الملخص
Objective:To investigate the levels of interleukin-7 (IL-7), T helper cell 9 (Th9), and cytotoxic T cell 9 (Tc9), and to assess the regulation of exogenous IL-7 to Th9 in septic cardiomypathy patients.Methods:A cross-sectional study was conducted. Septic cardiomypathy patients, septic patients, and controls were enrolled in the 7th People’s Hospital of Zhengzhou between June 2018 and January 2022. Plasma and peripheral blood mononuclear cells (PBMCs) were isolated. PBMCs were stimulated with recombinant IL-7. Plasma IL-7, IL-9, and soluble CD127 levels were measured by enzyme-linked immunosorbent assay. CD127 expression on T cells and Th9/Tc9 percentageswere measured by flow cytometry. mRNAexpressions of purine-rich nucleic acid binding protein 1 (PU.1) and forkhead box protein O1 (Foxo1) were semi-quantified by real-time PCR. IL-7-stimulated Th9 cells from septic cardiomypathy patients were co-cultured with autologous CD8 + T cells and human umbilical vein endothelial cells. Target cell death, cytotoxic molecules and cytokines secretions were investigated. Kruskal-Wallis test was used for comparison among groups, while Dunns multiple test was used for comparison between the two groups. Paired t test or Wilcoxon paired test was used for comparison before and after stimulation. Results:A total of 21 septic cardiomypathy patients, 56 septic patients, and 31 controls were enrolled. IL-7 level was lower in septic cardiomypathy patients compared with in septic patients and controls [75.71(42.28, 135.59) pg/mL vs. 118.47(60.18, 171.73) pg/mL vs. 168.42(105.41, 232.30) pg/mL, P<0.05]. Soluble CD127 level was higher in septic cardiomypathy patients and septic patients compared with in controls [96.70(56.76, 147.04) pg/mL vs. 69.75(43.19, 111.28) pg/mL vs. 29.13(19.33, 42.11) pg/mL, P<0.001]. There was no significant difference of CD127 +CD8 + percentage among three groups ( P=0.477). Th9 percentage and Foxo1 mRNA expression was lower in septic cardiomypathy patients compared with in septic patients and controls ( P<0.001). Tc9 percentage, PU.1 mRNA expression and IL-9 level was lower in septic cardiomypathy patients and septic patients compared with in controls ( P<0.001). Th9 percentage, PU.1/Foxo1 mRNA expression, and IL-9 secretion was up-regulated in response to IL-7 stimulation in PBMCs from septic cardiomypathy patients ( P<0.05). IL-7 stimulated Th9 cells from septic cardiomypathy patients promoted CD8 + T cell-mediated target cell death ( P<0.05). Perforin, granzyme B, and interferon-γ levels were also increased in the supernatants ( P<0.05). Conclusion:IL-7 could enhance Th9 cell activity in septic cardiomypathy patients.
الملخص
To develop a method based on immunoreactions for detection of Ectropis obliqua Nucleopolyhedrovirus (EoNPV), the polyhedra of the virus were purified and used to immunize the mouse BALB/c. The spleen cells from the immunized mice were then fused with the myeloma cell line Sp2/0. A hybridoma cell line which can stably secrete the monoclonal antibody against EoNPV was achieved by using indirect ELISA screening and cloning methods, and was named as 7D3. Meanwhile, the polyhedrin gene was cloned from EoNPV and expressed in E. coli. Western blotting analysis showed that the monoclonal antibody prepared from 7D3 could specifically react with the recombinant polyhedrin. An indirect ELISA method based on this monoclonal antibody for detecting EoNPV in infected tea looper was developed.