[Efficacy of Activated Autologous Hematopoietic Stem Cell Transplantation in the Treatment of Acute Myeloid Leukemia].

OBJECTIVE: To compare the efficacy of activated autologous bone marrow and peripheral blood hematopoietic stem cell transplantation (Auto-HSCT) and matched sibling donor allogeneic hematopoietic stem cell transplantation (MSD-HSCT) for the first complete remission of adult acute myeloid leukemia (AML-CR1). METHODS: For 86 adult patients with first complete remission of AML who underwent auto-HSCT (41 cases) and MSD-HSCT (45 cases) in our hospital from June 2012 to June 2020, the patients were treated with modified MAC ï¼»Malflane 160 mg/(m2·d), -3 days, Ara-C 2 g/(m2·2), -3 days 21∶00, -2 days 9∶00, CTX 60 mg/(kg·d),-3 d, -2 dï¼½, the stem cells were activated by IL-2 (1 000 U/ mL), IFN-α (100 U/ mL) and IFN-γ (100 U/ml). The overall survival (OS), leukemia free survival (LFS), cumulative incidence of recurrence (CIR) and non-recurrence mortality (NRM) of patients with different types of transplantation were compared. RESULTS: The 3-year OS rates of Auto-HSCT group and MSD-HSCT group were 75% and 69.5%, and the 3-year LFS rates were 70.6% and 82.4%, respectively. There was no statisticaly significant difference in the 3-year OS rates of low risk, medium risk and high risk patients in the Auto-HSCT and MSD-HSCT group (90.2% vs 87.5%, 68.4% vs 68.8%, 28.6% vs 53.3%), the LFS rates of low risk, medium risk and high risk patients in the auto-HSCT and MSD-HSCT group were 90.2% and 87.5%(P=0.838), 71.8% and 91.7%(P=0.184), 0 and 67.5%(P=0.027), respectively. The NRM of Auto-HSCT and MSD-HSCT group were 4.9% and 20% (P=0.036), and CIR were 24.4% and 13.3% (P=0.188). Univariate analysis showed that the survival time of patients was significantly correlated with the number of CR courses and disease risk stratification (P=0.005, P=0.000). Cox multivariate analysis showed that disease risk stratification was an independent risk factor affecting OS (P=0.001). CONCLUSION: For adult patients with primary AML-CR1, Auto-HSCT is safe and effective. In the absence of sibling donor, Auto-HSCT can be regarded as an effective post-remission treatment for patients with intermediate risk AML-CR1.

The impact of second-donor lymphocyte infusion on secondary graft failure after allogeneic hematopoietic stem cell transplantation through activation of Foxp3 and regulatory T cells.

Secondary graft failure (SGF) is a fatal complication of allogeneic hematopoietic stem cell transplantation without effective treatment methods, especially after haploidentical transplantation. This study aimed to analyze the efficacy of donor lymphocyte infusion (DLI) from a second donor in treating SGF and the underlying immune mechanisms. A second donor is a candidate donor who did not initially provide stem cells for HLA-matched sibling donor or HLA-haploidentical donor transplantation. We conducted a retrospective study of 237 patients with a median age of 38 years (range 9-56) for whom the degree of mixed chimerism (MC) and complete donor chimerism (CC), mRNA expression levels of Forkhead box P3 (Foxp3), and the proportion of regulatory T cells (Tregs) were regularly assessed. The median time to SGF was 62 days (range 41-117) after transplantation. Twenty-one patients with SGF received DLI, including 12 patients who initially received DLI from a second donor (i.e., a donor other than the transplantation [first] donor) and 9 patients who initially received DLI from the first donor but showed no response. Three of those 9 patients subsequently received DLI from a second donor. The incidence of acute GVHD and chronic GVHD induced by DLI from the second donor was significantly higher than that of DLI from the first donor (P = 0.006). Twenty-one patients with SGF exhibited synchronous MC, and the overall MC rate after transplantation was 65% (range 42%-85%).The proportion of Tregs significantly decreased in SGF patients, from a median of 2.61% ± 0.88% to 0.92% ± 0.23% at the indicated time point after transplantation (P = 0.03). Second-donor DLI resulted in a complete response (CR) in 13 patients, and MC gradually converted into CC; simultaneously, there was a significant increase in the mRNA level of Foxp3 and the proportion of Tregs (baseline, 0.92% ± 0.23% versus CR, 3.61% ± 0.82%; P = 0.01). For the patients who did not respond to DLI from either donor type, there was no significant change in donor chimerism, Foxp3 expression level or Treg proportion. Overall survival and disease-free survival 2 years after DLI were 66.7% ± 3.08% and 59.8% ± 4.11%, respectively. DLI from a second donor may be an effective treatment for SGF, and the mechanism is related to MC-to-CC conversion and activation of Foxp3 and Tregs.

Effect of Donor Lymphocyte Infusion From Two Types of Donors on Mixed Chimerism With Secondary Graft Failure After Allogeneic Hematopoietic Stem Cell Transplantation.

Mixed chimerism (MC) and secondary graft failure (SGF) with recipient- or donor-type chimerism is a major obstacle in allogeneic hematopoietic stem cell transplantation (HSCT). Donor lymphocyte infusion (DLI) can eradicate minimal residual disease or be used to rescue a hematologic relapse, being able to induce durable remissions after HSCT. This study aimed to analyze the efficacy and immune mechanism of DLI from the original and alternative donor for patients of mixed donor chimerism with SGF. The alternative donor refers to the candidate relative donor who did not initially provide stem cells and includes HLA-matched sibling donor or HLA-haploidentical donor. We conducted a retrospective study of 246 patients with a median age of 37 (9-58) years who had regularly detected MC, complete donor chimera (CC), and regulatory T cells (Treg). The median diagnosis time of SGF was 69 (39-141) days after transplantation. Sixteen patients with SGF received DLI from the alternative donor, including 3 patients who chose DLI from the original donor with no initial response and 13 patients who directly chose DLI from the alternative donor. Sixteen patients with SGF existed mixed chimerism synchronously and the rate calculated overall chimerism of MC was 63% (range 42%-85%) after transplantation. The proportion of Treg decreased significantly in SGF patients from a median of 2.66% ± 0.80% to 0.93% ± 0.57% at a time point after transplantation (P = .02). The DLI of the alternative donor in 14 patients achieved complete response and MC gradually convert to CC state, simultaneously there was significant increase in the Treg fraction (SGF versus complete response: 0.93% ± 0.57% versus 3.61% ± 0.82% [P = .01]). For the clinical nonresponders from 2 types of donors, there was no significant change in MC and Treg cells. The OS and disease-free survival at 2 years after DLI were 69.7% ± 3.19% and 61.3% ± 4.80%, respectively. DLI from the alternative donor may be an effective treatment for MC with SGF, and the mechanism is closely related to the activation of Treg cells level.

[The Prognostic Value of Early Relapse after Autologous Hematopoietic Stem Cell Transplantation in Newly Diagnosed Multiple Myeloma].

OBJECTIVE: To assess the impact of early relapse (ER) after autologous hematopoietic stem cell transplan-tation (AHSCT) on overall survival (OS) for multiple myeloma (MM) patients. METHODS: Clinical data of 37 patients with MM undergoing AHSCT in department of hematology of Shanxi Bethune Hospital from January 2012 to December 2017 were retrospectively analyzed. The effect of ER on OS of patients was analyzed. The effects of international staging system (ISS) staging, cytogenetics, pre-transplant efficacy, minimal residual disease, and age on OS of the patients were also analyzed respectively. RESULTS: Among the 37 patients, 13 cases (35.1%) had ER, and 24 cases (64.9%) had non-ER. 3 patients with ER had extramedullary disease, but none with non-ER showed extramedullary disease. More than or equal to very good partial rate (VGPR) in patients with ER and without ER were 3 cases (23.1%) and 15 cases (62.5%), respectively, and the curative effect of the former was significantly lower than that of the latter (P<0.05). The median follow-up time was 31 (12-96) months, and median OS time was 93 months in all the patients. The median survival time of patients with ER was 17 months, and the median progression free survival was 7 months, both were significantly shorter than 93 months and 38 months of patients with non-ER (P<0.05). Univariate analysis showed that the OS was affected by ER, cytogenetic abnormalities (FISH), and ≥VGPR before transplantation. Multivariate analysis showed that ER was an independent prognostic factor. CONCLUSION: The prognosis of patients with ER after AHSCT in newly diagnosed MM is poor. ER is an independent prognostic factor of survival.

[Prophylactic effect of TLR5 agonist flagellin on acute graft versus host disease after allogeneic hematopoietic stem cell transplantation and its mechanism].

This study was aimed to investigate the prophylactic effect of Toll like receptor (TLR)5 agonist flagellin on acute graft versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT) and its possible mechanism. The animal model with allo-HSCT aGVHD was established by using purebred mice (male mouse C57BL/6 as donor, female mouse BALB/c as recipient) with complete-unidentical major histocompatibility antigen. The recipient mice were randomly divided into 3 groups: group 1 in which mice were injected with high purity (95%) flagellin before and after allo-HSCT respectively, group 2 in which mice received allo-HSCT without injection of flagellin, group 3 in which mice were radiated alone. The aGVHD features of mice in group 1 and 2 were observed and compared. The results showed that the typical symptoms of aGVHD appeared in transplanted mice. The death peak of mice in group 2 appeared at day 4-5 after transplantation. The aGVHD symptoms were obviously alleviated and the mean survival time was prolonged significantly in mice group 1 as compared with mice in group 2 (P < 0.05). The comparison of WBC count in peripheral blood of mice in 3 groups before transplantation showed no significant difference (P > 0.05), while WBC count of mice in group 1 and 2 showed the significant difference at days 14 and 21 after transplantation (P < 0.05). The pathological appearances of aGVHD in mice of group 1 were obviously reduced as compared with mice in group 2. The flow cytometric detection of Treg cell/CD4(+) T cell levels at different time before and after transplantation demonstrated that the Treg cell level in mice of group 1 at weeks 2-4 after transplantation significantly increased as compared with mice in group 2 (P < 0.05). It is concluded that flagellin can effectively prevent the aGVHD occurrence after allo-HSCT, reduce the symptoms and pathological changes of aGVHD, obviously prolong mean survival time of mice in group 1. The mechanism of flagellin effect may be associated to increase of Treg cell level in mice after allo-HSCT.

[Effect of bortezomib on the drug sensitivity of imatinib resistant K562/G01 cells].

OBJECTIVE: To explore the effect of bortezomib (BOR) on the drug sensitivity of imatinib-resistant chronic myeloid leukemia cell line K562/G01 cell and its mechanism. METHODS: MTT assay was used to detect the inhibition effect of cell growth, flow cytometry to cell cycle, and real time-PCR to the expression of COX-2 and mdr1 mRNA. RESULTS: Combination of 10 and 20 nmol/L BOR with imatinib could significantly enhance the sensitivity of K562/G01 to imatinib, the reverse factor was 1.83 and 2.72-fold respectively. Cell cycle arrested at G(2)/M phase could be observed by flow cytometry on BOR treatment. The over-expression of COX-2 and mdr1 could be down-regulated by BOR. CONCLUSIONS: BOR can enhance the imatinib sensitivity of imatinib resistant K562/G01 cell. The mechanism may be related to cell cycle phase arrested at G2/M and down-regulation of COX-2 and mdr1 expression.

[Relationship between CD4(+)CD25(+) regulatory T cells, IL-2, TGF-beta and acute graft-versus host disease after allogeneic hematopoietic stem cell transplantation].

This study was aimed to investigate the relationship between CD4(+)CD25(+) regulatory T cells, IL-2, TGF-beta and acute graft-versus host disease (aGVHD) after allogeneic hematopoietic stem cell transplantation (allo-HSCT). The percentage of peripheral blood CD4(+)CD25(+) Treg cells in CD4(+) T cells of 13 patients with hematological malignancies after allo-HSCT were detected by flow cytometry; serum levels IL-2 and TGF-beta in these patients were measured by ELISA. The results indicated that all the patients achieved engraftment. 5 patients developed aGVHD of grade I-II, 4 patients developed aGVHD of grade III-IV. The percentage of peripheral blood CD4(+)CD25(+) Treg cells out of CD4(+) T cells in patients without aGVHD was higher than that in patients with aGVHD (p < 0.05); the serum level of IL-2 in patients without aGVHD was lower than that of patients with aGVHD (p < 0.05); the serum level of TGF-beta in patients without aGVHD was higher than that of patients with aGVHD (p < 0.05). It is concluded that CD4(+)CD25(+) Treg cell level and the serum level of IL-2 and TGF-beta all are related to incidence and severity of aGVHD. These factors may be used as indicators for early evaluating and monitoring aGVHD after allo-HSCT.