| Part ?.Retrospective clinical observation of peripheral blood stem cells(G-PBSCs)± decitabine in the treatment of patients with MDSObjective:To analyze the efficacy and safety of peripheral blood stem cells(G-PBSCs)±DNA methyltransferase inhibitor-decitabine(micro-transplantation)in the treatment of myelodysplastic syndrome.Materials and Methods:A retrospective analysis of 22 cases of myelodysplastic syndromes treated with conventional doses of decitabine or peripheral blood stem cells(G-PBSCs)±decitabine in the Department of Hematology,Guangdong Provincial People's Hospital from September 2009 to September 2016.The clinical data of patients with MDS are mainly diagnosed by bone marrow biopsy,bone marrow cytology,karyotype analysis,and genetics,and are scored according to the latest version of the International Prognostic Scoring System(IPSS-R)>3,each MDS patient completed at least 2 courses of decitabine demethylation or at least 2 courses of micro-transplantation.Twenty-two patients were given a regular dose of decitabine 20 mg/m2/d for 1 hour intravenous,infusion for 5 days and 4 weeks for 1 cycle.Statistical analysis and evaluation were made on the clinical therapeutic effect and adverse events of micro-transplantation therapy on MDS,and make a comparison of therapeutic effects on MDS with the same dose of decitabine alone.The related factors affecting the therapeutic effect were analyzed,the survival time of the patients were followed up,and the survival time of the two treatments were compared.Results:Nine patients(9/11,81.8%)in the MST group responded to treatment.Six(6/11,54.5%)patients in the DAC group responded and there was no significant differe nce between the two groups(P=0.18),but the ratio of overall response rate(ORR)?m arrow complete remission(mCR)and hematological improvement(HI)coexistence in th e micro-transplantation group was higher than that in the DAC group(81.8%vs.54.5%,P=0.09;63.6%vs.18.2%,P=0.18).The incidence of AML conversion was lower in the MST group than in the DAC group within 12 months(0.0%and 27.3%,P=0.07).At the same time,the 24-month mortality rate was lower in the MST group t han in the DAC group(45.5%vs.81.8%,P=0.08).The median OS of the MST grou p was 24 months,and the median OS of the DAC group was 14.3 months;significan t differences were observed between the two groups(HR 0.34;95%CI 0.12-1.01;P=0.04).The median PFS was 24.0 months in the MST group and 17.9 months in th e DAC group(HR 0.47;95%CI 0.12-1.83;log-rank test,P=0.25).No significant stati stical difference was observed between the two groups.Safety data were evaluated for 22 patients treated with decitabine or micro-transplantation.In the first 12 months of treatment,1 patient died in the MST group(1/11,9.1%)and 4 patients died in the DAC group(4/11,45.4%)(P=0.14).The 24-month mortality rate in the MST group w as 5/11(45.5%).9/11(81.1%)in the DAC group(P=0.08).During the entire treatmen t period,except for thrombocytopenia(64%in the MST group and 27.0%in the DA C group,P=0.09),the incidence of hematologic adverse events(AEs)was lower in th e MST group(Anemia:27.2%in the MST group vs.55.0%in the DAC group,P=0.20;Leukopenia:MST group 54.5%vs.73.0%in the DAC group,P=0.39;Neutropeni a:MST group 55.0%vs.DAC group 64.0%,P=0.67).The same results were found in the non-hematology AEs in the MST group.At the same time,the fluctuation tren d of up-regulation and decline of tumor burden index WT1 was consistent with the tr eatment response.Conclusion:The micro-transplantation therapy in MDS which was evaluated intermediate?high-risk and very high-risk can significantly improve patient survival,reduce the incidence of h ematologic AEs and non-hematologic AEs,and reduce leukemia conversion rate and m ortality ratePart ?.Preliminary Study on Mechanism of Micro-transplantation for MDS Patients-Analysis of ?? T Cell Subsets and Preliminary Study on Expression of BTLA/HVEM in MDS PatientsObjective:There is evidence that T cell immune disorder plays an important role in MDS.The purpose of this study is to investigate the expression of ?? T cell subpopulation and its surface protein BTLA/HVEM in MDS patients.Materials and Methods:Bone marrow and peripheral blood samples of different MDS patients before micro-transplantation treatment and normal control samples were collected.The levels of peripheral blood T lymphocytes,y8 T cells,V?1 cells,V?2 cells,CD27+and CD27-?? T cells as well as the expression of BTLA and its ligand HVEM on the surfaces of T lymphocytes and ?? T cells were detected by polychromatic flow meter.The expression levels of BTLA and HVEM mRNA in different MDS patients were detected by real-time quantitative PCR,and the relationship between abnormal y8 T cells and BTLA expression in MDS patients was analyzed.In vitro culture,amplification and functional detection of ?? T cells:The peripheral blood mononuclear cells of patients were isolated,and an ?? T cell culture system in vitro was established to induce the amplification of ?? T cells,and the proliferation ability and killing activity of ?? T cells in MDS patients were detectedResults:1.Flow cytometry was used to detect the expression of ?? T cells and their subsets in peripheral blood of MDS patients;1.1 The level of ?? T cells in peripheral blood of RAEB patients was 2.94(0.68-7.37)%,significantly lower than the normal control level of 5.74(1.38-10.04)%.V82 T cell subpopulation showed an increasing trend compared with normal control(P=0.052).The CD27+ and CD27-cell subsets in ?? T cells showed a decreasing trend compared with the normal control(P=0.062,P=0.058),while the CD27+cell ratio in RAEB patients was slightly higher than that of CD27-cells.The proportion of CD27+ cells in normal control is slightly lower than that of CD27-cells.1.2 T lymphocyte level in peripheral blood of CMML patients was 32(7.14-76.38)%,significantly lower than that of the control group(P<0.05).V?1T cells showed an increasing trend(P=0.063).2.The expression of BTLA and its ligand HVEM on peripheral blood T cells and y8 T cells was detected by polychromatic flow:BTLA expression in peripheral blood ?? T cells of CMML patients was significantly decreased(P=0.029),and the expression of its ligand HVEM showed an increasing trend(P=0.118),with no significant difference.3.Real-time quantitative PCR detection of BTLA and HVEM mRNA expression in different MDS patients:3.1 The expression levels of BTLA and HVEM mRNA in peripheral blood of 32 MDS patients at the same time were significantly higher than that in bone marrow(P<0.01).3.2 mRNA Expression Levels of BTLA and HVEM mRNA in different MDS;(1)Expression of BTLA:Compared with the normal control,the expression level of BTLA mRNA in the peripheral blood of RCMD patients is significantly higher than that of the normal control(P=0.003),and the expression level of BTLA mRNA in the peripheral blood of RAS patients is significantly lower than that of the normal control(P=0.011).The expression level of BTLA mRNA in the peripheral blood of RCMD patients was significantly higher than that of other MDS patients(P<0.05),while the expression level of BTLA mRNA in the peripheral blood of RAS patients was significantly lower than that of other MDS patients(P<0.01).The expression level of BTLA mRNA in bone marrow of patients with RAEB-1 and RAEB-2 was significantly lower than that of normal controls(P<0.05).(2)Expression of HVEM:the expression level of HVEM mRNA in peripheral blood of patients with RAEB-1 and RAEB-2 was significantly lower than that of the normal control(P<0.05).there was no significant difference between RCMD patients and the normal control(P=0.297),but it was significantly higher than that of patients with RAEB-2(P=0.018).3.3 The expression levels of BTLA and HVEM in peripheral blood of CMML patients were significantly lower than those of normal controls(P<0.05).4.Study on proliferation and function of peripheral blood ?? T cells from MDS patients in vitro4.1 Expansion of ?? T Cells from MDS Patients in Vitro:The ?? T cells in five of 12 patients can be successfully effectively amplified to 80%after peripheral blood mononuclear cells being stimulated by zoledronic acid,and all the normal control group can be successfully amplified.(P=0.002)4.2 There was no significant difference in killing activity of ?? T cells that successfully expanded in vitro between normal people and MDS patients(P>0.05)Conclusion:1.Compared with normal donors,the proportion of peripheral blood ??T cells in MDS-RAEB and CMML patients decreased,suggesting that they have certain immune defects.2.The expression of BTLA and its ligand HVEM mRNA in peripheral blood of CMML patients is lower than that of normal donors,and the proportion of ??T cells in peripheral blood is also lower,which indicates that BTLA/HVEM is impaired in regulating ??T cells.Meanwhile,??T cells in peripheral blood of CMML patients cannot be successfully expanded in vitro,while normal donors can be successfully expanded,which indicates that the proliferation function in vitro is decreased.3.The killing function of ??T cells that successfully expanded by MDS in vitro is not significantly different from that of normal control group... |
PDF Full Text Request