Molecular genetics research of a Chinese family carrying triple HLA-A alleles / 中国输血杂志

【Objective】 To determine molecular basis of a rare HLA-A typing results carrying triple A alleles in potential allo-HSCT donor and her family. 【Methods】 HLA-A, -B, -C, -DRB1, -DQB1, -E, -F, -G of 5 members in the family were genotyped at a high-resolution level using next-generation sequencing (NGS). HLA-A of probosita was re-checked using polymerase chain reaction-sequence-based typing (PCR-SBT), and SNP oligonucleotide probes (SNP-array)were scanned with genomic DNA of probosita. 【Results】 There was 162.9Kb duplication in 6p22.1(29, 803, 377-29, 966, 301)of probosita who carried triple A alleles A*02∶01∶01, A*11∶01∶01, A*24∶02∶01. Other two family members were found to carry this haplotype: A*02∶01∶01, A*24∶02∶01, B*54∶01∶01, C*01∶02∶01, DRB1*04∶05∶01, DQB1*04∶01∶01, E*01∶01∶01∶03, F*01∶01∶01, G*01∶01∶01∶01, which as a Mendelian gene was segregated and stably transmitted through two generations. 【Conclusion】 Tiny gene duplication induces one haplotype carries two HLA-A alleles in a potential healthy donor for allo-transplantaion and stably transmits through two generations.Routine HLA typing laboratories should pay more attention to this situation and accurately report.

Clinical outcomes and early immune reconstitution in patients with acute lymphoblastic leukemia underwent allogeneic hematopoietic stem cell transplantation / 中国输血杂志

【Objective】 To investigate the correlation between early immune reconstitution and clinical outcomes in patients with acute lymphoblastic leukemia (ALL) underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT). 【Methods】 The basic information and treatment data of 99 patients with ALL undering allo-HSCT from December 2018 to February 2022 were collected. The proportions of CD3+ T, CD3+CD4+ T, CD3+CD8+ T and CD3-CD16+CD56+ NK cells were detected before and 30, 60 and 90 days after transplantation using flow cytometry. The correlation between early cellular immune reconstitution and neutrophil engraftment, platelet engraftment, infection, and acute and chronic graft-versus-host disease (GVHD) was analyzed. 【Results】 Among 99 ALL patients, the median time of neutrophil engraftment was day +11 (range, 8-28), and the median time of platelet engraftment was day +14 (range, 10-120). The cumulative incidence of blood stream infection (BSI) was 11.10% and the cumulative incidence of CMV within 100 days of transplantation was 40.40%. The cumulative incidence of EBV within 100 days was 7.10%. The cumulative incidence of acute graft-versus-host disease (aGVHD) was 22.30%. The cumulative incidence of chronic graft-versus-host disease (cGVHD) within 1 year of transplantation was 16.20%. 1 -year cumulative relapse rate was 13.84%. The 1 -year cumulative disease-free survival (DFS) for all patients was 80.60% and the 1-year overall survival (OS) was 90.30%. The CD4+/CD8+ ratio was positively associated with the development of aGVHD at 30 days post-transplant (OR 1.21, 95CI 1.01-1.45, P<0.05). The proportion of CD16+ CD56+ NK cell were higher in the group without BSI than that in the BSI group before and 30 days after transplantation (P < 0.05). The proportion of CD4+ T-cell were lower in the CMV infection group than that in the group without CMV infection at 60 and 90 days post-transplant(P<0.05). The higher level of CD4+ T-cells at 60 days post-transplant was a protective factor for CMV infection within 100 days (HR 0.91, 95CI 0.84-0.99, P<0.05). 【Conclusion】 Early immune reconstitution after allo-HSCT in patients with ALL is associated with aGVHD, CMV and BSI.

Effect and mechanism of extracorporeal photopheresis on preventing acute graft versus host disease in ice / 中国输血杂志

【Objective】 To investigate the preventive effects of early apoptotic splenic mononuclear cells induced by extracorporeal photopheresis (ECP) on acute graft versus host disease (aGVHD) in mice and explore the underlying mechanisms. 【Methods】 1) Splenic mononuclear cells were extracted from C57BL/6 mice and treated with different concentrations of 8-MOP (50 ng/mL, 100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL). After treatment, irradiate the cells with 2 J/cm2 of ultraviolet light. Then, use the Annexin V-FITC/PI apoptosis detection kit to assess the early apoptosis rate of the cells and determine the optimal concentration of 8-MOP for the experiment.2) There were 35 SPF-grade female BALB/C mice (H-2Kd) aged 6-8 weeks. After whole-body irradiation with 8Gy X-rays, the mice were divided into five groups: sham irradiation group received intravenous infusion of 0.2 mL of normal saline, the syngeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of BALB/C mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the allogeneic bone marrow transplantation group received intravenous infusion of 0.2 mL of C57BL/6 mouse bone marrow nucleated cell suspension (including a cell count of 1×107), the aGVHD group received intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107) in 0.2 mL, the ECP prevention group received pre-transplant intravenous infusion of 0.2 mL of ECP-treated splenic mononuclear cells of C57BL/6 mice (including a cell count of 1×107 ) 48 hours before transplantation, and on the day of transplantation, intravenous injection of a mixture of C57BL/6 mouse bone marrow nucleated cells (including a cell count of 1×107) and splenic mononuclear cells (including a cell count of 1×107 ) in 0.2 mL.The preventive effects of ECP on aGVHD were observed, and the concentrations of IFN-γ, IL-2, TNF, IL-4 and IL-6 in mouse serum were measured using CBA. Th1 cell counts were determined by flow cytometry. 【Results】 Different concentrations of 8-MOP (50 ng/mL,100 ng/mL, 200 ng/mL, 300 ng/mL, 600 ng/mL) were used to treat mouse splenic mononuclear cells. The early apoptosis rates (%), observed after treatment were as follows: (14.18±0.865) vs (16.76±0.407) vs (18.83±0.404) vs (19.27±0.404) vs (14.5±0.529). The appropriate concentration of 8-MOP was determined to be 200 ng/mL.In vivo experiment, the results showed that the aGVHD group had decreased survival rate, reduced body weight, and increased clinical scores compared to the syngeneic and allogeneic bone marrow transplantation groups (P<0.01), and the chimerism of bone marrow cells in mice after transplantation was over 90%. ECP significantly improved the survival rate of mice after transplantation, reduced clinical scores (P<0.05), and decreased the concentrations of Th1 cell cytokines in serum (P<0.05) and the counts of Th1 cells in the spleen (P<0.05). 【Conclusion】 ECP-induced early apoptotic single nuclear cells from the spleen can prevent the occurrence of aGVHD by reducing the Th1 response in mouse.

Clinical Analysis of SET-NUP214 Fusion Gene Positive Patients with Acute Leukemia / 中国实验血液学杂志

OBJECTIVE@#To analyze the characteristics and prognosis of acute leukemia(AL) with SET-NUP214 fusion gene.@*METHODS@#The clinical data of 17 patients over 14 years old newly diagnosed with SET-NUP214 positive AL admitted in Institute of Hematology and Blood Diseases Hospital from August 2017 to May 2021 were analyzed retrospectively.@*RESULTS@#Among the 17 SET-NUP214 positive patients, 13 cases were diagnosed as T-ALL (ETP 3 cases, Pro-T-ALL 6 cases, Pre-T-ALL 3 cases, Medullary-T-ALL 1 case), AML 3 cases (2 cases M5, 1 case M0) and ALAL 1 case. Thirteen patients presented extramedullary infiltration at initial diagnosis. All 17 patients received treatment, and a total of 16 cases achieved complete remission (CR), including 12 cases in patients with T-ALL. The total median OS and RFS time were 23 (3-50) months and 21 (0-48) months, respectively. Eleven patients received allogeneic hematopoietic stem cell transplantation(allo-HSCT), with median OS time of 37.5 (5-50) months and median RFS time of 29.5 (5-48) months. The median OS time of 6 patients in chemotherapy-only group was 10.5 (3-41) months, and median RFS time of 6.5 (3-39) months. The OS and RFS of patients with transplantation group were better than those of chemotherapy-only group (P=0.038). Among the 4 patients who relapsed or refractory after allo-HSCT, the SET-NUP214 fusion gene did not turn negative before transplantation. While, in the group of 7 patients who have not relapsed after allo-HSCT till now, the SET-NUP214 fusion gene expression of 5 patients turned negative before transplantation and other 2 of them were still positive.@*CONCLUSION@#The fusion site of SET-NUP214 fusion gene is relatively fixed in AL patients, often accompanied by extramedullary infiltration. The chemotherapy effect of this disease is poor, and allo-HSCT may improve its prognosis.

Major molecular events of reactivation of human cytomegalovirus after allogeneic hematopoietic stem cell transplantation / 器官移植

Immune deficiency of the host caused by allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the initial factor of reactivation of latent human cytomegalovirus (HCMV). The risk factors of reactivation of HCMV in allo-HSCT recipients consist of the serological status of HCMV in donors and recipients, the matching degree of human leukocyte antigen (HLA) and pretreatment patterns, etc. The reactivation of HCMV is associated with the expression of a series of viral cleavage and proliferation proteins induced by the overexpression of major immediate early promoter/enhancer (MIEP) in the viral genome. In this article, the risk factors of reactivation of HCMV after allo-HSCT, the molecular changes related to maintaining latent infection of HCMV, the key role of MIEP overexpression in reactivation of HCMV, and the molecular pathways involved in reactivation of HCMV after allo-HSCT were reviewed and the major molecular events of reactivation of HCMV after allo-HSCT were elucidated, aiming to provide reference for the prevention and treatment of cytomegaloviral disease (CMVD) after allo-HSCT.

New progress on clinical characteristics and related mechanisms of chronic ocular graft-versus-host disease / 器官移植

Allogeneic hematopoietic stem cell transplantation (allo-HSCT) could effectively treat multiple hematological diseases. At present, with persistent improvement of transplantation techniques and rapid development of economy, more and more patients with hematological diseases are able to survive for a long time due to allo-HSCT treatment. Chronic ocular graft-versus-host disease (coGVHD) is the most common ocular complication after allo-HSCT, which is primarily manifested with refractory dry eye. In severe cases, it may cause imbalance of ocular surface homeostasis and limbal stem cell insufficiency, further leading to a series of complications that threaten the visual function and eye health, such as corneal perforation and symblepharon, etc. It is highly difficult to cure these symptoms. At present, relevant studies of clinical manifestations, diagnostic criteria, treatment specification and pathogenesis of coGVHD have been gradually deepened within the international community. However, related research and the establishment of clinical specification are still in the primary stage in China. In this article, research progress on clinical characteristics and related mechanisms of coGVHD was reviewed, aiming to deepen the understanding of this disease by ophthalmologists, especially specialists in corneal and ocular surface diseases, and provide novel ideas for subsequent in-depth research.

Decitabine for Relapsed Acute Lymphoblastic Leukemia after Allogeneic Hematopoietic Stem Cell Transplantation / 华中科技大学学报（医学）（英德文版）

Relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) remains a main question on treatment failure.Current strategies for management that usually include salvage chemotherapy,donor lymphocytic infusion and second transplantation.Our study assessed the efficacy of decitabine (DAC) for treating patients with acute lymphoblastic leukemia (ALL) who relapsed after allogeneic hematopoietic stem cell transplantation (allo-HSCT).We retrospectively analyzed the outcomes of 12 patients with relapsed ALL after allo-HSCT who received DAC therapy.Nine patients received DAC combined with chemotherapy and donor stem cell infusion,and 3 patients received single-agent DAC.Ten of the 12 patients achieved complete remission (CR),1 achieved a partial remission (PR),and 1 had no response (NR) after treatment at the latest follow-up (LFU),the median survival was 11.2 months (range,3.8-34,7 months).The 1-and 2-year overall survival (OS) rates were 50％ (6/12) and 25％ (3/12),respectively.Five patients were still alive;4 had maintained CR and 1 was alive with disease.Patients with Philadelphia chromosome-positive ALL had higher survival rate than patients with Philadelphia chromosome-negative ALL (57.1％ vs.20％).No aggravated flares of graft-versus-host disease (GVHD) were observed during DAC treatment.Therefore,DAC may be a promising therapeutic agent for ALL recurrence after allo-HSCT.

Relationship between the methylation status of STAT3 promoter DNA in peripheral blood CD4+ T cells from patients after allo-HSCT and aGVHD / 中南大学学报(医学版)

Objective:To study the relationship between acute graft versus host disease (aGVHD) and the methylation status of the STAT3 promoter in peripheral blood CD4+ T cells from patients after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods:We collected 40 patients who underwent allo-HSCT from HLA-identical sibling donors.Serum IL-10,TGF-β1,IL-17A and IL-17F levels were detected by ELISA.Foxp3 cytotoxic T-lymphocyte-associated protein 4 (CTLA4),IL-10,TG F-β 1,RO Rγt,IL-17A and IL-17F mRNA levels in CD4+ T cells were measured by real-time PCR.STAT3 expression levels were detected by real-time PCR and Western blot,and promoter DNA methylation was analyzed by bisulfite sequencing PCR (BSP).Results:IL-10 and TGF-β1 levels were significantly down-regulated,while IL-17A and IL-17F levels were significantly up-regulated in patients with aGVHD compared with patients without aGVHD.Foxp3,CTLA4,IL-10,TGF-β1 mRNA levels were significantly down-regulated,while RORγt,IL-17A,IL-17F mRNA levels were significantly up-regulated in patients with aGVHD compared with patients without aGVHD.STAT3 expression was increased,while STAT3 promoter DNA was hypomethylated in patients with aGVHD compared with those without aGVHD.The STAT3 mRNA level was negatively correlated with STAT3 promoter DNA methylation.Conclusion:The imbalance of Treg/fh17 in CD4+ T cells from patients after allo-HSCT is a key factor for triggering aGVHD,and the DNA hypomethylation of STAT3 promoter could promote its expression in CD4+ T cells and contribute to the imbalance.

Transplantation time and detection method of allo-HSCT and the DNA of the recipients and its significance ;in forensic medicine( / 中国法医学杂志

There were many factors that affect the chimerism of donor DNA, such as the type of disease, the treatment of the disease, the severity of the disease, the source of hematopoietic stem cells, HLA, GVHD, and so on. This paper mainly discussed the two factors that affect forensic science, including the posttransplantation intervals and sensitivity of the detection method. Forensic workers in forensic practice and paternity and individual identification of allo HSCT recipients, should pay attention to the time after transplantation and the detection method of sensitivity of chimerism. When female recipients received HSCT in the case of male donors, Y chromosome DNA of the donor can be detected from the recipient's hair follicles. The case of biological specimens from the crime scene can not only detect Y chromosome speciifc genetic markers, so as to avoid wrong identiifcation.

Correlation between cytokine gene polymorphism and aGVHD in allo-HSCT recipients / 中国免疫学杂志

Objective:To investigate the relationship between gene polymorphisms of disease-relevant multiple cytokines including TNF-α,IL-6,IL-10,TGF-β1,IFN-γand acute graft versus host disease(aGVHD) in allogeneic hematopoietic stem cell trans-plantation ( allo-HSCT ) . Methods:32 cases of recipients received allo-HSCT and 36 cases of normal groups in January 2014 to December 2015 were selected as objects of study. We detected genotypes on specific SNP of target genes by polymerase chain reation ( PCR) combined with gene sequencing and observed the occurrence of aGVHD in postoperative recipients. The influence of cytokine gene polymorphisms on prognosis of allo-HSCT patients was analyzed,and the potential relationship between specific SNP mutation of the disease-relevant cytokine genes and severity of aGVHD was discussed. Results:Distribution of cytokines gene polymorphism including TNF-α-308(G/A),IL-6-174(G/C),IL-10-1082(A/G),TGF-β1+915(G/C),IFN-γ(T/A) had no significant differences with incidence of severe aGVHD(P>0. 05). However,the occurrence of severe aGVHD in allo-HSCT recipients with C/T genotype was significantly higher than C/C and T/T in SNP of TGF-β1+869(P<0. 01). Conclusion:Gene polymorphism of TGF-β1+869(C/T) in allo-HSCT patients was closely related to the occurrence of severe aGVHD. The research show allo-HSCT patients with C/T genotype occurred severe aGVHD more frequently, which is an important potential risk factor to induce the incidence of severe aGVHD. Therefore,detecting gene polymorphism of TGF-β1+869 ( C/T ) in allo-HSCT recipients and developing the appropriate therapeutic regimen may be helpful to reduce the incidence of aGVHD.

Medication Analysis of Therapeutic Drug Monitoring for Tacrolimus in Allogeneic Hematopoietic Stem Cell Transplantation / 中国药房

OBJECTIVE:To investigate blood concentration monitoring data,therapeutic efficacy,toxic reaction and combi-nation of tacrolimus in patients with Allo-HSCT,and to provide reference for clinical application of tacrolimus. METHODS:The blood concentration of tacrolimus in 16 inpatients were monitored with EMIT 3 months after Allo-HSCT. The occurrence of Graft-versus-Host disease(GVHD),ADR and drug combination were analyzed and discussed. RESULTS:A large individual dif-ferences were found in blood concentration of tacrolimus. When blood concentration 20 ng/ml,the occurrence of diabetes,kidney damage and other side effects seems increased. Tacrolimus had a good effect on treatment and prevention of GVHD,and could have an interaction with other drugs. CONCLUSIONS:Blood concentration monitoring of tacrolimus plays an important role on the prevention and treatment of GVHD,and the decrease of side effects such as diabetes and neurotoxicity. The satisfactory blood concentration is 8-20 ng/ml,when there seems a smaller possibility of GVHD and ADR 3 months after Allo-HSCT.

Characteristics of peripheral blood CD45RA+and CD45RO+T lymphocyte subsets in patients with cGVHD and its clinical significance / 中国免疫学杂志

To characterize the CD45RA+and CD45RO+T lymphocyte subsets in the peripheral blood of patients with cGVHD induced by allogeneic haematopoietic stem cell transplantation ( allo-HSCT ) and to explore their relations with the disease.Methods:The peripheral blood was collected from 64 patients after allo-HSCT,including 21 non-cGVHD patients,15 light grade cGVHD patients,18 mild grade cGVHD patients and 10 severe grade cGVHD patients,then CD4+CD45RA+,CD4+CD45RO+, CD8+CD45RA+and CD8+CD45RO+T lymphocyte subsets were detected by flow cytometry ( FCM).Results: Compared with the control,the percent of CD4+CD45RA+T lymphocyte in patients with light,mild and severe grade cGVHD decreased markedly (P<0.05),the percent of CD4+CD45RO+T lymphocyte increased markedly (P<0.05).But there were not obviously change in the patient with different grade cGVHD.The percent of CD8+CD45RA+,CD8+CD45RO+T lymphocyte did not change obviously.Conclusion:CD4+CD45RA+and CD4+CD45RO+may play an important role in the pathogenesis of cGVHD.

Research advances of CD+4CD+25 Treg cell and it's function in GVHD / 肿瘤研究与临床

CD+4 CD+25 regulatory T cell (Treg) has immune incompetent and immune suppression functions, and is a kind of suppressor T-cell subsets. It can inhibit the activation and proliferation of CD+4 T cells and CD+8 T cells, so as to effectively suppress the immune system to foreign organ and reduce the graftversus-host disease (GVHD) after hematopoietic stem cell transplantation (HSCT), without affecting the role of graft-versus-leukemia (GVL), which plays an important role in the transplantation immune tolerance.

A case of donor-derived granulocytic sarcoma after allogeneic hematopoietic stem cell transplantation

The occurrence of granulocytic sarcoma as a pattern of relapse after allogeneic hematopoietic stem cell transplantation (allo-HSCT) is rare. In this paper, we report a rare case of acute myeloid leukemia (AML) relapsed as a granulocytic sarcoma of the donor type. The patient was diagnosed as having AML and underwent an allo-HSCT from his matched sibling donor. Fifty-seven months after allo-HSCT, he developed granulocytic sarcomas of duodenum, jejunum, and left sterno-cleido-mastoid muscle. The bone marrow was normal with 100% donor chimerism. A Y chromosome PCR was performed on the patient's duodenum specimen as well as bone marrow aspirate in order to check the patient-origin cells. The duodenal specimen was found to contain 41.2% SRY-positive cells (from the donor). Repeat endoscopy on day 2 of chemotherapy showed that the granulocytic sarcoma had shrunk dramatically. The patient died of sepsis during the nadir state 35 days after starting salvage chemotherapy.