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Cardiac Fibroblast Functionality Under the Influence of TH2 Cytokines

Undergraduate #13
Discipline: Biological Sciences
Subcategory: Cell and Molecular Biology

Jelonia Rumph - Florida Memorial University
Co-Author(s): William Bracamonte-Baran and Daniela Cihakova, Johns Hopkins University, Baltimore, MD



Background: Recent studies have found that the cytokine IL-33 activates Th2 response via Innate Lymphoid Cells (ILCs). IL-33 also induces eosinophilic pericarditis and is involved in the pathology of autoimmune myocarditis. Furthermore, exogenous IL-33 induces its endogenous production by cardiac fibroblast (CFs). However, CFs do not have ST2, which is the surface receptor for IL-33. Aims/Hypothesis: This project aimed to investigate the role of CFs and innate lymphoid cells in a pro-inflammatory loop involved in pericarditis and myocarditis. In this loop we hypothesize: CF-derived IL-33 activates ILC type 2, resulting in the release of cytokines that can further activate CFs; in turn creating a pro-inflammatory cycle. Methods/Results: Sca1+ CFs were FACS sorted from mice hearts (WT Balb/c) and cultured in vitro for 6 days with focal ILC type 2-derived cytokines: IL5, IL9 and IL13. The readout of IL33 production was determined by ELISA, and the phenotype of CFs were analyzed through flow cytometry. A constitutive expression of IL33 by Sca1 CFs (27.55±1.35 pg/mL) was found. The production of IL-33 was not enhanced by IL5, IL9 or IL13 (29.30±1.20; 28.25±1.65; 33.2±2.90 pg/mL, respectively). Conclusion: The project showed that stimulation by IL-5, IL-9 and IL-13 were not the direct source of the release of IL-33 by CFs. As consequence, further investigation is required to determine if other ILC2-derived cytokine can stimulate CFs, or if another ST2+ cell, like mast cells, can mediate the ILC-CFs cross-talk.

Funder Acknowledgement(s): NHLBI RO1HL 118183; NHLBI RO1HL 113008; Catalyst Award JHU; American Autoimmune Related Diseases Association Inc.

Faculty Advisor: Marilyn Sherman, msherman@fmuniv.edu

Role: I performed the entire project on my own, but was overseen by William Bracamonte-Baran. My duties included sacrificing the mice by injecting them with an anesthetic and cervical dislocation. I also harvested and purified the heart and liver (centrifugation and lysis of RBCs) until pure cardiac fibroblasts were obtained. I then stained the samples with antibodies. I was also responsible for running the samples through flow cytometry and analyzing the results.

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This material is based upon work supported by the National Science Foundation (NSF) under Grant No. DUE-1930047. Any opinions, findings, interpretations, conclusions or recommendations expressed in this material are those of its authors and do not represent the views of the AAAS Board of Directors, the Council of AAAS, AAAS’ membership or the National Science Foundation.

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