Identification of prognostic molecular subtypes and model based on CD8+ T cells for lung adenocarcinoma

Background:Cytotoxic T lymphocytes(CD8+T)cells function critically in mediating anti-tumor immune response in cancer patients.Characterizing the specific functions of CD8+T cells in lung adenocarcinoma(LUAD)could help better understand local anti-tumor immune responses and estimate the effect of immunotherapy.Methods:Gens related to CD8+T cells were identified by cluster analysis based on the single-cell sequencing data of three LUAD tissues and their paired normal tissues.Weighted gene co-expression network analysis(WGCNA),consensus clustering,differential expression analysis,least absolute shrinkage and selection operator(LASSO)and Cox regression analysis were conducted to classify molecular subtypes for LUAD and to develop a risk model using prognostic genes related to CD8+T cells.Expression of the genes in the prognostic model,their effects on tumor cell invasion,and interactions with CD8+T cells were verified by cell experiments.Results:This study defined two LUAD clusters(CD8+0 and CD8+1)based on CD8+T cells,with cluster CD8+0 being significantly associated with the prognosis of LUAD.Three heterogeneous subtypes(clusters 1,2,and 3)differing in prognosis,genome mutation events,and immune status were categorized using 42 prognostic genes.A prognostic model created based on 11 significant genes(including CD200R1,CLEC17A,ZC3H12D,GNG7,SNX30,CDCP1,NEIL3,IGF2BP1,RHOV,ABCC2,and KRT81)was able to independently estimate the death risk for patients in different LUAD cohorts.Moreover,the model also showed general applicability in external validation cohorts.Low-risk patients could benefit more from taking immunotherapy and were significantly related to the resistance to anticancer drugs.The results from cell experiments demonstrated that the expression of CD200R1,CLEC17A,ZC3H12D,GNG7,and SNX30 was significantly downregulated,while that of CDCP1,NEIL3,IGF2BP1,RHOV,ABCC2 and KRT81 was upregulated in LUAD cells.Inhibition of CD200R1 greatly increased the invasiveness of the LUAD cells,but inhibiting CDCP1 expression weakened the invasion ability of LUAD cells.Conclusion:This study defined two prognostic CD8+T cell clusters and classified three heterogeneous molecular subtypes for LUAD.A prognostic model predictive of the potential effects of immunotherapy on LUAD patients was developed.

外周血CD8+CD28-CD57+T淋巴细胞对老年脓毒症患者预后的预测价值

目的 探讨外周血CD8+CD28-CD57+T淋巴细胞对老年脓毒症患者预后的预测价值。方法 采用回顾性队列研究,选取2015年2月—2016年8月西安交通大学第二附属医院重症医学科住院的年龄≥60岁的脓毒症患者75例,依据ICU结局分为存活组(n=54)及死亡组(n=21)。收集患者一般资料,诊断脓毒症当天进行急性生理和慢性健康评分Ⅱ(APACHEⅡ)评分及序贯器官衰竭(SOFA)评分,检测外周血液标本TNF-a、IL-10及CD8+CD28-CD57+T淋巴细胞。结果 死亡组平均年龄大于存活组(P<0.05)。死亡组较存活组有更高的APACHEⅡ评分、SOFA评分及休克比例,差异均有统计学意义(P<0.05)。死亡组外周血CD8+CD28-CD57+T淋巴细胞、TNF-a、IL-10较存活组更高(P<0.05)。存活组的耐药菌感染比例低于死亡组,但差异无统计学意义(P>0.05)。无论单因素还是对一系列协变量进行调整的多因素Logistic回归分析均显示,较高的外周血CD8+CD28-CD57+T淋巴细胞比例与高的ICU死亡率相关(OR, 1.21;95%CI, 1.10~1.33)(OR, 1.30;95%CI, 1.07~1.58);APACHEⅡ评分预后预测的AUC为0.78(95%CI 0.67~0.90),将最适诊断界点22.5分作为预测死亡可能的临界点,敏感性和特异性分别为61.9%和75.2%。SOFA评分预后预测的AUC为0.80(95%CI,0.68~0.92),将最适诊断界点9.5分作为预测死亡可能的临界点,敏感性和特异性分别为71.4%和77.8%。外周血CD8+CD28-CD57+T淋巴细胞预后预测的AUC为0.91(95%CI,0.83~0.99),将最适诊断界点60.2%作为预测死亡可能的临界点,敏感性和特异性分别为81.0%和92.6%。结论 老年脓毒症患者外周血高CD8+CD28-CD57+T淋巴细胞比例与ICU死亡率增加相关,一定程度上可用于评估此类人群的病情严重程度及预测预后。

Predicting the Prognosis and Immunotherapeutic Response of Triple-Negative Breast Cancer by Constructing a Prognostic Model Based on CD8+T Cell-Related Immune Genes

Objective Triple-negative breast cancer(TNBC)poses a significant challenge for treatment efficacy.CD8+T cells,which are pivotal immune cells,can be effectively analyzed for differential gene expression across diverse cell populations owing to rapid advancements in sequencing technology.By leveraging these genes,our objective was to develop a prognostic model that accurately predicts the prognosis of patients with TNBC and their responsiveness to immunotherapy.Methods Sample information and clinical data of TNBC were sourced from The Cancer Genome Atlas and METABRIC databases.In the initial stage,we identified 67 differentially expressed genes associated with immune response in CD8+T cells.Subsequently,we narrowed our focus to three key genes,namely CXCL13,GBP2,and GZMB,which were used to construct a prognostic model.The accuracy of the model was assessed using the validation set data and receiver operating characteristic(ROC)curves.Furthermore,we employed various methods,including Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway,immune infiltration,and correlation analyses with CD274(PD-L1)to explore the model's predictive efficacy in immunotherapeutic responses.Additionally,we investigated the potential underlying biological pathways that contribute to divergent treatment responses.Results We successfully developed a model capable of predicting the prognosis of patients with TNBC.The areas under the curve(AUC)values for the 1-,3-,and 5-year survival predictions were 0.618,0.652,and 0.826,respectively.Employing this risk model,we stratified the samples into high-and low-risk groups.Through KEGG enrichment analysis,we observed that the high-risk group predominantly exhibited enrichment in metabolism-related pathways such as drug and chlorophyll metabolism,whereas the low-risk group demonstrated significant enrichment in cytokine pathways.Furthermore,immune landscape analysis revealed noteworthy variations between(PD-L1)expression and risk scores,indicating that our model effectively predicted the response of patients to immune-based treatments.Conclusion Our study demonstrates the potential of CXCL13,GBP2,and GZMB as prognostic indicators of clinical outcomes and immunotherapy responses in patients with TNBC.These findings provide valuable insights and novel avenues for developing immunotherapeutic approaches targeting TNBC.

Tumor-derived DEFB1 induces immune tolerance by inhibiting maturation of dendritic cell and impairing CD8+T cell function in esophageal squamous cell carcinoma

Objective:CD8+T cells are the key effector cells in the anti-tumor immune response.The mechanism underlying the infiltration of CD8+T cells in esophageal squamous cell carcinoma(ESCC)has not been clearly elucidated.Methods:Fresh ESCC tissues were collected and grouped according to the infiltration density of CD8+T cells.After the transcriptome sequencing on these samples and the combined analyses with The Cancer Genome Atlas(TCGA)ESCC data,a secreted protein DEFB1 was selected to explore its potential role in the infiltration of CD8+T cells.Bioinformatics analyses,histological verification and in vitro experiments were then performed.Results:DEFB1 was highly expressed in ESCC,and the high expression of DEFB1 was an independent risk factor for overall survival.Since the up-regulation or down-regulation of DEFB1 did not affect the proliferation,migration and apoptosis of ESCC cells,we speculated that the oncogenic effect of DEFB1 was achieved by regulating microenvironmental characteristics.Bioinformatics analyses suggested that DEFB1 might play a major role in the inflammatory response and anti-tumor immune response,and correlate to the infiltration of immature dendritic cell(imDC)in ESCC.Histological analyses further confirmed that there were less CD8+T cells infiltrated,less CD83+mature DC(mDC)infiltrated and more CD1a+imDC infiltrated in those ESCC samples with high expression of DEFB1.After the treatment with recombinant DEFB1 protein,the maturation of DC was hindered significantly,followed by the impairment of the killing effects of T cells in both 2D and 3D culture in vitro.Conclusions:Tumor-derived DEFB1 can inhibit the maturation of DC and weaken the function of CD8+T cells,accounting for the immune tolerance in ESCC.The role of DEFB1 in ESCC deserves further exploration.

CD8^(+)CD28^(-)T细胞对单倍型造血干细胞移植后急性移植物抗宿主病的影响

目的:探讨CD8^(+)CD28^(-)T细胞对单倍型造血干细胞移植(haplo-HSCT)后急性移植物抗宿主病(aGVHD)的影响。方法:回顾性分析60例行haplo-HSCT的血液病患者移植后CD8^(+)CD28^(-)T细胞绝对计数与aGVHD的关系,并比较不同CD8^(+)CD28^(-)T细胞绝对计数组间慢性移植物抗宿主病(cGVHD)、感染及预后的差异。结果:60例行haplo-HSCT患者中有40例发生aGVHD,发生率为66.67%,中位发生时间为32.5(20-100)天。Haplo-HSCT后30天,aGVHD组CD8^(+)CD28^(-)T细胞绝对计数显著低于无aGVHD组(P=0.03)。ROC曲线表明移植后30天CD8^(+)CD28^(-)T细胞绝对计数在一定程度上可预测aGVHD的易感性。移植后30天低细胞计数组(CD8^(+)CD28^(-)T细胞绝对计数<0.06/μl)患者的aGVHD发生率显著高于高细胞计数组(CD8^(+)CD28^(-)T细胞绝对计数≥0.06/μl,P=0.011)。多因素Cox回归分析进一步验证了移植后30天CD8^(+)CD28^(-)T细胞绝对计数是aGVHD发生的独立风险因素,低细胞计数组aGVHD的发生风险是高细胞计数组的2.222倍(P=0.015)。不同CD8^(+)CD28^(-)T细胞绝对计数组间cGVHD、真菌感染、EB病毒感染、巨细胞病毒感染的发生无统计学差异。不同CD8^(+)CD28^(-)T细胞绝对计数组的总生存率、非复发相关死亡率、复发率没有显示出明显的统计学差异。结论:行haplo-HSCT后30天CD8^(+)CD28^(-)T细胞延迟重建的患者更易发生aGVHD,并且CD8^(+)CD28^(-)T细胞绝对计数在一定程度上可预测其易感性。单倍型造血干细胞移植后CD8^(+)CD28^(-)T细胞绝对计数与cGVHD、真菌感染、EB病毒感染、巨细胞病毒感染无关,且与移植后的生存预后无显著相关。

外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期宫颈癌患者腹腔镜根治术后预后的预测价值

目的分析外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期宫颈癌(CC)患者腹腔镜根治术后预后的预测价值。方法招募2018年9月至2020年9月于儋州市人民医院接受腹腔镜根治术治疗的早期CC患者204例,根据患者术后随访期间预后情况分为预后不良组(43例)和预后良好组(161例)。比较两组临床资料。采用Spearman秩相关分析外周血CD4^(+)CD25^(+)调节性T细胞水平与CD8^(+)CD28^(+)调节性T细胞水平的相关性。采用多因素logistic回归分析早期CC患者腹腔镜根治术后预后不良的影响因素。采用受试者工作特征(ROC)曲线评估外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平对早期CC患者腹腔镜根治术后预后不良的预测价值。结果预后不良组CD4^(+)CD25^(+)调节性T细胞、癌胚抗原(CEA)、糖类抗原125(CA125)水平,术后切缘阳性占比以及术中宫旁浸润占比高于预后良好组,CD8^(+)CD28^(+)调节性T细胞水平低于预后良好组,差异有统计学意义(P<0.05)。Spearman秩相关分析结果显示,早期CC患者外周血CD4^(+)CD25^(+)调节性T细胞水平与CD8^(+)CD28^(+)调节性T细胞水平呈负相关(r_(s)=-0.478,P<0.05)。多因素logistic回归分析结果显示,较高的CEA、CA125、CD4^(+)CD25^(+)调节性T细胞水平是促进早期CC患者腹腔镜根治术后预后不良发生的独立危险因素(P<0.05),较高的CD8^(+)CD28^(+)调节性T细胞水平是抑制早期CC患者腹腔镜根治术后预后不良发生的独立保护因素(P<0.05)。ROC曲线分析结果显示,外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平能有效预测早期CC患者腹腔镜根治术后预后不良(P<0.05),两项指标联合可进一步提高预测效能[AUC(95%CI)=0.939(0.898~0.979),P<0.001],灵敏度和特异度分别为86.00%、88.20%。结论外周血CD4^(+)CD25^(+)、CD8^(+)CD28^(+)调节性T细胞水平与早期CC患者腹腔镜根治术后预后不良有关,二者能有效预测早期CC患者腹腔镜根治术后预后不良。

Analysis of CD8^+CD28^-T-suppressor cells in living donor liver transplant recipients

BACKGROUND:Human CD8 + CD28 - T-suppressor(Ts) cells have been considered to indicate a reduced need for immunosuppression in pediatric liver-intestine transplant recipients and recipients of deceased heart-kidney transplants.However,in adult-to-adult living donor liver transplantation(A-A LDLT)little information is available and the clinical significance is still unknown. METHODS:Flow cytometry was used to detect the population of CD8+CD28 -Ts cells present in peripheral blood in A-A LDLT recipients(n=31),patients with end- stage liver disease(n=24)and healthy controls(n=19). Meanwhile,we tested the graft function and trough levels of immunosuppression in recipients.The clinical and follow- up data of 31 transplant recipients were analyzed. RESULTS:Compared with diseased controls(P=0.007) and healthy individuals(P=0.000),a notable expansion of CD8 + CD28 - Ts cells was found in recipients of A-A LDLT.This was associated with graft function,levels of immunosuppression and rejection episodes. CONCLUSIONS:To monitor the CD8 + CD28 - Ts cells levels is important to evaluate the immune state of recipients. Meanwhile,it is also important to promote expansion of CD8+CD28 -Ts cells in recipients of A-A LDLT,not only to sustain good graft function and decrease the dosage of immunosuppressants,but also to reduce the occurrence of rejection.

Changes of CD8+CD28-T regulatory cells in rat model of colitis induced by 2,4-dinitrofluorobenzene

AIM:To determine the changes of CD8+ T subsets especially CD8+CD28-T regulatory cells in rat model of experimental colitis induced by 2,4-dinitrofluorobenzene (DNFB). METHODS:The rat model of experimental colitis was induced by enema with DNFB.Ten days later,colonic intraepithelial and splenic lymphooltes were isolated from colitis animals (n=16) and controls (n=8).The proportion of CD8+ T cells,CD8+CD28+ T cells and CD8+CD28-T regulatory cells were determined by flow cytometry. RESULTS:The model of experimental colitis was successfully established by DNFB that was demonstrated by bloody diarrhea,weight loss and colonic histopathology.The proportion of CD8+ T cells in either splenic or colonic intraepithelial lymphocytes was not significantly different between colitis animals and controls (spleen:34.6±7.24 % vs 33.5±9.41%, colon:14.0±8.93 % vs 18.0±4.06 %,P>0.05).But CD8+CD28- T regulatory cells from colitis animals were significantly more than those from controls (spleen:11.3±2.26 % vs 5.64±1.01%, colon:6.50±5.37 % vs 1.07±0.65 %,P<0.05).In contrast, CD8+CD28+ T cells from colitis animals were less than those from controls (spleen:23.3±6.14 % vs 27.8±9.70 %,P=0.06; colon:7.52±4.18 % vs 16.9±4.07 %,P<0.05).The proportion of CD8+CD28-T regulatory cells in splenic and colon intraepithelial CD8+ T cells from colitis animals was higher than that from controls (spleen:33.3±5.49 % vs 18.4±7.26 %, colon:46.0±14.3 % vs6.10±3.72 %,P<0.005). CONCLUSION:Experimental colitis of rats can be induced by DNFB with simplicity and good reproducibility.The proportion of CD8+CD28-T regulatory cells in rats with experimental colitis is increased,which may be associated with the pathogenesis of colitis.

The Mechanisms of CD8+ T Cells Exhaustion in the Tumor Microenvironment and Immune Therapy

In the tumor immune microenvironment, CD8+ T cells differentiate towards functional failure. The exhaustion of CD8+ T cells (Tex) showed varying degrees of effect dysfunction, loss of proliferation ability, and sustained high expression of a variety of inhibitory receptors, with metabolic and epigenetic changes. Tex cells are heterogeneous, including several subsets with different characteristics at different stages of differentiation. Immune checkpoint inhibitors (ICIs) can restore the effect or function of Tex cells, indicating that this T cell subset plays a key role in tumor immunotherapy. The understanding of the mechanism of CD8+ T cell exhaustion will be helpful to the implementation of tumor immunotherapy. This article reviews the production, differentiation and functional characteristics of Tex cells and their relationship with tumor immunotherapy.

Peripheral CD4^(+)CD8^(+) double positive T cells:A potential marker to evaluate renal impairment susceptibility during systemic lupus erythematosus

Lupus nephritis(LN) has a high incidence in systemic lupus erythematosus(SLE) patients, but there is a lack of sensitive predictive markers. The purpose of the study was to investigate the association between the CD4^(+)CD8^(+)double positive T(DPT) lymphocytes and LN. The study included patients with SLE without renal impairment(SLE-NRI), LN, nephritic syndrome(NS), or nephritis. Peripheral blood lymphocyte subsets were analyzed by flow cytometry. Biochemical measurements were performed with peripheral blood in accordance with the recommendations proposed by the National Center for Clinical Laboratories. The proportions of DPT cells in the LN group were significantly higher than that in the SLE-NRI group(t=4.012, P<0.001), NS group(t=3.240,P=0.001), and nephritis group(t=2.57, P=0.011). In the LN group, the risk of renal impairment increased significantly in a DPT cells proportion-dependent manner. The risk of LN was 5.136 times(95% confidence interval, 2.115–12.473) higher in cases with a high proportion of DPT cells than those whose proportion of DPT cells within the normal range. These findings indicated that the proportion of DPT cells could be a potential marker to evaluate LN susceptibility, and the interference of NS and nephritis could be effectively excluded when assessing the risk of renal impairment during SLE with DPT cell proportion.

The changes and clinical significance of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke

Objective:To study the changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke and their correlation with neuronal damage markers, inflammatory cytokines and plaque stability indicators.Methods: The patients who were diagnosed with acute ischemic stroke in our hospital between June 2014 and December 2017 were selected as the stroke group of the research, and healthy volunteers who received physical examination during the same period were selected as the control group. Peripheral blood was collected to determine the contents of CD4+CD25+ and CD4+CD28-T cells, and serum was collected to determine the contents of neuron damage markers, inflammatory cytokines and plaque stability indicators.Results: Peripheral blood CD4+CD25+ cell content as well as serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents of stroke group was lower than those of control group whereas peripheral blood CD4+CD28-T cell content as well as serum NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents was higher than those of control group;peripheral blood CD4+CD25+ cell content of stroke group was positively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and negatively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents;peripheral blood CD4+CD28-T cell content was negatively correlated with serum BDNF, IGF-1, IL-10, TGF-β1, TIMP2 and Vaspin contents, and positively correlated with NSE, VILIP-1, ET-1, IL-6, CXCL12, VCAM-1, P-selectin, ox-LDL, CatS, ICTP and VEGF contents.Conclusion: The changes of CD4+CD25+ and CD4+CD28-T cells in peripheral blood of patients with stroke can aggravate the neuron damage and promote the inflammatory response activation and plaque stability decline.

骨髓间充质干细胞通过上调CD8^+CD28^-T细胞抑制T细胞的增殖

本研究旨在探讨CD8+ CD2 8- T细胞在间充质干细胞 (MSC)抑制T细胞增殖中的作用。应用尼龙毛柱分离T淋巴细胞 ,再经CD8磁珠分选出CD8+ T淋巴细胞 ;用植物血凝素 (PHA)刺激与或未与MSC共培养 3天的CD8+ T细胞 ,应用MTT法测定T细胞的增殖 ;应用流式细胞术分别检测与或未与MSC共培养 8天的CD8+ T细胞亚群的变化及用PHA刺激与或未与MSC共培养 3天的CD8+ T细胞亚群的变化。结果表明 ,MSC抑制PHA刺激引起的CD8+ T细胞的增殖 ,且这种抑制作用是剂量依赖性的 ;流式细胞术结果显示 ,与未经MSC处理的CD8+ T细胞相比 ,MSC显著上调共培养 8天的CD8+ T细胞中的CD8+ CD2 8- 亚群 ,及经PHA作用 72小时后的CD8+ T细胞中的CD8+ CD2 8- 亚群。结论 :MSC通过上调CD8+ CD2 8- T细胞发挥抑制作用。

CD8^+CD28^+/CD8^+CD28^-T细胞平衡预测炎症性肠病患者并发消化道出血的价值

目的评价CD8^+CD28^+/CD8^+CD28-T细胞平衡在预测炎症性肠病(IBD)患者并发消化道出血(GH)的作用与价值。方法收集IBD患者49例,其中溃疡性结肠炎(UC)30例,克罗恩病(CD)19例,使用流式细胞术检测外周血CD8^+CD28^+及CD8^+CD28-T细胞T细胞的百分含量,对患者进行为期1年的随访,使用受试者工作特征(ROC)曲线法评价CD8^+CD28^+/CD8^+CD28-T细胞平衡(比值)在预测IBD患者出现GH的效能,并使用Kaplan-Meier生存分析法比较不同因素下的持续缓解时间(LTR)差异,并对相关指标进行相关性分析。结果 (1)CD组的免疫抑制剂、激素及生物制剂(BA)使用率均显著高于UC组(P=0.003、0.043及0.002);(2)UC组患者的CD8^+CD28^+T细胞显著高于CD组(t=3.022,P=0.004);(3)ROC结果显示CD8^+CD28^+T细胞、CD8^+CD28-T细胞及CD8^+CD28^+/CD8^+CD28-比值三者在预测GH方面均具有良好的效能(均为P<0.01),但以CD8^+CD28^+/CD8^+CD28-最优[曲线下面积(AUC)为0.977,P=0.000],截值分析显示当CD8^+CD28^+/CD8^+CD28-比值取值为1.14时(13.95%/12.24%),其对应的敏感度达93.3%,特异度为91.2%;(4)未使用BA及未行手术治疗的IBD患者算术及中位LTR均显著长于使用BA及已行手术的IBD患者(分别为χ2=9.730,P=0.002;χ2=15.981,P=0.000);(5)Spearman分析显示CD8^+CD28^+/CD8^+CD28-与BA及手术均成显著相关性(P=0.009、0.038)。结论外周血CD8^+CD28^+T细胞降低或CD8^+CD28-T细胞升高与IBD患者出现GH密切相关,CD8^+CD28^+/CD8^+CD28-平衡预测GH的敏感度及特异度均高,尤其是在比值为1.14时;该平衡与生物制剂及手术存在显著相关性。

CD8^+CD28^-Ts、CD3^+CD56+NKT细胞在B细胞非霍奇金淋巴瘤患者外周血中分布的分析

背景及目的:目前认为B细胞非霍奇金淋巴瘤(B-NHL)常伴随免疫抑制,CD8+CD28-Ts(Ts)细胞和CD3+CD56+NKT(NKT)是新鉴定的新型免疫抑制性调节细胞,在肿瘤的免疫抑制及免疫逃逸机制中起重要作用,但它们在B细胞淋巴瘤患者外周血的分布情况及其免疫抑制中的作用目前尚不清楚。本文通过分析两者在化疗前及化疗后B-NHL患者外周血中比例及变化规律,初步探讨它们在B-NHL的免疫抑制作用及其影响因素,为有效干预患者的免疫功能提供参考。方法:应用流式细胞仪检测79例治疗前的B-NHL患者、经4～6周期化疗后完全缓解(CR)的18例患者、30例健康志愿者外周静脉血中NKT及Ts细胞的比例。结果:在79例治疗前B-NHL患者的外周血中,Ts细胞比例为(18.19±5.03)%,较正常对照组(11.20±3.49)%明显增高(P<0.01);NKT的比例为(6.08±3.29)%,亦较正常对照组的(3.52±1.56)%明显增高(P<0.01)。Ts在不同临床分期的患者之间无显著性差异P>0.05;Ⅰ期为(17.56±4.10)%、Ⅱ期为(18.05±5.64)%、Ⅲ期为(18.14±5.58)%、Ⅳ期为(18.95±4.64)%;在不同恶性程度的患者之间亦无显著性差异P>0.05;低度恶性为(17.81±5.24)%、中度恶性为(18.37±4.83)%、高度恶性为(18.31±5.93)%;在治疗前(18.64±4.55)%和CR后(19.42±4.95)

哮喘患儿CD8^+CD28^+、CD8^+CD28^-T淋巴细胞检测及其临床意义

目的 通过对哮喘患儿CD4、CD8和CD2 8的联合检测 ,探讨哮喘患儿淋巴细胞免疫功能状态及其临床意义。方法 采用流式细胞术检测哮喘患儿外周血的总T细胞 (CD3+ )、辅助 /诱导T淋巴细胞 (CD4+ )、抑制 /细胞毒T淋巴细胞(CD8+ )、细胞毒T细胞 (CD8+ CD2 8+ )、抑制T细胞 (CD8+ CD2 8 )。结果 哮喘患儿组与对照组比较 ;CD3+ 、CD4+ 、CD8+CD2 8 细胞均低于对照组 (P <0 .0 1,P <0 .0 5 ) ,CD8+ CD2 8+ 细胞增高 (P <0 .0 5 ) ,CD4+ /CD8+ 比值、CD8+ 与对照组比较均无显著性差异 (P >0 .0 5 )。结论 哮喘患儿存在T淋巴细胞亚群免疫功能紊乱 ,而CD8+ CD2 8+ 、CD8+ CD2 8 T细胞失衡可能是导致机体免疫功能紊乱的主要因素。

咳嗽合剂对老年感染后咳嗽病人外周血CD8^+CD28^-T细胞Foxp3 mRNA表达的影响

目的探讨咳嗽合剂对感染后咳嗽疾病老年病人外周血CD8^+CD28^-T细胞Foxp3 mRNA的表达作用。方法选取2016年11月至2017年10月感染后咳嗽病人50例,随机分成2组。对照组不予干预治疗;试验组口服咳嗽合剂20 mL,3次/d,疗程7 d;分别检测治疗前1 d与治疗7 d后的CD8^+CD28^-T细胞含量、占单核细胞的百分比及CD8^+CD28^-T细胞Foxp3 mRNA的表达。结果对照组CD8^+CD28^-T细胞含量、CD8^+CD28^-T细胞占单核细胞的百分比及CD8^+CD28^-T细胞Foxp3 mRNA表达在治疗前后比较差异无统计学意义。试验组CD8^+CD28^-T细胞含量、CD8^+CD28^-T细胞占单核细胞的百分比及CD8^+CD28^-T细胞Foxp3 mRNA的表达在治疗前后比较差异具有统计学意义。结论咳嗽合剂抗炎作用可能与CD8^+CD28^-T细胞Foxp3 mRNA表达有关,可能通过抑制CD8^+CD28^-T细胞产生,减少免疫抑制,加快炎症消退。

肝星状细胞通过诱导产生CD8^+CD28^-T淋巴细胞抑制T细胞增殖

【目的】研究肝星状细胞(HSC)是否通过诱导产生CD8+CD28-T淋巴细胞抑制T细胞增殖。【方法】流式细胞仪分选出CD3+T细胞,用羧基荧光素双乙酸盐琥珀酰亚胺酯(CFSE)标记后与人肝星状细胞系LX2细胞共培养,PHA刺激72 h后,流式细胞仪检测T细胞增殖的百分比;为了研究肝星状细胞调控T细胞增殖的机制,将CD3+T细胞与LX2细胞共培养3 d,流式细胞仪检测CD8+CD28-T淋巴细胞的百分比。进一步将CD3+T细胞与LX2细胞共培养3 d,通过流式细胞仪分选出CD8+CD28-T淋巴细胞与CFSE标记的CD3+T细胞共培养,PHA刺激72 h后,流式细胞仪检测T细胞增殖的百分比。【结果】LX2细胞明显抑制了T细胞增殖,且抑制作用呈剂量依赖性;LX2剂量依赖性上调CD8+CD28-T淋巴细胞百分比。和LX2细胞共培养后的CD8+CD28-T淋巴细胞能够显著抑制T细胞增殖。【结论】HSC细胞通过诱导CD8+CD28-T调节细胞比例抑制T淋巴细胞增殖。

支气管哮喘小鼠外周血CD_8^+CD_(28)^-T细胞群和CD_8^+CD_(56)^+T细胞群作用机制研究

目的研究哮喘小鼠外周血CD8+CD28-T细胞与CD8+CD56+T细胞间的关系及两种细胞群在哮喘中的作用。方法 30只BALB/c小鼠随机分为对照组和哮喘组,每组各15只。测定小鼠的气道反应性,对支气管肺泡灌洗液行细胞学分类,观察肺组织的病理变化,流式检测小鼠外周血CD8+CD28-T细胞及CD8+CD56+T细胞的比例,分析哮喘中两种细胞之间的相关性。结果成功建立小鼠哮喘模型。①哮喘组小鼠BALF中细胞总数和嗜酸性粒细胞(%)较对照组明显增高;②哮喘组小鼠外周血CD8+CD28-T细胞及CD8+CD56+T细胞比例均较对照组升高;③CD8+CD28-T细胞与CD8+CD56+T细胞数量上呈正相关(r=0.737,P=0.002)。结论哮喘时外周血CD8+CD56+细胞和CD8+CD28-细胞数目异常,两种细胞正相关,可能在哮喘发病中起重要作用。

初发过敏性紫癜患儿外周血CD_4^+CD_^(28)-和CD_8^+CD_(28)^-T细胞水平变化及意义

目的观察初发过敏性紫癜(HSP)患儿外周血T淋巴细胞亚群(CD4+CD2-8T细胞和CD8+CD2-8T细胞)的表达变化,并探讨其临床意义。方法采用流式细胞技术(FCM)检测89例初发HSP患儿(HSP组)和30例健康儿童(对照组)外周血T淋巴细胞免疫表型。结果 HSP组CD4+CD2+8、CD8+CD2+8、CD3+CD4+T淋巴细胞表达水平及CD4+/CD8+比率较对照组显著降低(P均<0.05);HSP组CD3+和CD3+CD8+T淋巴细胞表达水平与对照组相比无显著性差异(P>0.05);HSP组CD4+CD2-8T细胞和CD8+CD2-8T细胞较对照组明显增高(P均<0.05),但在皮肤型、关节型、腹型、混合型和紫癜性肾炎五种临床类型间无明显差别(P均>0.05)。结论 HSP患儿存在T淋巴细胞亚群的紊乱,CD4+CD2-8T细胞及CD8+CD2-8T细胞的异常增高可能参与了其发病过程,但与HSP临床类型无关。

CD8^+CD28^-T细胞在原发性肝癌患者外周血的表达及意义

目的探讨原发性肝癌(PHC)患者外周血中CD8+CD28-T细胞在PHC细胞免疫中所发挥的作用。方法采用流式细胞技术对38例PHC患者及40例健康对照人群外周血中的CD8+T细胞表面CD28分子的表达进行检测。结果38例PHC患者与40例正常对照组外周血测定结果比较发现:PHC患者外周血中CD8+测定值较正常对照组显著增高;CD8+CD28-较正常对照组明显增高;而PHC患者外周血中CD8+CD28+与正常组比较明显下降,统计学差异均具有显著性。结论CD8+CD28-T细胞在PHC患者外周血中增生活跃,提示高表达的CD8+CD28-T细胞在肝癌免疫应答中具有重要意义。