Function And Polarization Of Peritoneal Macrophages In Interleukin-33 Knock-out Mouse With Acute Toxoplasma Gondii Infection

Objective:To investigate the effect of IL-33 knockout on the polarization of peritoneal macrophages from mouse with acute toxoplasma infection,in order to uncover the function of IL-33 knockout in mice macrophages with acute toxoplasma infection.Methods:1.pM?was isolated from wild-type C57BL/6 and IL-33-deficient mice and was infected with toxoplasma tachyzoites for 30 minutes.Then the phagocytosis of pM?was detected;2.pM?was isolated from wild-type C57BL/6 and IL-33 deficient mice and was divided into infected group and control group respectively.The following experiments were performed based on the mice model infected with 1.2×10⁶tachyzoites/mouse.;3.The mRNA of iNOS,Arg-1,IL-1,IL-10 and IL-12 were detected by real-time PCR.4.IL-12,TNF-?and IL-10 were detected by ELISA;5.NO was detected by Griess;6.The surface molecules?CD80,CD86,CD206,TLR4,TLR2,MHC??were analyzed by flow cytometry;7.All the data were analyzed with Split-plot design of two-factor in SPSS software.Results:1.The phagocytic percentage of pM?from IL-33^-/-mice and wild type mice infected with Toxoplasma was 0.95±0.01 and 0.978±0.011 respectively with significant difference?P<0.05?;2.There was no significant difference in mRNA expression level of IL-1,IL-10,IL-12,iNOS and Arg-1 from IL-33^-/-mice infected and control group,wild type mice infected and control group?P>0.05?.3.IL-10 expression level from IL-33^-/-mice infected and control group was6229.70±3673.16 pg/ml and 5811.18±3484.13 pg/ml respectively with significant difference?P<0.05?;TNF-?expression level was 1732.49±325.17pg/ml and 1602.31±247.94 pg/ml respectively with significant difference?P<0.01?;NO expression level was 31.62±14.47 uM and 28.46±12.60 uM respectively with significant difference?P<0.01?;IL-10 expression level from wild type mice infected and control group was4637.62±1645.85 pg/ml and 4395.06±1469.16 pg/ml respectively with significant difference?P<0.05?;TNF-?expression level was 1488.35±757.99pg/ml and 1324.04±623.64 pg/ml respectively with significant difference?P<0.01?;NO expression level was 15.77±9.16 uM and 11.94±9.62 uM respectively with significant difference?P<0.01?;There was no significant difference in IL-10 and TNF-?expression between IL-33^-/-and wild type mice infected group?P>0.05?;NO expression was increased in IL-33^-/-mice infected group compared with that in wild type mice infected group?P<0.01?.IL-12 was undetectable with applied reagent in this study.4.The pM?proportion of CD86 was 90.53±6.67%and 61.69±8.08%respectively from IL-33^-/-mice infected and control group with significant difference?P<0.01?;The pM?proportion of CD86 was 50.49±22.99%and 37.86±12.74%respectively from wild type mice infected and control group with significant difference?P<0.01?;There was no significant difference between IL-33^-/-and wild type mice infected group?P>0.05?.The pM?proportion of CD80 from IL-33^-/-mice infected and control group,wild type mice infected and control group was 78.72±12.22%,70.75±15.02%,77.74±8.24%,77.90±8.98%respectively with no significant difference?P>0.05?.The pM?proportion of CD206 from IL-33^-/-mice infected and control group,wild type mice infected and control group was 12.00±3.88%,12.03±2.96%,14.96±2.56%,17.38±6.41%respectively with no significant difference?P>0.05?.The CD206 expression was decreased in IL-33^-/-mice infected group compared with that in wild type mice infected group?P<0.01,Independent-Samples T Test?.5.The pM?proportion of TLR2 was 54.44±9.88%and 57.75±10.49%respectively from IL-33^-/-mice infected and control group without significant difference?P>0.05?;The pM?proportion of TLR2 was 64.20±17.77%and 42.29±0.23%respectively from wild type mice infected and control group without significant difference?P>0.05?;The TLR2 was decreased in IL-33^-/-mice infected group than that in wild type mice infected group?P<0.01?.6.The pM?proportion of MHC?was 58.07±10.76%and 53.97±13.83%respectively from IL-33^-/-mice infected and control group with significant difference?P<0.01?;The pM?proportion of MHC?was 43.20±13.73%and34.99±11.20%respectively from wild type mice infected and control group with significant difference?P<0.01?;There was no significant difference in MHC?expression between IL-33^-/-and wild type mice infected group?P>0.05?7.The pM?proportion of TLR4 was 1.84±0.43%and 1.26±0.57%respectively from IL-33^-/-mice infected and control group with significant difference?P<0.01?;The pM?proportion of TLR4 was 1.06±0.27%and 0.63±0.28%respectively from wild type mice infected and control group with significant difference?P<0.01?;The TLR4 was increased in IL-33^-/-mice infected group than that in wild type mice infected group?P<0.05?.Concludes:IL-33 knockout promote the secretion of NO and the expression of MHC?,CD86 and TLR4 via driving polarization of M1 macrophages,thereby enhancing the immune protection in acute toxoplasma infection.