PMID- 32458608 OWN - NLM STAT- MEDLINE DCOM- 20200604 LR - 20200604 IS - 1005-6661 (Print) IS - 1005-6661 (Linking) VI - 32 IP - 2 DP - 2020 Mar 25 TI - [Polarization of bone marrow-derived macrophages induced by recombinant Trichinella spiralis cysteine protease inhibitors in vitro]. PG - 181-186 LID - 10.16250/j.32.1374.2019245 [doi] AB - OBJECTIVE: To investigate the regulatory role of recombinant Trichinella spiralis cysteine protease inhibitors (rTs-Cys) in induction of polarization of bone marrow-derived macrophages (BMDMs) in vitro. METHODS: BMDMs were captured and cultured in conditioned medium for 7 days. Then, mature BMDMs were harvested and assigned into four groups. Cells in Group A (negative control) were given 10 ng/mL IFN-gamma combined with 100 ng/mL LPS, cells in Group B (positive control) were treated with IL-4 and IL-10 (at 10 ng/mL both), and cells in Group C (recombinant protein alone) were stimulated with 1 mug/mL rTs-Cys, while cells in Group D (protein co-culture) were simultaneously treated with 1 mug/mL rTs-Cys, 10 ng/mL IFN-gamma and 100 ng/mL LPS. Cells and culture supernatant were collected 24 hour post-treatment, and the proportions of F4/80(+), CD11b(+), CD206(+) and CD11c(+) cells were detected by flow cytometry. The levels of interleukin IL-6 (IL-6), tumor necrosis factor-alpha (TNF-alpha), IL-10 and transforming growth factor-beta (TGF-beta) in the cell culture supernatant were measured by ELISA and the CD86(+) and CD206(+) phenotypes were identified by immunofluorescent staining. RESULTS: Flow cytometry detected no significant difference in the proportion of F4/80(+) CD11b(+) CD11c(+) cells among the four groups (F = 46.184, P < 0.001), and a lower proportion of F4/80(+) CD11b(+) CD11c(+) cells was seen in groups C and D than in group A (all P values < 0.001). There was a significant difference in the proportion of F4/80(+) CD11b(+) CD206(+) cells among the four groups (F = 11.032, P < 0.001), and a greater proportion of F4/80(+) CD11b(+) CD206(+) cells was seen in groups C and D than in group A (all P values < 0.01). Immunofluorescent staining showed higher CD206(+) expression and lower CD86(+) expression in groups C and D than in Group A. There were significant differences in the IL-6 and (F = 3.950, P < 0.001) and TNF-alpha (F = 205.827, P < 0.001) levels in the cell culture supernatants among the four groups, and significantly lower IL-6 and TNF-alpha levels were measured in groups C and D than in Group A (both P < 0.05). There were significant differences in the IL-10 and (F = 8.274, P < 0.001) and TGF-beta (F = 13.559, P < 0.01) levels in the cell culture supernatants among the four groups, and greater IL-10 and TGF-beta levels were measured in Group C than in Group A (both P values < 0.01). In addition, the TGF-beta level was significantly higher in Group D than in Group A (P < 0.05); however, there was no significant difference in the IL-10 level between groups D and A (P > 0.05). CONCLUSIONS: rTs-Cys may induce the polarization of BMDMs to antiinflammatory M2 macrophages in vitro and inhibit the activation of M1 macrophages. FAU - Xie, H AU - Xie H AD - Basic Medical College, Bengbu Medical College, Bengbu 233000, China. AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Chu, L AU - Chu L AD - The Second Affiliated Hospital of Bengbu Medical College, China. FAU - Wu, L Q AU - Wu LQ AD - Anhui Key Laboratory of Infection and Immunity, China. AD - The First Affiliated Hospital of Bengbu Medical College, China. FAU - Fan, X Y AU - Fan XY AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Wang, P AU - Wang P AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Ma, S Y AU - Ma SY AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Zheng, D X AU - Zheng DX AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Li, K L AU - Li KL AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Chen, X Z AU - Chen XZ AD - Basic Medical College, Bengbu Medical College, Bengbu 233000, China. AD - Anhui Key Laboratory of Infection and Immunity, China. FAU - Yang, X D AU - Yang XD AD - Basic Medical College, Bengbu Medical College, Bengbu 233000, China. AD - Anhui Key Laboratory of Infection and Immunity, China. LA - chi GR - gxbjZD15/Science Foundation of Anhui Province/ GR - 2018H174/Science Foundation of Anhui Province/ GR - KJ2019A0383/Program of Natural Science Foundation of the Anhui Higher Education Institutions/ GR - Byycx1806/Postgraduate Scientific Research Innovation Program of Bengbu Medical College/ GR - Byycx1903/Postgraduate Scientific Research Innovation Program of Bengbu Medical College/ GR - 201810367012/National University Students' Innovation and Entrepreneurship Training Program/ GR - 2016-40/Scientific Research Innovation Platform Team of University/ GR - BYTM2019002/Science Foundation of Bengbu Medical College/ PT - Journal Article PL - China TA - Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi JT - Zhongguo xue xi chong bing fang zhi za zhi = Chinese journal of schistosomiasis control JID - 101144973 RN - 0 (Cysteine Proteinase Inhibitors) RN - 0 (Tumor Necrosis Factor-alpha) SB - IM MH - Animals MH - Cells, Cultured MH - Cysteine Proteinase Inhibitors/pharmacology MH - *Macrophage Activation/drug effects MH - Macrophages/drug effects MH - *Trichinella spiralis/genetics/metabolism MH - Tumor Necrosis Factor-alpha/metabolism OTO - NOTNLM OT - Cysteine protease inhibitor OT - Immune regulation OT - Macrophage OT - Trichinella spiralis EDAT- 2020/05/28 06:00 MHDA- 2020/06/05 06:00 CRDT- 2020/05/28 06:00 PHST- 2020/05/28 06:00 [entrez] PHST- 2020/05/28 06:00 [pubmed] PHST- 2020/06/05 06:00 [medline] AID - 10.16250/j.32.1374.2019245 [doi] PST - ppublish SO - Zhongguo Xue Xi Chong Bing Fang Zhi Za Zhi. 2020 Mar 25;32(2):181-186. doi: 10.16250/j.32.1374.2019245.