卵巢早衰患者外周血CD4~+CD25~+Treg细胞的变化及意义

目的:探讨CD4+CD25+调节性T细胞(即CD4+CD25+Treg细胞)在卵巢早衰发病机制中的作用。方法:流式细胞仪定量检测卵巢早衰(premature ovarian failure,POF)患者、卵巢储备功能下降(diminished ovarian reserve,DOR)患者及健康对照组外周血CD4+T、CD8+T细胞及CD4+CD25+Treg细胞数量,应用3H-thymidine掺入法测定POF患者及对照组外周血CD4+CD25+Treg细胞对效应性T细胞的增殖抑制功能。结果:与对照组相比,POF患者及DOR患者CD4+CD25+Treg细胞比例降低(P<0.01)、POF患者CD4+T/CD8+T细胞比值增高(P<0.05),DOR患者CD4+T/CD8+T细胞比值无明显变化(P>0.05);POF患者免疫抑制功能无明显降低(P>0.05)。结论:CD4+CD25+Treg细胞比例降低与T细胞亚群失衡可能是POF的发病机制。     

宫颈癌患者外周血CD4+CD25+调节性T细胞及其Foxp3表达的临床意义

目的评价宫颈癌患者外周血CD4+CD25+调节性T细胞(Regulatery T cell,Treg)及其Foxp3表达的临床意义。方法收集中国医科大学附属盛京医院自2012年1月至2013年6月手术治疗的早期宫颈癌患者(FIGO分期≤Ⅱ期)34例,高级别宫颈上皮内瘤变(CIN2/3)患者30例以及20例健康女性作为正常对照。流式细胞技术检测各研究组手术前、后及对照组外周血Treg及Foxp3+Treg细胞占CD4+T细胞比例,应用t检验和单因素方差法进行统计学分析。结果比较宫颈癌、CIN2/3及对照组外周血Treg及Foxp3+Treg细胞所占CD4+T细胞比例,各组间差异有统计学意义(P0.05);术后早期(7d),宫颈癌及CIN2/3患者外周血Foxp3+Treg细胞所占比例较术前显著下降(P0.05),但Treg细胞所占比例无显著性变化(P0.05);术后恢复期(1个月),宫颈癌及CIN2/3患者外周血Foxp3+Treg细胞所占比例较术后早期无显著变化,但Treg细胞所占比例较术后早期显著性降低(P0.05)。结论宫颈癌患者外周血Treg细胞及其Foxp3的表达均与病变程度相关;Treg细胞Foxp3的表达具有不稳定性;宫颈癌肿瘤微环境可能是维持Treg细胞分化增殖及Foxp3稳定表达的外在客观条件。     

复发性宫颈癌免疫治疗的研究进展

宫颈癌是女性常见的妇科恶性肿瘤之一,与人乳头瘤病毒（HPV）感染密切相关。尽管该疾病通常可以通过单独手术或手术及辅助放化疗在早期阶段治愈,但对于晚期及复发性转移性宫颈癌尚无有效的疗法。免疫反应是对抗HPV感染和相关癌症的关键因素。近期随着免疫学的发展,免疫治疗已成为晚期及复发性宫颈癌治疗的新策略,这些治疗策略旨在调节免疫反应以抑制肿瘤。现就复发性宫颈癌免疫治疗的临床研究新进展作一综述。     

宫颈上皮内瘤变及宫颈癌患者外周血Treg/Th17细胞及相关细胞因子的检测与临床意义

目的:探讨Treg/Th17平衡失调在宫颈上皮内瘤变(CIN)及宫颈癌病情进展中的作用及其参与CIN及宫颈癌免疫逃避的具体机制。方法:研究对象分为3组:宫颈癌组(34例),CIN组(47组),对照组(30例)。采用流式细胞术检测CIN及宫颈癌患者外周血单个核细胞(PBMC)中Th17细胞及Treg细胞的表达频率,采用酶联免疫吸附试验(ELISA)方法检测相关细胞因子(IL-23、IL-10、IL-17、IL-6、TGF-β)的浓度。结果:与对照组相比,Treg细胞及其相关细胞因子(TGF-β、IL-10)在CIN及宫颈癌患者外周血中的表达均明显增高(P<0.05),且宫颈癌组的表达亦明显高于CIN组(P<0.05)。与对照组相比,Th17细胞及其相关细胞因子(IL-23、IL-17、IL-6)在CIN及宫颈癌患者外周血中的表达均明显降低(P<0.05),且宫颈癌组的表达亦明显低于CIN组(P<0.05)。结论:随着CIN向宫颈癌的进展,Treg/Th17平衡向Treg细胞偏离,这一失衡参与了CIN的进展及宫颈癌的免疫逃避,其具体机制可能与二者在肿瘤免疫内环境相关的细胞因子调节有关。     

卵巢肿瘤患者外周血中T细胞亚群及调节T细胞含量变化的临床意义

目的 探讨T淋巴细胞亚群及调节T细胞含量变化在卵巢肿瘤评价中的临床意义.方法 应用流式细胞仪检测了155例卵巢恶性肿瘤患者及100例卵巢良性肿瘤患者及50例健康人群外周血T淋巴细胞亚群及调节T细胞含量.结果 与健康对照(1.81±0.45)相比,卵巢癌患者外周血中CD4/CD8比值明显降低(0.91±0.31),二者差异有统计学意义(P＜0.01),CD4/CD8比值随卵巢癌分级的升高而下降,但卵巢癌组间差异无统计学意义(P＞0.05).与健康对照组(4.67±1.84)％及卵巢良性肿瘤(5.32±0.83)％相比,卵巢癌患者外周血中Treg比例明显增加(11.86±2.41)％,差异有统计学意义(P＜0.01),Treg比例随卵巢癌分级的升高而升高,卵巢癌组间差异无统计学意义(P＞0.05).结论 卵巢癌患者外周血CD4/CD8比值明显降低,而CD4+ CD25+ Treg细胞水平明显升高,并且与肿瘤的分级有关,可能提示卵巢癌患者机体免疫杀伤功能低下而免疫耐受增加,从而使肿瘤发生免疫逃逸,促使肿瘤的发生发展.     

复发性流产患者外周血T淋巴细胞亚群及CD4^＋辅助性T淋巴细胞亚型的变化

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IROT对宫颈癌Ⅱb患者CD4+/CD8+的影响及血清TNF-α水平的变化与其临床意义

目的　探讨术中大剂量放射治疗 (intraoperationradiationtherapy ,IROT)对宫颈癌Ⅱb患者免疫功能的影响及其临床意义。方法　 2 0 0 3年 6月至 2 0 0 4年 1月西安交通大学第一医院应用流式细胞仪、放射免疫分析法对6 1例宫颈癌Ⅱb患者 (33例行单纯放疗 ,2 8例行IROT)外周血T细胞亚群 (CD4 、CD8 、CD4 CD8 )、血清中肿瘤坏死因子 α(TNF α)的水平进行检测。结果　单纯放疗组、IROT组在放疗前CD4 、CD8 、CD4 CD8 、TNF α差异无显著性 ;放疗后单纯放疗组CD4 、CD4 CD8 、TNF α明显低于IROT组 ,差异有显著性 (P <0 0 5 ) ,CD8 差异无显著性。结论　IROT与单纯放疗相比 ,对患者CD4 CD8 的影响较轻 ,血清中TNF α水平变化较小 ,有利于机体免疫功能的恢复。     

卵巢恶性肿瘤患者外周血自然杀伤细胞活性及T淋巴细胞亚群的改变

目的了解卵巢恶性肿瘤患者外周血自然杀伤细胞（ＮＫ细胞）活性及Ｔ淋巴细胞亚群的改变,并探讨其与肿瘤的组织学类型、细胞分化程度及临床分期的关系。方法采用放射性同位素掺入法及ＯＫＴ单克隆抗体,分别检测卵巢恶性肿瘤患者外周血ＮＫ细胞活性及Ｔ淋巴细胞亚群,并采用妇科良性肿瘤患者作对照。结果卵巢恶性肿瘤患者外周血ＮＫ细胞活性较妇科良性肿瘤患者显著降低（Ｐ＜００１）,ＯＫＴ４细胞显著减小（Ｐ＜００５）,而ＯＫＴ８细胞显著增多（Ｐ＜００５）,ＯＫＴ４／ＯＫＴ８比值显著减少（Ｐ＜００５）。ＮＫ细胞活性及Ｔ淋巴细胞亚群的改变与肿瘤的组织学类型、肿瘤细胞分化程度均无关（Ｐ＞００５）,而与临床分期有关（Ｐ＜００５）,临床分期越晚,其ＮＫ细胞活性越低,ＯＫＴ４细胞越少,ＯＫＴ８细胞越多,ＯＫＴ４／ＯＫＴ８比值越小。结论卵巢恶性肿瘤患者免疫功能是低下的,且临床分期越晚,其免疫功能越低。提示患者的免疫功能与自身体内肿瘤负荷有关。     

原发性痛经患者外周血T淋巴细胞亚群的研究

目的 :探讨原发性痛经患者月经周期中外周血T淋巴细胞亚群的变化 ,分析原发性痛经患者的免疫状况。方法 :原发性痛经患者及正常对照组各 10例 ,分别于月经期、卵泡期、排卵期、黄体期取静脉血 ,应用流式细胞仪测定血浆CD4 + 、CD8+ 百分比值。结果 :原发性痛经患者月经期、卵泡期CD4 + 百分比值及CD4 /CD8的比值显著低于正常对照组 (P <0 0 5 ) ,原发性痛经患者整个月经周期CD8+ 百分值均明显高于正常对照组 (P <0 .0 5 )。结论 :原发性痛经患者存在免疫功能低下     

复发性流产患者外周血T淋巴细胞亚群及CD_4~ 辅助性T淋巴细胞亚型的变化

目的　探讨不明原因的复发性流产 (URSA)与T淋巴细胞免疫功能的关系。方法　采用酶联免疫斑点 (ELISPOT)法对 30例URSA患者和 2 4例正常妇女 (对照 )的外周血T淋巴细胞亚群及CD 4 辅助性T淋巴细胞 (Th细胞 )亚型进行检测。结果　与对照相比 ,URSA患者外周血中CD 4 辅助性T淋巴细胞百分率增多 (分别为 45 %、6 0 % ,P <0 0 5 ) ,Th1细胞的百分率升高 (14%与 2 6 % ,P<0 0 1) ,Th2细胞的百分率降低 (2 0 %与 7% ,P <0 0 5 ) ,Th1/Th2细胞比值增大 (0 73与 3 80 ,P<0 0 1)。结论　URSA的发生可能与CD 4 辅助性T淋巴细胞的增多及Th1细胞介导的细胞免疫功能的增强有关。     

调节性T细胞和Notch1信号通路在原因不明复发性自然流产中的作用

目的探讨调节性T细胞(Treg)和Notch1信号通路在原因不明复发性自然流产(URSA)中的作用。方法流式细胞仪检测URSA患者(URSA组)及正常妊娠妇女(对照组)蜕膜CD4~+CD25~+T细胞Treg表达比例,real time RT-PCR及Western blotting检测蜕膜中CD4~+T细胞中Notch1信号通路和叉头转录因子家族3(Foxp3)表达情况。结果 URSA组CD4~+CD25~+T细胞/淋巴细胞、CD4~+Foxp3~+T细胞/淋巴细胞和CD4~+Foxp3~+T细胞/CD4~+T细胞比例均低于对照组(P0.05)。URSA组CD4~+T细胞中Notch1-Ic、RBPJκ、Foxp3 m RNA及蛋白表达均低于对照组。结论 URSA患者蜕膜CD4~+T细胞中Notch1信号通路和Foxp3表达下调,CD4~+CD25~+T细胞表达比例下降,提示URSA患者Notch1信号通路和Foxp3表达下调可能阻碍CD4~+T细胞转化为CD4~+CD25~+T细胞,进而诱发免疫排斥,诱导流产。     

子宫颈上皮内病变及子宫颈癌患者外周血及局部免疫功能分析#br#

Objective?To investigate the distribution of T cell subsets in peripheral blood of different cervical lesions and the level of cytokines secreted, and it’s difference in the location and density of immune cell distribution in the epithelium and stroma of cervical lesion tissues. Methods?We used CD series cell detection slide quality control kit to detect the distribution of T cell subsets in peripheral blood of 55 patients with different cervical lesions, analyzed the changes of peripheral blood cytokines, and analyzed the localization and density distribution of T lymphocytes (CD4+T, CD8 +T) in the tissue microenvironment of 64 patients with different cervical lesions by immunohistochemistry. Results?The level of CD4+T and CD8+T in peripheral blood of patients with low grade lesion (LSIL) were lower, with 479.13±229.65 unit and 378.00±231.74 unit respectively. The level of CD4+T and CD8+T in high grade cervical squamous intraepithelial lesion(HSIL)was 816.30±284.65 unit and 668.75±268.92 unit respectively, and the level of CD4+T and CD8+T in carcinoma was 824.80±330.65 unit and 564.00±58.31 unit. Meanwhile the concentration of IL-17 in the serum of LSIL patients was higher than that of HSIL (P<0.05). CD8+T cells in epithelial and stroma tissues of cervical cancer were higher than those in control, LSIL and HSIL tissues, with statistical significance (P<0.05). However, CD4+T cells in the stroma of cancer tissues were higher than those of control, LSIL and HSIL tissues, with statistical significance (P<0.001). Conclusion?The patients with cervical lesion have abnormal immune function in the systemic immunity and local microenvironment of the lesion, both CD8+T cells and CD4+T cells play a key role in eliminating HPV-infected cervical epithelial cells. The development of cervical lesions is related to the cellular inflammatory factors.     

Less circulating mucosal-associated invariant T cells in patients with cervical cancer

Objective

Mucosal-associated invariant T cells (MAITs) are important for immune defense against infectious pathogens and regulation of various inflammatory diseases. However, their roles in cancer are rarely reported. Since cervical cancer is one of the diseases involving mucosal tissue, we try to investigate the association between circulating MAITs and cervical cancer.

Differentiation of metastatic from non-metastatic lymph nodes in patients with uterine cervical cancer using diffusion-weighted imaging

Objective

This study aims to determine the diagnostic value of diffusion-weighted imaging (DWI) in the differentiation of metastatic lymph nodes from non-metastatic lymph nodes in uterine cervical cancer.

Methods

In 42 patients who underwent lymph node dissection for uterine cervical cancer, conventional MRI and DWI examinations were performed before surgery. Of the 1109 total dissected pelvic lymph nodes, 188 enlarged nodes with a short-axis diameter of 5 mm or greater were included for further analysis. Each of the size-based criteria (i.e., short-axis diameter and long-axis diameter) and ADC-based criteria (i.e., mean ADC, minimum ADC, mean rADC (relative ADC) and minimum rADC) were compared between metastatic lymph nodes and non-metastatic lymph nodes.

Results

There were statistically significant differences between metastatic and non-metastatic lymph nodes in the short-axis diameter, long-axis diameter, mean ADC, minimum ADC, mean rADC and minimum rADC (P < 0.001). The Az of the minimum ADC (0.990) was greater than that of the other ADC-based criteria (0.974, 0.939, 0.976 for mean ADC, mean rADC and minimum rADC, respectively) and all size-based criteria (0.878 for short-axis diameter and 0.858 for long-axis diameter) (P < 0.05). Using the minimum ADC criteria (≤ 0.881 × 10^− 3mm²/s), the sensitivity and specificity for differentiating metastatic from non-metastatic lymph nodes were 95.7% and 96.5%, respectively.

Conclusions

DWI is feasible for differentiating metastatic from non-metastatic pelvic lymph nodes in patients with uterine cervical cancer and minimum ADC could be served as a representative marker.     

Re-classification of uterine cervical cancer cases treated with radical hysterectomy based on the 2018 FIGO staging system

ObjectiveWe re-classified patients with stage IB–II disease (based on the 2008 system) and compared the outcomes with those obtained after using the 2018 International Federation of Gynecology and Obstetrics (FIGO) staging system.Materials and methodsWe reviewed the data of 154 patients with cervical cancer who underwent radical hysterectomy at our hospital during 2006–2016. Pathological, histological, and radiographic data were used to re-classify the cases based on the 2018 FIGO system. We compared these outcomes to those obtained after using the 2008 FIGO assignments. Overall survival (OS) was calculated from primary therapy initiation until death or the last follow-up examination.ResultsThe histological types were squamous cell carcinoma (108 cases) and others (46 cases). The 2008 FIGO system assignments were stage IB1, IB2, IIA1, IIA2, and IIB (87, 27, seven, five, and 28 patients, respectively). The new 2018 FIGO system assignments were stage IB1, IB2, IB3, IIA1, IIA2, IIB, and IIIC1 (52, 26, 16, six, three, 21, and 30 patients, respectively). Re-classification to stage IIIC1 disease was observed for previously assigned stage IB1, IB2, IIA1, IIA2, and IIB cases (10, seven, two, two, and nine cases, respectively). The median OS durations based on the 2018 FIGO system were 71.7, 61.1, and 62.3 months for patients with stage IB1, IB2, and IB3 (p = 0.04) disease, respectively. The new stage IB3/IIA2/IIB cases had longer OS than the old stage IB2/IIA2/IIB cases. A positive computed tomography (CT) finding of nodal involvement was observed in 37% of cases with pathological confirmation of pelvic lymph node (LN) involvement. Using CT to identify pelvic LN metastasis had a sensitivity of 37% and specificity of 93%.ConclusionThe 2018 FIGO staging system for cervical cancer after radical hysterectomy showed a better ability to differentiate survival outcomes. However, the image evaluation method should be reconsidered.     

人正常宫颈脱落细胞及宫颈癌细胞的AFM观察研究

目的:探讨原子力显微镜(AFM)在人宫颈脱落细胞及宫颈癌细胞微观形貌表征方面的应用。方法:利用液基薄层细胞学检测技术制备细胞样品,应用原子力显微镜分别观察人宫颈脱落细胞及宫颈癌细胞的微观形态。结果:癌变宫颈细胞核独立分布,尺寸变大,且易于聚集,细胞质分散在细胞核周围;正常宫颈细胞的细胞核被纳米条状的细胞质覆盖,成为一个整体。结论:采用AFM检测液基薄层细胞学检测技术制备的细胞样品的微观形貌能反映出与宏观光学显微镜检测获得相同的结论,是一种简易的检测方法。癌变宫颈细胞和正常宫颈细胞在微观上存在明显不同:癌变宫颈细胞核的尺寸增大、聚集在一起;而正常宫颈细胞被纳米条状的细胞质所覆盖。     

复发性流产外周IL-10~+Tim-3~+ T细胞降调节

目的探讨外周血CD3+T细胞中T细胞免疫球蛋白黏蛋白-3(Tim-3)及程序性死亡因子-1(PD-1)联合细胞因子在复发性流产(RSA)中的诊断价值。方法用流式细胞术检测19例RSA患者及17例正常早孕者外周血CD3~+T细胞表面Tim-3及PD-1含量,以及破细胞膜后细胞内因子干扰素(IFN)-γ和白介素(IL)-10在Tim-3~+PD-1~+、Tim-3~-PD-1~-、Tim-3~-PD-1~+和Tim-3~+PD-1~-4群细胞内的表达情况。结果 RSA患者Tim-3~+PD-1~+T细胞比例(0.57%±0.26%)明显低于早孕组(1.24%±0.77%)(P0.001)。RSA组4群T细胞中IL-10阳性细胞所占比例分别为33.55%±16.27%、0.92%±0.88%、1.61%±1.35%、16.36%±13.98%;早孕组4群T细胞中IL-10阳性细胞占比分别为45.92%±17.89%、0.49%±0.27%、0.92%±0.68%、33.43%±16.98%。RSA组和正常组Tim-3~+PD-1~+T细胞群中IL-10的含量均显著高于其他3群(P0.05);RSA组Tim-3~+PD-1~+和Tim-3~+PD-1~-T细胞中IL-10阳性细胞含量(33.55%±16.27%,16.36%±13.98%)显著低于正常早孕组(45.92%±17.89%,33.43%±16.98%)(P0.05,P0.01)。而IFN-γ在RSA组和正常组4群细胞中的表达无统计学差异。结论 RSA患者IL-10~+Tim-3~+T细胞显著降低,可作为判断RSA的新的参考指标。     

Generation of tumor-specific cytolytic T lymphocytes from peripheral blood of cervical cancer patients by in vitro stimulation with a synthetic human papillomavirus type 16 E7 epitope

OBJECTIVE: Approximately 90% of squamous carcinomas of the cervix harbor the human papillomavirus and type 16 has been detected in nearly 50% of cases. Recent studies in mice have shown that the human papillomavirus type 16 E7 oncoprotein contains peptide epitopes that are processed and presented in association with a major histocompatibility antigen for recognition by cytolytic T lymphocytes. We investigated whether an epitope from human papillomavirus type 16 E7 could be used to generate specific human cytolytic T lymphocytes in patients with cervical carcinoma. STUDY DESIGN: After radiation therapy, three patients with antigen HLA-A2 and with locally advanced cervical cancer underwent leukapheresis. Epitope-specific cytolytic T lymphocytes were generated from the peripheral blood mononuclear cells by in vitro stimulation with autologous peripheral blood mononuclear cells pulsed with a human papillomavirus type 16 E7, HLA-A2–restricted, synthetic peptide, E7_11-20 (YMLDLQPETT). RESULTS: In two patients cytolytic T lymphocytes were capable of E7_11-20–specific, HLA-A2–restricted cytolysis of the peptide-pulsed, HLA-matched, T2 target cell line. Cytolytic T lymphocytes from one of these patients also demonstrated specific cytolysis against the HLA-A2⁺, HPV-16⁺ CaSki cervical cancer cell line but did not lyse either HLA-A2⁺, HPV-16^– MS-751 cells or HLA-A2^-, HPV-16^– HT-3 cells. CONCLUSIONS: These experiments demonstrate that novel cytolytic T lymphocytes that recognize a human papillomavirus type 16 E7 epitope can be generated by using the peripheral blood mononuclear cells from irradiated patients with cervical cancer. In addition, because CaSki cells were specifically lysed by the cytolytic T lymphocytes, these data indicate that the peptide E7_11-20 is endogenously processed and presented on the cell surface of the CaSki cells. The demonstration of epitope-specific lysis of cytolytic T lymphocytes of HPV-16⁺ cervical cancer cells supports further efforts to develop human papillomavirus peptide-based vaccines or antigen-specific adoptive immunotherapy for the prevention and treatment of cervical carcinoma. (Am J Obstet Gynecol 1996;175:1586-93.)