| Background:Trichinosis is a globally parasitic disease that not only is harmful to human health,but also seriously affects the productions and developments of animal husbandry.The immune response elicited by Trichinella spiralis is an interactive multifactorial process.Now,the researches on the immune response elicited by Trichinella spiralis have been studied on the level of cells and cytokines,but the kinetics of CD4~+T cell subsets secreting related cytokines in infection with Trichinella Spiralis have not been systematically reported.Objective:To investigate the kinetics of Th1,Th2,Th9,Th17,Treg,CD4~+IL-22~+T cells and their related cytokines in Trichinella spiralis infection,and to provide experimental basis to further study the host immune mechanism after Trichinella Spiralis infection and the pathogenic mechanism of trichinosis.Methods:1.To establish a mouse model of Trichinella infection:taking KM mice infected with Henan pig-derived Trichinella spiralis in laboratory,collecting muscle larvaes by artificial pepsin digestion,and experimental group mice was gastrointestinally administrated 300 muscles larvae and the control group was given the same amount of physiological saline.2.The percentage of Th1,Th2,Th9,Th17,Treg,CD4~+IL-22~+T cells in total CD4~+T cells detected by flow cytometry:the spleens and mesenteric lymph nodes of mice infected with Trichinella spiralis were taken at 0day,2 days,5 days,7 days,14 days,21 days,28 days,42 days,56 days after infection(dpi),respectively.The proportion of Th1,Th2,Th9,Th17,Treg,CD4~+IL-22~+T cells in CD4~+T cells detected by flow cytometry.3.ELISA was used to detect the level of cytokine in serum:Serum level of IFN-?,IL-4,IL-9,IL-17,IL-10,TGF-?and IL-22 was detected by ELISA.Result:1.The kinetics of Th1 cells in mice infected with Trichinella spiralis:The proportion of Th1 cells in spleens from the infected group reached the peak at the 7dpi,and significantly decreased at 14dpi,then,it remained at a stable level,and decreased to the lowest value at 42dpi,and rose again at 56 dpi.Compared with the normal group,the proportion of Th1 cells in the infected group was increased from 7dpi to 56dpi(P<0.05).The proportion of Th1 cells in the mesenteric lymph nodes from the infected group began to increase at 2dpi,then gradually increased to the peak at 7dpi,and decreased to the lowest value at 14dpi.It rose again at 21dpi,then,remained at a stable level,decreased at 42dpi and remained at normal level.Compared with the normal group,the proportion of Th1 cells in the mesenteric lymph nodes from the infected group significantly increased from 2dpi to 28dpi(P<0.05),and there was no significant difference from 42dpi to 56dpi(P>0.05).2.The kinetics of Th2 cells in mice infected with Trichinella spiralis:The proportion of Th2 cells in spleens from the infected group began to increase at 2dpi,peaked at 7dpi,and then gradually decreased to normal levels.Compared with the normal group,the proportion of Th2 cells in the infected group was increased from 2 dpi to 14 dpi(P<0.05),and there was no difference from 21dpi to 56dpi(P>0.05).The proportion of Th2 cells in the mesenteric lymph nodes from the infected group increased at 2dpi,and remained at a higher level than normal group at 5 dpi to 14 dpi,reached at the highest value at 21dpi.Then,it dropped dramatically and returned to normal level at 28dpi(P>0.05),and then increased at 42dpi,but it returned to normal level again at 56dpi(P>0.05).3.The kinetics of Treg cells in mice infected with Trichinella spiralis:The proportion of Treg cells in spleens from the infected group rose to the peak at 5 dpi,and then decreased,remained stable at 7 dpi to 42 dpi,decreased to normal level at 56dpi(P>0.05).The proportion of Treg cells in the mesenteric lymph nodes from the infected group increased at 2 dpi,reached the highest value at 5dpi,then decreased,and maintained stable at 14 dpi to 42 dpi,dropped to normal level at56 dpi(P>0.05).4.The kinetics of Th17 cells in mice infected with Trichinella spiralis:The proportion of Th17 cells in the spleens from the infected group began to increase at 7 dpi,and rose to the highest value at 14 dpi,then began to decrease,and remained stable from 21dpi to 56dpi.Compared with the normal group,the proportion of Th17 cells in the spleens from the infected group significantly increased at 7dpi to 56dpi(P<0.01).The proportion of Th17 cells in the mesenteric lymph nodes from the infected group increased to the peak at 2 dpi,and then dramatically decreased to the normal level at 7dpi(P>0.05).Then,the proportion of Th17 cells increased again and reached the peak at 28dpi and then decreased rapidly,dropped to the normal level at 56dpi(P>0.05).5.The kinetics of Th9 cells in mice infected with Trichinella spiralis:The proportion of Th9 cells in spleens from the infected group gradually increased from 2 dpi to 21 dpi,and rose to the highest value at 21 dpi,then decreased dramatically,and returned to normal level at 28 dpi(P>0.05),then increased again.Compared with the normal group,the proportion of Th9 cells in spleens from the infected group was significantly increased at 42 dpi to 56 dpi(P<0.05).The proportion of Th9 cells in the mesenteric lymph nodes from the infected group increased at 14 dpi,reached the peak at 21dpi,and returned to the normal level at 28 dpi(P>0.05),rose again at 42dpi,and the difference was statistically significant compared with the normal group(P<0.01),and decreased to the normal level at 56 dpi(P>0.05).6.The kinetics of CD4~+IL-22~+T cells in mice infected with Trichinella spiralis:The proportion of CD4~+IL-22~+T cells in the spleens from the infected group began to increase at 5 dpi,and decreased to the normal level at 7dpi(P>0.05),then increased,and remained at a stable level at 14dpi to 56dpi.Compared with the normal group,the proportion of CD4~+IL-22~+T cells in the spleens from the infected group was significantly increased at 14 dpi to 56 dpi(P<0.05).The proportion of CD4~+IL-22~+T cells in the mesenteric lymph nodes from the infected group increased dramatically at 14dpi,reached the peak at 21dpi,then decreased rapidly,and remained at normal level(P>0.05).7.The kinetics of cytokines levels in serum in mice infected with Trichinella spiralis:IFN-?levels in Serum increased rapidly to the peak at 7dpi,then began to decline,and remained stable at 14 dpi to 21 dpi,and gradually decreased,but still higher than normal level(P<0.05).IL-4 levels in Serum increased dramatically to the peak at 2 dpi,then gradually decreased,and returned to the normal level at 28dpi(P>0.05),increased again at 42dpi,and decreased to normal level at 56 dpi(P>0.05).IL-10 levels in Serum increased at 2 dpi,rose to the peak at 14dpi,and then gradually decreased,and dropped to the normal level at 28dpi(P>0.05).Then,it gradually increased and rose to a level higher than the normal group at 56dpi.TGF-?levels in Serum began to decrease at 2dpi,declined to the lowest value at 5dpi.Then,it began to increase and reached to the normal level at 7dpi(P>0.05),and continued to rise,reached the peak at 21dpi,but serum TGF-?dorpped to normal level at 28dpi(P>0.05),and rose again at 42dpi,and higher than a normal level(P<0.01),then maintained a stable level.IL-17A levels in Serum increased gradually at 14dpi to 42dpi,and reached the peak at 42 dpi,and decreased to normal level at 56dpi(P>0.05).IL-9 levels in Serum began to rise at 5 dpi,reach the peak at 21 dpi,and dramatically decreased at 28 dpi,but still higher than the normal level(P<0.01).It rose again at 42dpi and then remained at a stable level.Although IL-22 levels in Serum fluctuated after Trichinella infection,there was no significant difference at each point(P>0.05).Conclusion:1.We preliminarily understand the kinetics of the percentage of Th1,Th2,Th9,Th17,Treg,CD4~+IL-22~+T cells in spleens and mesenteric lymph nodes and related cytokines in serum in mice infected with Trichinella spiralis.2.Except for Th9,CD4~+IL-22~+T cell in mesenteric lymph nodes began to rise in the middle and late stages of Trichinella infection,the remaining CD4~+T cell subsets began to increase significantly in the early stage of Trichinella infection(within one week),but the dynamic characteristics were different,indicating that the immunopathological process of Trichinella infection is the result of the combined effects of CD4~+T cell subsets such as Th1,Th2,Th9,Th17,Treg,CD4~+IL-22~+T cells.3.The dynamic changes of CD4~+T cell subsets in spleens and mesenteric lymph nodes were different after Trichinella infection,indicating that there is a difference between systemic and locally intestinal immunity,which may play an important role in the immunopathological process of Trichinella spiralis. |
PDF Full Text Request