage of PMN binding aIgG decreased after activation by rboIfn-gamma. Interferon-gamma treatment did not affect binding or LMFC of IgM. However, binding of IgM was reduced by treatment with actinomycin D. Binding of fluoresceinated IgG2 was inhibited by unlabeled IgG1, IgG2, and aIgG. Results indicate that bovine PMN Fc receptors (FcR) for IgG1 and IgG2 were rboIfn-gamma inducible, that induction required de novo transcription and translation, that a heterogeneous population of FcR exist on bovine PMN, and that IgG1 and IgG2 share a common FcR. Further, bovine PMN are capable of gene activation and are responsive to changes in their environment, thus being amenable to modulation for effective pathogen destruction.
Worku
M
M
Department of Animal Sciences, University of Maryland, College Park 20742.
Paape
M J
MJ
Marquardt
W W
WW
D000818:Animals; D001667:Binding, Competitive; D002417:Cattle; D003609:Dactinomycin / Q000494:pharmacology; D005260:Female; D005434:Flow Cytometry; D015870:Gene Expression / Q000187:drug effects*; D007074:Immunoglobulin G / Q000378:metabolism; D066298:In Vitro Techniques; D007371:Interferon-gamma / Q000494:pharmacology*; D007700:Kinetics; D009504:Neutrophils / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism; D011691:Puromycin / Q000494:pharmacology; D017452:Receptors, IgG / Q000096:biosynthesis* / Q000187:drug effects / Q000378:metabolism; D011994:Recombinant Proteins; D014158:Transcription, Genetic / Q000187:drug effects*
D007074:Immunoglobulin G; D017452:Receptors, IgG; D011994:Recombinant Proteins; D003609:Dactinomycin; D011691:Puromycin; D007371:Interferon-gamma