| Objective: To compare the changes of T?B and NK cell lymphocyte subsets ratio in patients with acute myeloid leukemia(AML)before and after the treatment of micro-transplantation(MST),and further explore the relationship between the changes of the ratio,the composition of the peripheral blood stem cells and clinical efficacy.Methods: 1.Research objects and grouping: retrospective analysis of 24 cases of AML patients receiving micro-transplantation in Hematology Department of 307 PLA Hospital from January 2015 to December 2017.According to the treatment,24 patients were divided into two groups: 14 patients in the initial MST group with no hematological remission(induction therapy group)and 10 patients in the initial MST group with hematological remission(consolidation tehrapy group).The changes of lymphocyte subsets in AML patients from each group before and after micro-transplantation were observed.The induction therapy group was divided into: Induced complete remission(CR)and induced non-complete remission(NR).The changes of lymphocyte subsets in CR group and NR group were observed,and the differences of lymphocyte subsets between the two groups were analyzed.The fect of the number and component of peripheral hematopoietic stem cells from donors after mobilized by G-CSF infusion on the changes of lymphocyte subsets after MST induction therapy in AML patients,as well as the effect of lymphocyte subsets changes on hematological remission were demonstrated.In the consolidation group,the patterns of lymphocyte subsets changes before and after three consecutive cycles of micro-transplantation were observed in AML patients with hematological remission after chemotherapy.All patients were assessed by MICM,which is morphological,immunological,cytogenetic and molecular genetic diagnostics,in line with FAB diagnostic criteria.2.Sample collection and detection methods: The venous blood were taken from all the patients,and the ratio of T,B lymphocyte subsets were detected using flow cytometry.3.Statistical methods: The normal distribution was not suitable for the continuous variables.After the logarithm,the mean ± standard deviation was used for statistical analysis.The mean of two samples were compared by t test.The Kaplan-Meier method was used to map the leukemia-free survival(DFS time)of patients with elevated NK cells and those with reduced NK cells before and after MST treatment in the induction therapy group.SPSS24.0 software was used for statistical analysis,and P ? 0.05 indicates statistical differences.Results: 1.Induction therapy Group(14 cases): There are 11 patients with remission and 3 patients without remission,with a remission rate of 78.6%.In the induction therapy group,CR patients were analyzed before and after MST: in 11 patients with remission,the ratio of CD8~+ T cells increased in 9 cases,while decreased in 2 cases;the ratio of total B lymphocytes increased in 2 cases,decreased in 9 cases;the ratio of NK cells increased in 8 cases,decreased in 3 cases.The ratio of CD8~+ T cells(inhibitory/killer T lymphocytes)after micro-transplantation was significantly higher than that before treatment(1.48±0.17 Vs.1.34±0.24,P=0.019).Compared with before treatment,the ratio of total B lymphocytes was significantly decreased after micro-transplantation(0.40 ± 0.59 Vs.1.05 ± 0.36,P = 0.004);whereas,the percentage of NK cells was significantly upregulated after treatment(0.71 ± 0..43 Vs.0.29 ± 0.67,P = 0.034).The comparison of NR patients before and after treatment: In 3 patients without remission,the ratio of CD8~+ T cells increased,while total B lymphocytes and NK cell ratio decreased.We observed the effect of hematopoietic stem cells on lymphocytes after G-CSF during infusion during MST.In the 14 patients from induction group,received more CD3~+cells in hematopoietic stem cells after G-CSF mobilization is relevant to increased ratio of NK after MST treatment(P=0.038)and decreased CD4~+ T cell ratio(P=0.009).There was no significant correlation between the number of MNC,CD34~+ and NK cells in stem cells and the increased ratio of NK cells after MST treatment.We observed the relationship between hematopoietic stem cells and DFS after MST: Patients received a higher dose of donor CD3~+ T cells have longer survival time without leukemia than those received a lower dose of donor CD3~+ T cells(P=0.005).Relationship between NK cell changes and DFS after MST: Patients with increased NK cell percentage have longer survival time without leukemia than those with decreased NK cell percentage(P=0.005).2.Consolidation therapy group: 10 patients received 30 hematopoietic stem cells MST treatment.The mean value of total T cell ratio in all 10 patients was higher than normal before treatment,and returned to normal and stable after the first MST.The mean value of total B-cell ratio was lower than normal before treatment,and returned to normal after the first MST;After the second and third treatments,the volatility decreased: the mean value of total B cell ratio was lower than the normal value but higher than that before treatment.CD4~+ T cells,CD8~+ T cells,NK cells,NKT cells,and resting T cells were all in the normal range before and after treatment,with little fluctuation.Activated T cells were higher than the normal range before and after treatment,and increased significantly after the first treatment,subsequently decreased gradually.We observed the effect of hematopoietic stem cells on lymphocytes after G-CSF infusion during MST.The higher NK ratio after MST treatment goes with the higher CD3 content in hematopoietic stem cells after infusion of G-CSF in the MST of the consolidation group(P=0.016);There was no significant correlation between the increase in NK cells ratio after MST treatment and the infusion of stem cells before the second and third MST treatment.Conclusion: 1.MST treatment resulted in the changes of lymphocyte subsets ratio in the patients with acute myeloid leukemia.2.Among the patients with MST-induction therapy the ratio of lymphocyte recovery before and after treatment was different between CR patients and NR patients.The increase of NK cell ratio indicated its associated with long-term survival and disease-free survival after induction therapy.3.The CD3 content in hematopoietic stem cells mobilized by G-CSF may affect the ratio of NK cells.4.The observation of dynamic changes in the ratio of NK cells,B cells,and T cell subsets can help to understand the immune status of AML patients,provide suggestion for the prognosis and efficacy,but still need to be further studied by multi-center large samples. |
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