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Human fibroblasts maintain the viability and augment the functional response of human neutrophils in culture
C J Ling1, W F Owen, K F Austen
1Department of Medicine, Harvard Medical School, Boston, Massachusetts.
Abstract:
When human neutrophils were co-cultured for 72 h with nontransformed human fibroblasts, 69 +/- 3% (n = 13) survived, as compared with survival levels of 2 +/- 1% (n = 15) and 26 +/- 6% (n = 7), respectively, for neutrophils cultured for the same time period in enriched medium alone or supplemented with 10 pM recombinant human granulocyte/macrophage colony-stimulating factor (rh GM-CSF). Conditioned medium from the human fibroblast cultures enhanced neutrophil survival in a dose-dependent fashion to the same level achieved with neutrophil/fibroblast co-cultures, and its soluble viability-sustaining activity was not inhibited by preincubation with neutralizing antiserum against rh GM-CSF. As compared with freshly isolated replicate samples, neutrophils co-cultured with human fibroblasts for 72 h exhibited augmented FMLP-stimulated superoxide production without spontaneous superoxide generation. This striking extension of survival and associated priming for a ligand response by neutrophils co-cultured with human fibroblasts suggests that fibroblasts may contribute to the proinflammatory properties of neutrophils in tissues.
Insights
Human fibroblasts significantly enhance neutrophil survival and function. Fibroblast-conditioned medium also boosts neutrophil viability, suggesting fibroblasts play a role in inflammation.
Area of Science:
- Immunology
- Cell Biology
Background:
- Neutrophils are key immune cells involved in inflammation.
- Understanding factors that regulate neutrophil survival and function is crucial for studying inflammatory diseases.
Purpose of the Study:
- To investigate the effect of human fibroblasts on neutrophil survival and function.
- To determine if fibroblast-conditioned medium can mimic the survival-enhancing effects of co-culture.
Main Methods:
- Human neutrophils were co-cultured with human fibroblasts for 72 hours.
- Neutrophil survival was assessed in enriched medium alone, with rh GM-CSF, or with fibroblast-conditioned medium.
- Neutrophil function, specifically FMLP-stimulated superoxide production, was measured.
Main Results:
- Co-culture with fibroblasts significantly increased neutrophil survival (69%) compared to medium alone (2%) or rh GM-CSF (26%).
- Fibroblast-conditioned medium enhanced neutrophil survival in a dose-dependent manner.
- Neutrophils co-cultured with fibroblasts showed enhanced FMLP-stimulated superoxide production without spontaneous generation.
Conclusions:
- Human fibroblasts provide a potent survival-promoting factor for neutrophils, independent of rh GM-CSF.
- Fibroblasts prime neutrophils for enhanced inflammatory responses.
- These findings suggest fibroblasts contribute to neutrophil-mediated inflammation in tissues.