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Effects of interleukins on connective tissue type mast cells co-cultured with fibroblasts
F Levi-Schaffer1, V Segal, M Shalit
1Department of Pharmacology, School of Pharmacy, Hadassah Medical School, Hadassah University Hospital, Hebrew University of Jerusalem, Israel.
Abstract:
We investigated the effects of interleukin-2 (IL-2), interleukin-3 (IL-3) and interleukin-4 (IL-4) on mouse and rat peritoneal mast cells (MC) co-cultured with 3T3 fibroblasts (MC/3T3). The continuous presence of these cytokines for 7-9 days in the culture media was neither toxic nor caused proliferation of MC, as determined by the stability of MC numbers in culture. Long-term incubation of mouse MC/3T3 with IL-2 (100 U/ml), IL-3 (50 U/ml), IL-4 (50 U/ml) or a mixture of IL-3 and IL-4 (25 U/ml) induced an increase in basal histamine release of 79.3 +/- 19.0%, 41.0 +/- 17.3%, 25.2 +/- 10.4% and 30.2 +/- 3.2%, respectively, over control cells incubated with medium alone. When rat MC/3T3 were incubated for 7 days with the various interleukins an enhancement in histamine release similar to that observed with mouse MC/3T3 was found. Preincubation (1 hr) of rat MC/3T3 with interleukins prior to immunological activation with anti-IgE antibodies enhanced histamine release. The highest effect was observed with IL-3 + IL-4 (60.4 +/- 10.8% increase) followed by IL-2 (51.5 +/- 4.5%), IL-4 (28.6 +/- 10.3%) and IL-3 (13.2 +/- 4.2%). This study demonstrates that when mouse and rat peritoneal MC are cultured with fibroblasts in the presence of interleukins they do not proliferate, suggesting that they preserve their connective tissue type MC phenotype. Moreover, interleukins display a pro-inflammatory effect on these cells by enhancing both basal and anti-IgE-mediated histamine release.
Insights
Interleukin-2, -3, and -4 do not cause mast cell proliferation but increase histamine release in mouse and rat models. These interleukins (ILs) enhance both basal and anti-IgE-mediated histamine release, indicating a pro-inflammatory effect.
Area of Science:
- Immunology
- Cell Biology
- Cytokine Signaling
Background:
- Mast cells (MC) are key players in allergic and inflammatory responses.
- Interleukins (ILs) are crucial signaling molecules in the immune system.
- The role of specific interleukins (IL-2, IL-3, IL-4) on peritoneal mast cells co-cultured with fibroblasts requires further elucidation.
Purpose of the Study:
- To investigate the effects of IL-2, IL-3, and IL-4 on mouse and rat peritoneal mast cells co-cultured with 3T3 fibroblasts (MC/3T3).
- To determine if these interleukins induce mast cell proliferation or toxicity.
- To assess the impact of interleukins on basal and antigen-induced histamine release from mast cells.
Main Methods:
- Co-culture of mouse and rat peritoneal mast cells with 3T3 fibroblasts for 7-9 days.
- Incubation of mast cell cultures with IL-2, IL-3, IL-4, or a combination of IL-3 and IL-4.
- Measurement of basal histamine release.
- Preincubation with interleukins followed by activation with anti-IgE antibodies to measure histamine release.
Main Results:
- Continuous presence of IL-2, IL-3, and IL-4 was not toxic and did not induce proliferation of mast cells.
- Incubation with interleukins significantly increased basal histamine release in both mouse and rat MC/3T3 cultures.
- Preincubation with interleukins enhanced anti-IgE-mediated histamine release, with IL-3 + IL-4 showing the strongest effect.
Conclusions:
- Interleukins (IL-2, IL-3, IL-4) do not alter the phenotype of connective tissue-type mast cells in co-culture with fibroblasts.
- Interleukins exert a pro-inflammatory effect on mast cells by enhancing both spontaneous and antigen-stimulated histamine release.
- These findings highlight the modulatory role of interleukins in mast cell-mediated inflammatory processes.