碳离子(^(12)C^(6+))抑制JAK2/STAT3通路促进肺癌中CD8^(+)T细胞浸润的研究北大核心CSCD

目的探索碳离子(^(12)C^(6+))照射后JAK2/STAT3通路的改变及下游蛋白FOXP3调控的肺癌中CD8+T细胞的浸润差异。方法基于C57BL/6小鼠Lewis荷瘤模型的RNA测序分析,筛选出碳离子照射后肺癌中显著改变的JAK2/STAT3通路及相关的差异表达基因及蛋白如FOXP3。利用R软件“GSVA”中ssGSEA免疫浸润算法,探索FOXP3与肺癌免疫微环境中主要免疫细胞浸润的相关性并基于碳离子联合STAT3抑制途径(氯硝柳胺)对肺癌中CD8+T细胞浸润进行分析。结果碳离子照射后,肺癌中JAK2/STAT3通路被抑制,相关基因和蛋白表达下调。基于ssGSEA算法的免疫评分显示,FOXP3表达与肺癌免疫微环境中CD8+T细胞浸润呈显著负相关。通过碳离子照射联合STAT3抑制剂氯硝柳胺,进一步明确了靶向JAK2/STAT3通路对于增加肺癌中CD8^(+)T细胞浸润的协同作用。结论碳离子(^(12)C^(6+))可以通过靶向JAK2/STAT3通路与免疫治疗发挥协同增效的作用。     

干细胞样CD8+T细胞在抗肿瘤免疫治疗中的地位与演进效应

以抗细胞程序性死亡-1/细胞程序性死亡-配体1(programmed cell death-1/programmed cell death-ligand 1,PD-1/PD-L1)为代表的免疫检查点抑制剂(immune checkpoint inhibitors,ICIs)治疗显现了CD8⁺T细胞在治疗和可能治愈恶性肿瘤方面的潜力。但仅约20%的患者对ICIs治疗获益,因此阐明CD8⁺T细胞在免疫微环境中的有效抗肿瘤功能,及其分子和空间的决定因素尤为重要。具有自我更新、增殖分化和杀伤肿瘤细胞能力的干细胞样CD8⁺T细胞亚群,在介导抗肿瘤免疫效应方面扮演极为重要的角色。本文就干细胞样CD8⁺T细胞在抗肿瘤免疫循环中的地位与演进效应进行综述。     

肺腺癌浸润CD8+T细胞表达CD39的机制

目的：探讨肿瘤浸润CD8⁺T细胞表达CD39的可能机制。方法：通过TCGA数据库的肺腺癌(LUAD)组织及正常肺组织转录组数据分析CD39在LUAD组织和正常肺组织中的表达差异及其对患者预后的影响,分析CD39表达与T细胞浸润、激活的关系。用小鼠LUAD Lewis细胞建立小鼠皮下移植瘤模型,FCM检测淋巴结、脾脏以及移植瘤组织中CD8⁺CD39⁺T细胞。收集Lewis细胞培养上清液作为条件培养基(CCM),免疫磁珠法(MACS)分选CD8⁺T细胞、CD11b⁺细胞;在培养基中分别加入CCM、IL-6和采取非接触或接触培养方式进行培养,探索CD8⁺T细胞表达CD39的可能机制。结果：CD39在LUAD组织中呈低表达(P<0.01),其表达水平与LUAD患者OS、T细胞浸润和激活水平均呈正相关(P<0.05或P<0.001)。FCM检测结果显示,在移植瘤组织中CD8+CD39⁺T细胞的比例明显高于淋巴结及脾脏(P<...     

CD8+T细胞耗竭发生和特征及其与肿瘤免疫治疗耐药的研究进展

免疫检查点抑制剂(immune checkpoint inhibitors,ICIs)是治疗多种肿瘤的重要手段,但耐药成为其最大难题。肿瘤免疫治疗耐药与肿瘤微环境(tumor microenvironment,TME)密切相关,TME中CD8⁺T细胞耗竭不仅持续性高表达抑制性受体(inhibitory receptors,IRs),同时也是导致ICIs耐药的关键环节,靶向IRs为克服免疫治疗耐药提供了新思路。本文将重点对CD8⁺T细胞耗竭发生和特征及其与肿瘤免疫治疗耐药性相关的研究进行综述。     

CD4+CAR-T细胞亚群在肿瘤治疗中作用的研究进展

嵌合抗原受体T(CAR-T)细胞疗法是目前治疗癌症最有效的一种免疫治疗方法,但CAR-T细胞功能障碍很大程度限制其自身对癌症治疗的效果。T细胞功能的差异以及记忆和效应T细胞的作用被证明在CAR-T细胞治疗中极为重要。CD8⁺T细胞作为发挥抗癌作用的主要效应细胞一直是研究焦点,而对CD4⁺T细胞的关注较少。CD4⁺T细胞不仅可以通过激活CD8⁺T细胞使其杀伤肿瘤细胞,还可以独立发挥抗肿瘤作用。现有研究发现,细胞因子、共刺激域和细胞代谢等因素均可影响CD4⁺CAR-T细胞亚群的增殖和分化。本文主要综述了CD4⁺CAR-T细胞亚群在治疗肿瘤中的重要性以及影响其增殖分化因素的研究进展,为进一步研发高效的CAR-T细胞提供新思路。     

食管鳞癌组织Snail与PDL-1和CD8+T表达及预后分析

目的 探讨食管鳞癌(ESCC)组织中Snail与细胞程序性死亡配体1(PDL-1)、CD8⁺T之间的相关性及预后观察。方法 收集222例2014-02-01-2015-12-30就诊于新乡医学院第一附属医院,手术后经病理确诊为ESCC的癌组织。应用免疫组织化学法检测Snail、PDL-1和CD8⁺T表达,Spearman相关分析三者的相关性,Kaplan-Meier法绘制生存曲线,Cox回归模型进行预后因素分析。结果 Spearman相关分析显示,Snail与PDL-1蛋白表达呈低度正相关,r=0.210,P=0.002;与CD8⁺T浸润呈低度负相关,r=-0.174,P=0.009。Cox单因素分析显示,远处转移(HR=4.060,95%CI为2.472～6.668,P<0.001)、G₂+G₃(HR=1.562,95%CI为1.080～2.260,P=0.018)、T₃+T₄(HR=2.053,95%CI为1.339～3.1...     

结直肠癌及腺瘤患者CD4+、CD8+和CD28+T淋巴细胞的亚群变化及临床意义

背景与目的：结直肠癌（colorectal cancer,CRC）严重影响患者生存。探讨肿瘤微环境（tumor microenvironment,TME）T细胞亚群在CRC和腺瘤中的表达及意义。方法：用免疫组织化学法和流式细胞术对51例健康人（对照组）、46例结直肠腺瘤（腺瘤组）、100例CRC（癌症组）和15例CRC术后（癌术后组）患者进行T细胞亚群检测。结果：① 对照组、腺瘤组及癌症组3组中CD4⁺T细胞的阳性率分别是90.00%、43.75%及32.65%,CD8⁺T淋巴细胞的阳性率分别是30.00%、56.25%及75.51%,CD28⁺T淋巴细胞的阳性率分别是42.86%、30.00%及20.00%。② 对照组、腺瘤组及癌症组3组中CD4⁺、CD4⁺/CD8⁺、CD28⁺、CD8⁺CD28⁺和CD8^－CD28⁺逐渐降低,CD8⁺、CD8⁺CD28^－逐渐增加（P＜0.05）;癌术前术后T细胞亚群差异有统计学意义（P＜0.05）。结论：① CRC微环境T细胞亚群中CD4⁺、CD4⁺/CD8⁺、CD28⁺、CD8⁺CD28⁺和CD8^－CD28⁺呈递减趋势,CD8⁺、CD8⁺CD28^－呈递增趋势,且在癌前病变腺瘤中已逐步出现上述趋势变化。② CRC患者行肿瘤切除术后,其T细胞亚群有所恢复,故在一定程度上,CRC中T细胞亚群的变化可以早期预测结直肠疾病的发展。     

CD+T细胞及Treg细胞表面PD-1水平与肺癌患者术后预后的关系

目的 探讨CD⁺T细胞及调节性T(Treg)细胞表面程序性死亡受体1(PD-1)水平与肺癌患者术后预后的关系。方法 选取行肺癌根治术的肺癌患者126例,统计术后2年内预后不良发生率,并根据其预后情况将其分为良好组和不良组,单因素分析肺癌患者术后预后不良的影响因素,Logistic回归分析肺癌患者术后预后不良的影响因素,受试者工作特征曲线(ROC)分析CD⁺T细胞及Treg PD-1水平对肺癌患者术后预后的预测价值。结果 两组肿瘤淋巴结转移(TNM)分期、CD3⁺、CD3⁺CD4⁺、CD3⁺CD8⁺及Treg PD-1水平比较,均有显著差异(P<0.05);Logistic回归分析显示,CD3⁺、CD3⁺CD4⁺是肺癌患者术后预后不良的独立保护因素,CD3⁺CD8⁺、Treg PD-1是肺癌患者术后预后不良的独立危险因素(...     

CD4+/CD8+ T淋巴细胞在肝细胞癌组织中的浸润及与预后的相关性研究

目的 检测CD4⁺/CD8⁺ T淋巴细胞在肝细胞癌（hepatocellular carcinoma,HCC）组织中的浸润程度,并分析其与预后的相关性。方法 收集行肝切除术的HCC患者215例,采用免疫组化技术检测CD4⁺/CD8⁺ T淋巴细胞在HCC癌组织中的浸润程度,根据浸润情况比较患者肝切除术后无瘤生存率和总生存率。结果 CD4⁺ T淋巴细胞高浸润和低浸润比例分别为60.9%和39.1%。CD4⁺ T淋巴细胞高浸润组患者总生存率和无瘤生存率均显著高于低浸润组（P=0.015,P=0.038）。CD8⁺ T淋巴细胞高浸润和低浸润比例分别为34.9%和65.1%。CD8⁺ T淋巴细胞高浸润组患者的总生存率和无瘤生存率亦显著高于低浸润组患者（P=0.033,P=0.047）。结论 CD4⁺或CD8⁺ T淋巴细胞低浸润可能与HCC患者术后不良预后相关。     

乳腺浸润性导管癌组织中ALDH1+/CD133+干细胞样细胞与血管生成的关系

目的 探讨乳腺浸润性导管癌组织中肿瘤干细胞标志物ALDH1、CD133的表达及其与肿瘤血管生成的关系。方法 应用免疫组织化学双染法检测120例乳腺浸润性导管癌组织中ALDH1⁺/CD133⁺ 干细胞样细胞,单染法检测血管性标记CD34、CD105及VEGF的表达情况。统计ALPH1⁺/CD133⁺干细胞样细胞与临床病理因素;CD34、CD105与VEGF的关系。结果 25.83％（31/120）的病例存在ALDH1⁺/CD133⁺干细胞样细胞,ALDH1⁺/CD133⁺干细胞样细胞与ER、VEGF的表达及MVD均相关（P＜0.05）,但与年龄、肿瘤直径、PR、Her-2、组织学分级及淋巴结转移均无关（P＞0.05）。结论 乳腺浸润性导管癌组织中ALDH1⁺/CD133⁺干细胞样细胞可能通过调节VEGF的表达促进肿瘤新生血管的生成。     

Regulatory (FOXP3+) T cells as target for immune therapy of cervical intraepithelial neoplasia and cervical cancer

Regulatory (FOXP3⁺) T cells (T_regs) comprise a subpopulation of CD4⁺ T cells that suppress autoreactive immune cells, thereby protecting organs and tissues from autoimmunity. T_regs have also been detected in human malignancies and their depletion or inactivation substantially improves cellular antitumor immunity in preclinical studies. Novel therapeutic strategies for cervical cancer and precancerous cervical intraepithelial neoplasia (CIN) focus on immune-modulatory and cancer vaccination approaches. In this context, the frequency of T_regs in cervical cancer and precancerous CIN could influence therapeutic strategies. We determined the frequency of infiltrating CD4⁺ and CD8⁺ T cells as well as FOXP3⁺ T_regs in high-grade CIN lesions (CIN III) and cervical carcinoma compared to colon carcinoma, skin melanoma, and bronchial carcinoma. We show that human papilloma virus-derived lesions have a significantly higher number of infiltrating lymphocytes and FOXP3⁺ T_regs compared to three other common tumor entities. In addition we explored the therapeutic effect of agonistic anti-glucocorticoid-induced tumor necrosis factor receptor family-related protein antibodies that, by single systemic application, inactivate T_regs and induce strong intratumoral invasion of CD8⁺ T cells and complete tumor eradication in 70% of treated animals. The large number of T_regs in human papilloma virus-derived lesions suggests a pivotal role of T_regs for counteracting the host immune response. We therefore regard CIN and cervical cancer as prime targets for new immune-based non-invasive therapies. ( Cancer Sci 2009; 100: 1112–1117)     

胆固醇代谢调控CD8+T细胞抗肿瘤作用的研究进展

CD8+T细胞又名细胞毒性T淋巴细胞(cytotoxic T lymphocyte,CTL),具有直接杀死病原体感染细胞和癌细胞的作用.然而,CD8+T细胞常常丧失其效应功能,继而限制肿瘤微环境中的抗肿瘤免疫,因此,如何重新激活CD8+T细胞的抗肿瘤效力是目前需要解决的问题.最近研究发现,胆固醇代谢在肿瘤中发挥重要作用...     

Human epithelial ovarian carcinoma cell-derived cytokines cooperatively induce activated CD4+CD25−CD45RA+ naïve T cells to express forkhead box protein 3 and exhibit suppressive ability in vitro

Regulatory T cells play an important role in tumor escape from host antitumor immunity. Increased frequencies of CD4⁺CD25⁺ regulatory T cells have been documented in the tumor sites, malignant effusions, and peripheral blood of patients with ovarian carcinoma. However, the mechanism involved remains unclear. In the present study, we collected high-purity human CD4⁺CD25⁻CD45RA⁺ naïve T cells by microbead cell separation. These cells did not express FOXP3 by single-cell analysis, and few cells expressed FOXP3 when they were activated with anti-CD3/CD28 dual signal. However, more cells expressed FOXP3 when the supernatant of human epithelial ovarian carcinoma cell culture was added, yet not the supernatant of normal human ovarian surface epithelia cell culture. Neutralization assays revealed that neutralizing antibody against transforming growth factor β (TGF-β), interleukin-10, and interleukin-4 did not abrogate elevated FOXP3 expression induced by carcinoma cell culture supernatant, whereas neutralizing leukemia inhibitory factor (LIF) partially abrogated FOXP3 expression, but LIF alone could not increase FOXP3 expression in activated naïve T cells. Further, an in vitro coculture suppression assay showed that these cells could suppress the proliferation of autologous CD4⁺CD25⁻CD45RA⁻ T cells. In summary, our findings show that ovarian carcinoma cells are able to induce expression of FOXP3 and exhibit suppressive ability in activated naïve T cells by producing soluble substances, and multiple cytokines involve in the induction of FOXP3 expression. ( Cancer Sci 2009)     

CD4+ T lymphocytes: a critical component of antitumor immunity

Both prophylactic and therapeutic vaccines targeting a wide variety of cancers are being developed. Because of the potency of cell-mediated immunity, many vaccine strategies are focused on activating tumor-specific cytotoxic CD8⁺ T lymphocytes. CD4⁺ T lymphocytes are a key element in optimal activation of CD8⁺ T cells and in the maintenance of immune memory, and therefore their activation is critical for cancer vaccine efficacy. This article reviews the mechanisms by which CD4⁺ T cells facilitate tumor immunity and the vaccine strategies that enhance CD4⁺ T cell activity.     

Murine Asialo GM1+CD8+ T Cells as Novel Interleukin-12-responsive Killer T Cell Precursors

Freshly isolated CD8⁺ T cells, but not CD4⁺ T cells, contained 20–30% of asialo GM1⁺ (ASGM1⁺) T cells which were distinct from ASGM1⁺NK1.1⁺ natural killer cells. This novel ASGM1⁺CD8⁺ T cell subpopulation showed a strong proliferative response to interlenkin-12 (IL-12) in the presence of IL-2. Culture of ASGM1⁺CD8⁺ T cells with IL-12 plus IL-2 allowed the generation of anomalous killer T cells concomitantly with the accumulation of cytolytic molecules. Moreover, ASGM1⁺CD8⁺ T cells produced high levels of interferon-γ (IFN-γ), but not IL-4, upon stimulation with IL-12 plus IL-2. Such immune responses were not observed in ASGM1⁻ CD8⁺ T cell snbpopulations constituting the majority of CD8⁺ T cells. These results demonstrated that ASGM1⁺CD8⁺ T cells are a novel subpopulation of IL-12-responsive and IFN-γ-producing killer T cell precursors.     

The Roles of CD8+ and CD4+ Cells in Tumor Rejection

In vivo administrations of anti-Lyt-2.2 (CDS) mAb and anti-L3T4 (CD4) mAb selectively eliminated CD8⁺ cells amd CD4⁺ cells, respectively. The relative potencies of CD8⁺ cells and CD4⁺ cells and their roles in primary tumor rejections were studied by investigating the effects of these mAbs on tumor growth. CD8⁺ cells were themselves fully capable of mediating rejection in 5 different tumor rejection systems: two radiation leukemia virus (RadLV)-induced leukemias, B6RV2 and BALBRVD, a radiation-induced leukemia BALBRL♂1, and a plasmacytoma BALBMOPC-70A in CB6F₁ mice, and a Friend virus-induced leukemia B6FBL-3 in B6 mice. On the other hand, CD4⁺ cells were capable of resisting tumor growth of B6FBL-3, but not of the other four tumors. Furthermore, for efficient rejection of CB6F₁UV+^˚l sarcoma by CB6F₁ mice, synergy of CDS⁺ and CD4⁺ cells was necessary. Blocking of UV+^˚ 1 rejection was abrogated by delayed administration of anti-L3T4 (CD4) mAb but not anti-Lyt-2.2 (CDS) mAb, indicating the involvement of CD4⁺ cells in only the initial phase of rejection.     

Cytotoxic Activity of CD4+ T Cells against Autologous Tumor Cells

The ⁵¹Cr-release assay is mostly applied to detecting the cytotoxic activity of CD8⁺ T cells, and little is known about the activity of CD4⁺ T cells. Therefore, the correlation between the cytotoxic activity of CD4⁺ or CD8⁺ T cells and the incubation period with autologous tumor cells was analyzed by two methods. The incubation periods were 4 and 20 h (4 h and 20 h assay) for the ⁵¹Cr-release assay. Eight pairs of tumor cells and T cells were assayed. T cells were fractionated into CD4⁺ and CD8⁺ T cells by using magnetic beads and panning methods, and those cells were activated by culture with recombinant interleukin-2 and immobilized anti-CD3 monoclonal antibody. In 6 out of 8 cases, no cytotoxic activity of CD4⁺ T cells was detected by the 4 h assay, whereas cytotoxic activity was detected in all cases in the 20 h assay. The cytotoxic activities in 20 h assay of CD4⁺ T cells were increased 67-fold in comparison with the activities in 4 h assay (range: 5–197). In the case of CD8⁺ T cells, cytotoxic activities were detected in 6 out of 8 cases in the 4 h assay. The lytic unit ratio of CD4⁺ and CD8⁺ T cells was calculated as 1.5 in the 20 h assay (range: 0.2->7.2) versus 0.4 in the 4 h assay (range: < 0.1–1.3). Cytotoxic activities in colorimetric assay using Crystal Violet with a 24 h incubation were similar to those in the 20 h ⁵¹Cr-release assay in all eight cases. These results indicate that CD4⁺ T cells have cytotoxic activity as strong as that of CD8⁺ T cells towards autologous tumor cells.