Effects of gamma-interferon on human neutrophils: protection from deterioration on storage

Abstract

Human polymorphonuclear leukocytes (PMN) preincubated overnight with 100 U/mL gamma-interferon (IFN-gamma) had an increased metabolic response, as measured by iodination and/or superoxide production, to stimulation by tumor necrosis factor (TNF), granulocyte-macrophage colony-stimulating factor (GM-CSF), formylmethionyl-leucyl- phenylalanine (FMLP), opsonized zymosan, and lipopolysaccharide (LPS), as compared with cells comparably preincubated in the absence of IFN- gamma. The decline in the staphylocidal activity of the stored PMN was also prevented in part by IFN-gamma, as was the depressed adherence of PMN stimulated with phorbol myristate acetate (PMA), FMLP, TNF, GM-CSF, and LPS. This protective effect of IFN-gamma on PMN function was associated with the prolonged surface expression of the complement receptor three (CR3) alpha-chain (CD11b), CR3 beta-chain (CD18), FcRII (CD32), and FcRIII (CD16), and the appearance of surface FcRI (CD64). The polymerase chain reaction (PCR) was used to amplify neutrophil RNA- derived cDNA recognized by synthetic oliogonucleotides designed from published nucleotide sequences for specific proteins. Using this procedure, mRNA for gp91-phox, p67-phox, p47-phox, CD64, two forms of CD32, CD16, CD11b, CD18, and actin were found to be depressed after overnight storage of neutrophils, and this decrease in steady-state mRNA levels was in part or totally prevented by IFN-gamma. CD64 and gp91-phox mRNA were generally increased by IFN-gamma to a level greater than that of freshly isolated neutrophils. Northern analysis of CD64 and p47 phox mRNAs confirmed the findings with the PCR method. These findings suggest that storage of PMN in a functionally active state is favored by the presence of IFN-gamma.

IFN-γ alters the expression of diverse immunity related genes in a cell culture model designed to represent maturing neutrophils

“…Thus, based on the PLB-985 cell model used in this study, up regulation of integrins is another potential mechanism by which PMN transmigration could be enhanced by IFN-γ. Strikingly, the IFN-γ induced upregulation of ITGAM which we noted is consistent with a previous study looking at the protective effects of INF-γ on stored neutrophils [ 19 ]. In this work it was found that incubation with IFN-γ slowed the decline of the oxidative burst, bactericidal activity and adherence in stored neutrophils.…”

“…It should also be noted that Fc receptors can also activate processes such as the respiratory burst and degranulation, the observed upregulation of Fc receptors seen in our PLB-985 based model thus suggests a potential mechanism by which IFN-γ might enhance neutrophil microbial activity through phagocytosis-independent mechanisms. It is noteworthy that the IFN-γ induced upregulation of FCGRIA and FCGRIIA mRNAs and FCGRIA protein ( Fig 2 ) which we observed is consistent with the protective effects of INF-γ on stored neutrophils [ 19 ]. This earlier work described IFN-γ mediated protection of the oxidative burst, bactericidal activity and adherence in stored neutrophils.…”

Differential Constitutive and Cytokine-Modulated Expression of Human Toll-like Receptors in Primary Neutrophils, Monocytes, and Macrophages

“…Neutrophils were isolated from EDTA-anticoagulated blood by sequential sedimentation in dextran (Sigma, St. Louis, MO) in 0.9% sodium chloride, centrifugation over Histopaque-1077 (Sigma), and hypotonic lysis of erythrocytes, as previously described. 43 Cells were cultured in RPMI 1640 (BioWhittaker, Walkersville, MD) supplemented with 10% heat-inactivated fetal bovine serum (BioWhittaker), 1% L-glutamine, 1% HEPES, and 0.5% penicillin-streptomycin (all from BioWhittaker). Isolated monocytes were used immediately or were incubated at 37˚C in a 5% CO 2 -controlled incubator for the designated time period without stimuli or in the presence of recombinant human IFN-γ (1000 units/ml) (R&D Systems Minneapolis, Minnesota), recombinant human GM-CSF (100 ng/ml) (Immunex, Seattle, WA), recombinant human M-CSF (100 ng/ml) (R&D Systems), or LPS (1 μg/ml) (Sigma).…”

Up-regulation of the dendritic cell marker CD83 on polymorphonuclear neutrophils (PMN): divergent expression in acute bacterial infections and chronic inflammatory disease

“…Prolongation of lifespan is also achieved by cultivating PMN, for example with cytokines such as interferon‐gamma (IFN‐ γ ), granulocyte/macrophage colony stimulating factor (GM‐CSF) or interleukin (IL) ‐3 [8–11] or, as shown more recently, with low doses of tumour necrosis factor alpha (TNF‐ α ) [12]. Prolongation of lifespan depends on de novo protein synthesis and is linked to numerous phenotypical and functional changes [13–17], a process we refer to as transdifferentiation [18,19].…”