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.

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.

Natural course of chronic hepatitis B is characterized by changing patterns of programmed death type-1 of CD8-positive T cells

AIM:To investigate if and how programmed death type-1(PD-1)expression affects the natural course of hepatitis B virus(HBV)infection. METHODS:Sixty-four patients in different natural stages of chronic HBV infection were enrolled in this study.PD-1 expression in total T cells was detected by flow cytometry.Levels of total CD8+T cell responses and proliferation in relation to PD-1 expression levels were analyzed with intracellular staining and PD-1/ PD-L1 blockage. RESULTS:The PD-1 expression in T cells was dynamically changed during the natural course of chronic HBV infection,did not significantly increase in the immune tolerance phase,and returned to normal in the inactive virus carrier stage.Blockage of the PD-1/PD-L1 pathway could not affect the T-cell response in the immune tolerance and inactive virus carrier stages of chronic HBV infection.However,it could significantly restore the T-cell response in the immune clearance stage of chronic HBV infection.Furthermore,the PD-1 expression level in T cells was associated with the alanine aminotransferase level during the immune clearance stage of chronic HBV infection. CONCLUSION:The PD-l/PD-L1 pathway plays a different role in T-cell response during the natural course of chronic HBV infection.

Super-resolution immunohistochemistry study on CD4 and CD8 cells and the relation to macrophages in human cochlea

Recently, the human cochlea has been shown to contain numerous resident macrophages under steady-state. The macrophages accumulate in the stria vascularis, among the auditory nerves, and are also spotted in the human organ of Corti. These macrophages may process antigens reaching the cochlea by invasion of pathogens and insertion of CI electrode. Thus, macrophages execute an innate, and possibly an adaptive immunity. Here, we describe the molecular markers CD4 and CD8 of T cells, macrophage markers MHCⅡ and CD11b, as well as the microglial markers TEME119 and P2Y12, in the human cochlea. Immunohistochemistry and the advantageous super-resolution structured illumination microscopy(SR-SIM) were used in the study. CD4^+ and CD8^+ cells were found in the human cochleae. They were seen in the modiolus in a substantial number adjacent to the vessels, in the peripheral region of the Rosenthal's canal, and occasionally in the spiral ligament. While there are a surprisingly large number of macrophages in the stria vascularis as well as between the auditory neurons,CD4^+ and CD8^+ cells are hardly seen in these areas, and neither are seen in the organ of Corti. In the modiolus,macrophages, CD4^+ and CD8^+ cells appeared often in clusters. Interaction between these different cells was easily observed with SR-SIM, showing closely placed cell bodies, and the processes from macrophages reaching out and touching the lymphocytes. Otherwise the CD4^+ and CD8^+ cells in human cochlear tissue are discretely scattered. The possible roles of these immune cells are speculated.

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.

Prognostic potential of an immune score based on the density of CD8＋ T cells, CD20＋ B cells, and CD33＋/p-STAT1＋ double-positive cells and HMGB1 expression within cancer nests in stage ⅢA gastric cancer patients

Objoctive： There is heterogeneity in the prognosis of gastric cancers staged according to the tumornodes-metastasis （TNM） system. This study evaluated the prognostic potential of an immune score system to supplement the TNM staging system. Mothodsg An immunohistochemical analysis was conducted to assess the density of T cells, B cells, and myeloid-derived suppressor cells （MDSCs） in cancer tissues from 100 stage IIIA gastric cancer patients; the expression of the high-mobility group protein B1 （HMGB1） was also evaluated in cancer cells. The relationship between the overall survival （OS）, disease-free survival （DFS）, and immunological parameters was analyzed.Results： An immune score system was compiled based on the prognostic role of the density ofT cells, B cells, MDSCs, and the expression of HMGB1 in cancer tissues. The median 5-year survival of this group of patient was 32%. However, the 5-year survival rates of 80.0%, 51.7%, 0%, 5.8%, and 0% varied among the patients with an immune score of 4 to those with an immune score of 0 based on the immune score system, respectively. Similarly, differences in DFS rates were observed among the immune score subgroups. Concluslons： An immune score system could effectively identify the prognostic heterogeneity within stage IliA gastric cancer patients, implying that this immune score system may potentially supplement the TNM staging system, and help in identifying a more homogeneous group of patients who on the basis of prognosis can undergo adjuvant therapy.

Glutamine deprivation impairs function of infiltrating CD8^(+)T cells in hepatocellular carcinoma by inducing mitochondrial damage and apoptosis

BACKGROUND The functions of infiltrating CD8^(+)T cells are often impaired due to tumor cells causing nutrient deprivation in the tumor microenvironment.Thus,the mechanisms of CD8^(+)T cell dysfunction have become a hot research topic,and there is increased interest on how changes in metabolomics correlate with CD8^(+)T cell dysfunction.AIM To investigate whether and how glutamine metabolism affects the function of infiltrating CD8^(+)T cells in hepatocellular carcinoma.METHODS Immunohistochemical staining and immunofluorescence were performed on surgically resected liver tissues from patients.Differentially expressed genes in infiltrating CD8^(+)T cells in hepatocellular carcinoma were detected using RNA sequencing.Activated CD8^(+)T cells were co-cultured with Huh-7 cells for 3 d.The function and mitochondrial status of CD8^(+)T cells were analyzed by flow cytometry,quantitative real-time polymerase chain reaction,and transmission electron microscopy.Next,CD8^(+)T cells were treated with the mitochondrial protective and damaging agents.Functional alterations in CD8^(+)T cells were detected by flow cytometry.Then,complete medium without glutamine was used to culture cells and their functional changes and mitochondrial status were detected.RESULTS There were a large number of infiltrating PD-1+CD8^(+)T cells in liver cancer tissues.Next,we cocultured CD8^(+)T cells and Huh-7 cells to explore the regulatory effect of hepatoma cells on CD8^(+)T cells.Flow cytometry results revealed increased PD-1 expression and decreased secretion of perforin(PRF1)and granzyme B(GZMB)by CD8^(+)T cells in the co-culture group.Meanwhile,JC-1 staining was decreased and the levels of reactive oxygen species and apoptosis were increased in CD8^(+)T cells of the co-culture group;additionally,the mitochondria of these cells were swollen.When CD8^(+)T cells were treated with the mitochondrial protective and damaging agents,their function was restored and inhibited,respectively,through the mitochondrial damage and apoptotic pathways.Subsequently,complete medium without glutamine was used to culture cells.As expected,CD8^(+)T cells showed functional downregulation,mitochondrial damage,and apoptosis.CONCLUSION Glutamine deprivation impairs the function of infiltrating CD8^(+)T cells in hepatocellular carcinoma through the mitochondrial damage and apoptotic pathways.

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。

Neutrophils inhibit CD8^(+)T cells immune response by arginase-1 signaling in patients with sepsis

BACKGROUND:Patients with sepsis often exhibit an acute inflammatory response,followed by an immunosuppressive phase with a poor immune response.However,the underlying mechanisms have not been fully elucidated.METHODS:We sought to comprehensively characterize the transcriptional changes in neutrophils of patients with sepsis by transcriptome sequencing.Additionally,we conducted a series of experiments,including real-time quantitative polymerase chain reaction(RT-qPCR)and flow cytometry to investigate the role of arginase-1 signaling in sepsis.RESULTS:Through the analysis of gene expression profiles,we identified that the negative regulation of T cell activation signaling was enriched,and the expression of arginase-1 was high in neutrophils from patients with sepsis.Furthermore,we conducted flow cytometry and found that the function of CD8^(+)T cells in septic patients was impaired.Moreover,neutrophils from septic patients inhibited the percentage of polyfunctional effector CD8^(+)T cells through arginase-1.Additionally,the proportions of granzyme B^(+)IFN^(-)γ^(+)CD8^(+)T and TNF^(-)α^(+)IFN^(-)γ^(+)CD8^(+)T cells increased after inhibition of arginase-1 signaling.CONCLUSION:The impaired effector function of CD8^(+)T cells could be restored by blocking arginase-1 signaling in patients with sepsis.

白细胞介素-38对乳腺癌患者CD8^(+)T淋巴细胞功能的影响

目的探讨白细胞介素-38(IL-38)在乳腺癌患者中的表达及其对CD8^(+)T细胞功能的调控作用。方法纳入2020年7月—2022年9月在新乡医学院第一附属医院就诊的44例乳腺癌患者、25例乳腺良性肿瘤患者和20例对照者。分离所有受试者的血浆和外周血单个核细胞,分离乳腺癌患者肿瘤组织中的肿瘤浸润淋巴细胞,纯化CD8^(+)T细胞。应用酶联免疫吸附试验(ELISA)检测血浆IL-38蛋白水平,应用实时定量PCR检测组织IL-38 mRNA相对表达量。使用重组人IL-38刺激乳腺癌患者外周血和肿瘤组织分离的CD8^(+)T细胞,建立CD8^(+)T细胞与乳腺癌细胞系MCF-7的共培养系统,通过测定乳酸脱氢酶水平计算靶细胞死亡比例,ELISA法检测培养上清中穿孔素、颗粒酶B、干扰素-γ和肿瘤坏死因子-α(TNF-α)水平,流式细胞术检测CD8^(+)T细胞的免疫检查点分子表达。结果乳腺癌患者血浆IL-38水平(74.23±19.88 pg/mL)高于乳腺良性肿瘤患者(62.87±16.27 pg/mL,P=0.018)和对照者(61.77±12.75 pg/mL,P=0.013)。乳腺癌患者肿瘤组织中IL-38 mRNA相对表达量显著高于癌旁组织(1.57±0.22 vs.1.00±0.18,P<0.001)。外周血和肿瘤浸润CD8^(+)T细胞诱导靶细胞死亡比例、穿孔素和颗粒酶B分泌在直接接触共培养组中的水平高于间接接触共培养组(P<0.05),但干扰素-γ和TNF-α分泌水平在直接接触共培养组和间接接触共培养组之间的差异无统计学意义(P>0.05)。在直接接触共培养组内,靶细胞死亡比例、穿孔素、颗粒酶B、干扰素-γ、TNF-α在IL-38刺激组中的水平低于无刺激组(P<0.05)。在间接接触共培养组内,靶细胞死亡比例、干扰素-γ、TNF-α在IL-38刺激组中的水平亦低于无刺激组(P<0.05),但穿孔素和颗粒酶B水平在间接接触共培养组内的IL-38刺激组和无刺激组之间的差异无统计学意义(P>0.05)。CD8^(+)T细胞中免疫检查点分子表达水平在无刺激组和IL-38刺激组之间的差异均无统计学意义(P>0.05)。结论乳腺癌患者中高表达的IL-38可能参与诱导CD8^(+)T细胞功能衰竭。

肿瘤微环境中与CD8^(+)T细胞耗竭相关的分化及代谢重编程的研究进展

CD8^(+)T细胞耗竭是导致机体免疫应答减弱及恶性肿瘤复发转移的核心原因。T细胞不仅在迁移进入肿瘤微环境(tumor microenvironment,TME)时需要突破重重屏障,而且在浸润至肿瘤微环境的T细胞的分化及代谢过程依然受到多因素的调控,往往造成CD8+T细胞抗肿瘤活性受到抑制,进一步形成耗竭性T细胞,导致免疫逃逸。因此,研究导致T细胞耗竭的相关因素及发生机制是逆转机体低免疫应答的关键,为抗肿瘤免疫治疗提供坚实的基础及依据。该文综述了近年来关于CD8^(+)T细胞在肿瘤微环境中的浸润分化、代谢重编程及耗竭性T细胞形成的相关过程,以期为改善T细胞抗肿瘤疗效提供潜在策略。

慢性HBV感染者外周血可诱导共刺激分子在CD8+ T淋巴细胞的表达及其临床意义

目的 探讨慢性乙型肝炎病毒(HBV)感染者外周血可诱导共刺激分子(ICOS)在CD8+T淋巴细胞的表达特点及临床意义。方法 选取2017年5月—2018年10月就诊的慢性乙型肝炎(CHB)100例、HBV-肝硬化(LC)25例和HBV-慢加急(或亚急)性肝衰竭(ACLF)26例分别纳入CHB组、HBV-LC组和HBV-ACLF组,健康对照(NC)组35例来自同期门诊体检健康者。采用流式细胞仪检测各组ICOS在CD8+T淋巴细胞的表达情况;分析CD8+T淋巴细胞ICOS表达水平与慢性HBV感染者疾病严重程度、HBV-ACLF预后及并发症的相关性;动态观察HBV-ACLF患者治疗过程中CD8+T淋巴细胞ICOS表达变化。结果 HBV-ACLF组外周血CD8+T淋巴细胞ICOS表达比率及平均荧光强度(MFI)均高于CHB组、HBV-LC组和NC组(P<0.05)。慢性HBV感染者ICOS的MFI与白蛋白、胆碱酯酶、总胆固醇、血红蛋白呈负相关(r=-0.263、-0.269、-0.273、-0.302,P=0.003、0.003、0.011、0.004);与直接胆红素、天冬氨酸转氨酶、凝血酶原时间、国际标准化比值呈正相关(r=0.248、0.208、0.331、0.315,P=0.005、0.020、0.003、0.009);与HBV-DNA、腹水及感染无明显相关性(P>0.05)。治疗过程中,HBV-ACLF患者ICOS的MFI无明显改变。但生存组ICOS的MFI在治疗第1周时较治疗前上升(P<0.05)。结论 HBV-ACLF患者外周血CD8+T淋巴细胞ICOS的表达水平明显升高,并与肝脏合成功能、炎症程度及预后相关,治疗早期ICOS水平变化有助于预测慢性HBV感染者的预后。

LncRNA LINC01137通过诱导CD8^(+)T细胞耗竭促进非小细胞肺癌进展的机制研究

目的研究长链非编码RNA(long non-coding RNA,LncRNA)LINC01137在非小细胞肺癌(nonsmall cell lung cancer,NSCLC)免疫逃逸中的生物学功能及其潜在的调节机制。方法采集24例健康志愿者和24例NSCLC患者血液样本,并收集NSCLC肿瘤组织和癌旁组织检测LINC01137水平。利用Starbase数据库预测LINC01137与miR-22-3p的结合位点,荧光素酶报告基因分析进行验证。采用A549细胞来源的外泌体和/或sh-LINC01137干扰序列转染A549细胞,检测细胞增殖和侵袭能力;收集转染后的细胞上清液培养CD8^(+)T细胞,检测CD8^(+)T细胞耗竭标志物干扰素-γ(interfereron-γ,IFN-γ)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)、颗粒霉素B(granzyme B)和白细胞介素-2(interleukin-2,IL-2)水平,以及PD-1+Tim3^(+)CD8^(+)T细胞百分比。采用外泌体和/或miR-22-3p模拟物(miR-22-3p mimic)转染CD8^(+)T细胞,检测PD-1蛋白水平。结果与癌旁组织相比,NSCLC肿瘤组织中LINC01137表达(3.357±0.548 vs 1.011±0.371)明显升高;与健康志愿者相比,NSCLC患者外周血LINC01137表达(3.216±0.342 vs 1.007±0.313)亦明显升高,差异具有统计学意义(t=-17.367,-17.147,均P<0.001)。肿瘤组织LINC01137表达与外周血中LINC01137表达呈正相关(r=0.755,P<0.05)。在A549细胞来源的外泌体中LINC01137显著富集。与Exo+sh-NC组相比,Exo+sh-LINC01137组细胞活力(65.852%±4.715%vs 100.153%±11.934%)及细胞侵袭(21.464%±3.481%vs 43.126%±1.447%)能力显著降低,差异具有统计学意义(t=4.630,9.953,均P<0.01)。NSCLC患者外周血中LINC01137表达和CD8^(+)T细胞百分比呈负相关(r=-0.520,P<0.05)。与Exo+sh-NC组相比,Exo+sh-LINC01137组IFN-γ(3865.314±543.852 pg/ml vs 1786.971±105.982 pg/ml),TNF-α(4631.930±510.715pg/ml vs 1973.242±379.623pg/ml),Granzyme B(3876.496±312.438pg/ml vs 1879.439±287.584pg/ml)和IL-2 mRNA水平(3.286±0.437 vs 1.015±0.314)升高,PD-1+Tim3^(+)CD8^(+)T细胞百分比(7.680%±2.185%vs 18.952%±3.216%)降低,差异具有统计学意义(t=-6.497,-7.237,-8.146,-7.310,5.021,均P<0.01)。miR-22-3p是LINC01137的靶基因。与Exo+NC mimic组相比,Exo+miR-22-3p组PD-1蛋白水平(0.384±0.087 vs 1.003±0.147)显著降低,差异有统计学意义(t=6.277,P<0.01)。结论NSCLC患者肿瘤组织及外周血中LINC01137表达显著上调;NSCLC细胞来源的外泌体中LINC01137通过靶向CD8^(+)T细胞中miR-22-3p并抑制其表达,诱导CD8^(+)T细胞耗竭,促进NSCLC细胞免疫逃逸。

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.

TRANSFORMING-GROWTH FACTOR-PI PREFERENTIALLY INHIBITS THE INDUCTION OF CYTOTOXICITY IN HUMAN T CELLS STIMULATED VIA CD28

Generally, TGF-βs are held to down- regulate the growth of immune cells and to inhibit the development of certain differentiated functions, such as the induction of LAK activity by IL-2. In the present study, the effects of TGF-β1 on the proliferation and cytotoxicity of human PBMC activated by anti-CD3 or and-CD3 plus anti-CD28 was investigated. The results demonstrated that TGF- β1 clearly inhibits the induction of cytotoxic ability in human PBMC stimulated via CD3 or CD3 and CD28 ( P<0. 01) , without significantly altering the proliferative response to these stimuli, at the tested doses of TGF-β1. Co-stimulation with IL-2 was hardly altered, suggesting that TGF-β1 action is affected by the nature of the costimulatory signals.

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.

抑制细胞毒性CD8^(+)T细胞治疗白癜风研究进展

白癜风是一种色素脱失性皮肤病,易诊治难根治,致病机制可能涉及遗传、自身免疫、氧化应激、黑色素细胞破坏等方面,最终致病环节为细胞毒性CD8^(+)T细胞异常增殖靶向损伤黑素细胞。综述了阻断细胞毒性CD8^(+)T细胞应激损伤黑素细胞途径治疗白癜风研究进展。

CD4^(+)和CD8^(+)T细胞在结核病免疫应答中的作用

结核分枝杆菌是世界范围内单一感染源致死亡的主要原因,并且已有大量人群被感染后处于长期潜伏感染状态。机体清除结核分枝杆菌主要依赖于特异性免疫应答,其中T淋巴细胞作为参与机体细胞免疫的主要细胞,其增殖和活化在机体特异性免疫应答中发挥着至关重要的作用。笔者分析了结核分枝杆菌感染机体后,T淋巴细胞亚群中的CD4^(+)T和CD8^(+)T细胞在结核分枝杆菌感染免疫应答中的作用,并且对机体抗结核分枝杆菌感染免疫应答的机制进行总结,为进一步深入探索适应性免疫的抗结核感染网络、结核病的诊断及临床研制新型疫苗提供借鉴。

TARGETED ACTIVATION OF CD_(8)^(+) CELLS AND INFECTION OF β_2-MICROGLOBULIN-DEFICIENT MICE FAIL TO CONFIRM A PRIMARY PROTECTIVE ROLE FOR CD_(8)￣(+) CELLS IN EXPERIMENTAL LEISHMANIASIS

CD8+ T cells play an important role in the immunologic control of intracellular pathogens,particularly viruses,Leishmania are obligate intracellular parasites of macrophages in the mammalian host,and previous studies using deletion of CD8+ cells by administration of mAb to infected animals have suggested a protective role for these cells.Two complementary approaches were used to define more carefully the role of CD8+ cells in leishmaniasis.In BALB/c mice susceptible to Leishmania major(L. major)infection, targeted activation of CD8+ cells was attempted by immunization with nonapeptides derived from the conserved major outer surface protein of the organism,8p63, that contained the consensus binding motif for MHC class 1 H-2kd molecules.Two of the nonapeptides induced CTL activity in subsequently infected BALB/c mice that could be elicited against p815 cells pulsed either with peptide or lysates of L.major.purified CD8+ T cells from immunized mice had elevated levels of IFN-γ mRNA transcripts as compared to unimmunized mice.Despite evidence for activation of CD8+ cells,none of the mice immunized with nine different peptides alone or in combination were protected from progressive disease. In a second series of experiments, β2-microglobulin deficient mice that lack CD8+ cells were infected with L. major and the course of infection monitored.These mice cured disease as'rapidly as β2-m +/- and +/+ littermates,and cure was associated with comparable levels of IFN-Y mRNA in the draining lymph node population. Neither of these approaches was able to confirm a substantive role for CD8+ T cells in the primary protective response to L.major.