Application of Chimeric Antigen Receptor-Modified NK Cells in Multiple Myeloma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the killing effect of CAR (CD138-CD28-CD3ζ)-NK cells on myeloma cells through construction of CAR(CD138-CD28-CD3)-NK cells.</p><p><b>METHODS</b>The antiCD138scFv-CD28-CD3 zeta plasmid pcDNA3.1 was constructed, which then together with 3 plasmid lentiviral packaging system were transfected into 293T cells, the virus was collected. Furthermore, in order to get the stably transfected cell line, the NK92MI cell line was infected by the virus, then the positive cells were screened by puromycin. The expression of the CARNK cells were verified by RT-PCR and Western blot. At last the ability of secreting cytokine CD107a was detected by flow cytometry, and the statistical analysis was carried out to verify the anti-myeloma effect of CAR-NK cells.</p><p><b>RESULTS</b>Gene fragment of the CAR(antiCD138scFv-CD28-CD3ζ) was constructed successfully by gene engineering technique in vitro, and the gene sequence was verified to be correct by sequencing. By virus packaging technology, the virus expressing the protein of the CAR was obtained. PCR and Western blot verified the expression of CAR fusion protein on the sufurce of NK cells. The cell killing experiment confirmed that the CAR-NK cells possessed the ability to secrete cytokine CD107a superior to control cells and showed the obvious killing effect on multiple myeloma cells.</p><p><b>CONCLUSION</b>The CAR can be constructed in vitro, and express on NK92 cells. The CAR-NK cells can kill the multiple myeloma cells expressing CD138 antigen, thereby plays an antimyeloma effect.</p>

Effect of rhG-CSF Mobilization on S1P5 Expression in T Lymphocyte Subsets of Allo-HSCT Donors / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the effect of recombinant human granulocyte colony-stimulating factor (rhG-CSF) mobilization on S1P5 expression in T lymphocyte subsets of allo-HSCT donors.</p><p><b>METHODS</b>The peripheral blood was collected from 10 allo-hematopoietic stem cell transplantation (allo-HSCT) donors before and after mobilization with rhG-CSF for 4 days. The flow cytometry was used to detect S1P5 expression in T lymphocyte subsets.</p><p><b>RESULTS</b>There was no S1P5 expression on the surface of T-lymphocytes both before and after rhG-CSF mobilization. After fixation with permeabilization agent, S1P5 expression could be detected in lymphocytes after rhG-CSF mobilization, which indicates S1P5 may be located in cells. Compared with level before rhG-CSF mobilization, S1P5 expression was significantly increased in T lymphocyte subsets after rhG-CSF mobilization, CD3(+)T cells (57.92±2.32)% vs (7.94±1.47)%(P<0.05）, CD4(+)T cells (72.58±1.73)% vs （5.48±0.82）%(P<0.05）, CD8(+)T cells（51.79±3.57）% vs （6.46±1.01）%（P<0.05）,CD3-/CD56(+)NK cells（40.00±1.47）% vs（4.97±0.74）%（P<0.05）. The up-regulated level of S1P5 expression in CD4(+)T cells was most high(P<0.05).</p><p><b>CONCLUSION</b>S1P5 expression significantly increases in T lymphocyte subsets after rhG-CSF mobilization, and the up-regulated level of S1P5 expression in CD4(+)T cells is the most high.</p>

Expression Profile of lncRNA NONHSAT040475 in Peripheral Blood Leukocytes of Healthy Persons / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the expression profile of lncRNA NONHSAT040475 in peripheral blood leukocytes of healthy persons.</p><p><b>METHODS</b>The peripheral blood mononuclear cells were collected from 10 healthy volunteers, the CD3(+) T cells,CD19(+) B cells,CD56(+) NK cells and granulocytes were purified and sorted by flow cytometry. Then, the expression of lncRNA NONHSAT040475 in peripheral blood leukocyte subsets was detected by real-time quantitative PCR.</p><p><b>RESULTS</b>the expression levels of lncRNA NONHSAT040475 were various in lymphocyte subsets, its expression in B lymphocytes was significantly higher, as compared with T lymphocytes, while the expressions of lncRNA NONHSAT040475 in NK cells and granulocytes were relative low（P<0.01）.</p><p><b>CONCLUSION</b>lncRNA NONHSAT040475 is widely expressed in human peripheral blood leukocytes, and mainly expressed in B lymphocytes, therefore, laying the foundation for further study of B lymphocyte-related diseases. This study has brought a new opportunity for diagnosing and illustrating the molecular mechanism of hematologic disorder or autoimmune disease.</p>

Therapeutic Efficacy Analysis of Allogeneic Peripheral Blood Hematopoietic Stem Cell Transplantation for 14 Adult Patients with T Lymphoblastic Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To investigate the therapeutic efficacy of allogeneic peripheral blood hematopoietic stem cell transpdantation (allo-HSCT) for T lymphoblastic lymphoma (T-LBL).</p><p><b>METHODS</b>The clinical data of 14 adult patients with T-LBL treated with allo-HSCT were collected, the hematopoietic reconstruction, survival and relapse, as well as overall survival (OS) rate, event-free survival (EFS) rate of 1, 3 and 5 years were analysed retrospectively.</p><p><b>RESULTS</b>All the patients were engrafted with neutrophil successfully, the median time of absolute neutrophil count >0.5 × 10(9)/L was 13 (10-19) d; 13 patients were engrafted with platelets successfully, the median time of Plt count >20 × 10(9)/L was 17 (12-62) days. The acute GVHD occurred in 6 patients, but among them only 1 case with 3 grade of aGVHD; out of 14 patients, 5 developed chronic GVHD. The transplant-related mortality at 100 days was 7.1% (1/14), mainly from coronary heart disease and pulmonary infection. The median follow-up time was 26.5 months, the estimated 1, 3 and 5 year OS rate was 85.7%, 47.6% and 38.1%, respectively, and estimated 1, 3 year EFS rate was 85.7%, 34.4% and 34.1%, respectively. The relapse rate was 42.8% (6/14) and the median relapse time was 22.5% months after transplantation. Up to now, 7 patients still survive, 1 patient out of them have survived for 103 months.</p><p><b>CONCLUSION</b>The allo-HSCT is a safe and effective method for treatment of T-LBL.</p>

Role of Mesenchymal Stem Cells in Preventing GVHD: A Meta-Analysis / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To evaluate the efficacy of mesenchymal stem cells (MSC) in the prevention of graft versus host disease (GVHD) after hematopoietic stem cell transplantation (HSCT).</p><p><b>METHODS</b>Randomized controlled trials (RCT) were identified from PubMed (1950.1-2014.3), EMbase (1970.1-2014.3), Cochrane Central Register of Controlled Trials (CENTRAL, issue 4, 2014) of the Cochrane Library, China Biological Medicine (CBM, 1978.1-2014.3). References of retrieved articles were also identified. The quality of each RCT was evaluated by the Cochrane collaboration's tool for assessing the risk of bias. Data analysis was performed with Review Manager 5.1 to evaluate the efficacy of MSC in the prevention of GVHD after HSCT.</p><p><b>RESULTS</b>A total of 3 English articles involving 117 patients were included. Meta-analysis indicated that MSC did not reduce the incidence of acute GVHD and chronic GVHD (RR:0.44, 95% CI: 0.08 to 2.51, P = 0.35; RR:0.85, 95% CI: 0.54 to 1.33, P = 0.47). However, MSC did not increase occurrence of relapse and cytomegalovirus infection (RR:1.52, 95% CI:0.63 to 3.68, P = 0.35;RR:1.05, 95% CI:0.72 to 1.53, P = 0.78). Finally, MSC did not improve overall survival rate of patients received HSCT (RR:1.06, 95% CI:0.79 to 1.43, P = 0.71).</p><p><b>CONCLUSION</b>MSC may have a preventive effect on GVHD in patients undergoing HSCT. However, the evidence is weak due to the small sample sizes. Thus, a reliable conclusion about the preventive effect of MSC on GVHD at the moment has not been made, further larger, high quality, randomized and controlled trials are warranted.</p>

Expression and Prognostic Value of BCL-2 Protein in Diffuse Large B-cell Lymphoma / 中国实验血液学杂志

<p><b>OBJECTIVE</b>To explore the value of BCL-2 protein for evaluating the prognosis of patients with diffuse large B cell lymphama (DLBCL).</p><p><b>METHODS</b>The clinical data of 128 patients with DLBCL including clinical features, BCL-2 protein expression, therapeutic outcome and so on were analyzed retrospectively in departenent of hematology, Chinese PLA general hospital from January 2008 to December 2010, and the prognosis of DLBCL patients with different expression levels of BCL-2 protein was compared, including overall survival (OS) and progression-free survival (PFS) rates.</p><p><b>RESULTS</b>The BCL-2 expression postive was found in 83 cases (64.8%), while BCL-2 expression negative was observed in 45 cases (35.2%). The OS rates in BCL-2 expression positive and negative groups were 76.6% vs 76.8% in 3 years (P >0.05), and the PFS rates in BCL-2 expression positive and negative groups were 57.1% vs 70.5% (P >0.05) in 3 years, suggesting that BCL-2 expression level had no significant impact on OS and PFS rates in all DLBCL patients. However, among the older patients aged ≥ 60 years, there was singnificant different of 3 year OS rates in BCL-2 expression positive and negative groups (66.7% vs 76.4%, P >0.05), while 3-year PFS rate in BCL-2 expression positive group was obviosusly lower than that in BCL-2 expression negative group (35.8% vs 83.3%, P < 0.05).</p><p><b>CONCLUSION</b>The positive expression of BCL-2 protein is a poor prognostic factor for older patients aged ≥ 60 years, thus this indicator possesses the prognostic value for these patients with DLBCL.</p>

Analysis of prognosis in acute myeloid leukemia treated with allogeneic hematopoietic stem cell transplantation / 中国实验血液学杂志

<p><b>OBJECTIVE</b>The study was to investigate the prognosis factors in acute myeloid leukemia (AML) patients treated with allogeneic hematopoietic stem cell transplantation (allo-HSCT).</p><p><b>METHODS</b>The clinical information of 60 patients in our hospital was retrospectively analyzed and the prognosis factors of survival and relapse were explored by COX's proportional hazard model.</p><p><b>RESULTS</b>The elderly (HR = 4.530, P = 0.012), cGVHD (HR = 0.023, P = 0.003) and infection fungal disease (IFD) (HR = 4.019, P = 0.017) were influence factors for 2 year cumulative overall survival (OS). Response status (high risk vs low risk: HR = 3.465, P = 0.028), preconditioning regimens (TBI/Cy vs Bu/Cy: HR = 0.071, P = 0.012; FB vs Bu/Cy: HR = 7.547, P = 0.025) and cGVHD (HR = 0.088, P = 0.004) were influence factors for 2 year cumulative relapse rate (RR). cGVHD (P = 0.017) and IFD (P = 0.000) had an effect on OS after 2 years since allo-HSCT.</p><p><b>CONCLUSION</b>Age, response status, preconditioning regimens, cGVHD and IFD are closely associated with the prognosis of AML patients treated with allo-HSCT, and different patients need the individualized treatment.</p>