环孢菌素A和1,2 5(OH)2D3防治实验性自身免疫性甲状腺炎的研究

将猪的甲状腺球蛋白（pTG)100μg/只分别于第0d,第14d皮下注入CBA小鼠体内，制作实验性自身免疫性甲状腺炎（EAT）的动物模型。免免疫干预组从0－28d，治疗组从10－38d单独或者联合应用环孢菌素A（CsA,10mg/kg)灌胃和（或）1,25(OH)2D3(0.2μg/kg）腹腔注射，pTG免疫后第28d，第38d处死小鼠，取甲状腺组织作病理学检查，并检测血清中猪的甲状腺球蛋白抗体（pTGAb)、猪的甲状腺微粒体抗体（pTMAb)。免疫干预组和治疗组联合应用小剂量CsA和1，25（OH）2D3分别使EAT发病率降低44．44％和37．50％。严重病例分别降低71．43％和60．32％，免疫干预组的血清pTGAb,pTMAb的值均降低。提示：小剂量免疫抑制剂CsA和1，25（OH）2D2联合防治EAT有效，并具有协同作用。     

强直性脊柱炎患者外周血CD62P的表达

目的探讨剖宫产指征的变化情况，分析影响剖宫产率的因素。方法 回顾性分析1393例剖宫产患者的临床资料。结果2年的平均剖宫产率为66．7％，其中2004年的剖宫产率为59．9％，2005年的剖富产率为72．2％，差异有统计学意义（P〈0．05）。剖宫产率的主要影响因素依次为社会因素（29．43％）、胎儿宫内窘迫（25．13％）、胎位异常（9．62％）、巨大胎儿（7．25％）、中至重度妊娠高血压综合征（6．25％）。结论剖宫产率的上升不只是一个纯医学问题，社会因素使剖宫产指征相对扩大，只有合理掌握剖宫产指征，通过医患双方、社会的共同努力，才能更好地降低剖宫产率。     

红细胞调控白细胞免疫功能新的自然实验研究体系

目的用血液免疫反应路线图实验体系评估红细胞在白细胞免疫活性中的作用。方法将0·3ml血浆加入0·2ml全血细胞悬液(全血细胞组)或0·2ml白细胞悬液(白细胞组)中,37℃温育1h,用免疫酶联法测定IL-8和IL-12水平,流式细胞仪测定白细胞膜CD4、CD8、CD35和CXCR4表达量。结果全血细胞组IL-8和IL-12水平(分别为5·96±4·26、9·84±2·23ρB·pg-1·ml-1)明显低于白细胞组(分别为15·09±9·86、13·59±3·69ρB·pg-1·ml-1,P<0·05),淋巴细胞CD4、CD35、CXCR4表达量(分别为37·79±12·00、154·66±70·00、34·40±20·45)明显高于白细胞组(分别为18·54±11·32、83·26±35·99、16·69±11·09,P<0.01),粒细胞CD35表达量(603·63±257·64)明显高于白细胞组(384·86±174·16,P<0.01)。成人全血细胞组淋巴细胞和粒细胞CD35和CXCR4表达量明显高于脐血全血细胞组(P<0·05或P<0·01)。结论红细胞是白细胞(包括T淋巴细胞、B淋巴细胞、NK细胞、树突状细胞、粒细胞等)免疫功能的调控者和指导者,脐血红细胞免疫调节功能明显下降;本研究为红细胞免疫调控活性测定提供了新的近似自然的方法。     

调节性CD4+T细胞在大鼠自发肝移植耐受中的作用

目的 探讨在肝移植的自发耐受模型中，调节性CD4^＋T细胞的免疫抑制作用机制。方法 利用近交系大鼠从Lewis（LEW）到Wistar Furth（WF）的肝移植组合，对移植后不同时期的宿主注射抗CD4的单克隆抗体（Anti-CD4mAb），然后抽血检测丙氨酸氨基转氨酶（ALT）的动态变化；并结合细胞毒性T淋巴细胞（CTL）试验了解宿主脾细胞中T细胞亚群的动态改变。结果 对肝移植自然生存的宿主注射Afiti—CD4mAb后，术后第21天、42天均能够诱导出肝损害（排斥反应），但第56天、100天以上的则未能诱导出来，且该损害能被抗CD8单克隆抗体阻断。另外CTL试验显示宿主的脾细胞中，初始型CTL前体细胞在移植56d后未能检测出来。结论 在自发性肝耐受模型中。宿主术后早期存在由CD4^＋T细胞介导的下调原始效应性T细胞的作用机制。     

Triptolide promotes generation of FoxP3+ T regulatory cells in rats

Ethnopharmacological relevance

Triptolide (TPT), a component of the Chinese herb Triptergium wilfordii, has potent immunosuppressive and anti-inflammatory activity and is used clinically in recipients of kidney transplantation.

Aim of the study

This work aimed to investigate the effect of TPT on the differentiation of regulatory T lymphocytes (Tregs) from CD4+ cells in rats.

Materials and methods

MACS-purified rat CD4+ cells were costimulated with anti-CD3 and anti-CD28 in the presence of TGF-β to induce the expression of FoxP3, which was detected by flow cytometry. TPT and cyclosporine A (CsA) were separately added into the cultures to observe the effect on the expression of FoxP3. Kidney transplantation was performed in rats that either received no treatment or were treated with TPT after transplantation.

Results

TPT treatment enhanced the expression of FoxP3 in CD4+ cells, whereas CsA inhibited the FoxP3 expression. In the rat kidney transplantation model, the recipient rats treated with TPT survived longer than the control rats (18–19.83 vs 6.83 days, P < 0.05). Meanwhile, the FoxP3+ T cells in the spleens of treated rats were higher than those from the untreated rats (12.4% vs 4.7%, P < 0.05).

Conclusions

These data suggest that TPT may promote the differentiation of CD4+ cells to FoxP3+ Tregs. This would be at least one of the pathways responsible for the immunosuppressive activity of TPT.     

Immunomodulatory activity of Asparagus racemosus on systemic Th1/Th2 immunity: Implications for immunoadjuvant potential

Ethnopharmacological relevance

Roots of Asparagus racemosus Willd (Shatavari in vernacular) are widely used in Ayurveda as Rasayana for immunostimulation, galactogogue as also in treatment of conditions like ulcers and cancer. Various studies have indicated immunomodulatory properties of Shatavari root extracts and formulations.

Aim of the study

To study the effect of standardized Asparagus racemosus root aqueous extract (ARE) on systemic Th1/Th2 immunity of SRBC sensitized animals.

Materials and methods

We used HPTLC to quantify steroidal saponins (Shatavarin IV, Immunoside^®) and flow cytometry to study effects of ARE on Th1/Th2 immunity. SRBC specific antibody titres and DTH responses were also monitored as markers of Th2 and Th1 responses, respectively. We also studied lymphocyte proliferation. Cyclosporin, cyclophosphamide and levamisole were used as controls.

Results

Treatment with ARE (100 mg/(kg b.w. p.o.)) resulted in significant increase of CD3⁺ and CD4/CD8⁺ percentages suggesting its effect on T cell activation. ARE treated animals showed significant up-regulation of Th1 (IL-2, IFN-g) and Th2 (IL-4) cytokines suggesting its mixed Th1/Th2 adjuvant activity. Consistent to this, ARE also showed higher antibody titres and DTH responses. ARE, in combination with LPS, Con A or SRBC, produced a significant proliferation suggesting effect on activated lymphocytes.

Conclusion

The study suggests mixed Th1/Th2 activity of ARE supports its immunoadjuvant potential.     

针灸及其血清对荷瘤小鼠CD4~+CD25~+Treg细胞体外增殖的影响

目的:观察针灸治疗对H_(22)荷瘤小鼠CD4~+CD25~+Treg细胞(CD4~+CD25~+regulatory T cells)体外增殖的影响和针灸血清刺激下的CD4~+CD25~+Treg细胞的体外增殖变化,探讨针灸及其血清对荷瘤小鼠免疫调节细胞的干预作用。方法:48只小鼠随机分为电针治疗组、艾灸治疗组、肿瘤对照组、正常对照组。采用H_(22)肿瘤细胞移植性实体瘤模型,电针和艾灸"大椎"治疗后,磁珠分离CD4~+CD25~+Treg细胞,氚标记胸腺嘧啶核苷掺入法观察不同组小鼠CD4~+CD25~+Treg细胞体外增殖能力和针灸血清对CD4~+CD25~+ Treg细胞体外增殖的影响。结果:肿瘤对照组CD4~+CD25~+Treg细胞增殖水平比正常对照组明显增高(P<0.05);电针治疗组和艾灸治疗组CD4~+CD25~+Treg细胞增殖水平与肿瘤对照组比较均明显降低(P<0.01)。电针治疗组和艾灸治疗组1:1、1:8稀释度血清刺激正常小鼠CD4~+CD25~+Treg细胞体外增殖,较正常对照、肿瘤对照组血清显著增高(P<0.05),各组1:16、1:32稀释度血清对Treg细胞作用的差别无统计学意义(P>0.05)。结论:针灸治疗能下调荷瘤小鼠CD4~+CD25~+Treg细胞体外增殖能力。不同浓度针灸血清体外刺激CD4~+CD25~+Treg细胞增殖表现出不一致的效应。     

CD44v7/8基因表达与宫颈癌关系的探讨

目的探讨宫颈癌组织中CD44v7/8基因的异常表达及临床意义。方法1986～2000年沈阳医学院附属中心医院采用免疫组化的方法随机测定59例宫颈癌、18例尖锐湿疣和18例慢性宫颈炎组织中CD44v7/8基因的表达情况。结果宫颈癌、尖锐湿疣及慢性宫颈炎3种疾病中CD44v7/8表达阳性率分别为76.27%、11.11%和11.11%,宫颈癌组的CD44v7/8表达分别高于慢性宫颈炎组及尖锐湿疣组,且差异非常显著(P<0.01)。慢性宫颈炎组及尖锐湿疣组比较差异无显著性(P>0.05)。结论CD44v7/8蛋白的表达增强与宫颈癌的发生、发展及和局部浸润转移有关。     

Rational design of hypoallergens applied to the major cat allergen Fel d 1

BACKGROUND: Allergen-specific immunotherapy is the only treatment for allergic disease providing long-lasting symptom relief. Currently, it is mainly based on the use of crude allergen extracts. The treatment may be improved by the use of genetically engineered allergens, hypoallergens, aiming at a more effective and safer therapy. OBJECTIVE: The aim of this study was to provide a rational design of hypoallergen candidates for immunotherapy by using structural information and knowledge of B and T cell epitopes of an allergen. METHODS: The three-dimensional structure of the major cat allergen Fel d 1 was systematically altered by duplication of selected T cell epitopes and disruption of disulphide bonds. Seven Fel d 1 derivatives were generated and screened for allergenic reactivity in comparison with recombinant Fel d 1 in competition-ELISA. The allergenicity was further evaluated in basophil activation experiments and T cell reactivity was assessed in a lymphoproliferation assay. RESULTS: Three out of seven Fel d 1 derivatives, with two duplicated T cell epitopes and one or two disulphide bonds disrupted, were carefully evaluated. The three derivatives displayed a strong reduction in allergenicity with 400-900 times lower IgE-binding capacity than recombinant Fel d 1. In addition, they induced a lower degree of basophil activation and similar or stronger T cell proliferation than recombinant Fel d 1. CONCLUSION: By a rational approach, we have constructed three Fel d 1 hypoallergens with reduced IgE-binding capacities and retained T cell reactivities. This strategy may be applied to any well-characterized allergen to improve immunotherapy for allergic patients.     

CD80mAb协同imDC诱导同种异体大鼠胰十二指肠移植免疫耐受

目的 观察共刺激分子阻断剂CD80单克隆抗体（CD80mAb）在协同未成熟树突细胞（imDC）诱导同种异体大鼠胰十二指肠移植免疫耐受中的作用。方法 建立糖尿病大鼠胰十二指肠移植动物模型；4E5杂交瘤细胞株BABIMC小鼠腹腔注射，抽取腹水，分离纯化后获得CD80mAb；分离供体大鼠骨髓来源DC细胞前体，经GM—CSF、IL-4体外刺激后。再加入IL-10共培养，鉴定为imDC；移植前7d，将2×10^6imDC经静脉途径注射至受体体内，同时分别给予生理盐水1ml、CD80mAb5mg连续14d。结果 四组受体大鼠移植后中位生存时间分别为12．7d、32．4d、50．2d、92．0d，实验组存活时间明显延长；组织学观察发现移植后7dCD80mAb＋imDC组移植物形态尚完整，淋巴细胞浸润减少；混合淋巴细胞反应证实移植后7dCD80mAb＋imDC组供受体间呈低反应性。结论 共刺激分子阻断剂CD80mAb能够协同imDC诱导受体T细胞对移植物的免疫耐受，降低宿主对移植物的急、慢性排斥反应，延长移植物的存活时间。