白细胞介素-35对肝细胞癌患者CD4+CD25+CD127dim/-调节性T细胞的调控作用

目的 观察肝细胞肝癌(HCC)患者外周血白细胞介素-35(IL-35)水平和CD4⁺CD25⁺CD127^dim/-调节性T细胞(Treg)比例,评估IL-35对Treg数量和功能的影响。方法 共有51例HCC患者和25例对照者,分离外周血单个核细胞(PBMC)和血浆,实时定量PCR法检测PBMC中IL-35亚基mRNA相对表达量,酶联免疫吸附试验(ELISA)法检测外周血IL-35水平,流式细胞术检测PBMC中CD4⁺CD25⁺CD127^dim/-Treg比例。使用重组人IL-35(1 ng/ml)刺激HCC组PBMC 24 h,观察Treg比例和EBI3、IL-12p35 mRNA相对表达量的变化。分选17例HCC患者的Treg,经重组人IL-35刺激后与自体PBMC共培养48 h,通过检测细胞增殖和细胞因子表达评估IL-35对Treg功能的影响。组间比较采用t检验或配对t检验。结果 HCC组血浆IL-35水平显著高于对照组[(1024±218.0)pg...     

贲门癌患者外周血CD+4 CDhi25 CDlow127 调节性T细胞水平及其临床意义

 目的　探讨贲门癌患者的免疫功能异常及其临床意义。方法　采用流式细胞术（FCM）和酶联免疫吸附（ELISA）方法分别检测 56例贲门癌患者、15名健康人外周血中CD+4 CDhi25 CDlow127 调节性T细胞（Treg细胞）和血清IL-10、TGF-β1水平,结合临床资料进行分析。结果　56例贲门癌患者外周血中CD+4 CDhi25 CDlow127 Treg细胞占CD+4淋巴细胞的比例为（5.73±1.56）%,与健康对照组的（4.45±1.06）%相比,差异有统计学意义（P＜0.01）;血清中IL-10和TGF-β1含量均明显高于健康对照（P＜0.05）。贲门癌患者外周血CD+4 CDhi25 CDlow127 Treg细胞水平与血清IL-10和TGF-β1含量呈正相关。贲门癌患者外周血CD+4 CDhi25 CDlow127 Treg 细胞数量与患者临床分期、淋巴结转移有关。结论　贲门癌患者外周血中CD+4 CDhi25 CDlow127 Treg细胞表达增高,与临床分期有关,提示Treg水平异常与贲门癌的发生发展密切相关。     

DC-CIK对胃癌合并腹水患者外周血CD4+CD25+调节性T细胞增殖及功能的影响

目的:探讨DC-CIK对胃癌合并腹水患者外周血CD4+CD25+调节性T胞(Treg细胞)比例及功能的影响。方法:60例胃癌合并腹水患者,于输注DC-CIK前1天及DC-CIK治疗结束后1周分别采集外周血。流式细胞术检测外周血Treg细胞的比例,RT-PCR法检测其Foxp3mRNA表达情况;将分选出的Treg细胞和CD4+CD25-T细胞分为单纯Treg细胞组（A组）、1∶1混合培养（B组）、单纯CD4+CD25-细胞组（C组）进行培养,3H-TdR掺入法检测Treg细胞抑制CD4+CD25-细胞增殖的能力。结果:治疗后外周血Treg细胞占CD4+T细胞的比例较疗前显著下降［(6.21±1.37)% vs (9.38±1.06)%,P＜0.05］。治疗后Treg细胞Foxp3mRNA表达水平较治疗前显著下降［(56.18±13.25)% vs (85.26±11.58)%,P＜0.05］。治疗后Treg对CD4+CD25-T细胞抑制增殖能力较治疗前明显下降［(37.31±4.16)% vs (48.92±5.25)%,P＜0.05］。结论:输注DC-CIK免疫治疗,可显著降低胃癌合并腹水患者外周血Treg细胞比例,下调Foxp3mRNA表达水平,降低Treg细胞免疫抑制功能,有利于诱导抗肿瘤免疫效应。     

贲门癌患者外周血CD4+CD25hiCD127low调节性T细胞水平及其临床意义

目的 探讨贲门癌患者的免疫功能异常及其临床意义.方法 采用流式细胞术(FCM)和酶联免疫吸附(ELISA)方法分别检测56例贲门癌患者、15名健康人外周血中CD4+CD25hiCD127lowc调节性T细胞(Treg细胞)和血清IL-10、TGF-β1水平,结合临床资料进行分析.结果 56例贲门癌患者外周血中CD4+CD25hiCD127lowTreg细胞占CD4+淋巴细胞的比例为(5.73±1.56)%,与健康对照组的(4.45±1.06)%相比,差异有统计学意义(P<0.01);血清中IL-10和TGF-β1含量均明显高于健康对照(P<0.05).贲门癌患者外周血CD4+CD25hiCD127low Treg细胞水平与血清IL-10和TGF-β1含量呈正相关.贲门癌患者外周血CD4+CD25hiCD127low Treg细胞数量与患者临床分期、淋巴结转移有关.结论 贲门癌患者外周血中CD4+CD25hiCD127low Treg细胞表达增高,与临床分期有关,提示Treg水平异常与贲门癌的发生发展密切相关.     

原发性肝癌患者CD4+CD25+CD127low调节性T细胞的检测及意义

探讨CD4+CD25+CD127low调节性T细胞（Tregs）在原发性肝细胞性肝癌（HCC）患者外周血中的变化及其临床意义。方法:采集40例乙型肝炎病毒（HBV）相关的HCC患者［巴塞罗那临床肝癌（BCLC）分期A期患者7例、B期患者8例、C期患者20例、D期患者5例］、35例慢性乙型肝炎（CHB）患者及28例正常健康人的外周抗凝血,应用CD4（PE-CY5）、CD25（FITC）、CD127（PE）三种特异性荧光抗体标记后,通过流式细胞术对CD4+CD25+CD127lowTregs水平进行三色荧光抗体检测。结果:HCC患者外周血CD4+CD25+CD127lowTregs占CD4+T细胞的百分比显著高于正常健康人（P<0.001）和CHB患者（P=0.017）,CHB患者外周血CD4+CD25+CD127lowTregs占CD4+T细胞的百分比高于正常健康人（P=0.035）;HCC中BCLC分期为C期的患者外周血CD4+CD25+CD127lowTregs占CD4+T细胞的百分比显著高于A期患者（P=0.020）和B期患者（P=0.019）。结论:CD4+CD25+CD127lowTregs水平异常增高可能是HCC免疫逃逸的一个重要机制,且其变化水平与临床病情的进展存在一定的相关性。      

CD+4CD+25调节性T细胞在小儿急性白血病发病中的作用

 目的　观察CD+4 CD+25 调节性T（Treg）细胞在急性白血病患儿及非白血病患儿外周血中的变化,研究CD+4 CD+25 Treg细胞在小儿急性白血病发病中的作用。方法　采用流式细胞术检测急性白血病初诊患儿组20例及非白血病患儿对照组20例外周血CD+4 CD+25 Treg细胞的数量及比例。结果　初诊患儿组及对照组外周血CD+4 CD+25 CD-127 Treg细胞占CD+4 T细胞的比例分别为（11.57±1.04）%和（6.75±0.75）%,在初诊患儿组高于对照组（t＝16.808,P＜0.001）。结论　急性白血病患儿外周血中CD+4 CD+25 CD-127 Treg细胞数量升高,提示CD+4 CD+25 Treg细胞可能在白血病的发生、发展中起一定作用。     

CD+4 CD+25 调节性T细胞与急性髓细胞白血病关系的研究

 目的　探讨CD+4 CD+25 调节性T细胞（CD+4 CD+25 Treg细胞）在急性髓细胞白血病（AML）患者化疗前后外周血中的表达水平及其临床意义。方法　采用流式细胞术检测AML初发组（A组）、缓解3年内组（B组）、缓解3年以上组（C组）及正常对照组外周血中CD+4 CD+25 CDlow127 T细胞所占比例。结果　A组和B组外周血中CD+4 CD+25 Treg细胞比例均高于对照组（P＜0.05）;C组外周血CD+4 CD+25 Treg细胞比例与对照组相比差异无统计学意义（P＞0.05）;B组外周血中CD+4 CD+25 Treg细胞比例高于C组（P＜0.05）;A组外周血中CD+4 CD+25 Treg细胞比例明显高于B、C两组（P＜0.05）。结论　CD+4 CD+25 Treg细胞与AML发生有关,尤其与3年内容易复发密切相关。     

肺癌患者外周血中CD+8 CD-28及CD+4 CDhigh25 CDlow127调节性T细胞的表达及其临床意义

 目的　观察肺癌患者外周血CD+8CD+28、CD+8CD-28及CD+4CDhigh25CDlow127调节性T（Treg）细胞的表达水平,并对CD+8CD-28和CD+4CDhigh25CDlow127 Treg细胞行相关性分析,探讨其在肺癌免疫中的作用及临床意义。方法　采用流式细胞术检测60例初治肺癌患者外周血CD+8CD+28、CD+8CD-28及CD+4CDhigh25CDlow127 Treg细胞的表达率,以60名健康体检者作为对照,分析各指标与肺癌生物学及临床特征之间的关系。结果　肺癌组CD+8CD-28 T细胞［（58.430±15.749）%］和CD+4CDhigh25CDlow127 Treg细胞［（7.365±2.025）%］表达均分别显著高于对照组的（41.057±15.436）%、（6.648±1.669）%,差异有统计学意义（t＝6.102,P＜0.05;t＝2.115,P＜0.05）。肺癌组CD+8CD+28 T细胞表达［（41.570±15.739）%］低于对照组［（58.700±15.298）%］,差异有统计学意义（t＝-6.043,P＜0.05）。上述三个指标的表达与性别、年龄及病理学类型无关（P＞0.05）。随着TNM分期增加,CD+4CDhigh25CDlow127 Treg细胞表达逐渐升高,其中Ⅳ期和ⅢA期之间差异有统计学意义（t＝3.898,P＜0.05）。而CD+8CD-28、CD+8CD+28 T细胞的表达与临床分期无关（P＞0.05）。CD+8CD-28与CD+4CDhigh25CDlow127Treg细胞的表达无线性相关（r＝-0.169,P＞0.05）。结论　CD+8CD-28、CD+4CD25high CD127low Treg细胞在肺癌患者外周血呈高表达,CD+8CD+28T细胞呈低表达,这可能是肺癌患者免疫功能低下的原因之一。CD+8CD-28与CD+4CDhigh25 CDlow127 Treg细胞之间无相关性,但两者联合检测可对了解肺癌患者免疫功能,并为肺癌寻找特异性免疫治疗及预后评价提供新的参考。     

甲状腺乳头状癌患者外周血CD4+CD25+CD127low/-Treg的检测及其临床意义

目的:检测CD4+ CD25+ CD127low/-调节性T细胞(regulatory T cell,Treg)在甲状腺乳头状癌(papillary thyroid carcinoma,PTC)患者外周血的比例,探讨其临床意义。方法:选择2010年8月至2011年2月在上海交通大学医学院附属仁济医院进行手术治疗的PTC患者40例及甲状腺腺瘤患者30例,以CD4+ CD25+ CD127low/-为标志,采用流式细胞术检测PTC患者及甲状腺腺瘤患者外周血中CD4+ CD25+ CD127low/-Treg的比例,分析其与PTC临床病理特征的关系。结果:与甲状腺腺瘤患者比较,PTC患者外周血中CD4+ CD25+ CD127low/- Treg比例明显升高[(7.836±1.668)% vs(5.365±1.156)%,P<0.05];外周血CD4+ CD25+ CD127low/-Treg比例与PTC临床分期、颈部淋巴结转移相关(均P<0.05),与年龄、性别及肿瘤大小无关(P>0.05)。结论:PTC患者外周血CD4+ CD25+ CD127low/-Treg比例显著增加,且与PTC临床分期、颈部淋巴结转移相关。     

卵巢癌患者腹水及外周血CD4+ CD25+调节性T细胞含量及抑制功能的研究

  目的   研究卵巢癌患者腹水及外周血单个核细胞（PBMC）中CD4+ CD25+调节性T细胞（Treg）的表达差异、免疫调节功能及其含量与化疗、复发的关系,探究其在卵巢癌腹腔微环境中发挥免疫调节的具体作用及意义。   方法   采用流式细胞术分别检测27例卵巢癌患者腹水及28例卵巢癌患者PBMC中CD4+ CD25+/CD4+ T细胞百分比,并根据收集标本时患者临床特征进行分组,比较处于初治（PD）、化疗后（AC）及复发（RD）3个阶段卵巢癌患者腹水及PBMC中Treg含量的差异。应用免疫磁珠分选卵巢癌患者腹水及外周血Treg,与经羧基荧光素二醋酸盐琥珀酰亚胺酯（carboxyfluorescein succinimidylester,CFSE）标记的自CD4+CD25- T细胞按不同比例（0:1,1:1,1:2及1:4）共培养,检测Treg免疫抑制功能。   结果   卵巢癌患者腹水中CD4+ CD25+/CD4+ T细胞百分比（28.25±14.06）%较PBMC中（14.6±4.74）%显著增高（P < 0.000 1）。卵巢癌患者PD、AC及RD 3个阶段腹水及PBMC中Treg含量均显示为AC>RD>PD,且均有显著统计学意义（P < 0.000 1）。体外实验结果显示,卵巢癌患者腹水中CD4+ CD25+ Treg可有效抑制CFSE标记的自体CD4+ CD25- T细胞增殖,且抑制功能较外周血显著增强（P < 0.01）。   结论   卵巢癌腹腔微环境中存在CD4+ CD25+Treg,且其含量及免疫抑制功能较外周血PBMC显著增高,提示卵巢癌腹腔内更易发生免疫逃逸作用。卵巢癌患者化疗后及复发阶段腹水及PBMC中CD4+ CD25+ Treg含量均大于原发阶段,提示化疗可能促进卵巢癌患者体内Treg含量升高,而Treg升高可能参与促进肿瘤复发。       

CD4+CD25+调节性T细胞与CD4+T、CD8+T细胞在结直肠癌组织中的分布

 目的 分析CD4+CD25+ FOXP3+调节性T细胞（Treg）与CD4+T、CD8+T在结直肠癌（colorectal carcinoma, CRC）组织中的分布及其与临床病理特征之间的关系。方法 收集42例CRC新鲜手术标本,应用冰冻切片、免疫组织化学SP法检测肿瘤组织和癌旁组织中FOXP3+、CD4+T和CD8+T阳性细胞数。结果 CRC患者肿瘤组织中FOXP3表达水平显著升高,与癌旁组织相比差异有统计学意义（P<0.01）;中低分化组Treg细胞数明显高于高分化组（P<0.01）;淋巴结转移组Treg细胞数明显高于无淋巴结转移组（P<0.05）;癌巢内CD4+、CD8+T细胞数及CD4+/CD8+值显著低于间质（P<0.01）;Ⅲ+Ⅳ期、淋巴结转移组癌巢内CD4+/CD8+比值显著低于Ⅰ+Ⅱ期及无淋巴结转移组（P<0.05）;CRC中Treg数量与癌巢内CD4+/CD8+比值显著负相关（r=-0.605, P<0.01）。结论 CRC的发生发展可能与其癌组织局部微环境中Treg数量变化相关,肿瘤局部Treg数量的增多与T淋巴细胞亚群比例失调可能成为肿瘤免疫逃逸的机制之一。     

外周血CD3+、CD4+、CD8+、CD19+水平与喉癌患者预后的相关性

目的 探讨喉癌患者免疫水平的变化与病情进展的关系.方法 选取56例喉癌患者,用流式细胞仪检测不同分组的喉癌患者外周血淋巴细胞和NK细胞的表达水平.结果 手术后患者CD19+显著性下降(P＜0.05),而NK细胞显著上升(P＜0.05);有淋巴结转移的患者比无淋巴结转移的患者NK细胞上升,CD4+下降,二者均有统计学差异(P＜0.05);病理分期中Ⅲ～Ⅳ期的患者比Ⅰ～Ⅱ期表现出CD19+显著性下降(P＜0.05),CD4+则显著上升(P＜0.05).在术后随访18个月中,有复发或转移的患者比无转移且无复发患者CD4+下降,而NK细胞和CD19+上升,均具有统计学差异(P＜0.05).结论 喉癌患者免疫状态和病情之间有一定的联系.     

Epithelial mesenchymal transition correlates with CD24+CD44+ and CD133+ cells in pancreatic cancer

The epithelial-mesenchymal transition (EMT) has been linked to induction of a stem-cell like phenotype, characterized by altered cell surface marker expression and increased tumor formation. The aim of this study was to investigate whether EMT correlates with CD24+CD44+ and CD133+ cells in pancreatic cancer. The morphology of untreated and gemcitabine-treated SW1990 gemcitabine-resistant cells and normal SW1990 cells were compared. NF-κB p65 expression was knocked down using siRNA. Vimentin and E-cadherin expression were analyzed using western blotting, and CD24+CD44+, CD133+ cells were quantified by FACS. Additionally, immunohistochemistry of EMT-associated markers and stem cell-associated markers were performed in 41 cases of human pancreatic ductal adenocarcinoma. In SW1990 gemcitabine-resistant cells, gemcitabine induced a mesenchymal cell phenotype, expression of EMT-related molecular markers and increased CD24+CD44+ and CD133+ cells compared to untreated SW1990 gemcitabine-resistant and SW1990 cells. Knockdown of NF-κB p65 inhibited the ability of gemcitabine to increase the proportion of CD24+CD44+ or CD133+ cells and expression of EMT-related molecular markers. In human pancreatic ductal adenocarcinoma, significant correlations were observed between expression of the EMT-associated markers vimentin and E-cadherin, and stem cell-associated markers CD24, CD133 and CD44. This study demonstrated that EMT correlated with CD24+CD44+ and CD133+ cells in pancreatic cancer. This study also suggests that EMT may induce cancer stem-like cells in pancreatic cancer, with different degrees of EMT probability inducing different proportions of CD24+CD44+ and CD133+ cells.     

Immune surveillance: both CD3+ CD4+ and CD3+ CD8+ T cells control in vivo growth of P815 mastocytoma.

The aim of this study was to examine whether a spontaneous immune response controls neoplastic growth in P815-bearing DBA/2 mice, and to characterize the cells involved in tumor resistance in vivo. Several cell lineages such as T-cell-receptor (TcR)-bearing T cells, NK cells and macrophages mediate some anti-tumor activity in vitro. P815 was chosen as a model because it is weakly immunogenic and is a good target both for tumor-specific, MHC-restricted CTL-mediated lysis and for MHC-unrestricted lysis exerted by long-term cultured lymphocytes or activated macrophages. Since most "NK-like activity" in freshly isolated populations appears to be associated with CD3- cells, whereas antigen-specific, MHC-restricted T cells mostly express CD3 determinants, CD3 was a good marker for evaluating the role of T cells and "NK" cells in tumor resistance in vivo. The survival of anti-CD3-treated animals that were inoculated with tumor cells was strongly reduced (mean survival time: 17 days vs. 40 days for the control group) and was associated with increased tumor growth rate. We followed the same approach to define the T-cell subset(s) that mediate(s) this immune response. Both CD4+ and CD8+ T cells were required for induction of immune control on neoplastic growth. The approach used has revealed the important role of CD4+ T cells in immune responses that control in vivo growth of a class-I-positive, class-II-negative tumor and suggests that these cells may play a central role in tumor resistance. Since CD4+ cells are activated by soluble, exogenous proteins, this finding may have important implications for immunotherapy.     

CD4 + CD25 + 调节性T细胞与肿瘤免疫治疗

 CD4＋CD25＋调节性T淋巴细胞（regulatory Tcell，Treg）是一类具有独特免疫调节功能的T淋巴细胞亚群，一般占人外周血CD4＋T细胞的5％～15％。近年来，国内外对这类调节性T细胞的研究已从自身免疫耐受、移植免疫逐渐扩展到肿瘤免疫，认为Treg是形成肿瘤免疫耐受的关键成分。     

CD+4CD+25调节性T细胞与肿瘤免疫

 近期研究发现一个有独特免疫调节功能的T细胞亚群：CD+4 CD+25调节性T细胞,不仅能抑制自身免疫性疾病发生,还参与肿瘤免疫的调节。这群细胞具有免疫无能和免疫抑制特性,与肿瘤免疫逃逸有密切的关系。肿瘤环境中CD+4 CD+25调节性T细胞增加,导致肿瘤免疫失调,去除这群细胞可有效诱导肿瘤免疫,为肿瘤治疗提供了一种新的思路。     

CD4^＋CD25^＋调节性T细胞与肿瘤的相关性研究

通过了解CD4^＋CD25^＋调节性T细胞（CD4^＋CD25^＋Treg）表面分子的特性和CD4^＋CD25^＋Treg在外周血和组织中的表达,认识CD4^＋CD25^＋Treg在肿瘤免疫调节中的作用,探索其作用的分子机制。     

The impact of CD4+CD25+ Treg on tumor specific CD8+ T cell cytotoxicity and cancer

There is sufficient evidence to suggest that tumor growth elicits specific immune responses, including CD8(+) and CD4(+) T cell responses that may delay tumor growth and could potentially be harnessed to eradicate cancer. Nevertheless the frequent outcome of cancer is lethality associated with uncontrolled growth and dissemination of tumor cells. The failure of the immune response may be naturally programmed and related to a specific subpopulation of CD4(+)CD25(+) regulatory T cells, whose function is to protect us against autoimmunity. Recent investigations have shed light on the in vivo behavior and functions of these cells. It is becoming evident that a major impact of these cells is on the cytolytic action of specific CD8(+) T cells that target the tumor. Inhibition of cytotoxicity is dependent on TGF-beta signaling by the effector cells. Thus, targeting immune regulation may provide a promising approach to the immune therapy of cancer. This approach however could also have unexpected deleterious consequences, as surprising new observations indicate that regulatory T cells can also delay tumor growth by independent mechanisms that relate to their cross talk with the innate immune response to cancer.     

Co-immunization with L-Myc enhances CD8+ or CD103+ DCs mediated tumor-specific multi-functional CD8+ T cell responses

Renal carcinoma shows a high risk of invasion and metastasis without effective treatment. Herein, we developed a chitosan (CS) nanoparticle-mediated DNA vaccine containing an activated factor L-Myc and a tumor-specific antigen CAIX for renal carcinoma treatment. The subcutaneous tumor models were intramuscularly immunized with CS-pL-Myc/pCAIX or control vaccine, respectively. Compared with single immunization group, the tumor growth was significantly suppressed in CS-pL-Myc/pCAIX co-immunization group. The increased proportion and mature of CD11c⁺ DCs, CD8⁺CD11c⁺ DCs and CD103⁺CD11c⁺ DCs were observed in the splenocytes from CS-pL-Myc/pCAIX co-immunized mice. Furthermore, the enhanced antigen-specific CD8⁺ T lymphocyte proliferation, cytotoxic T lymphocyte (CTL) responses, and multi-functional CD8⁺ T cell induction were detected in CS-pL-Myc/pCAIX co-immunization group compared with CS-pCAIX immunization group. Of note, the depletion of CD8 T cells resulted in the reduction of CD8⁺ T cells or CD8⁺CD11c⁺ DCs and the loss of anti-tumor efficacy induced by CS-pL-Myc/pCAIX vaccine, suggesting the therapeutic efficacy of the vaccine was required for CD8⁺ DCs and CD103⁺ DCs mediated CD8⁺ T cells responses. Likewise, CS-pL-Myc/pCAIX co-immunization also significantly inhibited the lung metastasis of renal carcinoma models accompanied with the increased induction of multi-functional CD8⁺ T cell responses. Therefore, these results indicated that CS-pL-Myc/pCAIX vaccine could effectively induce CD8⁺ DCs and CD103⁺ DCs mediated tumor-specific multi-functional CD8⁺ T cell responses and exert the anti-tumor efficacy. This vaccine strategy offers a potential and promising approach for solid or metastatic tumor treatment.     

CD4+CD25+调节性T细胞与抗肿瘤免疫

CD4^＋CD25^＋调节性T（CD4^＋CD25^＋Tr）细胞是一类维持机体自身耐受的T细胞亚群,分布广泛,但不同组织表型有所不同。它们可由胸腺自然产生,也可在外周血中诱导产生,其活化要依赖于特异性抗原的存在。CD4^＋CD25^＋Tr细胞发挥抑制效应是通过细胞接触依赖或分泌细胞因子这两种方式。去除CD4^＋CD25^＋Tr细胞或抑制其功能,重新募集效应性T细胞能够增强机体抗肿瘤作用,这将成为一种可行的肿瘤免疫治疗方法。