结直肠癌患者外周血CD4＋ CD25high调节性T细胞的作用

目的探讨结直肠癌患者外周血CD4＋CD25high调节性T细胞（Treg）的作用。方法采用流式细胞仪测定36例结直肠癌患者及30例健康对照者外周血中CD4＋CD25highFoxP3 Treg、CD4＋CD25highTreg、CD4＋CTLA-4 Treg及CD4＋T细胞水平。结果与健康对照组比较,结直肠癌组患者外周血CD4＋CD25highFoxP3 Treg显著增多（P〈0.01）,CD4＋CD25high Treg和CD4＋CTLA-4 T细胞也增多（P〈0.05）。结论CD4＋CD25highFoxP3 Treg可能在结直肠癌发展过程中起着重要作用。     

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。     

CD4+CD25+调节性T细胞在结直肠癌中的表达及其意义

背景与目的:CD4 CD25 调节性T细胞(Treg细胞)是体内自然发生的调节性T细胞的重要亚群,其在体内不仅参与自身免疫性疾病、移植排斥反应等,还在多种恶性肿瘤的发生、发展及免疫治疗中发挥重要作用.本研究探讨CD4 CD25 Treg细胞在结直肠癌中的表达及其意义.方法:收集30例结直肠癌患者的肿瘤组织及远离肿瘤部位组织,用流式细胞术和免疫组织化学方法对肿瘤组织中浸润的CD4 CD25 Treg细胞进行定量及定位分析.结果:CD4 CD25 Treg细胞含量:远离肿瘤部位组织为(5.5±1.3)%,肿瘤组织为(24.1±4.8)%,远离肿瘤部位组织与肿瘤组织相比,差异有显著性(t=5.155,P=0.002);有淋巴结转移者为(27.9±3.6)%,无淋巴结转移者为(20.3±1.3)%,差异有显著性(t=3.489,P=0.025).结论:结直肠癌患者肿瘤组织中浸润的CD4 CD25 Treg细胞的比率显著升高,且与结直肠癌的进展有关.     

结直肠癌患者外周血CD4+CD25high调节性T细胞分析

目的探讨结直肠癌患者外周血高表达CD25的CD4+调节T细胞(CD4+CD25highTr)的比率变化特点及其临床意义.方法采用流式细胞术检测37例结直肠癌患者外周血CD4+CD25high调节T细胞水平,并进行分层分析.结果结直肠癌患者外周血高表达CD25 T细胞水平占总CD4+T细胞的(4.59±1.62)%(n=37),与健康对照组比较(2.04±1.03)%(n=15),差异有统计学意义(t=2.007 582;P=0.000 001 2).该CD4+CD25highTr细胞低表达CD45RA,不表达CD69.Ⅰ～Ⅲ期结直肠癌患者外周血CD4+CD25highTr占总T细胞的(3.93±1.48)%(n=17),Ⅳ期结直肠癌患者外周血CD4+CD25highTr细胞占总T细胞的(5.18±1.54)%(n=20),明显高于Ⅰ～Ⅲ期患者,差异有统计学意义(t=2.080 91;P=0.015).结论结直肠癌患者外周血CD4+CD25highTr细胞比率明显升高,可能在结直肠癌的发生发展中起重要作用.     

结直肠癌患者外周血CD4+ CD25high调节性T细胞的作用

目的探讨结直肠癌患者外周血CD4+CD25high调节性T细胞(Treg)的作用。方法采用流式细胞仪测定36例结直肠癌患者及30例健康对照者外周血中CD4+CD25highFoxP3 Treg、CD4+CD25highTreg、CD4+CTLA-4 Treg及CD4+T细胞水平。结果与健康对照组比较,结直肠癌组患者外周血CD4+CD25highFoxP3 Treg显著增多(P<0.01),CD4+CD25high Treg和CD4+CTLA-4 T细胞也增多(P<0.05)。结论CD4+CD25highFoxP3 Treg可能在结直肠癌发展过程中起着重要作用。     

乳腺癌患者外周血CD4+CD25+调节性T细胞的检测及意义

目的探讨乳腺癌患者外周血中CD4+CD25+调节性T细胞的变化及意义.方法采用流式细胞技术检测64例乳腺癌患者外周血中CD4+CD25+调节性T细胞,采用ELISA法检测外周血中转化生长因子-β1(TGF-β1)的表达水平.结果乳腺癌患者外周血中CD4+CD25+调节性T细胞占T淋巴细胞的百分比为(5.1±2.9)%,高于乳腺良性肿物患者和健康志愿者(P均<0.05).乳腺癌患者外周血中CD4+CD25+T细胞水平与肿物大小、TGF-β1呈正相关(r分别为0.511和0.253),与CD8+CD28+T细胞和NK细胞呈负相关(r分别为-0.243和-0.301).结论乳腺癌患者外周血中具有免疫抑制活性的CD4+CD25+调节性T细胞水平较高,对乳腺癌患者具有免疫抑制作用.     

CD4+CD25+调节性T细胞在肿瘤免疫领域的研究进展

CD4^ CD25^ 调节性T细胞(Regulatory T cell，Treg)在维持自身免疫耐受中起重要作用。本文就其在肿瘤免疫领域的研究进展进行综述。     

肿瘤患者CD4+CD25+调节性T细胞流式细胞术分析

目的:分析比较肿瘤患者和健康人外周血CD4+CD25+调节性T细胞的特点,为肿瘤免疫学研究及治疗探索新方法.方法:收集并分离30例肿瘤患者和32例健康人的外周血单个核细胞(PBMCs),用荧光标记的抗人CD4及抗人CD25单抗标记肿瘤患者和健康人PBMCs细胞,FCM检测CD4+CD25+Treg细胞,分析CD4+CD25+Treg细胞在肿瘤患者和健康人中的差别.结果:肿瘤患者的CD4+CD25+Treg细胞百分数明显高于健康人(年龄<55者62.4 vs 40.4;年龄≥55者53.1 vs 31.0,P<0.05).结论:肿瘤患者的CD4+CD25+Treg细胞高于健康对照,为肿瘤免疫治疗提供新策略,通过删除CD4+CD25+Treg细胞,有可能增强抗瘤免疫反应.     

非小细胞肺癌患者外周血CD4+CD25+调节T细胞的检测及其临床意义

目的探讨非小细胞肺癌患者外周血CD4 CD25 调节T细胞水平的特点及其临床意义。方法采用流式细胞术检测61例非小细胞肺癌患者外周血CD4 CD25 调节T细胞水平,并进行分层分析。结果非小细胞肺癌组外周血CD4 CD25 调节T细胞水平(27.14±12.74)%,明显高于30例健康人群对照组CD4 CD25 T细胞水平(14.02±5.42)%(P<0.01),而且该CD4 CD25 调节T细胞CD45RA(-),CD69(-)。进一步分层分析显示与正常对照组比较,随肺癌进展,外周血CD4 CD25 T细胞水平显著升高,进展期尤其明显[Ⅲ期(25.97±5.94)%,n=20,P<0.01;Ⅳ期(35.13±13.27)%,n=26,P<0.001]。结论非小细胞肺癌患者外周血CD4 CD25 调节T细胞水平明显高于正常,且与肿瘤的发生发展密切相关。去除这群细胞可能有效诱导肿瘤免疫,为肿瘤治疗提供一种新的方法。     

胃癌患者外周血CD4+CD25+调节性T细胞的检测及其临床意义

目的探讨胃癌患者外周血CD4 CD25 调节性T细胞水平的特点及其临床意义。方法采用流式细胞术检测35例胃癌患者外周血CD4 CD25 调节性T细胞水平并进行分层分析。结果健康对照组CD4 CD25 调节性T细胞水平为(8.69±2.28)%,35例胃癌患者CD4 CD25 调节性T细胞比例为(17.77±8.45)%,统计学有显著差异(P<0.01);进一步分层分析显示随疾病进展外周血CD4 CD25 调节性T细胞水平升高,在III期、IV期尤为明显,统计学有极显著差异(P<0.01)。结论胃癌患者外周血CD4 CD25 调节性T细胞水平的升高与胃癌免疫功能低下及肿瘤的发生发展密切相关,去除这群细胞可有效诱导肿瘤免疫,为肿瘤治疗提供一种新的方法。     

T-Cell Membrane CD45RA (2H4) and CD45RO (UCHL1) Determinants: I, Diverse Patterns of Expression in Mature (Post-Thymic) T-Cell Proliferations

By simultaneous two- and three-colour flow cytometry, this study analysed the expression of membrane CD45RA (2H4) and CD45RO (UCHL1) determinants by normal thymocytes (n = 5) and peripheral blood lymphocyte subpopulations (CD4⁺, n = 21; CD8⁺, n = 12; CD8^dim+, n = 12) and compared these patterns with those of T-cells from representative CD4⁺CD8^- (n = 8), CD4⁺CD8⁺ (n = 2), CD4^-CD8⁺ (n = 10) and CD4^-CD8^- (n = 1) proliferations. These comprised cases of prolymphocytic leukaemia (T-PLL, n = 5), adult T-cell leukaemia-lymphoma (ATLL, n = 2), Sezary Syndrome (SS, n = 4), chronic lymphocytic leukaemia (T-CLL, n = 4), and lymphoproliferative disease of granular lymphocytes (LDGL, n = 5). Normal thymocyte fractions, of which a mean of 85% cells co-expressed membrane CD4 and CD8, were predominantly (mean 89%) 2H4^-UCHL1⁺ with the remaining cells consisting of 2H4^intUCHL1⁺ and 2H4⁺UCHL1^- components. Further analysis showed that virtually all CDla⁺ thymocytes were UCHL1⁺ whereas the CD1a^- fraction comprised similar proportions of both UCHL1^- and UCHL1⁺ subpopulations. Similarly, normal blood CD4⁺, CD8⁺ and CD8^dim+ lymphocytes showed reciprocal CD45RA/CD45RO expression and could be phenotypically grouped into 2H4⁺UCHL1^- 2H4^intUCHL1⁺ and 2H4^-UCHL1⁺ subpopulations. Mean proportions of 48% and 68%, for CD4⁺ and CD8⁺ lymphocytes respectively, showed a composite 2H4⁺UCHL1^- phenotype, whereas the percentage of NK-associated CD8^dim+ cells with this phenotypic pattern was considerably higher (mean, 85%). Normal lymphocyte subpopulations lacking both determinants (2H4^-UCHL1^-) were only rarely noted. Comparing normal patterns of CD45RA/CD45RO expression with those of the T-cell proliferations revealed diverse and abnormal patterns of staining for 3/6 of the CD4⁺CD8^- SS and ATLL, and for 5/5 of the T-PLL (CD4⁺CD8^-, n = 2; CD4⁺CD8⁺, n = 2; and CD4^-CD8⁺, n = 1) cases studied. In contrast, the nine cases of CD4^-CD8⁺ T-CLL and LDGL all showed CD45RA/CD45RO staining patterns similar to that of normal CD8⁺/CD8^dim+ blood lymphocytes (i.e. a predominance of 2H4⁺UCHL1^- cells). Although the variant CD45RA/CD45RO pattern types of the CD4⁺ proliferations did not appear to be related to either the diagnostic category or other phenotypic characteristics, the high proportion of abnormal patterns within this case group suggests that recognition of these abnormalities may be potentially relevant to the differentiation of benign and malignant CD4⁺ proliferations and, in addition, may be of aetiological importance with respect to the diverse acquired defects in immunity commonly seen in patients with such disorders.     

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

Objective To analyse the dysfunction of immunity and clinical significance in patients with cardiac cancer.Methods The level of CD4+ CD25hi CD127low Treg cells were detected by flow cytometry (FCM),and serum IL-10 and TGF-β1 levels were determined by enzyme linked immunosorbent assay (ELISA) kit in 56 patients with cardiac cancer.15 healthy volunteers were tested as normal controls.The clinical data of each patient were collected and analyzed. Results There was a significantly higher percentage of CD4+ CD25hi CD127low Treg cells in patients with cardiac cancer (5.73±1.56)% than that (4.45±1.06)% of healthy volunteers (P<0.01).The IL-10 and TGF-β1 levels in the serum of patients with cardiac cancer were also significantly higher than that of healthy volunteers (P<0.05).There was a positive correlation between levels of IL-10.TGF-β1 and the levels of CD4+ CD25hi CD127low Treg cells.The number of CD4+ CD25hi CD127low regulatory T cells in the peripheral blood of cardiac cancer patients were significantly correlated with clinical stages and metastasis lymph node.Conclusion The CD4+ CD25hi CD127low Treg cells in the peripheral blood of cardiac cancer patients is significantly increased in comparison with that in healthy volunteers,and was also correlated with different stages.The abnormal levels of CD4+ CD25hi CD127low Treg cells may be related to tumor progression in patients with cardiac cancer.     

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

Objective To analyse the dysfunction of immunity and clinical significance in patients with cardiac cancer.Methods The level of CD4+ CD25hi CD127low Treg cells were detected by flow cytometry (FCM),and serum IL-10 and TGF-β1 levels were determined by enzyme linked immunosorbent assay (ELISA) kit in 56 patients with cardiac cancer.15 healthy volunteers were tested as normal controls.The clinical data of each patient were collected and analyzed. Results There was a significantly higher percentage of CD4+ CD25hi CD127low Treg cells in patients with cardiac cancer (5.73±1.56)% than that (4.45±1.06)% of healthy volunteers (P<0.01).The IL-10 and TGF-β1 levels in the serum of patients with cardiac cancer were also significantly higher than that of healthy volunteers (P<0.05).There was a positive correlation between levels of IL-10.TGF-β1 and the levels of CD4+ CD25hi CD127low Treg cells.The number of CD4+ CD25hi CD127low regulatory T cells in the peripheral blood of cardiac cancer patients were significantly correlated with clinical stages and metastasis lymph node.Conclusion The CD4+ CD25hi CD127low Treg cells in the peripheral blood of cardiac cancer patients is significantly increased in comparison with that in healthy volunteers,and was also correlated with different stages.The abnormal levels of CD4+ CD25hi CD127low Treg cells may be related to tumor progression in patients with cardiac cancer.     

T-Cell Membrane CD45RA (2H4) and CD45RO (UCHL1) Determinants: II, Aberrant HLA-ABC Expression by CD45RA and CD45RO Cell Subpopulations of Mature CD4+ T-Cell Leukaemias

Six thymocyte suspensions, 10 normal blood CD4⁺ CD8^- lymphocyte-enriched fractions and leukaemic cells from 24 patients with CD4⁺ mature T-cell lymphoid malignancy (five Sezary Syndrome, six adult T-cell leukaemia-lymphoma and 13 cases of T-cell prolymphocytic leukaemia) were examined in this study for the expression of membrane HLA-ABC by CD45RA (2H4) and CD45RO (UCHL1) subpopulations. These analyses showed that the main increase in HLA-ABC expression by normal CD4⁺ CD8^- blood lymphocytes (mean 490 to 760 FITC units) paralleled the loss of membrane 2H4 whilst the acquisition of UCHL1 was not associated with any significant change in HLA-ABC staining intensity. The sequence of 2H4 differentiation by normal thymocytes, based on the observed increasing levels of HLA-ABC staining intensity appeared to be (a) CD1a ⁺ 2H4^- UCHL1⁺ (25 HLA-ABC fluorescent units), (b)CD1a^-2H4^intUCHL1⁺ (134 units), and (c) CD 1a^- 2H4 ⁺ UCHL1 ^- (197 units). Quantitative estimates of membrane HLA-ABC expression by leukaemic T-cells revealed marked heterogeneity between individual cases irrespective of diagnostic subgroup. Based on the lower observed limits for normal CD4⁺ 2H4⁺ (318 units) and CD4 ⁺ 2H4^- (478 units) fractions, 14% and 38% respectively of the leukaemic 2H4⁺ and 2H4^- components examined showed reduced HLA-ABC expression. Two cases showed very low membrane HLA-ABC levels that were within the range observed for normal CD1a^- thymocytes. In contrast, HLA-ABC staining intensities exceeding that of corresponding normal CD4⁺ 2H4⁺ (710 units) and CD4⁺ 2H4^- (1286 units) subpopulations were seen in a high proportion (65%) of leukaemic 2H4 ⁺ components, with only 14% of 2H4^- fractions showing raised levels and, in two cases, these staining intensities exceeded three times the normal observed limits. In addition to the quantitative differences in HLA-ABC expression, a remarkably consistent (81% of evaluable cases) feature of the leukaemic T-cells was that the 2H4^-UCHL1⁺ subpopulation in CD4⁺ malignancies had a lower HLA-ABC level than the 2H4⁺UCHL1 subpopulation. This was in marked contrast to normal post-thymic T-cells where increasing HLA-ABC expression was seen with increasing UCHL1 (or decreasing 2H4) staining. These results suggest that leukaemic T-cells have an aberrant intra-thymic and post-thymic sequence of 2H4/UCHL1 expression which has become 'uncoupled' from CD1a/HLA-ABC expression.     

B细胞非霍奇金淋巴瘤患者外周血CD4＋CD25high CD127low调节性T细胞水平及其临床意义

目的 探讨外周血CD4＋ CD25high CD127low调节性T细胞（Treg细胞）在B细胞非霍奇金淋巴瘤（B-NHL）患者外周血中表达水平及其临床意义.方法 采用流式细胞术检测100例初诊B-NHL患者及50名健康对照者外周血CD4＋ CD25high CD127low Treg细胞表达水平,进行统计学分析.结果 健康对照者外周血CD4＋ CD25high CD127low Treg细胞中位表达水平为5.00％,初诊B-NHL患者为7.20％,两者之间差异具有统计学意义（P＜ 0.001）.男性患者外周血CD4＋ CD25high CD 127low Treg细胞水平高于女性患者（P＜0.01）,乳酸脱氢酶（LDH）增高患者外周血CD4＋ CD25high CD127low Treg细胞水平高于LDH正常患者（P＜0.01）,Ⅲ～Ⅳ期患者外周血CD4＋C D25high CD 127low Treg细胞水平较Ⅰ～Ⅱ期患者增高（P＜0.01）,有B症状患者外周血CD4＋ CD25high CD127low Treg细胞水平高于无B症状患者（P＜0.01）.而不同年龄、国际预后指数评分、有无大包块患者之间外周血CD4＋ CD25high CD 127low Treg细胞水平差异均无统计学意义（均P＞0.05）.结论 B-NHL患者体内存在免疫抑制,在男性、LDH增高、有B症状及晚期患者中CD4＋ CD25high CD127low Treg细胞水平明显增高.检测其水平对于判断B-NHL的预后有一定价值.     

结直肠癌患者外周血CD4＋ CD25high调节性T细胞检测及其表型分析

目的分析CD4＋ CD25high T细胞（Treg）在50例结直肠癌患者外周血中的分布及其表型特征,初步探讨其临床意义。方法采用流式细胞术检测结直肠癌患者及健康人外周血中Treg的比例,探讨其表型特征并分析Treg比例与临床分期间的关系。结果与正常对照（3.1±1.46）%相比,结直肠癌患者外周血中Treg比例明显增加（11.22±5.47）%,二者差异有显著统计学意义（P〈0.01）;手术后,患者外周血中Treg的比例相比术前显著下降（P〈0.01）;患者外周血CD4＋ CD25high T细胞水平在疾病的临床分期间差异无显著性（P〉0.05）。表型分析显示：Treg高表达CD45RO、HLA-Ⅰ、Foxp3分子,较高水平表达OX40、GITR、CD152、CD95、CD95L、B7-H1,较低水平表达CD62L,低表达CD80、CD86、B7-H4分子,基本不表达CD45RA、CD69分子,HLA-DR、CD154的表达水平在个体间差异较大,同时大部分细胞CD127表达阴性。结论结直肠癌患者外周血CD4＋ CD25high T细胞水平明显升高,可能与患者肿瘤免疫功能低下密切相关。     

肺癌患者外周血白细胞介素-17+CD4+T细胞和白细胞介素-17+CD8+T细胞表达的意义

目的 检测白细胞介素(IL)-17+CD4+T(Th17)细胞和IL-17+CD8+ T(Tc17)细胞在肺癌患者外周血中的表达水平,探讨二者在肺癌免疫中的作用及临床意义.方法 采用流式细胞术(FCM)检测60例肺癌患者及40例健康对照者外周血中Th17和Tc17细胞占CD;T细胞的比例.结果 肺癌组外周血中Th17细胞[(1.795±0.623)％]和Tc17细胞[(0.865±0.357)％]比例分别高于对照组[(1.405±0.256)％、(0.640±0.204)％],(t=28.944,P＜ 0.001;t=14.051,P＜ 0.001).两组内Th17细胞与Tc17细胞的表达水平均呈正相关(肺癌组r=0.770,P＜0.05;对照组r=0.532,P＜0.05).Th7细胞和Tc17细胞表达均与临床分期有关(F值分别为4.882、3.633,均P＜0.05),但与病理类型无关(均P＞0.05).结论 肺癌患者体内Th17细胞和Tc17细胞表达升高,二者可能参与了肺癌的发生、发展;Th17与Tc17细胞的表达水平可作为评价肺癌患者免疫功能状态的新指标,能为病情监测提供参考.     

外周T细胞淋巴瘤患者中PD-1的表达对CD45RA+、CD45RO+T细胞的影响

目的:探讨外周T细胞淋巴瘤(PTCL)患者淋巴瘤组织中PD-1的表达与外周血中初始和记忆T细胞水平的关系.方法:20例PTCL患者采用免疫组化法检测淋巴瘤组织中PD-1的表达,采用流式细胞术检测CHOP方案化疗前后外周血中CD4+CD45RA+、CD4+CD45RO+、CD8+CD45RA+和CD8+CD45RO+T细胞的比例,分析PD-1的表达与T细胞亚群的关系.结果:PD-1、PD-L1蛋白在PTCL患者中表达升高,PD-1阳性患者疗效较差.PD-1阳性患者中CD4+T细胞、CD8+T细胞明显低于PD-1阴性患者;PD-1阴性组患者CD4+CD45RO+、CD8+CD45RA+、 CD8+CD45RO+T细胞明显高于PD-1阳性患者,差异有统计学意义.结论:PTCL患者中存在PD-1/PD-L1蛋白的表达异常,PD-1阳性患者存在更明显的T细胞免疫功能缺陷.     

CD45RA-Foxp3high but not CD45RA+Foxp3low suppressive T regulatory cells increased in the peripheral circulation of patients with head and neck squamous cell carcinoma and correlated with tumor progression

Background

T regulatory cells (Tregs) contribute to the progression of head and neck squamous cell carcinoma (HNSCC) by suppressing antitumor immunity. However, little is known regarding the functional heterogeneity of Tregs in HNSCC patients.

Methods

Using multicolor flow cytometry, the frequency of three Treg subsets, separated on the basis of CD45RA and Foxp3, from the peripheral circulation of newly-presenting HNSCC patients (19 oral cavity squamous cell carcinoma, 20 hypopharyngeal squamous cell carcinoma, 18 nasopharyngeal squamous cell carcinoma, 19 oropharyngeal squamous cell carcinoma, and 36 laryngeal squamous cell carcinoma) were assessed with regard to 31 healthy donors and clinicopathological features. Moreover, the functional capacity of each Treg subsets was evaluated based on CD45RA and CD25 expression.

Results

The frequency of Tregs in the peripheral circulation of HNSCC patients as a whole cohort was higher than in healthy donors (P < 0.0001). However, the frequency of Tregs was similar between patients with oral cavity squamous cell carcinoma and healthy donors (P = 0.269). Further dividing Tregs into three subsets based on Foxp3 and CD45RA expression revealed that the frequency of CD45RA^-Foxp3^high Tregs and CD45RA^-Foxp3^lowCD4⁺ T cells in patients with HNSCC developing from different subsites was higher than in healthy donors (P < 0.0001, P < 0.0001), whereas the frequency of CD45RA⁺Foxp3^low Tregs was lower than in healthy donors (P < 0.0001). Functionally study revealed that CD45RA^-CD25⁺⁺⁺ Tregs significantly inhibit the proliferation of CD4⁺CD25^- T cells (P < 0.001) and secrete lower levels of cytokines (P < 0.01) compared with CD45RA^-CD25⁺⁺CD4⁺ T cells. Importantly, the frequency of CD45RA^-Foxp3^high Tregs positively correlate with tumor stage (P < 0.0001) and nodal status (P < 0.0001).

Conclusions

CD45RA^-Foxp3^high Tregs increase in the peripheral circulation of HNSCC patients, and correlate with tumor stage and nodal status; suggesting a role in tumor progression which may be manipulated by future immunotherapy.