Peripheral T-lymphocyte subpopulations in different clinical stages of chronic HBV infection correlate with HBV load

AIM： To characterize the peripheral T-cell subpopulation profiles and their correlation with hepatitis B virus （HBV） replication in different dinical stages of chronic HBV infection. METHODS： A total of 422 patients with chronic HBV infection were enrolled in this study. The patients were divided into three stages： immune-tolerant stage, immune active stage, and immune-inactive carrier stage. Composition of peripheral T-cell subpopulations was determined by flow cytometry. HBV markers were detected by enzyme-linked immunosorbent assay. Serum HBV DNA load was assessed by quantitative real-time poiymerase chain reaction.RESULTS： CD8^＋ T-cells were significantly higher in patients at the immune-tolerant stage than in patients at the immune-active and -inactive carrier stages （36.87 ± 7.58 vs 34.37 ± 9.07, 36.87 ± 7.58 vs 28.09 ± 5.64, P 〈 0.001）. The peripheral blood in patients at the immune-tolerant and immune active stages contained more CD8^＋ T-cells than CD4^＋ T-cells （36.87 ± 7.58 vs 30.23 ± 6.35, 34.37 ± 9.07 vs 30.92 ± 7.40, P 〈 0.01）, whereas the peripheral blood in patients at the immune- inactive carrier stage and in normal controls contained less CD8^＋ T-cells than CD4^＋ T-cells （28.09 ± 5.64 vs 36.85 ±6.06, 24.02 ± 4.35 vs 38.94 ± 3.39, P 〈 0.01）. ANOVA linear trend test showed that CD8^＋ T-cells were significantly increased in patients with a high viral load （39.41 ± 7.36, 33.83 ± 7.50, 31.81 ± 5.95 and 26.89 ± 5.71, P 〈 0.001）, while CD4^＋ T-cells were significantly increased in patients with a low HBV DNA load （37.45 ± 6.24, 33.33 ± 5.61, 31.58 ± 6.99 and 27.56 ± 5.49, P 〈 0.001）. Nultiple regression analysis displayed that log copies of HBV DNA still maintained its highly significant coefficients for T-cell subpopulations, and was the strongest predictors for variations in CD3^＋, CD4^＋ and CD8^＋ cells and CD4^＋/CD8^＋ ratio after adjustment for age at HBV-infection, maternal HBV-infection status, presence of hepatitis B e antigen and HBV mutation.CONCLUSION： Differences in peripheral T-cell subpopulation profiles can be found in different clinical stages of chronic HBV infection. T-cell impairment is significantly associated with HBV load.

Bystanders or not?Microglia and lymphocytes in aging and stroke

As the average age of the world population increases,more people will face debilitating aging-associated conditions,including dementia and stroke.Not only does the incidence of these conditions increase with age,but the recovery afterward is often worse in older patients.Researchers and health professionals must unveil and understand the factors behind age-associated diseases to develop a therapy for older patients.Aging causes profound changes in the immune system including the activation of microglia in the brain.Activated microglia promote T lymphocyte transmigration leading to an increase in neuroinflammation,white matter damage,and cognitive impairment in both older humans and rodents.The presence of T and B lymphocytes is observed in the aged brain and correlates with worse stroke outcomes.Preclinical strategies in stroke target either microglia or the lymphocytes or the communications between them to promote functional recovery in aged subjects.In this review,we examine the role of the microglia and T and B lymphocytes in aging and how they contribute to cognitive impairment.Additionally,we provide an important update on the contribution of these cells and their interactions in preclinical aged stroke.

GZMB在乳腺癌和乳腺良性肿物患者中的表达及与T淋巴细胞水平的关系研究

目的:探讨颗粒酶B(granzyme B,GZMB)在乳腺癌和乳腺良性肿物患者中的表达及与T淋巴细胞水平的关系。方法:选取2022年1月至2023年8月南阳市第一人民医院收治的乳腺癌患者63例为观察组,根据肿瘤分期分为T_(1)N_(0)M_(0)组、T_(2)N_(0)M_(0)组、T_(2)N_(1)M_(0)组、T_(3)N_(0)M_(0)组及T_(3)N_(1)M_(0)组;选择同期治疗的乳腺良性肿物患者9例为对照组;采用流式细胞仪测定各组GZMB及T淋巴细胞水平;对乳腺癌患者GZMB与T淋巴细胞参数完成相关性分析。结果:观察组乳腺癌患者GZMB水平高于对照组(P<0.05);CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平低于对照组(P<0.05);乳腺癌不同分期下CD_(8)^(+)水平无统计差异(P>0.05);乳腺癌患者中T_(3)N_(1)M_(0)组GZMB水平高于其余4组(P<0.05),CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平低于其余4组(P<0.05);乳腺癌患者分期越高,GZMB水平越高,CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平越低;Pearson相关性结果表明,乳腺癌患者GZMB水平与CD_(3)^(+)、CD_(4)^(+)及CD_(4)^(+)/CD_(8)^(+)水平呈负相关(P<0.05)。结论:GZMB在乳腺癌患者中呈高表达,其表达水平在不同分期中存在差异,且与T淋巴细胞水平存在相关性,可指导临床诊疗。

伴单克隆B淋巴细胞和单克隆浆细胞增殖的AITL 1例

血管免疫母细胞性T细胞淋巴瘤(angioimmunoblastic T-cell lymphoma,AITL)是一种预后较差的侵袭性淋巴瘤。本文报道1例伴单克隆B淋巴细胞和单克隆浆细胞增殖AITL患者的诊疗经过,以提高对AITL的认识及诊疗水平,从而减少漏诊和误诊。

超声心动图参数及外周血T淋巴细胞亚群、NF-κB、CD64水平预测新生儿败血症预后的价值

【目的】探讨超声心动图参数及外周血T淋巴细胞亚群、核因子-κB(NF-κB)、簇分化抗原64(CD64)水平预测新生儿败血症预后的价值。【方法】选取2020年1月至2023年2月本院收治的95例新生儿败血症,根据预后情况将其分为存活组(n=80)和死亡组(n=15)。比较两组患儿超声心动图参数[左室短轴缩短率(FS)、升主动脉内径(AAO)、左室射血分数(LVEF)]、心肌损伤标志物[肌酸激酶(CK)、肌酸激酶同工酶(CK-MB)]、T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+))、NF-κB、CD64水平;采用Logistic多因素回归分析新生儿败血症预后不良的相关影响因素;采用受试者工作特征(ROC)曲线分析心电图指标及实验室指标预测新生儿败血症预后的临床价值。【结果】两组患儿肌酸激酶CK、CK-MB水平比较,差异无统计学意义(P>0.05);存活组FS、AAO及LVEF高于死亡组患儿,差异有统计学意义(P<0.05)。两组患儿CD8^(+)水平比较,差异无统计学意义(P>0.05);存活组患儿CD3^(+)、CD4^(+)水平高于死亡组,NF-κB、CD64水平低于死亡组,差异有统计学意义(P<0.05)。两组患儿中性粒细胞与淋巴细胞比值(NLR)、C反应蛋白(CRP)水平比较,差异无统计学意义(P>0.05);存活组患儿白细胞计数(WBC)低于死亡组,差异有统计学意义(P<0.05)。Logistic多因素回归分析显示:FS、LVEF、CD64是影响新生儿败血症预后的相关因素(P<0.05)。FS、LVEF、CD64联合预测新生儿败血症预后的ROC曲线下面积为0.948,明显高于各指标单独预测的0.827、0.895和0.877(P<0.05)。【结论】FS、LVEF联合CD64检测对新生儿败血症预后有良好的预测价值,临床可通过密切监测上述指标及时判断新生儿败血症预后,采取相关干预措施。

Observation of T Lymphocyte Subsets in the Liver of Patients with Advanced Schistosomiasis and Advanced Schistosomiasis Accompanied with Hepatitis B

T lymphocyte subsets in the liver were detected by Avidin-Biotin Complex (ABC) assay in 22 patients with advanced schistosomiasis (AS) and 5 cases of AS accompanied with hepatitis B. T lymphocytes in the liver of AS patients were distributed in the peripheral layer of egg granuloma or the area near eggs in non-granuloma. No infiltrative T lymphocytes were observed in area with extensive fibrosis. There was infiltration of many T cells in the portal tract, piecemeal and focal necro- sis area as well as in hepatic sinus in AS patients accompanied with hepatitis B. CD8+ T cells (sup pressor/cytotoxic T cells, Ts/Tc) in the liver were predominant in the two groups. In AS patients, marked hepatic fibrosis, a small number of T cell infiltration and slight hepatocellular degeneration and necrosis were observed. However, obvious hepatocellular degeneration and necrosis were seen in AS patients accompanied with hepatitis B, and 3 cases of them developed active liver cirrhosis. The results indicated immune response was weak in the liver in AS patients and T. cells might be predominant in the subset of CDS+ T lymphocytes. Cellular immune response was relatively strong in AS patients accompanied with hepatitis B and the infiltrative CD8+ T lymphocytes might be mainly Tc cells.

基于lncRNA PVT1表达分析银杏内酯B对ITP患者B淋巴细胞及Treg/Th17免疫失衡的影响

目的探讨银杏内酯B对原发性免疫血小板减少症(ITP)患者长链非编码RNA浆细胞瘤转化迁移基因1(lncRNA PVT1)、B淋巴细胞和调节性T淋巴细胞/辅助性T淋巴细胞17(Treg/Th17)免疫失衡的影响及其相关性。方法依据双盲法将2021年5月至2023年5月廊坊市中医医院收治的130例ITP患者分为对照组(65例)和观察组(65例)。对照组给予长春地辛治疗,观察组给予银杏内酯B治疗,两组均治疗4周。比较两组患者临床疗效;比较两组患者治疗前后lncRNA PVT1表达、B淋巴细胞亚群[CD19^(+)、B1淋巴细胞]比例、Treg/Th17免疫失衡细胞因子[白细胞介素-17(IL-17)、白细胞介素-21(IL-21)];分析lncRNA PVT1表达与CD19^(+)、B1淋巴细胞比例以及IL-17、IL-21水平的相关性。结果研究过程中对照组失访7例,观察组失访5例,最终分组为对照组58例,观察组60例。治疗后两组患者lncRNA PVT1表达、IL-17、IL-21水平以及CD19^(+)、B1淋巴细胞比例较治疗前均有所降低(P<0.05),观察组治疗后lncRNA PVT1表达、IL-17、IL-21水平以及CD19^(+)、B1淋巴细胞比例明显低于对照组(P<0.05)。与对照组相比,观察组临床疗效更优(P<0.05)。lncRNA PVT1表达与CD19^(+)、B1淋巴细胞、IL-17、IL-21水平均呈正相关(P<0.05)。结论银杏内酯B对ITP患者具有较高临床疗效,可以显著改善其lncRNA PVT1表达、B淋巴细胞比例、Treg/Th17免疫失衡细胞因子水平,值得临床推广。

Correlation analysis of TCM syndrome types with T lymphocytes and biochemical indices in patients with HBV-related primary liver cancer

Objective:To investigate the correlation between T lymphocytes and biochemical indices in patients with Primary liver cancer(PLC)associated with hepatitis B virus(HBV)and TCM syndrome differentiation.Methods:263 HBV-related PLC patients who were admitted to the First Affiliated Hospital of Henan University of Traditional Chinese Medicine from January 2018 to December 2019 were retrospectively collected.There were 127 cases of liver depression and spleen deficiency syndrome(48.3%),48 cases of spleen deficiency and dampness syndrome(18.3%),31 cases of liver and gallbladder dampness and heat syndrome(11.8%),35 cases of liver and blood stasis syndrome(13.3%),and 22 cases of liver and kidney Yin deficiency syndrome(8.4%).The general data,T cell subsets,oncology and virology indicators,oncology characteristics,biochemical indicators and other data were counted.Epidata and Excel were used to collect and summarize the data,and SPSS26.0 software was used for statistical analysis.Results:There was no significant difference in gender and age distribution among the five syndrome types(χ^(2)=5.462,F=1.979,ALL P>0.05).The differences among T lymphocyte count(χ^(2)=57.785,P<0.001),CD4(+)T cell count(χ^(2)=47.103,P<0.001)and CD8(+)T lymphocyte count(F=12.760,P<0.001)were statistically significant.The T lymphocyte count,CD4(+)T lymphocyte count and CD8(+)T lymphocyte explicit count in patients with liver and kidney Yin deficiency syndrome were significantly lower than those in the other four syndrome types.AFP(χ^(2)=89.986,P<0.001),CEA(χ^(2)=95.501,P<0.001),CA199(χ^(2)=30.044,P<0.001)of the five syndrome types increased successively from the syndrome of liver depression and spleen deficiency to the syndrome of liver and kidney Yin deficiency,and the difference was statistically significant.There were statistically significant differences in the inner diameter of main portal vein,portal vein cancer thrombin and extrahepatic metastasis among the five syndrome types(ALL P<0.001).The main symptoms of portal vein cancer thrombin and extrahepatic metastasis were liver-gallbladder dampness-heat syndrome and liver-blood stasis syndrome.The differences among PLT(χ^(2)=39.234,P<0.001),Alb(χ^(2)=75.171,P<0.001),TBil(χ^(2)=51.140,P<0.001),AST(χ^(2)=55.881,P<0.001),PT(χ^(2)=21.515,P<0.001)were statistically significant.PLT and Alb decreased successively from the syndrome of liver depression and spleen deficiency to the syndrome of liver and kidney Yin deficiency.PLT and Alb of the syndrome of liver depression and spleen deficiency were significantly higher than those of the other four groups,and TBil and AST of the syndrome of liver and gallbladder dampness and heat were significantly higher than those of the other four groups.PT of liver and kidney Yin deficiency was significantly higher than that of the other four groups.The lymphocyte count,CD4(+) lymphocyte count and CD8(+) lymphocyte count were negatively correlated with AFP,PT and TBil(ALL P<0.05),and positively correlated with PLT(P<0.05).T lymphocyte count was positively correlated with AIb(P<0.05).Conclusion:This study found that patients with liver depression and spleen deficiency syndrome have better cellular immune function,liver function and prognosis.Patients with liver and kidney Yin deficiency have lower cellular immunity,worse liver function,and worse prognosis.Portal vein carcinoma embolus and extrahepatic metastasis were mainly characterized by dampness and heat of liver and gallbladder and blood stasis of liver.Patients with lower lymphocyte counts have poorer blood clotting,worse the liver reserve,and the higher the risk of further cancer.

Profile of Plasma Galectin-3 Concentrations, Inflammatory Cytokines Levels and Lymphocytes Status in Breast Cancer under Chemotherapy

We investigated relationship between galectin-3 (Gal-3) levels and T lymphocytes apoptosis and the activation rates in breast cancer during chemotherapy. We used plasma samples from 112 women classified into two groups: 70 women with breast cancer (BC) and submitted to neoadjuvant chemotherapy (3 cycles) and 42 healthy women used as controls. In the group of BC, blood samples were taken before each cycle of chemotherapy and Gal-3 levels was evaluated by ELISA sandwich. Flow cytometry was used to study T cells apoptosis and activation. Before treatment, median value of Gal-3 was 6.31 ng/ml (range 1.07 - 50.74) in BC and 0.84 ng/ml (range 0.00 - 4.82) in HC. Gal-3 levels were highest in plasmas from BC (p p p = 0.010). In addition, we found a dynamic relationship between gal-3 levels, tumor size and T lymphocytes apoptosis rates during treatment depending to the cure efficiency. We suggest gal-3 plasma concentrations could be used as predictive biomarker for chemotherapy efficiency in breast cancer patients.

慢性乙型病毒性肝炎中Bcl-2及Bax的表达与T、B淋巴细胞数量变化的关系

目的:研究Bcl-2及Bax在慢性乙型病毒性肝炎(简称慢乙肝)患者肝组织中淋巴细胞的表达与T、B淋巴细胞数量变化的关系。方法:用免疫组织化学的方法,检测57例慢乙肝患者肝脏活检组织和20例肝血管瘤患者外科手术切除标本正常肝组织淋巴细胞中Bcl-2,Bax的分布及数量的变化,并同时进行CD3和CD20免疫组织化学染色,计算T和B淋巴细胞数量变化与表达Bcl-2、Bax的淋巴细胞数量变化的关系。阳性细胞的计数和分析数据均使用病理图像分析仪获得。结果:随着肝炎炎症活动度和纤维化程度的增加,慢乙肝患者肝组织中表达Bcl-2、Bax的淋巴细胞数量增加,同时T、B淋巴细胞数量也增多,均明显多于正常肝组织(P<0.01)。结论:Bcl-2、Bax在慢乙肝患者肝组织淋巴细胞中的表达与T、B淋巴细胞数量的变化有关,并影响慢乙肝患者炎症活动度和纤维化程度的进展。

临床中隐匿性HBV感染的病毒血清标志物和T淋巴细胞亚群特征

目的分析隐匿性乙型肝炎病毒感染(occult HBV infection,OBI)患者的病毒血清学标志物特征及外周T淋巴细胞亚群情况。方法将2020年5月至2021年12月就诊于首都医科大学附属北京佑安医院血清HBsAg阴性、HBV DNA阳性的292例OBI患者作为OBI组,240例同期入院的HBsAg和HBV DNA持续阳性(>6个月)的慢性HBV感染患者为HBsAg阳性组,收集录入入组532例患者的基本病历资料和实验室检查数据,如HBV血清学标志物、血清HBV DNA载量、肝功能指标等;在OBI组中T淋巴细胞亚群检测数据完整的48例患者作为OBI亚组,同时根据性别和年龄在HBsAg阳性组中匹配48例HBsAg阳性患者作为HBsAg阳性亚组,20例同期入院体检的健康人作为健康对照组,收集此116例患者的外周血T淋巴细胞亚群数量和比值的临床数据。结果OBI患者的主要血清学模式是:HBsAg阴性、抗-HBs阴性、HBeAg阴性、抗-HBe阳性、抗-HBc阳性,占总OBI组的48.97%;抗-HBs阳性率为45.21%,抗-HBe阳性率为85.62%,都明显高于HBsAg阳性患者,差异有统计学意义(P<0.05)。94.86%的OBI组患者的HBV DNA载量<200IU/mL。OBI亚组的淋巴细胞数、T淋巴细胞数、CD8^(+)T淋巴细胞/淋巴细胞、CD8^(+)T淋巴细胞数、CD4^(+)T淋巴细胞数低于健康对照组,且差异都有统计学意义(P<0.05);OBI亚组的淋巴细胞数、T淋巴细胞数、CD8^(+)T淋巴细胞/淋巴细胞、CD8^(+)T淋巴细胞数高于HBsAg阳性亚组患者,差异有统计学意义(P<0.05)。结论OBI患者的血清学标志物特征表明HBV DNA复制和转录水平低下;OBI患者的细胞免疫功能有一定程度的恢复,但与健康人相比仍处于较低水平,体内仍存在HBV感染,且具有HBV重新激活和发展为肝硬化和肝细胞癌的风险。

乙型肝炎相关性肝癌患者T淋巴细胞、白介素-28B及高尔基体蛋白73的表达及临床意义

目的:分析乙型肝炎相关性肝癌患者T淋巴细胞、白介素-28B (IL-28B)及高尔基体蛋白73 (GP73)的表达及临床意义。方法:选取2018年10月—2021年10月商丘市第一人民医院收治的119例乙型肝炎相关性肝癌患者作为观察组,选取同期体检的112例健康人群作为对照组。比较两组研究对象血清IL-28B、GP73及T淋巴细胞水平,分析IL-28B、GP73、T淋巴细胞与病理特征及预后的关系。结果:观察组血清IL-28B、GP73及CD8^(+)表达水平均高于对照组,CD3^(+)、CD4^(+)及CD4^(+)/CD8^(+)均低于对照组,差异有统计学意义(t=148.380、13.155、9.827、9.939、16.367、11.453,P<0.05)。不同年龄、性别、肿瘤直径的乙型肝炎病毒(HBV)相关性肝癌患者血清中IL-28B、GP73水平比较,差异无统计学意义(P>0.05)。有淋巴结转移、TNM分期Ⅲ~Ⅳ期、低分化、有血管浸润者血清中IL-28B、GP73水平高于无淋巴结转移、TNM分期Ⅰ~Ⅱ期、中/高分化、无血管浸润者,差异有统计学意义(P<0.05)。HBV相关性肝癌患者血清中CD3^(+)、CD4^(+)、CD8^(+)在不同年龄、性别、肿瘤直径中比较,差异无统计学意义(P>0.05),CD3^(+)、CD4^(+)、CD8^(+)水平在不同淋巴结转移情况、TNM分期、分化程度及血管浸润中比较,差异无统计学意义(P>0.05)。死亡者血清GP73、IL-28B水平均高于生存者,CD4^(+)/CD8^(+)指标低于生存者,差异有统计学意义(t=16.629、2.082、9.267,P<0.05)。结论:乙型肝炎相关性肝癌患者血清IL-28B、GP73及CD8^(+)表达水平明显升高,CD3^(+)、CD4^(+)及CD4^(+)/CD8^(+)指标下降,IL-28B、GP73及CD4^(+)/CD8^(+)水平与相关病理特征、预后具有密切的联系。

系统性红斑狼疮不同疾病时期外周血中ILC、B细胞、T淋巴细胞亚群水平变化及临床意义

目的探讨系统性红斑狼疮(SLE)不同疾病时期外周血中固有淋巴细胞(ILC)、B细胞、T淋巴细胞亚群水平变化及临床意义。方法前瞻性选择2020年1月至2023年1月神木市医院收治的90例SLE患者进行研究(观察组),采用系统性红斑狼疮疾病活动性指数(SLEDAI)评分评价所有SLE患者的病情活动程度,其中活动组(SLEDAI≥5分)32例,非活动组(SLEDAI<5分)58例,并选择同期在我院体检的90例健康者作为对照组。比较观察组与对照组,以及观察组不同疾病活动度患者的外周血ILC1、ILC2、ILC3、记忆B细胞、静止B细胞和CD4^(+)、CD8^(+)、CD4^(+)/CD8^(+)的表达水平。采用Pearson相关分析法分析ILC、B细胞、T淋巴细胞亚群水平与疾病活动度的相关性。结果观察组患者的ILC1、记忆B细胞、CD8^(+)水平分别为(20.35±2.51)%、(25.45±6.70)%、(36.20±2.93)%,明显高于对照组的(14.55±1.20)%、(21.79±6.53)%、(31.28±2.74)%,ILC2、ILC3、静止B细胞、CD4^(+)、CD4^(+)/CD8^(+)水平分别为(10.12±1.34)%、(6.20±1.12)%、(60.23±8.14)%、(30.85±3.30)%、0.81±0.07,明显低于对照组的(42.49±3.48)%、(14.36±2.41)%、(42.05±3.38)%、(42.05±3.38)%、1.53±0.10,差异均有统计学意义(P<0.05);活动组患者的ILC1、记忆B细胞、CD8^(+)水平分别为(22.40±2.53)%、(32.02±9.12)%、(39.36±3.16)%,明显高于非活动组的(18.64±2.04)%、(26.88±6.45)%、(34.90±2.20)%,ILC2、ILC3、静止B细胞、CD4^(+)、CD4^(+)/CD8^(+)水平分别为(8.66±0.81)%、(5.73±1.10)%、(56.86±10.42)%、(27.96±3.10)%、0.68±0.05,明显低于非活动组的(19.68±1.85)%、(8.50±1.84)%、(62.03±11.25)%、(34.50±3.62)%、0.98±0.12,差异均有统计学意义(P<0.05);经Pearson相关分析结果显示,ILC1、记忆B细胞、CD8^(+)水平与疾病活动度均呈正相关(r=0.726、0.549、0.500,P<0.05),LC2、ILC3、静止B细胞、CD4^(+)、CD4^(+)/CD8^(+)水平与疾病活动度均呈负相关(r=-0.503、-0.715、-0.705、-0.668、-0.734,P<0.05)。结论SLE患者外周血中ILC、B细胞、T淋巴细胞亚群水平与疾病活动度密切相关,对其监测有助于评估疾病进展和制定治疗方案。

CAR-T细胞免疫疗法对复发/难治性弥漫大B细胞淋巴瘤患者LDH、ALC、CD4+/CD8+、Ki67表达的影响

目的探讨嵌合抗原受体T细胞(CAR-T)免疫疗法对复发/难治性弥漫大B细胞淋巴瘤(DLBCL)患者乳酸脱氢酶(LDH)、淋巴细胞绝对值(ALC)、CD4+/CD8+、肿瘤增殖抗原(Ki67)表达的影响。方法回顾性选取复发/难治性DLBCL患者92例,均经CAR-T细胞免疫治疗。记录复发/难治性DLBCL患者临床疗效及不良反应,比较治疗前后复发/难治性DLBCL患者LDH、ALC、CD4+/CD8+、Ki67水平。观察不同疗效患者治疗前临床特征,包括性别、年龄、AnnArbor分期、ECOG评分、结外受累部位、国际预后指数(IPI)、CAR-T治疗线数,以及治疗前LDH、ALC、CD4+/CD8+、Ki67水平。结果CAR-T免疫治疗后CR 39例(42.39%),PR 24例(26.09%),SD 18例(19.57%),PD 11例(11.96%);总有效63例(68.48%)。不良反应细胞因子释放综合征(CRS)71例(77.17%),低免疫球蛋白血症73例(79.35%)。经CAR-T细胞免疫治疗后LDH、Ki67水平较治疗前下降,ALC、CD4+/CD8+较治疗前上升,差异具有统计学意义(P<0.05)。DLBCL治疗无效患者年龄、AnnArbor分期、结外受累部位数量、IPI、LDH、Ki67、CAR-T治疗线数均高于治疗有效组患者,ALC、CD4+/CD8+均低于治疗有效组患者(P<0.05)。Logistic回归分析显示高龄、AnnArborⅣ期、多结外受累部位数量、高IPI得分、二线以上CAR-T以及治疗前LDH、Ki67的水平升高和ALC、CD4+/CD8+的水平降低是影响DLBCL患者CAR-T细胞免疫治疗疗效的危险因素(P<0.05)。建立ROC曲线,LDH、ALC、CD4+/CD8+、Ki67 AUC分别为0.823、0.835、0.873、0.849,提示LDH、ALC、CD4+/CD8+、Ki67对CAR-T细胞免疫治疗疗效具有一定的预测价值。结论CAR-T细胞免疫疗法可降低LDH、Ki67水平表达,升高ALC和CD4+/CD8+表达,且治疗前LDH、ALC、CD4+/CD8+、Ki67表达对CAR-T细胞免疫疗法疗效具有预测价值。

Frequencies and Characterization of HBV-specific Cytotoxic T Lymphocytes in Self-limited and Chronic Hepatitis B Viral Infection in China

Hepatitis B virus （HBV）-specific cytotoxic T lymphocytes （CTLs） are believed to play a major role in viral clearance and disease pathogenesis during HBV infection. To clarify the differences in host immune responses between self-limited and chronic HBV infections, we constructed three HLA-A＊0201/HBV tetramers with immunodominant epitopes of core18-27, polymerase 575-583 and envelope 335-343, and analyzed the HBV-specific CTLs in peripheral blood mononuclear cells （PBMCs） from patients infected with HBV. The frequencies and expansion ability of HBV-specific CD8＋T cells in most self-limited HBV infected individuals were higher than those in chronic HBV-infected patients. HBV-specific CD8＋T cells could be induced by in vitro peptide stimulation from chronic patients with a low level of serum HBV-DNA but not from those with a high level of serum HBV-DNA. In chronic infection, no significant correlation was found either between the frequencies of HBV-specific CD8^＋ T cells and the viral load, or between the frequencies and the levels of alanine transaminase. Our results suggested that the frequencies of HBV-specific CTLs are not the main determinant of immune-mediated protection in chronic HBV infection and immunotherapeutic approaches should be aimed at not only boosting a HBV-specific CD8^＋T response but also improving its function.

基于BCMA突变体构建BCMA CAR-T细胞体外杀伤功能评价模型

目的:为解决野生型B细胞成熟抗原(BCMA)被γ分泌酶切割导致表达不稳定的问题,构建抵抗γ分泌酶切割的BCMA突变体并构建靶细胞,用于评价BCMA CAR-T细胞的杀伤功能。方法:将野生型BCMA的穿膜域替换为人CD8α穿膜域,构建抵抗γ分泌酶切割的BCMA突变体(BCMA-CD8αTM),构建过表达该突变体的U266(U266^(BCMA Mut))、K562(K562^(BCMA Mut))、SKOV3(SKOV3^(BCMA Mut))和CHO(CHO^(BCMA Mut))细胞;构建装载NFAT-EGFP报告基因的BCMA CAR Jurkat细胞(BCMA-CAR-Jurkat-Reporter)与U266^(BCMA Mut)细胞共培养,采用FCM检测该细胞中EGFP表达水平以指示NFAT激活水平,荧光素酶法检测BCMA CAR-T细胞对Luciferase标记的K562^(BCMA Mut)细胞的杀伤作用,实时无标记动态细胞分析技术(RTCA)检测BCMA CAR-T细胞对SKOV3^(BCMA Mut)和CHO^(BCMA Mut)细胞的杀伤作用。结果:应用γ分泌酶抑制剂LY411575抑制γ分泌酶活性,显著增强野生型U266细胞表面BCMA表达水平,平均荧光强度上调10倍以上;但撤除抑制剂后BCMA表达水平逐渐降低(P<0.01);BCMA-CD8αTM突变体可抵抗γ分泌酶的切割作用,在U266细胞表面稳定表达(P>0.05);U266细胞及过表达BCMA-CD8αTM的U266细胞与BCMA-CAR-Jurkat-Reporter细胞共培养后都可激活Reporter系统、增强EGFP表达,但该效应在BCMA-CD8αTM过表达的U266细胞中更显著(P<0.01);BCMA-CD8αTM在BCMA表达阴性的K562、SKOV3和CHO 3种靶细胞中成功过表达,且在LY411575处理下该突变体的表达水平仅有小幅度升高;荧光素酶法检测结果显示,不同效靶比下,BCMA CAR-T细胞均可特异、高效杀伤过表达BCMA-CD8αTM的K562细胞;RTCA结果显示,不同效靶比下,BCMA CAR-T细胞均可有效识别、杀伤过表达BCMACD8αTM的SKOV3和CHO细胞,但同等效靶比下的Mock-T细胞无此效应。结论:本实验构建的BCMA-CD8αTM突变体能够抵抗γ分泌酶的切割,在多种靶细胞表面稳定表达,为评价BCMA CAR-T细胞体外杀伤的有效性和特异性提供多种检测手段。

乙肝病毒感染后不同临床类型及HBV DNA载量与外周血T细胞亚群变化的相关性分析

目的探讨乙肝病毒（HBV）感染后不同临床类型乙型肝炎患者及HBV DNA载量与外周血T细胞亚群的关系。方法选取确诊的共193例急性乙型肝炎、慢性乙型肝炎、慢性重型肝炎、亚急性重型肝炎、急性重型肝炎、乙肝肝硬化、乙肝肝癌、HBV携带者的外周血,流式细胞仪检测T细胞亚群,荧光定量PCR法检测HBV DNA水平。结果对照组CD3＋、CD4＋细胞百分率、CD4＋/CD8＋细胞比值明显高于各临床类型组,随病情加重CD3＋、CD4＋细胞百分率、CD4＋/CD8＋细胞比值下降明显,除急性乙型肝炎组和慢性肝炎组CD4＋细胞百分率外与各组比较差异有统计学意义（P〈0.05及P〈0.01）;对照组CD8＋细胞百分率、CD4＋/CD25＋细胞比值明显低于各临床类型组,随病情加重CD8＋细胞百分率、CD4＋/CD25＋细胞比值升高明显,各组比较差异有统计学意义（P〈0.05及P〈0.01）。低水平HBV DNA载量组（≤103 Copies/ml）CD3＋、CD4＋细胞百分率明显高于其他各病毒组,随病毒载量增加CD3＋、CD4＋细胞百分率逐渐减少,与各组比较差异有统计学意义（P〈0.05及P〈0.01）;低水平HBV DNA载量组CD8＋细胞百分率最低,随病毒载量增加CD8＋细胞百分率升高明显,与各组比较差异有统计学意义（P〈0.05及P〈0.01）;低水平HBV DNA载量组CD4＋/CD8＋及CD4＋/CD25＋细胞比值最高,其比值随病毒载量增加下降明显,低水平组CD4＋/CD25＋细胞比值与各组比较差异有统计学意义（P〈0.05及P〈0.01）,低水平组CD4＋/CD8＋细胞比值与各组（103～105组除外）比较差异有统计学意义（P〈0.05及P〈0.01）。除乙肝肝硬化的CD4＋细胞百分率外,同病种肝病的HBV DNA（＋）组CD3＋、CD4＋、CD8＋细胞百分率、CD4＋/CD8＋及CD4＋/CD25＋细胞比值与HBV DNA（-）组比较,差异有统计学意义（P〈0.05及P〈0.01）。结论细胞免疫功能紊乱是HBV感染后发病机制之一,高水平HBV DNA载量可能诱导和加重细胞免疫功能紊乱,CD4＋/CD8＋及CD4＋/CD25＋比值变化可为了解病情变化和指导治疗提供依据。

南海蓖麻海绵Biemna fortis Topsent化学成分及其生物活性

目的:对采自我国南海的蓖麻海绵Biemna fortis Topsent的化学成分进行研究,从中寻找有生物活性的次生代谢产物。方法:用硅胶柱层析和凝胶柱层析对蓖麻海绵的甲醇提取物进行分离纯化,根据其化学性质,结合现代波谱技术(MS,NMR等),对得到的化合物进行结构鉴定。结果:分离得到9个甾体、2个邻苯二甲酸酯、1个长链脂肪醛类化合物,其结构分别鉴定为melithasterol B(1),(24R)-ergosta-7,22-diene-3,5,6-triol(2),(24R)-er-gosta-4,6,8(14),22-tetraene-3-one(3),(24R)-ergosta-4,7,22-triene-3-one(4),(24R)-ergosta-6,22-diene-5,8-epidioxy-3-ol(5),6-hydroxy-cholest-4-en-3-one(6),cholest-4-ene-3,6-dione(7),cholest-4-en-3-one(8),cholest-5,22-diene-3-one(9),bis-(2-ethylhexyl)phthalate(10),bis-diisobutyl phthalate(11)和hexaldehyde(12)。结论:化合物1-4,6-10,12系首次从蓖麻海绵中分离得到;化合物7不但对T和B淋巴细胞的增殖具有显著抑制活性,而且对II型糖尿病靶标分子蛋白质酪氨酸磷酸酯酶PTP1B具有显著的抑制活性,其IC50为1.6mol·L1。

综合需求在B淋巴细胞白血病CAR-T治疗患者自我效能感和生命质量间的中介效应

目的分析综合需求在B淋巴细胞白血病嵌合抗原受体T细胞(CAR-T)治疗患者自我效能感和生命质量间的中介效应。方法采取横断面研究法,选取2022年4月-2023年4月我院收治的B淋巴细胞白血病CAR-T治疗患者127例,采用一般资料问卷、一般自我效能感量表、中文版癌症患者综合需求评估量表(CNAT)、中文版癌症患者生命质量测定量表(FACT-G)进行问卷调查,分析综合需求与自我效能感、生命质量间的相关性,建立综合需求在自我效能感和生命质量间的中介模型并验证。结果回收有效问卷127份(97.69%)。127例患者自我效能感、生命质量和综合需求得分分别为(21.91±5.46)分、(68.18±17.23)分和(127.61±23.49)分。Pearson相关性分析结果显示,患者自我效能感与生命质量呈显著正相关(r>0,P<0.05),综合需求与自我效能感、生命质量呈显著负相关(r<0,P<0.05)。自我效能感对综合需求、生命质量有直接预测作用(P<0.05),综合需求在自我效能感和生命质量间具有部分中介作用,中介效应占总效应的24.35%。结论B淋巴细胞白血病CAR-T治疗患者综合需求处于较高水平,且综合需求、生命质量及自我效能感之间具有密切关系,自我效能感能够直接影响患者的生命质量,还可通过综合需求间接影响患者生命质量;临床应针对患者综合需求进行有效干预,影响患者自我效能感对生命质量的作用路径,提升生命质量。