Correlation Study Between T Lymphocyte Subsets and Rheumatoid Arthritis

Objective:To study the effect of Helicobacter pylori infection on rheumatoid arthritis and T-lymphocyte subpopulations in patients with rheumatoid arthritis and to provide a new method for the treatment of rheumatoid arthritis by removing Helicobacter pylori from patients.Methods:60 patients with rheumatoid arthritis admitted to the hospital from May 2022 to May 2023 were selected for the study,and all patients underwent a 13-carbon urea breath test to detect gastric H.pylori and the test results showed that 20 cases were negative and 40 cases were positive.The 40 positive patients were divided into the treatment group(n=20)and non-treatment group(n=20)by random number table method and the treatment group was given anti-Helicobacter pylori treatment,and the non-treatment group was given maintenance rheumatoid basic treatment,comparing the anti-cyclic citrulline peptide(CCP),DS28 score,peripheral blood T-lymphocyte subsets(CD4^(+)T-lymphocytes,CD8^(+)T-lymphocytes,CD4^(+)/CD8^(+)ratio)before and after the treatment of patients by 13-carbon urea respiration test(pylori-negative group,20 patients)and those who were positive for the treatment of H pylori(pylori-positive group,40 patients).Besides,the correlation of peripheral blood T-lymphocyte subsets and disease activity between treatment and non-treatment groups in the pylori-positive group was identified together with the correlation of DS28 scores,TNF-αlevels,sedimentation and immunoglobulin,lymphocyte subsets in the pylori-positive treatment group and positive non-treatment group as well as the level of globulin,lymphocyte subsets,and peripheral blood lymphocytes before and after treatment.Results:Before treatment,CCP,DS28 score,CD8^(+)T lymphocyte level of the pylori-negative group were lower than that of the positive group,and CD4^(+)T lymphocyte and CD4^(+)/CD8^(+)ratio were higher than that of the positive group(P<0.05);after treatment,the indexes of the pylori-positive group improved,and there was no significant difference in the comparison of the indexes with those of the pylori-negative group(P>0.05);the positive treatment group had a DS28(3.19±1.02)points,positive non-treatment group DS28(5.36±1.85)points,non-treatment group DS28 score and CD4^(+)T lymphocytes,CD4^(+)/CD8^(+)negative correlation with CD8^(+)T lymphocytes showed a positive correlation(P<0.05);before the treatment,pylori-positive treatment group and non-treatment group DS28 scores,TNF-αlevels,peripheral blood T lymphocyte subpopulation levels were not significantly different(P>0.05);after treatment,DS28 score,TNF-αlevel,CD8^(+)T of the treatment group were lower than those of the non-treatment group,and CD4^(+)T lymphocytes and CD4^(+)/CD8^(+)ratio were higher than those of the non-treatment group(P<0.05).Conclusion:H.pylori affects the level of T lymphocyte subsets in patients with rheumatoid arthritis,and there is a certain correlation between the two.Removal of H.pylori can improve the level of T lymphocyte subsets,which is important for the treatment of patients with rheumatoid arthritis.

Analysis of CD4^+CD25^+ Regulatory T Cells and Foxp3 mRNA in the Peripheral Blood of Patients with Asthma

The changes of CD4^＋CD25^＋ regulatory T cells （CD4^＋CD25^＋ Treg） and Foxp3 mRNA in peripheral blood mononuclear cells （PBMCs） from patients with asthma were investigated in order to elucidate the possible roles of CD4^＋CD25^＋ Treg in the development of asthma. The peripheral blood samples were collected from 29 healthy controls （normal control group） and 78 patients with asthma which included 30 patients in exacerbation group, 25 patients in persistent group, and 23 patients in remission group. By using flow cytometry and RT-PCR, the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs were detected. The CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA in PBMCs of exacerbation and persistent groups were lower than that of remission and normal control groups （P〈0.05）. Although the CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA of remission group were also lower than that of normal control group, there was no significant difference between them （P〉0.05）. As compared with persistent group, exacerbation group had lower CD4^＋CD25^＋ Treg ratio and Foxp3 mRNA （P〈0.05）. It was indicated that the decrease of CD4^＋CD25^＋ Treg ratio and its function in PBMCs may be responsible for pathogenesis of asthma.

Autologous Peripheral Blood Stem Cells and <i>γ</i>/<i>δ</i>T Cells May Improve Immunity in Treating Secondary Bacteremic Infection in HIV Infected Patient

Opportunistic bacteremia in adult HIV-infected patients is a normal co-infectious condition caused by Gram-negative bacilli. Respiratory infections, including cough, shortness of breath, and chest pain and skin infection with eruptions, pustules and itchiness, are common complaints in the setting of late HIV infection. The variety of infections ranges from mild, self-limited viral, bacteremia and fungal infections to severe, life-threatening demanding urgent care and hospitalization. Varicella pneumonia, for instance, is the most severe complication of chickenpox in HIV infected adults, potentially refractory, fulminant respiratory failure can ensue. Patients with impaired immune status and chronic lung disease are at an increased risk. In the United States as well as in Vietnam, bacterial/viral pneumonia and skin infection are the two most common HIV-associated conditions. While globally the incidence of opportunistic infection has decreased since the introduction of highly active antiretroviral therapy during the last 3 decades, HIV-associated diseases remain a significant source of mortality, thus any manifestation must be taken seriously. This study will present the most common HIV-related pulmonary and skin infections and provide an overview of the epidemiology, characteristic clinical and chest radiograph findings, diagnosis, treatment, and prevention globally as well in Vietnam. Though the extensive efforts of the Vietnamese Government during last decade contributed to a valuable decrease, yet epidemic in Vietnam still remains high, ranking Vietnam 5th in the South-East region. The second part of the study focuses on a unique and severe HIV case report of a 35-year-old man, with a rare form of pneumonia caused by Acitenobacter spp. concomitant with a prolonged and disseminating skin infection. The case has been treated with a combination of conventional anti-retroviral medication and autologous peripheral blood stem cells, the results showed that within 5 months there was an impressive amelioration of HIV viral activity together with a total recovery from pneumonia and skin infection.

Rapamycin increased development of CD4^+ CD25~h ighFoxp3^+ T cells of peripheral blood in liver transplant recipients

Objective To investigate the possible influence of immunosuppressive therapy,including sirolimus ( SRL) and calcineurin inhibitors ( CNI,tacrolimus) ,on level of Treg in liver allo - graft recipients. Methods Forty - seven liver transplant recipients with stable liver function were assessed for at least 2 years,and divided into

T淋巴细胞亚群、IL-6在胃癌患者中的表达及其与预后的相关性

目的分析外周血T淋巴细胞亚群及白细胞介素-6(interleukin-6,IL-6)在胃癌患者中的表达,探究其与预后的相关性。方法选取2021年6月至2023年6月连云港市第二人民医院收治的106例胃癌患者为试验组,入院次日检测其外周血T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+))及IL-6水平。选取同时段于连云港市第二人民医院进行体检的健康者80例为对照组,体检当日检测其外周血T淋巴细胞亚群及IL-6水平。比较两组T淋巴细胞亚群及IL-6水平。胃癌患者院外随访6个月,根据患者死亡、存活情况将其分为预后不良组(17例)、预后良好组(89例)。预后不良组男10例(58.82%),女7例(41.18%),年龄(60.47±5.89)岁;文化程度:中学及以上9例(52.94%),中学以下8例(47.06%)。预后良好组男55例(61.80%),女34例(38.20%),年龄(60.08±5.71)岁;文化程度:中学及以上32例(35.96%),中学以下57例(64.04%)。logistic回归分析影响胃癌患者预后的因素。采用受试者操作特征曲线(ROC)评估外周血T淋巴细胞亚群及IL-6预测胃癌患者预后的价值。采用t检验和χ^(2)检验。结果试验组CD3^(+)、CD4^(+)水平低于对照组[(55.27±8.72)%比(68.97±6.45)%、(26.42±10.21)%比(38.12±5.07)%],CD8^(+)、IL-6水平高于对照组[(33.36±6.02)%比(24.66±3.75)%、(6.97±3.29)ng/L比(3.62±1.01)ng/L],差异均有统计学意义(t=-11.821、-9.413、11.360、8.803,均P<0.05)。预后不良组CD3^(+)、CD4^(+)水平低于预后良好组[(45.12±10.06)%比(57.21±6.97)%、(20.01±10.12)%比(27.64±9.81)%],CD8^(+)、IL-6水平、肿瘤分期Ⅳ期患者占比高于预后良好组[(40.12±6.01)%比(32.07±5.12)%、(9.75±3.16)ng/L比(6.44±3.05)ng/L、70.59%(12/17)比32.58%(29/89)],差异有统计学意义(t=-6.068、-2.924、5.774、4.077,χ^(2)=8.692,均P<0.05)。logistic回归分析显示,肿瘤分期、CD3^(+)、CD8^(+)、IL-6水平是影响胃癌患者预后的因素(均P<0.05)。CD3^(+)、CD8^(+)、IL-6预测患者预后不良的ROC下面积分别为0.847、0.853、0.768。结论胃癌患者CD3^(+)、CD4^(+)低表达,CD8^(+)及IL-6高表达。CD3^(+)、CD8^(+)、IL-6水平是影响胃癌患者预后的因素。

儿童脐血移植后巨细胞病毒活化的危险因素及其与T细胞重建的相关性

目的 探讨儿童白血病接受脐带血移植后巨细胞病毒(CMV)活化的危险因素及其与T淋巴细胞重建的相关性。方法 回顾性分析2016年1月—2021年12月在苏州大学附属儿童医院血液科行脐带血造血干细胞移植的白血病患儿的临床资料及CMV活化的发生率,并分析单次及多次CMV活化对T淋巴细胞重建的影响,最后随访时间是2022年12月。结果 共纳入75例患儿,移植100d内53例发生CMV活化,18例表现为难治性。单因素分析示:预处理使用ATG是CMV活化的危险因素(P=0.043);移植时HLA配型不合、发生Ⅱ-Ⅳ度aGVHD是难治性CMV活化的危险因素(P<0.01);多因素分析示:发生Ⅱ-Ⅳ度aGVHD是难治性CMV活化的独立危险因素(P=0.021);发生多次CMV活化者移植后1年内CD4^(+)细胞重建延迟。移植后1个月内CMV活化对总体生存率无明显影响,CMV活化的AML患儿累积复发率下降。结论 预处理使用ATG是白血病儿童脐血移植后CMV活化的危险因素,供-受体HLA配型不合、发生Ⅱ-Ⅳ度aGVHD是发生难治性CMV感染的危险因素;多次CMV活化影响CD4^(+)细胞重建。

T-regulatory lymphocytes in peripheral blood of gastric and colorectal cancer patients

AIM: To assess the absolute number of T-regulatory cells (Tregs; CD4+CD25+Foxp3+) in the peripheral blood of gastric and colorectal cancer patients. METHODS: We enrolled 70 cancer patients (33 gastric cancer, 37 colorectal cancer) and 17 healthy volunteers. The CD3+CD4+ lymphocytes and CD4+CD25+Foxp3+ Tregs in the peripheral blood were analyzed with flow cytometry. The absolute numbers of Tregs were calculated based on the CD4+CD25+Foxp3+ cells percent-age of CD3+CD4+ cells and the absolute numbers of CD3+CD4+ cells per microliter. RESULTS: The mean number of CD4+CD25+Foxp3+ cells per microliter in colorectal cancer patients was 15.7 (SD: 21.8), for gastric cancer patients 12.2 (SD: 14.3), and for controls 17.5 (SD: 11.4). The absolute number of Tregs was significantly lower in gastric cancer patients than in controls (P = 0.026). There was no statistically significant difference for gastric vs colorectal cancer or colorectal cancer vs controls. The absolute number of Tregs was also significantly depressed in N+ vs Ncancer patients [22.0 (27.7) vs 10.1 (9.0), P = 0.013], and in the subgroup of gastric cancer patients [30.3 (27.6) vs 9.6 (8.0), P = 0.003]. No statistical difference was observed in the proportion of Tregs in the CD4+ population between the groups. CONCLUSION: The absolute number of Tregs in peripheral blood of gastric cancer but not colorectal cancer patients was significantly decreased in comparison with that in healthy controls.

肺癌患者外周血和胸腔积液T细胞受体组库特征分析

目的 比较肺癌患者外周血和胸腔积液的T细胞受体(TCR)组库的免疫学特征。方法 收集2020年7月至2021年1月在复旦大学附属中山医院就诊的5例合并恶性胸腔积液肺癌患者的外周血和胸腔积液,分离外周血单个核细胞(PBMC)和胸腔积液单个核细胞(PEMC),提取基因组DNA,采用Illumina MiSeq平台对TCR进行高通量测序,分析TCR组库的多样性等特征。结果 PBMC和PEMC中的TCR克隆型、VJ pairs和互补决定区3(CDR3)序列的多样性和相关性差异均无统计学意义。PBMC中检测到的TCR克隆数量高于PEMC(P<0.05),VJ pairs和CDR3数量与PEMC差异无统计学意义。PBMC和PEMC配对样本的共有VJ pairs数量占总VJ pairs的比例高于共有克隆型和CDR3序列的数量比例(P<0.05),后两者差异无统计学意义。PEMC中高扩增克隆(HEC)数量和占比均高于PBMC。同时鉴定了36个在PBMC和PEMC中差异表达的VJ pairs,其中25个在PEMC中表达升高、9个在PEMC中表达降低。结论 肺癌患者外周血和胸腔积液的TCR组库表现出不同的特征性变化,可能为肺癌重要的免疫病理学特征。

结核T细胞斑点试验联合外周血T淋巴细胞亚群检测在临床诊断中的价值

目的探讨结核T细胞斑点试验(T-SPOT.TB)联合外周血T淋巴细胞亚群检测在结核患者中的诊断价值。方法回顾性分析本院就诊的279例患者的临床资料。依据诊断标准分为结核组147例、非结核组132例,评估T-SPOT.TB与外周血T淋巴细胞亚群检测对结核病的诊断价值。结果结核组T-SPOT.TB阳性率高于非结核组(P<0.01);肺结核组患者CD3^(+)、CD4^(+)及CD8^(+)较非结核组相比有下降(P<0.05),CD4^(+)/CD8^(+)较非结核组变化不明显(P>0.05);两者联合检测受试者工作特征曲线下面积(AUC)为0.354(P<0.01),灵敏度90.48%,特异性38.64%,约登指数为0.291。结论T-SPOT.TB和外周血T淋巴细胞亚群检测诊断肺结核具有较高灵敏度,但在对结核和非结核的鉴别诊断中特异性降低,在临床中对结核的诊断价值有一定的意义,是对肺结核诊断的有效补充。

辐照红细胞与普通悬浮红细胞输注对肝癌患者CD4+T细胞亚群水平的影响

目的分析辐照红细胞与普通悬浮红细胞输注对肝癌患者CD4+T细胞亚群水平的影响。方法回顾性分析100例肝癌患者,根据输血红细胞类型,将以辐照红细胞输注治疗的65例患者纳入辐照组,以普通悬浮红细胞输注治疗的35例患者纳入普通组。比较输血前和输血后14 d时,2组患者外周血CD4+T细胞亚群水平(Th1、Th2、Th17、Treg)、细胞因子水平[白介素-2(IL-2)、白介素-10(IL-10)、白介素-17(IL-17)、干扰素(IFN-γ)]。比较输血后3个月内2组患者感染发生率。输血后随访3年,比较2组患者无进展生存期(PFS)。结果输血后14 d时,辐照组患者Th1、IL-2、IFN-γ水平显著高于普通组,Th2、Treg、IL-10、IL-17水平显著低于普通组(P<0.05),2组Th17水平比较差异无统计学意义(P>0.05)。输血后3个月内辐照组患者感染发生率低于普通组,比较差异无统计学意义(P>0.05)。输血后随访3年,2组患者PFS比较无统计学意义(P>0.05)。结论异体普通悬浮红细胞输注将抑制肝癌患者细胞免疫功能,而辐照红细胞输注可减轻异体输血引起的细胞免疫抑制反应,后者在肝癌手术中具有较高推广应用价值。

调节性T细胞在肝细胞癌患者外周血中的表达及临床意义

目的探讨在不同病理分期肝细胞癌(hepatocellular carcinoma,HCC)患者外周血中CD4^(+)CD25^(+)CD127^(LOW)Treg的含量变化及临床意义。方法收集60例HCC患者的外周抗凝血作为HCC试验组并进行Edmondson-steiner病理分期,20名健康体检者的外周抗凝血作为健康对照组,检测两组CD4^(+)CD25^(+)CD127^(LOW)Treg水平。结果HCC患者外周血中Treg占CD4^(+)T细胞的百分率显著高于健康对照组(P<0.01);且在HCC不同分期中,Ⅲ、Ⅳ期患者外周血中Treg占CD4^(+)T细胞的百分率显著高于Ⅰ、Ⅱ期患者(P<0.05)。结论在HCC患者外周血中,Treg随着疾病的发生发展而逐渐增多,且在Ⅲ、Ⅳ期快速增殖,其可能与肿瘤细胞免疫逃逸密切相关。

Establishment of a humanized mouse model using steady-state peripheral blood-derived hematopoietic stem and progenitor cells facilitates screening of cancer-targeted T-cell repertoires

Background:Cancer-targeted T-cell receptor T(TCR-T)cells hold promise in treating cancers such as hematological malignancies and breast cancers.However,approaches to obtain cancer-reactive TCR-T cells have been unsuccessful.Methods:Here,we developed a novel strategy to screen for cancer-targeted TCR-T cells using a special humanized mouse model with person-specific immune fingerprints.Rare steady-state circulating hematopoietic stem and progenitor cells were expanded via three-dimensional culture of steady-state peripheral blood mononuclear cells,and then the expanded cells were applied to establish humanized mice.The human immune system was evaluated according to the kinetics of dendritic cells,monocytes,T-cell subsets,and cytokines.To fully stimulate the immune response and to obtain B-cell precursor NAML-6-and triple-negative breast cancer MDA-MB-231-targeted TCR-T cells,we used the inactivated cells above to treat humanized mice twice a day every 7 days.Then,human T cells were processed for TCRβ-chain(TRB)sequencing analysis.After the repertoires had been constructed,features such as the fraction,diversity,and immune signature were investigated.Results:The results demonstrated an increase in diversity and clonality of T cells after treatment.The preferential usage and features of TRBV,TRBJ,and the V–J combination were also changed.The stress also induced highly clonal Science and Technology,Grant/Award Number:2021C03010;Zhejiang Provincial Natural Science Foundation of China,Grant/Award Numbers:LTGY24H080003,LY21H080004 expansion.Tumor burden and survival analysis demonstrated that stress induction could significantly inhibit the growth of subsequently transfused live tumor cells and prolong the survival of the humanized mice.Conclusions:We constructed a personalized humanized mouse model to screen cancer-targeted TCR-T pools.Our platform provides an effective source of cancer-targeted TCR-T cells and allows for the design of patient-specific engineered T cells.It therefore has the potential to greatly benefit cancer treatment.

获得性免疫缺陷综合征合并脓毒症患者外周血中T淋巴细胞亚群研究

目的 探讨获得性免疫缺陷综合征(AIDS)合并脓毒症患者外周血中T淋巴细胞亚群。方法 回顾性选取2021年10月至2023年9月海南医学院第二人民医院内AIDS合并脓毒症患者60例作为试验组,AIDS无脓毒症患者60例作为对照1组,健康献血员20名作为对照2组。应用免疫荧光和单克隆抗体标记技术检测试验组、对照1组和对照2组外周血T淋巴细胞亚群(CD3细胞数量、CD4细胞数量、CD8细胞数量、CD4/CD8比值),比较试验组、对照1组和对照2组T淋巴细胞亚群计数和比例,并统计分析试验组不同年龄、确诊时间患者的T淋巴细胞亚群计数和比例。结果 试验组患者的CD3细胞数量、CD4细胞数量分别为(864.72±184.22)、(145.37±25.37)个/μL,均明显少于对照1组[(971.80±135.14)、(218.11±36.24)个/μL]、对照2组[(1 098.12±156.23)、(296.23±41.25)个/μL],CD4/CD8比值为0.258±0.024,明显低于对照1组(0.344±0.034)、对照2组(0.458±0.045),差异均有统计学意义(P<0.05);对照1组患者的CD3细胞数量、CD4细胞数量均明显少于对照2组,CD4/CD8比值明显低于对照2组,差异均有统计学意义(P<0.05);但3组研究对象的CD8细胞数量比较,差异均无统计学意义(P>0.05)。不同年龄患者的CD3细胞数量、CD4细胞数量、CD8细胞数量、CD4/CD8比值比较,差异均无统计学意义(P>0.05)。不同确诊时间患者的CD4细胞数量比较,差异有统计学意义(P<0.05),其中,确诊时间0~1年、1~2年患者的CD4细胞数量均多于2~3年、3~4年、4~5年、≥5年患者(P<0.05);但不同确诊时间患者的CD3细胞数量、CD8细胞数量、CD4/CD8比值比较,差异均无统计学意义(P>0.05)。结论 AIDS合并脓毒症患者外周血中T淋巴细胞亚群降低,CD4细胞数量与确诊时间密切相关,不仅能拓宽临床对于AIDS机会性感染认识,更能为其诊治提供新的思路及方向。

脓毒症患者外周血T淋巴细胞亚群及炎症因子水平与预后的关系研究

目的:探讨脓毒症患者外周血T淋巴细胞亚群及炎症因子水平与预后的关系。方法:选取2020年7月至2023年5月在联勤保障部队第九八九医院治疗的70例脓毒症患者,纳入脓毒症组,入院后28d根据患者存活情况又分为存活组和死亡组,比较两组患者的外周血T淋巴细胞亚群包括CD_(4)^(+)、CD_(8)^(+)和CD_(4)^(+)/CD_(8)^(+)比值、降钙素原(PCT)和中性粒细胞与淋巴细胞计数比值(NLR)水平。随访所有脓毒症患者28d内预后情况,ROC分析相关指标对预后的预测价值。结果:随访28d内,脓毒症患者存活53例,死亡17例。死亡组患者的PCT、NLR、CD_(8)^(+)高于存活组,CD_(4)^(+)和CD_(4)^(+)/CD_(8)^(+)均较存活组低,差异有统计学意义(P<0.05)。ROC分析显示,PCT、NLR、CD_(4)^(+)、CD_(8)^(+)、CD_(4)^(+)/CD_(8)^(+)对脓毒症患者预后预测的曲线下面积为0.748、0.794、0.794、0.739和0.872,PCT预测敏感度和特异度为76.47%和94.34%,NLR预测敏感度和特异度为82.35%和92.45%,CD_(4)^(+)预测敏感度和特异度为64.71%和83.02%,CD_(8)^(+)预测敏感度和特异度为58.82%和94.34%,CD_(4)^(+)/CD_(8)^(+)预测敏感度和特异度为82.35%和86.79%。结论:外周血PCT、NLR及T淋巴细胞亚群在脓毒症患者中表达异常,其中NLR对脓毒症患者预后有较高预测价值,PCT和外周血T淋巴细胞亚群对脓毒症患者预后预测价值一般。

成人乙型肝炎疫苗接种后无应答的影响因素及与外周血滤泡T细胞的关联研究

目的分析成人乙型肝炎(乙肝)疫苗接种后无应答的影响因素,及与外周滤泡T细胞的关联。方法回顾性分析2022年1月至2022年12月至信阳市平桥区疾病预防控制中心接受乙肝疫苗接种的129例患者的临床资料,依据接种后乙型肝炎病毒表面抗体(HBsAb)水平,将接种后HBsAb≥10 mIU/mL的100例患者列为应答组,将接种后HBsAb<10 mIU/mL的29例患者列为无应答组。收集、对比两组患者的一般资料及临床资料,经统计学单因素分析、Logistic多因素回归分析归纳可导致成人乙肝疫苗接种后无应答的影响因素。比较两组患者外周血滤泡T细胞间的差异,经Spearman相关性系数检验外周血滤泡T细胞与乙肝疫苗接种后无应答的关联。结果统计学单因素分析结果显示,无应答组内男性、年龄≥60岁、体重指数(BMI)≥24 kg/m^(2)、有吸烟饮酒史、肌肉注射、按10μg 0-1-6月程序接种、有造血干细胞移植治疗史、合并人类免疫缺陷病毒(HIV)感染、Toll样受体(TLRs)高表达、外周血滤泡辅助性T细胞(Tfh)低表达、白细胞介素-12(IL-12)低表达、外周血滤泡调节性T细胞(Tfr)高表达、趋化因子受体5(CXCR5)高表达占比均高于应答组(P<0.05)。Logistic多因素回归分析结果显示,肌肉注射、按10μg 0-1-6月程序接种、Tfh低表达、IL-12低表达、Tfr高表达、CXCR5高表达为成人乙肝疫苗接种后无应答的主要影响因素。无应答组的Tfh、IL-12分别为(17.44±3.25)%、(70.33±10.21)ng/mL,均低于应答组[(19.25±3.16)%、(75.45±10.23)ng/mL];Tfr、CXCR5分别为(7.32±1.41)%、(33.49±5.18)ng/mL,均高于应答组[(6.65±1.24)%、(30.23±5.22)ng/mL],P<0.05。经Spearman相关性系数检验,Tfh、IL-12与HBsAb应答水平呈正相关,Tfr、CXCR5与HBsAb应答水平呈负相关。结论乙肝疫苗接种方式、程序不当以及个人免疫功能紊乱均是导致成人乙肝疫苗接种后无应答或应答低的重要因素。Tfh、IL-12与HBsAb应答水平呈正相关,Tfr、CXCR5与HBsAb应答水平呈负相关。

外周血T淋巴细胞亚群变化对于慢阻肺患者急性加重期相对危险性的评价

目的比较慢阻肺患者外周血T淋巴细胞亚群变化,分析引起慢阻肺患者急性加重期(AECOPD)相对危险因素并进行预测评价。方法选取2019年9月至2022年6月收治于我院的98例慢性阻塞性肺疾病(COPD)患者作为研究对象,根据患者病情是否急性加重分为两组,AECOPD组63例(男31例,女32例)和COPD组35例(男18例,女17例);经肺功能参数、血清上清液中炎症生物标志物和外周血中的淋巴细胞水平的检测,单因素以及多因素Logistic回归分析影响COPD患者急性加重的危险因素;构建人工神经网络模型进行预测和模型验证;采用Pearson分析T淋巴细胞亚群与患者肺功能指标的相关性。结果AECOPD组中有吸烟史、SII、NRS2002评分和WBC、NEUT、EOS、CRP、NLR水平显著高于COPD组,FEV_(1)/FVC、FEV_(1)%、PNI、BMI均显著低于COPD组(P均<0.05);AECOPD组的CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)、Treg水平显著低于COPD组,CD8^(+)、Th17、CD3^(-)CD56^(+)、CD3^(+)CD56^(+)水平高于COPD组(P<0.05);单因素以及多因素结果说明,CD8^(+)、Th17、CD3^(-)CD56^(+)、CD3^(+)CD56^(+)水平增加是引起急性加重的最主要危险因素(OR值均>1,P<0.05);人工神经网络预测模型具有良好的区分度、准确度和有效性;CD8^(+)、NEUT与AECOPD患者FEV_(1)%、FEV_(1)/FVC呈负相关,Th17与患者FEV_(1)%呈负相关,Treg与患者FEV_(1)%、FEV_(1)/FVC呈正相关,CD4^(+)/CD8^(+)与患者FEV_(1)%呈正相关(P<0.05)。结论分析外周血T淋巴细胞亚群变化可以对急性加重的机制有更清楚的了解,对患者的治疗和预后具有十分重要的预测价值,为AECOPD的临床诊断和免疫治疗提供研究思路。

外周血T淋巴细胞亚群、CD147及VWF水平与短暂性脑缺血发作患者早期临床转归的关系

目的研究外周血T淋巴细胞亚群、细胞外基质金属蛋白酶诱导子(CD147)及血管性血友病因子(VWF)水平对短暂性脑缺血发作(TIA)患者的临床意义。方法选择河北省沧州中西医结合医院2020年7月至2022年9月期间诊治的102例TIA患者作为实验组,根据卒中风险系数ABCD3⁃I评分表将TIA患者分为高危组、中危组、低危组,根据TIA患者发病72 h内是否发生脑梗死将患者分为脑梗死组(发生脑梗死)、无脑梗死组(未发生脑梗死),另选取98名健康志愿者作为对照组。分别比较实验组与对照组、不同危险程度TIA患者、不同早期临床转归TIA患者的外周血T淋巴细胞亚群、CD147及VWF水平;采用Spearman分析该些指标与TIA患者脑梗死发生率的相关性。结果实验组CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)明显低于对照组,CD8^(+)、CD147、VWF水平明显高于对照组,差异有统计学意义(P<0.05);三组不同危险程度TIA患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)呈现:高危组<中危组<低危组,CD8^(+)、CD147、VWF水平呈现:高危组>中危组>低危组,差异有统计学意义(P<0.05);脑梗死组TIA患者CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)明显低于无脑梗死组,CD8^(+)、CD147、VWF水平显著高于无脑梗死组,差异有统计学意义(P<0.05);Spearman分析结果显示,TIA患者脑梗死发生率与CD3^(+)、CD4^(+)、CD4^(+)/CD8^(+)呈负相关,与CD8^(+)、CD147、VWF水平呈正相关(P<0.05)。结论外周血T淋巴细胞亚群、CD147及VWF水平与TIA患者早期临床转归有明显的关联,可作为TIA患者是否会发生脑梗死的重要评估指标。

自制红细胞裂解液对流式细胞术测定外周血T淋巴细胞亚群的影响研究

目的比较实验室配制的红细胞裂解液的裂解时间对流式测定SD大鼠和食蟹猴外周血T淋巴细胞亚群的影响。方法(1)取15只食蟹猴的肝素钠抗凝血各50μl,血样与荧光检测抗体Per CP-CD3/FITC-CD8/PE-CD4孵育后分别经自制和市售红细胞裂解液裂解一段时间,比较样本经自制与市售裂解液处理不同时间后其CD3~+CD4~+,CD3~+CD8~+和CD3~+淋巴细胞的平均荧光强度(median fluorescence intensity,MFI)等数值的差异;(2)取15只SD大鼠的EDTA-K2抗凝血各50μl,血样与荧光检测抗体APC-CD3/FITC-CD4/PE-CD8a孵育后,分别经自制红细胞裂解液和市售裂解液裂解一段时间,比较样本经自制与市售裂解液处理不同时间后CD3~+CD4~+和CD3~+CD8~+淋巴细胞的MFI等数值的差异;(3)取15只大鼠EDTA-K2抗凝血各50μl,血样与荧光检测抗体APC-CD3/PE-CD4孵育后,分别经自制红细胞裂解液和市售裂解液裂解一段时间,比较样本经自制与市售裂解液处理不同时间后PE-CD4~+的MFI差异。结果(1)食蟹猴外周血经自制红细胞裂解液,裂解时间约15min和5min的FITC-CD8~+的MFI分别为1745±173和2439±253,淋巴细胞百分比分别为47.9%±7.2%和42.1%±7.6%;裂解时间约15min和5min的PE-CD4~+的MFI分别为12924±892和13695±984,淋巴细胞百分比分别为38.7%±7.2%和33.1%±7.2%。裂解时间约15min比裂解5min的FITC-CD8~+(t=8.779,P<0.0001)和PE-CD4~+(t=2.250,P<0.05)的MFI降低;淋巴细胞百分比占比(t=2.139,2.162,均P<0.05)增高,差异具有统计学意义。食蟹猴外周血经市售红细胞裂解液裂解15min,FITC-CD8~+,PE-CD4~+,Per CP-CD3~+的MFI分别为2413±240,12428±1208和1015±123。与自制裂解液裂解5min比较,市售裂解液裂解15min的PE-CD4~+和Per CP-CD3~+的MFI降低,差异具有统计学意义(t=3.150,4.862,均P<0.01)。(2)SD大鼠外周血经自制红细胞裂解液,裂解时间5min和2min的FITC-CD4~+的MFI分别为55.72±14.10和225.08±12.44;裂解5min与裂解2min的FITC-CD4~+的MFI比较差异有统计学意义(t=34.89,P<0.001);SD大鼠外周血经市售红细胞裂解液,裂解5min和15min的FITC-CD4~+的MFI分别为277.02±30.36和310.04±43.56,PE-CD8a~+的MFI分别为3453.43±443.19和3211.28±357.85;裂解时间5min与裂解15min的FITC-CD4~+和PE-CD8a~+的MFI比较差异有统计学意义(t=2.409,P<0.01)。(3)SD大鼠经自制红细胞裂解液,裂解5min和2min的PE-CD4~+的MFI分别为1766±67,1765±51,差异无统计学意义(t=0.045,P>0.05)。结论与市售裂解液比较,自制红细胞裂解液可缩短裂解时间。将FITC荧光抗体更换为PE荧光抗体后,在一定的裂解时间内可消除对大鼠测定淋巴细胞MFI的影响;但随着裂解时间延长会造成FITC和PE荧光抗体MFI值的下降。

传染性单核细胞增多症患儿外周血T淋巴细胞亚群变化及其临床意义

目的探究传染性单核细胞增多症(IM)患儿外周血T淋巴细胞亚群变化及其临床意义。方法选择2018—2020年宝鸡市妇幼保健院收治的105例IM患儿作为研究组,同期在医院体检的健康儿童100例作为对照组,应用流式细胞术检测两组儿童的外周血T淋巴细胞亚群(CD8^(+)、CD4^(+)、CD3^(+)、CD4^(+)/CD8^(+))水平,应用受试者工作特征曲线(ROC)曲线分析T淋巴亚群对IM诊断价值;依据病程和病情严重程度进行分组,采用Spearman相关分析T淋巴亚群与病程和病情严重程度之间的关系。结果研究组儿童的CD8^(+)、CD3^(+)分别为(40.33±7.26)%、(78.29±7.54)%,明显高于对照组的(21.59±6.28)%、(62.94±8.14)%,CD4^(+)、CD4^(+)/CD8^(+)分别为(20.53±5.19)%、(0.63±0.19),明显低于对照组的(35.94±6.57)%、(1.34±0.43),差异均有统计学意义(P<0.05);经ROC分析结果显示,CD8^(+)、CD4^(+)、CD3^(+)、CD4^(+)/CD8^(+)用于IM诊断曲线下面积(AUC)分别为0.850、0.848、0.810、0.771。急性期组儿童的CD8^(+)、CD3^(+)分别为(44.23±5.79)%、(80.55±7.22)%,明显高于恢复期组的(33.16±2.89)%、(74.14±6.31)%,CD4^(+)、CD4^(+)/CD8^(+)分别为(18.19±3.22)%、(0.59±0.17),明显低于恢复期组的(24.83±5.40)%、(0.70±0.20),差异均有统计学意义(P<0.05);重症组儿童的CD8^(+)、CD3^(+)分别为(44.76±4.67)%、(81.11±8.25)%,明显高于轻症组的(33.94±5.29)%、(33.94±5.29)%,CD4^(+)、CD4^(+)/CD8^(+)分别为(18.15±3.70)%、(0.56±0.17),明显低于轻症组的(23.96±5.14)%、(0.73±0.18),差异均有统计学意义(P<0.05)。经Spearman相关性分析结果显示,外周血CD8^(+)、CD3^(+)水平与病程呈负相关(r=-0.428,-0.625;P<0.05),与病情严重程度呈正相关(r=0.494,0.614;P<0.05);CD4^(+)、CD4^(+)/CD8^(+)水平与病程呈正相关(r=0.509,0.722;P<0.05),与病情严重程度呈负相关(r=-0.319,-0.747;P<0.05)。结论IM患儿外周血T淋巴细胞亚群水平异常,可用于患者病情诊断及病情严重程度评估。

子痫前期孕妇外周血T淋巴细胞亚群和线粒体损伤指数变化及其临床意义

目的 探讨子痫前期孕妇外周血T淋巴细胞亚群和线粒体损伤指数变化及其临床意义。方法 选取2021年2月至2023年2月绍兴市柯桥区妇幼保健院收治的58例行孕期管理并分娩的子痫前期孕妇作为研究对象,设为观察组。根据病情程度将其分为非重度子痫前期组(n=40)和重度子痫前期组(n=18)。另选取同期58例健康孕妇作为对照组。于孕中期(24~28周)分别检测各组外周血T淋巴细胞亚群(CD3^(+)、CD4^(+)、CD8^(+))和辅助性T细胞(Th)、抑制性T细胞(Ts)线粒体损伤指数,比较各组间指标差异,并关注妊娠结局,分析指标与妊娠结局的关系。结果 观察组CD3^(+)、CD4^(+)、CD8^(+)水平及Th、Ts线粒体损伤指数阳性率均高于对照组(P<0.05);非重度子痫前期组CD3^(+)、CD4^(+)、CD8^(+)水平及Th、Ts线粒体损伤指数阳性率均低于重度子痫前期组(P<0.05)。观察组总不良妊娠结局发生率为51.72%,其中妊娠结局良好者CD3^(+)、CD4^(+)、CD8^(+)水平及Th、Ts线粒体损伤指数阳性率均低于妊娠结局不良者(P<0.05)。受试者工作特征(ROC)曲线分析显示,CD3^(+)、CD4^(+)、CD8^(+)水平及Th、Ts线粒体损伤指数均具有预测新生儿不良妊娠结局的价值(P<0.05)。结论 子痫前期孕妇存在免疫功能紊乱和异常氧化应激,T淋巴细胞亚群和线粒体损伤指数的检测可作为病情评估和不良妊娠结局预测的指标。