Aggressive T-cell large granular lymphocytic leukemia represents a rare but highly fatal clinical entity, starkly contrasting with the typically indolent nature of classic large granular lymphocytic leukemia. This report details the case of a female in her early 60s who succumbed to the disease merely 3 days after hospitalization, highlighting a precipitous clinical course from unremarkable blood test results to death within approximately 4 months. The diagnosis was confirmed by peripheral blood morphology, immunophenotyping, and molecular cytogenetic studies. The leukemic cells demonstrated a natural killer T-cell phenotype, clonal T-cell receptor rearrangement, and a complex karyotype. A significant outcome of this case was the successful establishment of a stable cell line derived from the patient's peripheral blood mononuclear cells. This case underscores the need for early recognition of this aggressive T-cell large granular lymphocytic leukemia variant. Furthermore, the novel cell line provides an invaluable and unprecedented resource for elucidating the underlying pathogenesis and exploring potential therapeutic strategies for this devastating malignancy.

Hemophagocytic lymphohistiocytosis (HLH) is a group of hyperinflammatory disorders with a mortality rate exceeding 50%. We report a case of a female Asian patient who developed secondary HLH accompanied by aggressive natural killer cell leukemia and Epstein-Barr virus (EBV) infection, and was treated with tripterygium glycosides (TG), a Chinese patent medicine. Within 3 weeks of oral administration, the patient's recurrent high fever resolved, abdominal distension and splenomegaly improved, ascites diminished, serum soluble CD25 levels decreased, and hematopoietic and coagulation functions recovered. Triptolide, a major component of TG, exhibited cytotoxicity against the patient's ascitic cells and induced apoptosis in a dose-dependent manner ex vivo. Whole-genome and transcriptome sequencing of the patient's tumor cells revealed that TG regulated EBV-associated mutated genes such as PSMD7 and modulated inflammation-related pathways. Molecular docking further suggested direct targeting of PSMD7 by triptolide. Tripterygium glycosides quickly mitigated cytokine storm, alleviated symptoms of HLH, and showed no observed adverse effects with a good cost-benefit profile, thereby offering a potential bridge for follow-up hematopoietic stem cell transplantation.

Natural killer T cells (NKT), a unique subset of T lymphocytes, remain incompletely understood in terms of their development and functional regulation. Well-characterized immortalized cell lines are critical tools for investigating NKT cell biology in vitro. We established and maintained a novel NKT cell line, NKT617, derived from the peripheral blood of a patient with aggressive large granular lymphocyte leukemia. Cell morphology was assessed using scanning electron microscopy and transmission electron microscopy. Flow cytometry characterized the cell phenotype, whereas T-cell receptor (TCR) clonal gene rearrangement determined lineage. RNA sequencing compared gene expression profiles of NKT617 with other hematological tumor cell lines, such as natural killer (NK) cells, T cells, B cells, and monocytes. Tumorigenic potential was evaluated via in vitro colony formation assays and in vivo zebrafish xenograft models. NKT617 has been continuously cultured for >2 years, exceeding 100 passages, demonstrating monoclonal immortalization. It proliferates in a low-dose recombinant human interleukin-2-dependent manner and grows in suspension culture as single cells and aggregates. Flow cytometric analysis showed that NKT617 exhibits an NK-cell phenotype CD56+, CD158a+, NKp46+, membrane-bound CD3-, and T-cell features cytoplasmic CD3+, CD2+, CD4+, CD8+, CD45RA+, and CD45RO+, with clonal TCRβ rearrangement. NKT617 was negative for B-cell and myeloid markers and positive for the TCRα chain (Vα24-Jα18), confirming its identity as a type I NKT cell. Transcriptome analysis showed shared markers with NK-cell and T-cell lines but a gene expression profile closer to NK cells. In vitro and in vivo assays confirmed its tumorigenic capacity. We report the first Epstein-Barr virus-negative human type I NKT cell line, NKT617, which offers significant potential for studying NKT cell development and advancing chimeric antigen receptor-based therapies. This cell line serves as a valuable tool for exploring NKT cell biology and developing targeted immunotherapies.

Background Autoimmune modulators (Aire) mediate immune tolerance by inducing expression of tissue-restricted antigen (TRA) in thymic medullary epithelial cells. However, the function of Aire in peripheral B cells remains unknown. We aimed to investigate whether Aire is expressed in human peripheral B cells and how its expression is regulated. Methods Human thymus tissues were collected and firstly, islet-specific autoantibodies and insulinogen c-peptide were determined by luciferase immunoprecipitation (LIPS). Secondly, Aire expression in peripheral blood lymphocytes was identified by flow cytometry, Western Blot and immunofluorescent staining. Then, immunofluorescent staining was performed on cells cultured in vitro and surgically resected tissues. Meanwhile, Aire promoter methylation. Finally, insulinogen expression was quantified by reverse transcription qPCR and IF staining. Data were compared by Mann-Whitney U test between groups. Results Aire was expressed in CD19+ B cells but not in CD3+ cells, and the expression of Aire in B cells was much lower than that in thymus. Induction of Aire expression under BCR activation in B cells. The level of Aire expression in peripheral B cells of patients with T1D and healthy subjects showed no difference in Aire expression level between the two groups. Conclusions Aire expression in peripheral B cells may play a role in peripheral immune tolerance and protection of islet β-cell function in T1D patients.

OBJECTIVE:The leukemia cells from patients with T-cell acute lymphoblastic leukemia (T-ALL) were inoculated into NCG mice to establish a stable human T-ALL leukemia animal model.METHODS:Leukemia cells from bone marrow of newly diagnosed T-ALL patients were isolated, and the leukemia cells were inoculated into NCG mice via tail vein. The proportion of hCD45 positive cells in peripheral blood of the mice was detected regularly by flow cytometry, and the infiltration of leukemia cells in bone marrow, liver, spleen and other organs of the mice was detected by pathology and immunohistochemistry. After the first generation mice model was successfully established, the spleen cells from the first generation mice were inoculated into the second generation mice, and after the second generation mice model was successfully established, the spleen cells from the second generation mice were further inoculated into the third generation mice, and the growth of leukemia cells in peripheral blood of the mice in each group was monitored by regular flow cytometry to evaluate the stability of this T-ALL leukemia animal model.RESULTS:On the 10th day after inoculation, hCD45+ leukemia cells could be successfully detected in the peripheral blood of the first generation mice, and the proportion of these cells was gradually increased. On average, the mice appeared listless 6 or 7 weeks after inoculation, and a large number of T lymphocyte leukemia cells were found in the peripheral blood and bone marrow smear of the mice. The spleen of the mice was obviously enlarged, and immunohistochemical examination showed that hCD3+ leukemia cells infiltrated into bone marrow, liver and spleen extensively. The second and third generation mice could stably develop leukemia, and the average survival time was 4-5 weeks.CONCLUSION:Inoculating leukemia cells from bone marrow of patients with T-ALL into NCG mice via tail vein can successfully construct a patient-derived tumor xenografts (PDTX) model.

目的 检测初诊弥漫大B细胞淋巴瘤(DLBCL)采用R-CDOP(利妥昔单抗联合环磷酰胺、脂质体多柔比星、长春新碱及泼尼松)方案联合或不联合消癌解毒方治疗前后外周血调节性T细胞(Treg)百分比的变化,探讨Treg对于DLBCL发病及预后可能的影响以及消癌解毒方是否具有调控Treg表达的作用.方法 入组2018年1月至2022年1月南京中医药大学附属医院收治的初诊DLBCL病人共47例(治疗组24例,对照组23例)和健康对照45例,治疗组采用消癌解毒方联合R-CDOP方案,对照组单纯采用R-CDOP方案治疗,两组均化疗6个周期,采用流式细胞术检测两组病人治疗前后及健康对照的外周血CD4+CD25+CD127low Treg百分比,观察两组病人的临床疗效以及治疗前后CD4+CD25+CD127low Treg百分比的变化.结果 治疗前,治疗组及对照组两组病人和健康对照的CD4+CD25+CD127low Treg百分比中位数分别为6.01％、5.73％、7.32％,两组病人外周血Treg百分比均低于健康人(P<0.05).治疗6周期结束后,治疗组及对照组两组病人的CD4+CD25+CD127low Treg百分比中位数分别为7.33％和5.85％,治疗组Treg百分比较治疗前明显上升(P<0.05),对照组治疗前后Treg百分比变化差异不明显(P>0.05).治疗组和对照组相比,治疗组比对照组的CD4+CD25+CD127low Treg百分比明显上升(P<0.05).两组病人临床疗效评估结果比较,治疗组和对照组的总缓解率(ORR)分别为79.17％和69.56％,两组ORR差异无统计学意义(P>0.05).结论 消癌解毒方有可能通过上调DLBCL病人失衡的Treg百分比发挥调控肿瘤免疫环境的作用.

Abstract Background Cytokine release syndrome (CRS) is a life-threatening complication of chimeric antigen receptor T cell (CAR-T) therapy. Macrophages/ monocytes are mediators of CRS. Tripterygium glycosides is an immunomodulator which could inhibit macrophages/ monocytes in animal models. Methods Two patients with relapsed and refractory hematological malignancies developed CRS after receiving CAR-T therapy. They received short-term tripterygium glycosides orally. Results Both patients showed rapid mitigation of fever with evidently decrease in elevated inflammatory cytokines within 72 hours. The patients' monocytes diminished remarkably, while CAR-T cells were neglectably affected. Treatment of 30 ng/mL triptolide in ex vivo cultured patients' blood for 24 hours selectively deplete over half of monocytes. Single cell RNA sequencing suggested selective depletion of CD14+CD16+ monocytes with decreased pro-inflammatory cytokines. Conclusions The low-cost and orally available tripterygium glycosides could be a promising alternative for CAR-T induced CRS, as well as other diseases complicated with CRS, e.g., coronavirus disease 2019.

OBJECTIVE To investigate the effect of celastrol on the proliferation and apoptosis of human multiple myeloma (MM) cell lines, reveal the relationship between IRAK4/ERK/p38 signaling pathway and celastrol regulating the proliferation and apoptosis of H929 and ARP-1 cells, and explore whether celastrol combined with bortezomib has synergistic effect. METHODS CCK-8 method was used to detect the viability of MM cell lines H929 and ARP-1 treated by different concentrations of celastrol, bortezomib, and their combination, and the synergistic effect was determined by Kim's formula. The apoptosis rate of H929 cells and necrosis rate of ARP-1 were detected by Annexin V/PI method. The expression of key proteins and apoptosis proteins in IRAK4/ERK/p38 signaling pathway were detected by Western blot. RESULTS Celastrol could significantly inhibit the proliferation of H929 and ARP-1 cells (r=0.9018, r=0.9244) and induce apoptosis in a time-dependent manner. Compared with the control group, celastrol could significantly up-regulate the expression of PARP and cleaved caspase-3 while down-regulate the expression of p-IRAK4, p-ERK, and p-p38 in H929 and ARP-1 cells. Celastrol and bortezomib alone inhibited the proliferation of H929 and ARP-1 cells. Compared with celastrol and bortezomib alone, their combination had lower cell survival rate and higher apoptosis rate (P<0.05). CONCLUSION Celastrol can inhibit the proliferation and promote the apoptosis of H929 and ARP-1 cells, which may be related to inhibiting the phosphorylation of IRAK4 and blocking the activation of IRAK4/ERK/p38 signaling pathway. Celastrol combined with bortezomib has synergistic effect, which can more effectively inhibit the proliferation and induce apoptosis of H929 and ARP-1 cells.

PURPOSE:Cimicifuga dahurica (C. dahurica), which has been used in traditional oriental medicine for a long period, was reported to exert extensive antitumor activity, but the effect and molecular biological mechanism of C. dahurica on multiple myeloma (MM) has not been elaborated. Tumor-associated macrophages (TAMs) exhibit a sustained polarization between tumor killing M1 subtype and tumor supporting M2 subtype. And a lower ratio of M1/M2 is associated with tumor angiogenesis, proliferation and invasion. We explored the inhibitory effect of the aqueous extract of the root of C. dahurica (CRAE) on tumor growth by reprogramming macrophage polarization in the tumor microenvironment.METHODS:Mice bearing SP2/0 multiple myeloma were treated with CRAE. Western blotting (WB), immunohistochemistry (IHC) and immunofluorescence staining were utilized to assess tumor growth and TAM populations. Macrophages were depleted by injection of clodronate liposomes to determine and measure the role of CRAE as an anti-tumor agent by targeting macrophages. To simulate tumor microenvironment, MM cells H929 and TAMs were co-cultured using the transwell co-culture system. By using CRAE as an immunoregulator in M2-like macrophages, we analyzed CRAE-treated macrophage-associated surface markers and cytokines by flow cytometry and WB.RESULTS:The results indicated that CRAE treatment could reduce tumor burden of MM mice and a high degree of M1-like macrophages infiltration was detected in tumor tissues. In vitro co-culture system, CRAE significantly promoted the polarization of M2 to M1 phenotype, which led to the increase in apoptosis of myeloma cells. It was found that the M1 polarization induced by CRAE depended on the TLR4-MyD88-TAK1-NF-κB signal transduction.CONCLUSION:This study elucidated the anticancer mechanism of the aqueous extract of C. dahurica (CRAE) through reprogramming macrophage polarization and highlighted that CRAE could act as a potential novel option for cancer immunotherapy.

The prevalence of allergic rhinitis has exhibited an upward trend, and diabetes is a common endocrine metabolic disorder. Treatment of allergic rhinitis complicated with diabetes has been marginally explored. This study aimed to observe the effect of rupatadine fumarate combined with acupoint application in the treatment of allergic rhinitis complicated with diabetes and its effect on serum IgE levels. Totally 80 patients with allergic rhinitis complicated with diabetes admitted to our hospital from December 2019 to December 2020 were recruited and assigned to receive either rupatadine fumarate (control group) or rupatadine fumarate plus acupoint application (research group). The clinical observation indexes of the two groups of patients before and after treatment were analyzed, and the clinical efficacy of the two groups was evaluated. Rupatadine fumarate plus acupoint application was associated with a significantly higher efficacy (23 cases of markedly effective, 14 cases of effective, and 3 cases of ineffective) versus rupatadine fumarate alone (14 cases of markedly effective, 16 cases of effective, and 10 cases of ineffective) (χ 2 = 4.501, p = 0.034). The immunoglobulin E (IgE) and nasal mucosal eosinophils (EOS) levels of the two groups of patients after treatment decreased significantly, and the research group had lower results (p < 0.05). Patients in the research group showed significantly lower syndrome scores than those in the control group (p < 0.05). Rupatadine fumarate plus acupoint application resulted in significantly lower physical sign scores and interleukin-4 (IL-4) levels and higher levels of interferon-gamma (INF-γ) versus rupatadine fumarate alone (p < 0.05). The two groups showed a similar incidence of adverse events (p > 0.05). Rupatadine fumarate plus acupoint application may offer a viable alternative for the treatment of allergic rhinitis as it alleviates the clinical symptoms, improves the treatment efficiency, and enhances the anti-allergic effect of the drug, with a high safety profile.

Solasonine, the main active ingredient of Solanum nigrum L ., has been reported to exert extensive antitumor activity. However, the antitumor effects in acute monocytic leukemia and the exact mechanisms involved are unknown. In this study, we investigated the role of solasonine on inhibiting the progression of acute monocytic leukemia. Our findings showed that solasonine inhibited the proliferation of acute monocytic leukemic cell lines (THP-1 and MV4-11) in vitro . Solasonine promoted apoptosis and induced cell cycle arrest in the G2/M phase. Analysis of RNA-seq data suggested that solasonine correlated with increased expression of genes in the AMPK/FOXO3A pathway. Inhibition of AMPK with compound C followed by treatment with solasonine showed that solasonine reduced apoptosis, caused less cell cycle arrest, and inactivated the AMPK/FOXO3A axis in THP-1 and MV4-11 cells. Solasonine also inhibited tumor growth by the activation of the AMPK/FOXO3A axis. In conclusion, solasonine inhibited the progress of acute monocytic leukemia in vitro and in vivo and triggered the apoptosis and cell cycle arrest in the G2/M phase by upregulating the AMPK/FOXO3A pathway.

目的 观察消癌解毒方联合R-CHOP样方案治疗初诊弥漫大B细胞淋巴瘤的临床疗效.方法 将40例弥漫大B细胞淋巴瘤患者采用随机数字表法随机分为治疗组与对照组,每组20例,治疗组采用消癌解毒方联合一线R-CHOP样方案,对照组单纯采用R-CHOP样方案,两组均化学治疗6个周期.比较两组患者临床疗效、生活质量变化及中医临床证候改善情况.结果 两组实体瘤疗效比较,差异无统计学意义(P>0.05).与治疗前比较,两组患者治疗后卡氏功能状态(Karnofsky performance status,KPS)评分显著升高(P<0.05),中医证候积分显著降低(P<0.05),治疗组患者KPS评分升高程度和中医证候积分降低程度均显著大于对照组(P<0.05).两组患者不良反应发生率比较,差异无统计学意义(P>0.05).结论 消癌解毒方联合R-CHOP样方案治疗弥漫大B细胞淋巴瘤有助于改善患者临床症状,提高患者生活质量.

Review question / Objective: The benefits of Chinese herb in patients with multiple myeloma remain unclear.Condition being studied: We intend to evaluate Chinese herb applied

OBJECTIVE:To observe the changes of serum metabolites in patients with multiple myeloma (MM) by metabonomics, and explore the potential biomarkers for diagnosis, prognosis, and progression of MM.METHODS:Serum samples were collected from 26 patients with MM and 50 healthy controls. The data detected by liquid chromatography-mass spectrometry (LC-MS) was input into SIMCA-14.0 software for multivariate statistical analysis. Principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to analyze the changes of metabolites.RESULTS:The metabolic change of uric acid and trans-vaccenic acid in serum samples of MM patients was 9.39 times and 2.77 times of these in healthy people, respectively, which were significantly higher than those of healthy people, and the difference was statistically significant(P<0.01).CONCLUSION:Uric acid and trans-vaccenic acid are expected to be important metabolic indicators for the diagnosis, prognosis, and efficacy evaluation of MM, thus providing some clues for the pathogenesis of MM.

Background B lymphocyte activating factor (BAFF) is a growth factor regulating B lymphocytes survival and maturation. Serum BAFF levels were elevated in patients affected with autoimmune thyroid diseases (AITD), including Graves’ disease (GD) and Hashimoto’s thyroiditis (HT). The aim of this study is to explore the association of expression levels of BAFF and its receptors with AITD. Methods Fifty-two GD patients, 39 Hashimoto’s thyroiditis (HT) patients and 23 healthy controls (HC) were recruited in this study. Serum BAFF levels were measured by ELISA. Expression of BAFF receptors, including BAFF receptor 3 (BR3) and transmembrane activator and calcium-modulating and cyclophilin ligand interactor (TACI), on B lymphocytes were analyzed by flowcytometry. Effects of steroids on serum BAFF levels and expression of BR3 and TACI were also observed in 10 patients with Graves’ orbitopathy (GO) receiving steroids therapy. Results Serum BAFF levels were significantly elevated from 0.93 ± 0.24 ng/ml in HC to 1.18 ± 0.33 ng/ml in GD ( P = 0.0027) and 1.02 ± 0.24 ng/ml in HT ( P = 0.0331). BR3 expression on peripheral B lymphocytes were elevated in GD (mean MFI: 4.52 ± 2.06 in GD vs. 3.00 ± 0.87 in HC, P = 0.0015), while TACI expression on peripheral B lymphocytes were decreased in GD without significance (mean MFI: 7.96 ± 4.06 in GD vs. 9.10 ± 3.37 in HC, P = 0.1285). Expression of BR3 and TACI was not changed significantly in HT patients. Steroids significantly suppressed serum BAFF concentrations (from 1.18 ± 0.27 ng/ml to 0.97 ± 0.10 ng/ml, P = 0.0364) and BR3 expression in GO patients (mean MFI from 6.26 ± 4.91 to 4.05 ± 1.58, P = 0.0083). Conclusions Altered expression of BAFF and its receptor may mediate the autoimmunity in GD. Restoring the normal expression profile of receptors for BAFF could be a new strategy to treat GD.

Background Tumor associated macrophages (TAMs), a kind of inflammatory cells in the tumor microenvironment, are crucial for the occurrence and development of various tumors which increased the expression of CD163. Nevertheless, not much has been established regarding soluble CD163 and its connection to tumor diagnosis. In this case, a meta-analysis was conducted to determine the tumor diagnostic importance of serum sCD163. Methods In order to assess the correlation between sCD163 and the overall survival (OS) or progression-free survival (PFS) among tumor patients, a systematic perusal of literature published until June 2020 was conducted. Relevant data were primarily obtained from papers that have the following qualifications: 1) a confidence interval (CI) of 95%; 2) a report of the hazard ratios; and, 3) pooled by means of the Mantel-Haenszel random-effect representation. Results For the final meta-analysis, eight papers comprised of 1,236 cases were involved. Through pooled investigation, it was determined that a correlation exists between elevated serum sCD163 and worse OS (HR = 2.24, 95% CI: 1.50-3.35, P < 0.001) and PFS (HR = 3.90, 95% CI: 2.33-6.52, P < 0.001) among tumor cases. Subgroup analysis stratified by medium age at diagnosis demonstrated that patients over 60 years old with high sCD163 had worse OS (HR 2.28, 95% CI: 1.58-3.29, P < 0.001) than under 60 (HR 1.43, 95% CI: 1.15-1.77, P = 0.001). Subgroup analysis revealed that analysis method and medium age at diagnosis were the potential source of heterogeneity. Conclusions Overall, diagnosis of tumor cases can be adversely determined through substantial sCD163 levels. Consequently, it is encouraged that extensive researches regarding the rates of cancer survival be accomplished.

Myeloid-derived suppressor cells (MDSCs) are a significant obstacle for immunotherapy of cancer. It is of great clinical relevance to study the mechanism of MDSCs accumulation in mouse spleens and establish a stable method to obtain high-purity MDSCs in vitro for further research. Here, we established a new method for amplifying a large number of highly pure MDSCs in vitro. To mimic the microenvironment of MDSCs development in vivo, mouse splenic stroma feeder cells and serum-free medium containing granulocyte-macrophage colony stimulating factor (GM-CSF) were used to induce myeloid precursors in mouse bone marrow cells, which differentiate into MDSCs. Development and immunological functions of the cells were monitored both in vivo and in vitro. A total of 4 × 108 MDSCs could be obtained from the bone marrow from one mouse, the ratio of CD11b+Gr-1+ MDSCs could reach 93.8% ± 3.3% after nine days of culture in vitro. Cultured MDSCs maintained a similar immunophenotype with MDSCs found in tumor-bearing mice. Colony forming assay in vitro and in vivo demonstrated that these were myeloid precursor cells. These cells generated high levels of reactive oxygen species and arginase 1 to prevent proliferation of CD8+ T cells in vitro. These also increased regulatory T (Treg) cells in blood while promoting the growth of lymphoma in vivo. In addition, cultured MDSCs effectively inhibited acute graft-versus-host disease (aGVHD). Our findings suggest that mouse splenic stroma plays an important role in the generation of MDSCs and represent a preliminary mechanism for the accumulation of MDSCs in spleens, and thereby lay the foundation for basic research and the clinical application of MDSCs.

OBJECTIVE:To investigate the clinical prognostic factors of initially-treated AML children with t(8;21)/RUNX1-RUNX1T1+.METHODS:Clinical data of 41 initially-treated AML children with t(8;21)/RUNX1-RUNX1T1+ in our hospital in period from January 2009 to January 2017 were retrospectively analyzed. The baseline clinical characteristics, cumulative recurrence, event-free survival (EFS) and overall survival (OS) were recorded, and the influencing factors of prognosis were evaluated by χ2 test and Cox regression model.RESULTS:The complete remission (CR) rates in the first course and the second course of induction chemotherapy were respectively 82.93% (34/41) and 97.56% (40/41). The median EFS time and OS time were 30 months and 31 months respectively. The EFS rate and OS rate of children with CR after the first treatment course were significantly higher than those of children without CR (P＜0.05). The EFS rate of male children was significantly higher than that of female children (P＜0.05). The OS rate of children < 10 years old was significantly higher than that of children≥10 years old (P＜0.05). The expression level of RUNX1-RUNX1T1 gene after the second induction remission was the influencing factor of cumulative recurrence rate, EFS rate and OS rate in children (P＜0.05). Multivariate analysis by Cox regression model showed that the decreased levels of RUNX1-RUNX1T1 gene expression < 3 log after the second induction remission was the independent risk factor for EFS rate and OS rate in children (P＜0.05). The cumulative recurrence rate of children with RUNX1-RUNX1T1 gene expression increase for＞1 log after decreased 3 log was significantly higher than that of children with≤1 log (P＜0.05).CONCLUSION:Iuithally-treated AML children with t(8;21)/RUNX1-RUNX1T1+ show the fine clinical prognosis after standard chemotherapy. The expression level of RUNX1-RUNX1T1 gene should be closely relates with the recurrence and long-term survival of AML children.

Prognosis of patients with Philadelphia-positive acute lymphoblastic leukemia (Ph-ALL) relapsing after allogeneic hematopoietic stem cell transplantation (HSCT) is extremely poor. Therefore, effective alternative therapeutic measures are urgently needed. Recently, the use of antigen receptor-modified T cells holds great promise for relapsed and refractory ALL treatment. Prior to chimeric antigen receptor T-cell (CAR-T) infusion conditioning chemotherapy is used routinely to establish a favorable in vivo environment for CAR-T expansion, which has mostly involved fludarabine and cyclophosphamide. We report on a patient presented with extreme fatigue and anemia and was diagnosed with relapsed and refractory acute lymphoblastic leukemia (ALL) harbored T315I-BCR-ABL mutation, who had undergone allogeneic HSCT and multiple reinducing chemotherapy, but achieved complete hematologic remission (CHR) with CAR -T infusion as a later salvage treatment. Prior to CAR-T infusion there was no conditioning chemotherapy, but a bone marrow suppression period induced by ponatinib. CAR-T cell infusion was well tolerated and the patient achieved a CHR and maintained it for three months. At present, there is no relevant report on the use of tyrosine kinase inhibitors (TKI) as preconditioning protocols before CAR-T cells infusion. Our case indicated ponatinib not only reduces tumor burden but may also serve as a conditioning regimen for CAR-T therapy in the treatment of relapsed and refractory Ph-ALL.

SummaryBackgroundCytokine release syndrome (CRS) is a potentially life-threatening complication of chimeric antigen receptor T (CAR-T) cell therapy. Recent studies indicated critical roles of macrophages and monocytes in CAR-T induced CRS. Here, we report rapid dissipation of CAR-T induced CRS in two patients after receiving Tripterygium glycosides (TG). Effects of triptolide, the major active component of TG, on macrophages and monocytes were examined in animal models.MethodsTwo patients with CRS after CAR-T cell therapy (for hematological malignancy) received TG (50 mg, p.o.). Flow cytometry analysis and single cell RNA sequencing (scRNAseq) were conducted to examine the effects of TG on immune cells. Potential effects of triptolide were also examined ex vivo using patient-derived monocytes, as well as in mice.FindingsRapid alleviation of fever and cytokine storm was observed within 72 hours after TG treatment. Blood concentration of triptolide ranged from 21 to 154 ng/mL during treatment. Flow cytometry and scRNAseq showed selective depletion of monocytes with minimal impact on CAR-T cells in both patients. In ex vivo experiments with patient-derived monocytes, triptolide dramatically inhibited the synthesis of pro-inflammatory cytokines (e.g., IL-6, IL-10, and IP-10). Triptolide also rapidly and selectively depleted peritoneal concanavalin A activated macrophages and monocytes in mice.InterpretationTG could be a promising treatment for CAR-T induced CRS, as well as other diseases with similar mechanisms, e.g., hemophagocytic lymphohistiocytosis and COVID-19. Our preliminary findings require further verification with properly designed clinical trials.