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。     

妊娠期糖尿病患者母儿免疫水平变化的临床研究

目的:研究妊娠期糖尿病(GDM)患者母儿免疫球蛋白(Ig)、补体(C)、T淋巴细胞亚群以及NK细胞水平的变化,探讨GDM对母儿体液免疫和细胞免疫的影响。方法:选择58例不同糖耐量水平的GDM患者为研究对象根据是否需要胰岛素治疗进一步分为GDM1组和GDM2组,以30例健康孕妇做对照。免疫比浊法测定外周静脉血与脐血的免疫球蛋白、补体水平;流式细胞技术测定外周静脉血与脐血的T细胞亚群及NK细胞水平。结果:GDM组外周血CD4+、CD4+/CD8+、CD16+CD56+、IgG含量均下降,CD8+、C3、C4、IgE含量升高,差异有统计意义(P<0.05);CD3+、IgA、IgM含量下降,差异无统计学意义(P>0.05)。GDM组脐血中CD3+、CD4+、CD4+/CD8+、CD16+CD56+含量均下降,差异有统计学意义(P<0.05);IgM、IgG含量下降,CD8+、IgE含量升高,差异无统计学意义(P>0.05)。与GDM1组相比,GDM2的指标呈现更明显的变化趋势。结论:妊娠期糖尿病存在母儿体液免疫和细胞免疫的失衡,且病情越重,这种改变越显著。     

外阴白色病变患者T细胞亚群及脂质过氧化反应的变化

目的 :探讨外阴白色病变与免疫功能及脂质过氧化反应的关系。方法 :用FCM法检测患者静脉血CD3+、CD4 +、CD8+、CD16 +细胞 ,用亚硝酸盐法检测MDA ,用硫代巴比妥酸法检测SOD。结果 :患者组与对照组的CD3+、CD4 +、CD8+、CD16 +细胞水平无明显差异 ,患者组CD4 + /CD8+细胞比值及SOD明显低于正常对照组 (P <0 .0 5 ) ,而MDA较对照组明显增高 (P <0 .0 5 ) ;不同组织类型外阴白色病变患者之间的T细胞亚群水平和SOD及MDA无明显差异 ;直线相关分析显示 ,SOD变化与CD4 + /CD8+变化及CD 3+、CD4 +变化呈明显正相关 (P <0 .0 1) ,MDA与CD8+、CD16 +变化正相关 (P <0 .0 5 )。结论 :外阴白色病变患者体内存在免疫应答过度和过剩脂质过氧化反应 ,二者共同作用导致了机体自身组织的损伤。     

子痫前期患者外周血协同刺激分子CD28/CTLA-4的表达研究

目的:研究子痫前期患者外周血协同刺激分子CD28、CTLA-4、CD28/CTLA-4在CD3+CD4 +T细胞和CD3+ CD8+T细胞上的表达,探讨CD28/CTLA-4在子痫前期发病中的作用.方法:采集22例正常妊娠妇女(正常妊娠组),45例子痫前期患者(子痫前期组)外周血,用流式细胞技术分别检测外周血T淋巴细胞CD3+、CD4+、CD8+的表达和CD28、CTLA-4、CD28/CTLA-4在CD3+ CD4+T细胞和CD3+ CD8+T细胞上的表达.结果:子痫前期组外周血T淋巴细胞中CD3+及CD8+的表达低于正常妊娠组(P＜0.05),而CD4+及CD4 +/CD8+的表达高于正常妊娠组(P＜0.05).与正常妊娠组相比,子痫前期组CD28在CD3+ CD4+T细胞和CD3+ CD8+T细胞上的表达低于正常妊娠组,但差异无统计学意义(P＞0.05);CTLA-4在CD3+ CD4+T细胞和CD3+ CD8+T细胞上的表达高于正常妊娠组,且重度子痫前期组高于轻度子痫前期组,差异均有统计学意义(P＜0.05);CD28/CTLA-4比率在CD3+ CD4+T细胞和CD3+ CD8+T细胞上的表达低于正常妊娠组,且重度子痫前期组低于轻度子痫前期组,差异均有统计学意义(P＜0.05).结论:子痫前期患者外周血T细胞亚群明显偏移,且协同刺激分子CD28/CTLA-4表达异常,高表达的CTLA-4导致T淋巴细胞亚群失衡,这可能是促使子痫前期发生的重要原因之一.     

原因不明复发性流产与蜕膜CD4~+ CD25~+ T调节细胞和CD8~+ CD28~- T细胞的关系

目的:探讨原因不明复发性流产(unexplained recurrent spontaneous abortion,URSA)与蜕膜CD4+ CD25+ T细胞和CD8+ CD28-T细胞的关系。方法:采用流式细胞四色荧光法,检测原因不明复发性流产17例(URSA组)和正常早孕人流20例(对照组)的蜕膜CD4+ CD25+ T细胞及其FoxP3(+)表达,CD8+ CD28- T细胞及其表面CD95、CD95L表达。结果:两组蜕膜中CD4+ CD25+ T细胞比例无明显差异(P>0.05),URSA组蜕膜CD4+ CD25+ T细胞中FoxP3阳性率明显低于对照组(P<0.0001)。两组蜕膜CD8+ CD28- T细胞比例及其细胞表面CD95和CD95L的阳性表达率均无明显差异(P>0.05);结论:蜕膜CD4+ CD25+ FoxP3(+)T调节细胞明显减少,是导致URSA患者母胎界面免疫耐受异常的重要原因。     

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     

不明原因早期自然流产患者静脉血T细胞及T亚群变化的研究

目的探讨不明原因早期自然流产患者静脉血T细胞及T亚群变化.方法采用流式细胞仪与免疫荧光技术相结合,检测不明原因早期自然流产患者(病例组)和正常妊娠早期妇女(早孕组)外周血中CD3+,CD4+,CD8+细胞的百分含量.结果病例组31例早期自然流产妇女外周静脉血CD3+,CD4+,CD8+T细胞及T亚群百分含量以及CD4+/CD8+的比例分别为(67.11±5.17)%、(35.35±5.26)%、(23.76±4.92)%、1.49±0.59.早孕组33例正常妊娠12周以内妇女分别为(64.45±4.56)%、(36.07±5.78)%、(21.85±3.73)%、1.53±0.73.对两组结果进行统计学处理P＞0.05,无显著性差异.结论不明原因早期自然流产患者静脉血中T细胞及T亚群百分含量与正常妊娠早期妇女相同.     

宫颈癌患者外周血HPV16抗原特异性CD8~+细胞毒性T细胞的检测及意义

目的:初步探讨人乳头瘤病毒16型(HPV16)阳性宫颈癌患者细胞免疫状况,了解其外周血抗原特异性CD8~+细胞毒性T细胞的水平.方法:采取免疫荧光标染重组MHC Ⅰ类分子-肽五聚体技术,运用流式细胞术定量检测患者和正常人外周血CD8~+细胞毒性T细胞、记忆性细胞毒性T细胞、活化细胞毒性T细胞及HPV16抗原特异性CD8~+细胞毒性T细胞.结果:宫颈癌患者CD8~+细胞毒性T细胞计数、记忆性细胞毒性T细胞计数、体内抗原特异性CD8~+细胞毒性T细胞计数和对照组比较差异无统计学意义,但活化细胞毒性T细胞计数低于对照组(P=0.322),体外经抗原肽刺激后抗原特异性CD8~+细胞毒性T细胞计数高于对照组(P=0.0068).结论:HPV16 阳性宫颈癌患者外周血中活化细胞毒性T细胞数目减少,表明患者细胞免疫功能受到一定的影响,通过抗原表位有效的诱导是提高患者的特异性细胞免疫功能的重要途径,运用五聚体技术检测宫颈癌患者抗原特异性细胞毒性T细胞,可初步反映患者特异性细胞免疫状态并有利于进行复发和预后分析.     

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。     

卵巢癌患者术后NDV-ATV-ASI与化疗对免疫功能的影响

目的 :评价新城鸡瘟病毒修饰的自体肿瘤细胞疫苗特异性主动免疫治疗(NDV ATV ASI)及化疗后患者免疫功能的变化 ,为实施肿瘤生物治疗 +化疗联合方案提供依据。方法 :卵巢癌患者术后 3 4例给予NDV ATV ASI及化疗 ,采用流式细胞术免疫荧光双标记法检测患者治疗后外周血单个核细胞 (PBMC)中CD4+ 、CD8+ 细胞内IFN γ和IL 4的表达 ;体外实验以NDV修饰的自体肿瘤细胞与PBMC共培养 (MLTC) ,观察其早期活化分子CD2 5、CD69的表达。结果 :治疗后CD4+ 、CD8+ T淋巴细胞内IFN γ的表达明显增高 ,IL 4表达轻度增高 ,且增高的时间较晚。外周血CD4+ 、CD8+ T淋巴细胞CD2 5、CD69的表达增加。单纯化疗组CD4+ 、CD8+ 细胞内IFN γ表达降低。结论 :NDV ATV ASI可诱导机体产生抗肿瘤免疫应答 ,检测单个T细胞水平的细胞因子可作为一种简单且可靠的免疫治疗效果评价参数 ;而化疗对机体的免疫功能有抑制作用 ,二者联合应用应选择适当的时间。     

Vulvar lichen sclerosus: an immunologic study

OBJECTIVE: To investigate the seroimmunologic (CD3, CD4, CD8 lymphocytes, C3c and C4 complement fractions, and several autoantibodies) and immunohistochemical (T lymphocyte subpopulations, B lymphocytes, natural killer cells, macrophages, immunoglobulin [Ig] G, Ig M, and C3c complement fraction) characteristics of vulvar lichen sclerosus. METHODS: Serum samples from 68 women with histologically proven lichen sclerosus were compared with those from 53 healthy controls, and tissue samples from 14 of 68 women chosen at random were compared with those from 14 of 53 healthy controls. A scoring system was constructed to compare the number of cells in the tissue. RESULTS: Patients had significantly lower counts of circulating lymphocytes CD3 and CD4 than controls (P < .05) and a higher number of autoantibodies (P < .01). Analysis of the tissue samples confirmed a lower number of CD2 cells (two-tailed P = .002 in epidermis, .005 in dermis), CD3 cells (two-tailed P = .001 in epidermis and in dermis), CD4 cells (two-tailed P = .002 in epidermis, .011 in dermis), and CD8 cells (two-tailed P = .002 in epidermis, .051 in dermis) in subjects than in controls. Numbers of monocyte-macrophage cells were similar in the epidermis but different in the dermis (two-tailed P = .003). No natural killer CD56 cells or B lymphocytes (CD19-CD21) were detected in the affected areas. Deposits of IgG, IgM, and C3 were no greater in biopsy specimens of patients than in those of controls. CONCLUSION: Vulvar lichen sclerosus is not caused by a T cell-mediated response, and a viral origin is unlikely. The absence of CD19 and CD21 cells excludes local production of autoantibodies. Our data do not confirm an autoimmune pathogenesis for vulvar lichen sclerosus but help explain why systemic cortisone is of no benefit and justify the use of petroleum jelly to relieve pruritus.     

Distribution of transforming growth factor-beta isoforms TGF-beta 1, TGF-beta 2 and TGF-beta 3 and vascular endothelial growth factor in vulvar lichen sclerosus

OBJECTIVE: To study the distribution of transforming growth factor beta (TGF-beta) isoforms TGF-beta 1, TGF-beta 2 and TGF-beta 3 and vascular endothelial growth factor (VEGF) in vulvar lichen sclerosus. STUDY DESIGN: Biopsies were obtained from 10 patients with vulvar lichen sclerosus, snap frozen and stained with polyclonal antibodies to TGF-beta 1, TGF-beta 2, TGF-beta 3 and VEGF. Control tissues used were uninvolved thigh tissue from two of the lichen sclerosus patients and normal vulvar tissue obtained from eight patients during gynecologic procedures. Two specimens of morphea were also examined. RESULTS: Weak TGF-beta 1 staining was demonstrated in the epidermis of all the lichen sclerosus, morphea, thigh and five of the eight normal vulvar specimens. Slight increase in TGF-beta 1 staining was seen in the upper and middermis in 6 of the 10 lichen sclerosus specimens and in the morphea specimens as compared to the control tissue, and this staining was localized within cells. TGF-beta 2 staining was present throughout the epidermis in all the normal thigh, normal vulva, lichen sclerosus and morphea specimens. TGF-beta 2 staining was increased within cells in the upper and middermis of the lichen sclerosus and morphea specimens. TGF-beta 3 staining occurred in the basal half of the epidermis of all the control, lichen sclerosus and morphea specimens, and only slight upper dermal staining of a few individual cells was seen in 3 of the 10 lichen sclerosus specimens. VEGF staining was similar in the normal tissues, lichen sclerosus and morphea. CONCLUSION: These results suggest that TGF-beta may.     

聚焦超声波治疗女性外阴炎性疾病的疗效

目的探讨高强度聚焦超声治疗（HIFU）女性外阴炎性疾病（单纯外阴瘙痒症和外阴上皮内非瘤样病变）的疗效。方法应用高强度超声治疗仪对142例单纯外阴瘙痒症、267例不同病理类型外阴上皮内非瘤样病变（141例硬化苔藓和126例鳞状上皮细胞增生）,规范了对不同疾病的治疗方法以及疗效判别标准。结果本方法对单纯外阴瘙痒症组两次治疗后总有效率为90.4%;对外阴上皮内非瘤样病变4次治疗后,病变区皮肤面积由（14.26±4.79）cm2减少至（3.83±2.04）cm2,差异有统计学意义（P〈0.01）,其中对硬化苔藓型明显有效率为94.8%,对鳞状上皮细胞增生型患者明显有效率为92.0%。结论本组显示HIFU可使瘙痒症状显著减轻,外阴白色病变面积逐渐减少,并具有简便、无痛和安全无创等优点。     

Clinically identifying women with vulvar lichen sclerosus at increased risk of squamous cell carcinoma: a case-control study

OBJECTIVE: To identify clinical factors that might identify women with vulvar lichen sclerosus who are at increased risk of developing squamous cell cacinoma. STUDY DESIGN: A retrospective, case-control study compared 46 women presenting between 1992 and 2000 with clinical and histologic evidence of squamous cell carcinoma of the vulva arising in a background of lichen sclerosus and 213 new referrals with vulvar lichen sclerosus during the same period. RESULTS: The ages of the patients and presence of clinical hyperplasia were the only differences between the 2 groups. CONCLUSION: Women presenting with vulvar cancer arising within a background of lichen sclerosus are significantly older than women presenting with lichen sclerosus. In addition, clinical evidence of squamous hyperplasia is independently associated with vulvar carcinoma. Neither the presence nor duration of symptoms nor the loss of vulvar architecture is a useful indicator of potential cancer risk.     

Pimecrolimus for the treatment of vulvar lichen sclerosus: a report of 4 cases

BACKGROUND: Lichen sclerosus is a chronic cutaneous disorder with a predilection for the vulva. The etiology is unknown. Superpotent topical corticosteroids are the most effective treatment. Dermal atrophy, however, is a well-known complication associated with changes of lichen sclerosus superpotent topical corticosteroids. In addition, some women do not respond adequately to topical steroids. Therefore, a treatment regimen that does not rely on corticosteroids may be beneficial. As lichen sclerosus is a T-lymphocyte-mediated disorder, it has been suggested that a topical macrolide immunosuppressant, such as pimecrolimus, may be a safe and effective alternative treatment for lichen sclerosus. Since pimecrolimus does not affect collagen synthesis, it does not cause thinning of the dermis. CASES: Four patients with biopsy-proven lichen sclerosus were treated with 1% pimecrolimus cream twice daily. At the end of 3 months of treatment, 3 of the 4 patients reported complete resolution of their symptoms of vulvar itching and burning. Two patients had repeat vulvar biopsies at the end of treatment that showed reversal of the histologic changes of lichen sclerosus. CONCLUSION: In this small series, pimecrolimus appeared to be a safe and effective treatment of vulvar lichen sclerosus. A randomized, controlled trial comparing pimecrolimus and clobetasol propionate should be performed to determine which is the safer and more effective medication for the long-term treatment of lichen sclerosus.     

The epithelial changes associated with squamous cell carcinoma of the vulva: a review of the clinical, histological and viral findings in 78 women

Summary. Seventy-eight excised specimens of squamous cell carcinoma of the vulva were reviewed retrospectively for the presence of lichen sclerosus or vulvar intraepithelial neoplasia (VTN) at sites proximal to the tumour or more distant. Lichen sclerosus was evident in 61% and VIN alone in 31%. VIN III (differentiated) was associated with over 50% of the specimens with lichen sclerosus. HPV 16 was found in six of the 11 VIN lesions, investigated but in none of the six with lichen sclerosus.     

Guidelines for the follow-up of women with vulvar lichen sclerosus in specialist clinics

It is recommended that women with vulvar lichen sclerosus be followed in specialist clinics where difficulty exists with symptom control or where there is clinical evidence of localized skin thickening. Follow-up is also recommended for women who have previously been treated for squamous cell carcinoma of the vulva (arising in lichen sclerosus or vulvar intraepithelial neoplasia) or where the pathologist expresses concern and is unable to make a definitive diagnosis of differentiated vulvar intraepithelial neoplasia.     

Treatment of lichen sclerosus with topical dihydrotestosterone

Lichen sclerosus typically affects the vulva of postmenopausal women. Because serum levels of dihydrotestosterone are low in women with vulvar lichen sclerosus and because dihydrotestosterone is an effector androgen in vulvar skin, this double-blind cross-over study assessed five women with vulvar lichen sclerosus to determine the response to treatment with dihydrotestosterone. Objective gross and microscopic improvement in lichen sclerosus accompanied sustained treatment with topical dihydrotestosterone, but not with vehicle alone. However, there was no change in symptoms (itching and dyspareunia) in these women, although dihydrotestosterone did improve some of the features of vulvar lichen sclerosus and may represent a new treatment for this disease.     

The epithelial changes associated with squamous cell carcinoma of the vulva: a review of the clinical, histological and viral findings in 78 women

Seventy-eight excised specimens of squamous cell carcinoma of the vulva were reviewed retrospectively for the presence of lichen sclerosus or vulvar intraepithelial neoplasia (VIN) at sites proximal to the tumour or more distant. Lichen sclerosus was evident in 61% and VIN alone in 31%. VIN III (differentiated) was associated with over 50% of the specimens with lichen sclerosus. HPV 16 was found in six of the 11 VIN lesions, investigated but in none of the six with lichen sclerosus.