Reduced-intensity conditioning (RIC) haploidentical (haplo-) hematopoietic stem cell transplantation (HSCT) requires more hematopoietic progenitor and stem cells (HPSCs) to promote engraftment and immune reconstitution and needs a stronger graft-versus-leukemia effect. Peripheral blood stem cells (PBSCs) offer advantages over bone marrow; however, the use of higher-dose non-T cell-depleted (non-TCD) in vitro PBSCs may increase the occurrence of severe graft-versus-host disease (GVHD). This prospective, single-arm clinical study was performed to investigate using high-dose non-TCD in vitro PBSCs as the graft source, using fludarabine/Ara-C/busulfan (FAB) as the conditioning regimen, using rabbit antithymocyte globulin to remove T cells in vivo, and enhancing GVHD prophylaxis with an IL-2 receptor antagonist in RIC-haplo-HSCT in patients with hematologic malignancies age 50 to 70 years or <50 years with comorbidities (Hematopoietic Cell Transplantation Comorbidity Index score >= 2) classified as intermediate to high risk. The primary endpoint was day 100 acute GVHD (aGVHD). A total of 47 patients were enrolled; the median age was 52 years (range, 30 to 68 years), the median duration of follow-up was 34 months (range, 2 to 99 months), and the medium-infused doses of mononuclear cells, CD34(+) cells, and CD3(+) cells were 15.93 x 10(8)/kg, 8.68 x 10(6)/kg, and 5.57 x 10(8)/kg, respectively. The cumulative incidence of grade II-IV aGVHD at day 100 was 30.3% (95% confidence interval [CI], 15.9% to 44.8%), and that of grade III-IV aGVHD was 10.2% (95% CI,.6% to 19.8%). The 2-year cumulative incidence of chronic GVHD (cGVHD) was 34.9% (95% CI, 19.0% to 50.8%). The 2-year cumulative incidences of localized and extensive cGVHD were 26.1% (95% CI, 11.80% to 40.40%) and 8.7% (95% CI, 3.26% to 20.65%), respectively. The 2-year cumulative incidence of relapse was 17.3% (95% CI, 5.1% to 29.5%), the 2-year overall survival rate was 71.2% (95% CI, 57.9% to 84.5%), and the 2-year disease-free survival rate was 66.2% (95% CI, 52.1% to 80.3%). The incidence of aGVHD was not high, and the overall efficacy was good. This study demonstrates that this unique RIC-haplo-PBSC transplantation protocol was effective in treating hematologic malignancies. Nonetheless, larger prospective multicenter clinical trials and experimental studies should be performed to further confirm our findings. (c) 2023 The American Society for Transplantation and Cellular Therapy. Published by Elsevier Inc. This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/)

Background Although many reports have established the efficacy of post-transplantation cyclophosphamide (PTCy) or ATG for GVHD prophylaxis to in vitro non-T cell-depleted (non-TCD) haplo-HSCT and achieved a good clinical effect, the overall results of NRM, RI, DFS, and OS still need to be improved, especially for patients with high-risk or R/R hematologic malignancies. We speculated that if GVHD could be controlled well, higher doses of in vitro non-TCD peripheral blood stem cells (PBSCs) as a graft source for haplo-HSCT may be able to better balance engraftment, immune reconstitution, infection, and graft versus leukemia (GVL) effect to improve clinical outcomes. Apparently, due to the increased risk of GVHD as a major concern, the research of haplo-HSCT using ultra-high doses of in vitro non-TCD PBSCs as a graft source is now scarce. To this end, we constructed a unique haploidentical high-dose peripheral stem cell transplantation (haplo-HDPSCT) protocol. A prospective study was performed in our center using this protocol to treat patients with hematologic malignancies. Methods From January 2006 to December 2018, a total of 346 patients with hematologic malignancies less than 50 years old were enrolled. 205 patients who had no matched sibling donor (MSD) enrolled in the haplo-HDPSCT group, and 141 patients were enrolled in the MSD-PSCT group. Patients in two groups were comparable except for several characteristics such as more young and high-risk patients in the haplo-HDPSCT group. All patients were followed up until December 2022 with a median follow-up time of 66.1 months. The MSD-PSCT protocol was performed with the classical standard process. Our haplo-HDPSCT protocol was designed adopting high-dose in vitro non-TCD PBSCs as graft, Ara-C+Bu/Cy+rATG as the conditioning regimen, and Basiliximab plus short-term low-dose GCs added to standard GVHD prophylaxis. The long-term outcomes of the two groups were analyzed and compared. The primary endpoint was aGVHD, and the second endpoints were engraftment, infection, RI, DFS, and OS. Results In the haplo-HDPSCT group(n=205), the infused median mononuclear cells(MNCs), CD34+ cells, and CD3+ cells were 15.63×10 8/kg, 8.59×10 6/kg and 5.39×10 8/kg, respectively, which significantly higher than those in the MSD-PSCT group(n=141). Only 3 of 205 patients in the haplo-HDPSCT group developed primary graft failure (PGF) and the primary engraftment rate was 98.5%. The incidence of grades II-IV and III-IV aGVHD within 100 days was similar between the two groups (37.1% vs. 27.3%, P=0.109; and 11.7% vs. 8.7%, P=0.507). The 3-year cumulative incidence of localized cGVHD was similar between the two groups (50.3% vs 49.3%, P=0.357), but the extensive cGVHD in the haplo-HDPSCT group was significantly lower than that in the MSD-PSCT group (10.9% vs 26.9%, P=0.035). The 3-year cumulative RI and NRM were also similar between the two groups (RI: 31.1%vs 27.3%, P=0.625; NRM: 16.1% vs 11.5%, P=0.394). No significant differences in 3- and 5-year OS (69.6% and 66.9% vs. 74.9% and 73.2%, P=0.282) or 3- and 5-year DFS (67.2% and 63.7% vs. 72.0% and 69.4%, P=0.215) were observed in the haplo-HDPSCT group and the MSD-PSCT group. The CMV disease occurrence was 3.1% in the haplo-HDPSCT group. The EBV-DNAemia rate was 10.4% in the haplo-HDPSCT group and 5.7% in the MSD-PSCT group(P=0.836). Only two patients in the haplo-HDPSCT group developed PTLD. Conclusions This study demonstrates that aGVHD is not high in our haplo-HDPSCT protocol and the long-term follow-up shows that this protocol may bring benefits including high engraftment rate, less relapse and viral infection, which implies that higher doses of in vitro non-TCD PBSCs as a graft source for haplo-HSCT protocol is feasible if appropriate GVHD prophylaxis is adopted.

目的:了解异基因外周血造血干细胞移植后出血性膀胱炎(HC)的发生情况及BK与JC病毒感染情况.方法:选取2019年1-12月行异基因造血干细胞移植且发生出血性膀胱炎的患者54例,分析HC的发生情况.结果:迟发型出血性膀胱炎(LOHC)累计发生率为29.6％,单倍体42例LOHC累计发生率为34.6％.患者血及尿BK、JC病毒DNA检测,Ⅳ度为阴性1例,因新冠肺炎疫情影响未行病毒检测2例,其余14例患者均为阳性.结论:HC是异基因外周血造血干细胞移植后常见并发症,单倍体造血干细胞移植是其发生的独立危险因素,BK及JC病毒感染与HC发生直接相关.

Background: Aggressive natural killer cell leukemia (ANKL) is characterized by low incidence and high mortality. This study reported a case of aggressive ANKL treated with our unique, reduced-intensity conditioning regimen, haploid hematopoietic stem cell transplantation after complete remission (RIC-RHNT-PBSCT). Case presentation: A 51-year-old male had been diagnosed with acute cholecystitis and acute cholangitis due to jaundice and fever, and he received surgical treatment. After the operation, the patient reported a constant leukocyte rise, and he was finally diagnosed with ANKL based on a bone marrow puncture and leukemia immunophenotyping. After the diagnosis, with an active surgical incision and T-tube drainage, the very weak ANKL patient received the IAVD + asparaginase chemotherapy regimen and went into complete remission (CR) after two courses of the chemotherapy regimen. Our unique RIC-RHNT-PBSCT was performed to improve the long-term survival of this still weak patient. The hematopoietic stem cell donor was the patient’s son, who was haploid 3/6 matched. In order to reduce the transplantation-related mortality, the intensity conditioning regimen of FAB was adopted. Starting at day 15 after the stem cell transplantation, a complete donor chimerism was observed, with a chimeric rate of 100%. At the end of follow-up period, the bone marrow aspiration result was normal, and testing found the chimerism was a complete donor chimerism. Conclusion: A rare and weak ANKL patient achieved long-term survival with our RIC-RHNT-PBSCT.

Objective To explore the clinical outcome of HLA haploidentical vs HLA-matcbed peripheral blood hematopoietic stem cell transplantation (PBSCT) without in vitro T-cell depletion for malignant hematological diseases. Methods 111 patients with malignant hematological diseases underwent PBSCT without in vitro T-cell depletion between May 2004 and February 2009, including 51 patients with HLA-haploidentical and 60 patients with HLA-matched. All patients have received myeloablative conditioning regimen. A two-agent based graft-versus-host disease (GVHD) prophylaxis was used as cyclosporine A and a short course of methotrexate. Mycophenolate mofetile was added for the patients with one locus mismatch. Mycophenolate mofetile, antithymocyte globulin and CD25 monoclonal antibody were added for the patients with 2-3 loci mismatch. The grafts were granulocyte colony-stimulating factor-mobilized peripheral blood stem cells without in vitro T-cell depletion. Results 111 patients achieved sustained and full donor-type engraftment. The median time to reach an absolute neutrophil count above 0.5×10~9/L was 14 days and that to a platelet count exceeding 20×10~9/L was 15 days in 51 HLA-haploidentical patients, and that was 12 days and 13 days in 60 HLA-matched patients, respectively. In 51 HLA-haploidentical patients, 25 patients developed aGVHD, including 20 cases of grade Ⅰ aGVHD, and 5 cases of grade Ⅱ. Thirty-three patients developed cGVHD with limited in 30 and extensive in 3. The 4-year cumulative incidence of cGVHD was 70.4 %. The 3-year probabilities of leukemia-free survival (LFS) were 74.5% (77.3 % for standard risk patients and 68.2 % for high risk patients respectively). Seven patients had recurrence. In 60 HLA-matched patients, 14 patients developed aGVHD, including 10 cases of grade Ⅰ, 2 cases of grade Ⅱ and 2 cases of grade Ⅲ. Thirty-seven patients developed cGVHD with limited in 32 and extensive in 5. The 4-year cumulative incidence of cGVHD was 58.1%. The 3-year probabilities of LFS were 72.1% (77.6 % for standard risk patients and 52.7 % for high risk patients respectively). Ten patients had recurrence. The incidence of aGVHD in HLA-haploidentical cohort was significantly higher than in HLA-matched cohort (P<0.05). There was no significant difference in incidence of cGVHD, incidence of relapse and LFS between HLA-haploidentical and HLA-matched cohorts (P>0.05). Conclusion Haploidentical PBSCT is feasible and safe for malignant hematological diseases to use myeloablative conditioning regimen in combination with intensive immunosuppressants without in vitro T cell depletion.

OBJECTIVE:To analyze the clinical outcome of human leukocyte antigen (HLA) haploidentical peripheral blood stem cell transplantation (PBSCT) from related donors for hematological malignancies.METHODS:Thirty-six patients with hematological malignancies, with a median age of 25 (11-48) years, were transplanted with PBSC from an HLA-haploidentical family donors: 7 were 1 locus mismatched and 29 were 2-3 loci mismatched. The recipients received myeloablative conditioning regimen, in combination with different immunosuppressants according to the degree of HLA disparity followed by non-T-cell depleted PBSCT. The median number of CD34+ cells were 11 (4.16-21.00) x 10(6)/kg.RESULTS:All patients achieved sustained, full donor-type engraftment. Fifteen patients (41.7%) developed grade I-II aGVHD. Among 29 patients followed up more than 18 months, 17 (58.6%) developed cGVHD. There was no statistical difference in decrease and recovery of T, B and NK cell subsets after transplantation between HLA haploidentical group and HLA identical PBSCT group. The median follow-up duration was 15 (4 -69) months. Five patients (13.9% ) relapsed. The 2-year probability of leukemia-free survival (LFS) was 82.2%.CONCLUSION:Non-T-cell depleted HLA-haploidentical PBSCT is safe and feasible for patients with hematological malignancies after myeloablative conditioning regimen combined with intensive immunosuppressants.

Objective To investigate the correlation between immature granulocyte and CD34+ cells, mononuclear cells (MNC) in donor's peripheral blood by recombinant human granulocyte colony-stimulating factor (rhG-CSF). Methods The stem cell were mobilized by rhG-CSF 7.25-10 μg·kg-1·d-1 from 122 allo-PBSCT donors. Before and after mobilization, to test CD34+ cells of peripheral blood stem cell graft and number of MNC, immature granulocyte, CD34+ cell per patient' weight were calculated. Results White blood cell count and immature granulocyte gradually increased, and reached the peak on the frith day. There was a good relationship between increased immature granulocytes and increased CD34+ cells. The patients all achieved completed donor engraftment and achieved hematopoietic recovery. The chromosome, blood type and HLA type were transformed to be donor's type. Ph1 changed to be negative in CML patients. Conclusion rhG-CSF (7.25～10 μg·kg-1·d-1) had a good effect to mobilize PBSC. There was a good relationship between in-creased immature granulocytes and increased CD34+ cells after mobilization by rhG-CSF. The number of immature granulocytes can reflect indirectly the count of stem/progenitor, so the MNC and immature granulo-cytes can become a threshold of dosage standard.

从新疆维吾尔族健康人造血干细胞中提取总RNA,用反转录(reverse transcription,RT)和降落PCR(touchdoWn PCR,TD-PCR)相结合的方法扩增人细菌透性增加蛋白(human bactericidal/permeability-increasing protein,hBPI)的cDNA,将其克隆到pEGFP-N1载体上并进行DNA序列测定.结果表明,所克隆的hBPIcDNA全长为1,464个碱基,其序列与GenBank中另外4个序列进行了比对,有两个碱基与其它4个序列不同:其中第576位碱基其它序列为G,该位置碱基是C;第676位碱基其它序列为A,该位置碱基是G.其中第676位碱基的变化导致第185位氨基酸由Lys改变为Glu.

我院于1998年3月至2005年4月共施行异基因外周血干细胞移植（allo-PBSCT）45例,现报告如下。资料与方法一、一般资料45例患者,年龄14～47岁,年龄中位数为36岁,其中急性非淋巴细胞白血病（ANLL）18例（第1次缓解12例,第2次缓解3例,早期复发2例,难治型1例）,急性淋巴细胞白血病（ALL）5例（第1次缓解2例,第2次缓解3例）,慢性粒

Objective:To research the collecting methods of auto-peripheral blood stem cell and it's effect. Methods: According to the 7 lymphoma patient's MNC of peripheral blood after mobilization, turbidity of the collecting stem cell liquid and the patient's status, we used the Cobe-Spectra machine to collect the peripheral blood stem cell in a single apheresis session with 3 total blood volume(TBV) circles. MNC and CD34+ cells were calculated, and autologous peripheral blood stem cell transplantation (APBSCT) was underwent after the stem cells cryopreserved 10-14 days. The hematopoietic rebuilding situation was observed. Results: The 7 patient's average MNC and CD34+ cells were 7. 25×108/kg and 5. 57×106/kg respectively after peripheral blood stem cells harvest in a single large volume apheresis session. The collecting process was favoring. The hematopoietic rebuilt quickly. The average days of neutrophil≥0. 5×109/L and platelet≥20×109/L were +8. 9 and+8. 7 days respectively after APBSCT. Conclusions:The ade quate peripheral blood stem cell of partial lymphoma patients could be collected in a single large volume apheresis session. The collecting process is secure. The hematopoietic rebuild quickly after APBSCT.

目的:对ABO血型不合异基因外周血干细胞移植造血功能重建及移植后早期成分输血的选择进行探讨.方法:首先对供受者ABO血型及血清凝集效价进行鉴定.单采单个核细胞时控制红细胞污染量在10 mL以下,移植后检测受者血型转变、血清凝集素的动态变化和干细胞植活情况.结果:ABO血型不合移植与ABO血型相同移植后干细胞植活时间相同,移植时及移植后早期成分输血无1例发生溶血现象.结论:ABO血型不合不影响造血干细胞植活时间,移植后根据受者血型转变情况及凝集素动态变化选择移植后早期成分输血,可避免溶血性输血反应且不影响干细胞植活及血型转变时间.

近年来国内外报告异基因外周血干细胞移植(allo-PBSCT)的病例日渐增多。为探讨外周血干细胞(PBSC)的动员、检测及移植的剂量标准与临床效果，对6例恶性血液病患者进行了移植研究，报告如下。