The Effect And Mechanism Of Arsenic Trioxide On T Cell-mediated Immune Liver Injury And Acute GVHD

Background:Arsenic trioxide(ATO)has been proved effective in treatment of acute promyelocytic leukemia.The mechanism involved is mainly due to inducing differentiation and apoptosis.In recent years,many studies have shown that ATO has a therapeutic effect in a variety of immune diseases,such as systemic lupus erythematosus,ulcerative colitis,and immune rejection after solid organ transplantation.ATO can promote CD4+,CD8+T cell apoptosis,inhibit T cell proliferation,increase the proportion of CD4+CD25+ Foxp3+regulatory T cells and significantly reduce levels of cytokines such as IFN-y,TNF-?,IL-1?,IL-4,IL-6,IL-10 and IL-17.Graft versus host disease(GVHD)is a common complication after allogeneic hematopoietic cell transplantation(allo-HSCT).Since donor T cells are the main effector cells of GVHD,suppressing T cell responses is the standard treatment method for the prevention and treatment of GVHD.A number of studies have shown that cell metabolism affects the differentiation and function of T cells,and targeted T cell metabolism may become a new method for the treatment of GVHD.Some studies have found that ATO specifically binds to proteins in the glycolytic pathway through human proteomic chips.Therefore,we want to explore the therapeutic effect of ATO in aGVHD and further clarify whether the mechanism of therapeutic effect is related to T cell glucose metabolism.Purpose:1.To study the effect of ATO on T cell proliferation,activation,differentiation and apoptosis and the therapeutic effect of ATO in aGVHD;2.To study the mechanism of ATO treatment of aGVHD and its effect on T cell glucose metabolism.Methods:In vitro,ConA,CD3/CD28 stimulated beads and mixed lymphocyte culture were given to stimulate the proliferation of mice and donor T cells,different concentrations of ATO were used to perform T cell proliferation inhibition experiments.Flow cytometry was used to detect T cell apoptosis.We established a ConA-induced immune liver injury model in vivo,and performed liver function and pathological examinations on mice.Flow cytometry was used to detect the composition and activation of lymphocytes in theResults:1.In vitro,ATO can reduce ConA,CD3/CD28 stimulated beads and MLC-induced T cell proliferation in mice and donors,and the inhibitory effect of ATO on T cell proliferation is dose-dependent.In addition,1-2?mol/L ATO can significantly induce T cell apoptosis in a dose-dependent manner under the condition of T cell steady state and activated proliferation.As the concentration of arsenic trioxide increases,the proportion of surviving CD4+T cells gradually increased while the proportion of CD8+T cells gradually decreased.2.We used ConA(20mg/kg)to establish an immune liver injury model,the application of ATO can significantly increase the survival rate of mice,reduce liver function and liver cell necrosis and inflammatory cells shown in liver pathology infiltration.In addition,ATO treatment for 12 hours can significantly reduce the infiltration of T cells and NKT cells in the liver and spleen cells,and increase the proportion of treg and MDSC cells.ATO can also reduce the levels of IFN-?,TNF-?,IL-6,IL-4,IL-10 in mice with immune liver injury.Through virtual screening,we found that one of the binding targets of ATO is PFKFB3 in the glycolytic pathway.RT-PCR has also verified that ATO can reduce the expression of HK2,LDHa,LDHb,Pfkfb3,Glut1,MCT4,Tpi,PKM2,Pfkp,Pfkl,Pgaml,Enol,and Gpi in the glycolysis pathway of T cells in mice.At the same time,ATO also reduced the expression of HK2,Pfkfb3,MCT4,and Gpi in T cells proliferated in vitro.3.ATO can prolong the survival of aGVHD mice,reduce aGVHD score and reduce the pathological manifestations of aGVHD.14 days after transplantation,arsenic trioxide can reduce the CD3+ T cells of the donor in the spleen,inhibit the production of CD4+ and CD8+Tem cells,inhibit the differentiation of Th1 cells and promote the differentiation of treg cells,reduce the secretion of perforin by CD8+T cells and reduce IL-6 and IFN-y,TNF-? in serum.RT-PCR also verified that ATO can reduce the expression of T cells HK2,Pfkfb3,MCT4,Gpi,Glutl,LDHa in the glycolytic pathway of T cells in aGVHD mice.Conclusions:ATO can influence the secretion of cytokines,differentiation and activation of T cells,inhibit T cell proliferation and promote T cell apoptosis to reduce ConA-induced immune liver injury and aGVHD,which may be related to the inhibition of T cell glycolysis pathway by ATO.Objective:To compare the efficacy of autologous HSCT with matched sibling donor(MSD)HSCT in Ph+ALL and provide the basis for the choice of transplantation method.Methods:We retrospectively investigated the outcomes of 78 adult patients with Ph+ALL who underwent auto-HSCT(n=31)and MSD-HSCT(n=47)in our center from Jan 2008 to Dec 2017.The overall survival(OS)rate,leukemia-free survival(LFS)rate,cumulative incidence of relapse(CIR)rate,non-relapse mortality(NRM)rate and the impact of achievement of complete molecular response(CMR)within 3 months and sustaining CMR up to transplantation(s3CMR)on transplantation method were explored.Results:The median time of neutrophil and platelet reconstitution in auto-HSCT group and MSD-HSCT group were 12(10?29)d versus14(11?24)d(P=0.006)and 17.5(10?62)d versus 7(10?33)d(P=0.794)respectively.In the MSD-HSCT group,the incidence of ?-? and ?-? acute graft-versus-host disease(GVHD)were 27.7%(13/47)and 8.5%(4/47).The incidence of limited and extensive chronic GVHD werel7.0%(8/47)and 12.8%(6/47).The estimated CIR,NRM,LFS and OS at 3 years were not significantly different between auto-HSCT and MSD-HSCT group(P>0.05).For 44 patients who achieved s3CMR,3-year OS(84.0±8.6%vs 78.0±8.7%,P=0.612),LFS(70.3±10.3%vs 68.2±10.1%,P=0.970),CIR(24.9±10.0%vs 14.4±8.0%,P=0.286)and NRM(4.7±4.7%vs 17.4±8.1%,P=0.209)of the auto-HSCT group and MSD-HSCT group were not significantly different.But for 34 patients who did not reach s3CMR,3-year cumulative relapse rate of patients in the auto-HSCT group was significantly higher than MSD-HSCT group(80.0±14.7%vs 39.6±10.9%,P=0.057).Conclusions:Auto-HSCT with maintenance therapy after HSCT appears to be an attractive treatment option for patients with Ph+ALL especially for those s3CMR was kept up to transplantation.For non-s3CMR patients,allogeneic transplantation may be more effective from lower relapse.