Abstract
Although transcription of mast cell (MC) secretory granule neutral protease genes has been shown to distinguish MC subclasses in mucosal and serosal environments, the specific cytokines that regulate the expression of these genes have not been determined. To examine cytokine-mediated gene regulation, bone marrow-derived MC (BMMC) differentiated in vitro were obtained by culturing mouse bone marrow progenitor cells in the presence of WEHI-3 cell-conditioned medium, concanavalin A-stimulated splenocyte-conditioned medium (BMMCC), or recombinant (r) interleukin (IL)-3 (BMMCIL-3). All three populations of BMMC expressed the serosal MC-specific transcripts that encode mouse MC serine protease (MMCP)-5, MMCP-6, and MC carboxypeptidase A. However, only BMMCC contained MMCP-2 mRNA, a late expressed gene selectively transcribed by intestinal mucosal MC that proliferate during helminthic infestation in response to the T cell-derived cytokines IL-3, IL-4, and IL-10. When BMMCIL-3 were exposed to rIL-10 in the presence of either rIL-3 or rIL-4, they expressed MMCP-2 mRNA. Not only was the transcription of the MMCP-2 gene in BMMC dependent on continuous exposure of the cells to rIL-10, but the level of MMCP-2 mRNA in these cells could be down-regulated by rIL-3. These studies comparing the effects of two cytokines on the transcriptional regulation of secretory granule protease genes in MC demonstrate that rIL-10 induces BMMCIL-3 to express the mucosal MC protease MMCP-2, that rIL-3 attenuates the rIL-10-induced expression of this gene, and that transcription of the MMCP-2 gene is reversed in the absence of rIL-10.
Highlights
From the Departmentof Medicine, Harvard MedicalSchool and the Department of Rheumatology and Immunology, Brigham and Women’s Hospital, Boston, Massachusetts021I5
Granule neutral proteasegenes has been shown to dis- As assessed by their content of protein and/or RNA, mouse tinguish mast cell (MC) subclasses in mucosal and serosal envi- serosal MC express ronments,thespecificcytokines that regulate the expression of these genes have not been determined
BMMCc contained mouse MC protease (MMCP)-2 mRNA, a late expressed gene selectively transcribed by intestinal mucosal MC that proliferate during helminthic infestation in response to the T cell-derived cytokines IL-3, IL-4, and cyte-conditionedmedium (BMMCc) has not been determined
Summary
BMMC, bonemarrow-derivedMC;BMMCc, BMMC derived with These findings demonstrate the principle of cytokine regulaconcanavalin A-stimulated splenocyte-conditioned medium; BMMCIL.3, BMMC derived with rIL-3; BMMCw, BMMC derived with WEHI-3 cell-conditioned medium; MMCP, mouse MC protease; SG-PG, setion of transcription of secretory granule neutral proteases and illustrate the sequential interaction of IL-3 cretory granule proteoglycan peptide core; SDS, sodium dodecyl sul- followed byIL-10 in achieving expression of the mucosal MC, fate; r, recombinant. BMMC is reversible, mouse bone marrow cells were cultured for 2-3 weeks in the presenceof rIL-3, WEHI-3cell-conditioned medium, or concanavalin A-stimulated splenocyte-conditioned medium.
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have