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EFFECT OF ULINASTATIN ON REGULATORY T CELLS AND T HELPER 17 CELLS IN PATIENTS WITH SEVERE SEPSIS

GUOHUI LI 1, JILIN MA 2, LONG CAI 3, ZHANLI SHI 4, JING SUN 4, JIAYING ZHANG 4, ZHIHUI LI 4, LEIMIN SUN 4, HONGJUAN ZHOU 3, KUN FANG 4, SU YU 2*

1.Department of Occupational Medicine, Hang Zhou Red Cross Hospital, Hangzhou, 310003, China
2.Department of Rheumatology, Immunology and Nephrology, Hang Zhou Red Cross Hospital, Hangzhou, 310003, China
3.Central Laboratory, Hang Zhou Red Cross Hospital, Hangzhou, 310003, China
4.Department of Intensive Care Unit, Hang Zhou Red Cross Hospital, Hangzhou, 310003, China

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The aim of this study was to assess the effect of ulinastatin (UTI) on the levels of immune regulatory cells, pro-inflammatory mediators, and the expression of severe sepsis in regulatory T cells (Treg) and T helper 17 (Th17) cells monocytes. Peripheral blood was obtained from severe sepsis patients. The blood was divided in two groups, the experimental group treated with ulinastatin and the control group without any treatment. The rate of CD4+ CD25+ Foxp3+ Treg cells in CD4+ T cells was analysed by flow cytometry. CD4+ CD25+ Foxp3+ Treg cells were purified by magnetic cell labelling (MACS) and expanded in vitro by culture with addition of CD3/CD8 Dynal beads and IL 2. The cells number, activity and purity after expanding the cells for 0 day, 7 days, 14 days and 21 days were determined. The rate of CD4+ CD25+ Foxp3+ and CD8+ CD25+ Foxp3+ cells in the control cell group was (6.41 ± 0.37)% and (6.67 ± 0.48)%. The rate of CD4+ CD25+ Foxp3+ and CD8+ CD25+ Foxp3+ cells gradually decreased with the increase of UTI concentration. When the concentration of UTI reached 1600 U/mL, the percentage of CD4+ CD25+ Foxp3+ and CD8+ CD25+ Foxp3+ cells were (3.40 ± 0.16)% and (3.53 ± 0.22)% respectively. UTI inhibited the differentiation of CD4+ CD25- cells into CD4+ IL-17 cells and CD4+ CD25+ Foxp3+ cells and the differentiation of CD8+ CD25- cells into CD8+ IL-17 cells and CD8+ CD25+ Foxp3+ cells. The inhibitory effects observed were dose dependent.