桥本甲状腺炎患者血清IL-17、IL-35与维生素D的相关性

目的 探讨桥本甲状腺炎(HT)患者血清白细胞介素(IL)-17、IL-35与维生素D的相关性.方法 选取152例HT患者(HT组),按甲状腺激素水平分为甲状腺功能正常组(51例)、亚临床甲状腺功能减退(简称甲减)组(50例)、临床甲减组(51例),以体检健康者50名作为正常对照组.检测所有对象血清游离三碘甲状腺原氨酸(...     

细胞毒性T淋巴细胞抗原4拮抗剂基因治疗实验性自身免疫性甲状腺炎

目的 探讨利用携带人工合成的共刺激分子细胞毒性T淋巴细胞抗原4（CTLA-4）拮抗剂（CTLA4Ig）的真核表达载体,对自身免疫性甲状腺炎进行基因治疗的效果.方法 采用辅助性T淋巴细胞1型（Th1）倾向的C57BL/6J雌性小鼠,构建实验性自身免疫性甲状腺炎（EAT）动物模型20只,随机分为基因治疗组（治疗组）和模型对照组（模型组）,每组各10只,分别行甲状腺内质粒pCI/CTLA4Ig和空质粒pCI局部注射,另设非干预正常对照组（对照组）10只.获得血清和甲状腺、脾脏组织,通过实时定量PCR及酶联免疫吸附试验（ELISA）方法检测样本中甲状腺自身抗体及Th1、Th2相关细胞因子的表达.结果 与模型组比较,治疗组的甲状腺组织CTLA-4的表达升高近两倍（P=0.038）,Th1细胞因子干扰素γ（IFN-γ）及细胞间黏附分子1（ICAM-1）显著降低（P值分别为0.016及0.042）,而Th2细胞相关因子白细胞介素4（IL-4）的表达显著升高（P=0.044）.脾脏组织和小鼠血清中细胞因子水平显示了同样的改变.血清中抗甲状腺过氧化物酶抗体水平在治疗前后改变不显著.结论 甲状腺局部CTLA4Ig基因的注射可通过诱导Th1优势向Th2偏移而对EAT起治疗作用.     

IL-4及IL-8在桥本氏甲状腺炎发病机制中的作用

目的探讨白细胞介素4(IL-4)和白细胞介素8(IL-8)在桥本氏甲状腺炎(HT)发病过程中的作用。方法应用化学发光免疫分析法测定血清FT3、FT4、TSH含量,放射免疫分析法测定IL-4、IL-8含量,分析其与HT发病的关系。结果桥本氏甲状腺炎组FT3、FT4均值明显低于正常对照组,TSH水平明显高于正常对照组,IL-4水平明显低于正常对照组,IL-8水平明显高于正常对照组,IL-4、IL-8水平与FT3、FT4没有相关性。结论 IL-4、IL-8在HT发病机制和病理变化中发挥重要的作用,为临床诊断、治疗HT开辟了新的途径。     

IL-27对实验性自身免疫性心肌炎的干预作用研究

目的:探讨IL-27对实验性自身免疫性心肌炎(EAM)的干预作用.方法:用心脏肌球蛋白(MyHC-α614-629)诱导小鼠模型并用IL-27进行干预,苏木素-伊红染色分析心肌损伤程度,ELISA法测定细胞因子的水平,流式和ELISA法体外分析IL-27对EAM源的脾脏CD4+T细胞的作用.结果:IL-27干预后心肌损伤程度减轻,细胞因子TNF-α、IL-6和IL-17分泌水平降低,体内外研究证实IL-10分泌水平增加.结论:IL-27能够有效缓解EAM.     

阿托伐他汀对实验性自身免疫性脑脊髓炎治疗作用的研究

目的观察阿托伐他汀对实验性自身免疫性脑脊髓炎(EAE)的治疗作用并初步探讨其治疗机制。方法将大鼠分为正常组、模型组、阿托伐他汀组,每组6只。正常组不做任何处理;以豚鼠脑、脊髓为原料提取髓鞘碱性蛋白(MBP),免疫Lewis大鼠建立EAE模型。模型组注射抗原后以蒸馏水灌胃,阿托伐他汀组给予阿托伐他汀灌胃。建模后每日对大鼠神经症状进行评分,采用MTT法检测T淋巴细胞增殖,ELISA法检测大鼠脾细胞培养上清液中IFN-γ和IL-4水平。结果 (1)与模型组相比,阿托伐他汀组大鼠神经系统症状有不同程度减轻;(2)MTT法显示:与模型组大鼠相比,阿托伐他汀组大鼠的T细胞增殖被显著抑制(P<0.01);(3)与正常组相比,模型组大鼠13d时IL-4水平无明显变化,17、21d时,IL-4水平明显升高(P<0.01),与模型组相比,阿托伐他汀组13、17d时IL-4水平明显升高(P<0.01),IFN-γ水平在13、17、21d时均显著低于模型组(P<0.05)。结论阿托伐他汀能改善EAE大鼠的症状,其机制可能与抑制T淋巴细胞增殖,纠正Th1/Th2失衡有关。     

血清IL-4、INF-γ及1,25-(OH)2D3在婴幼儿哮喘检测中的临床意义

目的通过检测婴幼儿哮喘不同时期血清白细胞介素(IL)-4、γ-干扰素(INF-γ)、1,25二羟基维生素D3[1,25-(OH)_2D_3]的水平,探讨其临床意义。方法随机选取2014年1月至2015年12月在广元市中心医院确诊为支气管哮喘的婴幼儿63例为哮喘组,随访3年,失访3例,以随访3年后是否仍有哮喘发作为界,分为哮喘Ⅰ组(仍有哮喘发作,28例)、哮喘Ⅱ组(无哮喘发作,32例);随机选取同时期健康婴幼儿22例作为对照组,检测其不同时期血清IL-4、INF-γ、1,25-(OH)_2D_3水平。结果与对照组相比,哮喘组血清IL-4水平明显升高(P0.01),血清INF-γ水平明显降低(P0.01);哮喘Ⅰ组患儿血清1,25-(OH)_2D_3水平低于对照组(P0.01);哮喘Ⅰ组干预率高于哮喘Ⅱ组(P 0.01)。结论哮喘患儿血清IL-4水平持续升高,1,25-(OH)_2D_3水平持续降低,当IL-4/INF-γ比值持续升高时,应尽早进行预防干预。     

血清可溶性白细胞介素-2受体水平在预测高血压急性脑出血手术治疗中的意义

目的：探讨血清可溶性白细胞介素２受体（ＳＩＬ２Ｒ）水平在预测高血压急性脑出血（ＨＡＣＨ）患者手术治疗中的意义。方法：采用双抗体夹心酶联免疫吸附法测定２０例正常人及４９例ＨＡＣＨ急性期手术和非手术患者血清ＳＩＬ２Ｒ含量。结果：ＨＡＣＨ患者入院当日血清中ＳＩＬ２Ｒ含量〔（０．５９２±０．０３７）Ｕ／Ｌ〕显著高于正常对照组〔（０．２３７±０．０６９）Ｕ／Ｌ〕；手术组患者术前与术后第１天和第１０天血清ＳＩＬ２Ｒ含量比较分别为无显著性差异（Ｐ＞０．０５）和有显著性差异（Ｐ＜０．０１）；手术组患者术后第１日和第１０日与非手术组患者血清ＳＩＬ２Ｒ含量同步比较分别为无显著性差异（Ｐ＞０．０５）和有显著性差异（Ｐ＜０．０５）；死亡患者入院当日血清ＳＩＬ２Ｒ含量〔（０．７４３±０．０４９）Ｕ／Ｌ〕高于存活组〔（０．５９８±０．０４４）Ｕ／Ｌ，Ｐ＜０．０１〕，且显著高于对照组〔０．２３７±０．０６８）Ｕ／Ｌ，Ｐ＜０．０１〕。结论：ＳＩＬ２Ｒ与ＨＡＣＨ患者免疫功能紊乱有关，动态测定血清ＳＩＬ２Ｒ含量，可作为观察ＨＡＣＨ病情发展变化的一项指标，同时对判断ＨＡＣＨ的预后亦有一定临床意义。     

特发性血小板减少性紫癜患者T淋巴细胞亚群及 IL-2、IFN-γ、IL-4检测的临床意义

目的 检测特发性血小板减少性紫癜（ITP）患者T淋巴细胞亚群及白细胞介素（IL）-2、IL-4、γ-干扰素（IFN-γ）的水平与外周血血小板的关系及意义。方法 采用ITP患者和正常人的血清用改良碱性磷酸酶-抗碱性磷酸酶（APAAP）桥联酶免疫法检测T细胞亚群（CD3、CIM、CD8）,用酶联免疫吸附试验（ELISA）检测IL-2、IL-4、IFN-γ的水平。结果 ITP患者CD3、CIM、IL-2、IFN-γ的水平明显减少,IL-4、CD8水平增高,与正常对照组相比差异均有显著性。结论 ITP患者T细胞亚群比例失衡即CD4／CD8比例降低,IFN-γ和IL-2降低,IL-4升高,使机体体液免疫亢进,抗血小板抗体（PAIg）增多,导致外周血血小板减少。     

1,25二羟维生素D3对哮喘小鼠的Rac1-IL33-ILC2通路的作用

目的观察1,25二羟维生素D31,25-(OH)2-D3对哮喘小鼠Ras相关的C3肉毒素底物1(Ras-related C3 botulinum toxin substrate 1,Rac1)、白细胞介素-33(IL-33)和2型固有淋巴细胞(type 2 innate lymphocytes,ILC2)的影响。方法将30只7周龄BALB/c雌性小鼠随机分为对照组、哮喘组、干预组,每组各10只小鼠,卵清蛋白混合液致敏并雾化吸入建立哮喘小鼠模型,干预组每次激发前腹腔注射1,25(OH)2-D3混合液0.08 ml,对照组和哮喘组以生理盐水代替。末次激发24 h后对小鼠肺组织中IL-33、Rac1、ILC2进行检测并比较。结果与对照组比较,哮喘组Rac1下降,IL-33水平增高,ILC2数目升高。且Rac1与IL-33、ILC2呈明显的负相关性(r=-0.954、r=-0.957,P<0.05)。干预组IL-33及ILC2数目较哮喘组显著下降,Rac1较哮喘组显著上升,差异有统计学意义(P<0.05)。结论哮喘小鼠存在Rac1下降,IL-33水平增高,ILC2数目升高的免疫变化;1,25-(OH)2-D3可能通过上调哮喘小鼠Rac1的表达,导致IL-33等细胞因子水平产生减少,进而抑制ILC2的活化、减轻哮喘的免疫炎症。     

Development of spontaneous autoimmune peripheral polyneuropathy in B7-2-deficient NOD mice

An increasing number of studies have documented the central role of T cell costimulation in autoimmunity. Here we show that the autoimmune diabetes-prone nonobese diabetic (NOD) mouse strain, deficient in B7-2 costimulation, is protected from diabetes but develops a spontaneous autoimmune peripheral polyneuropathy. All the female and one third of the male mice exhibited limb paralysis with histologic and electrophysiologic evidence of severe demyelination in the peripheral nerves beginning at 20 wk of age. No central nervous system lesions were apparent. The peripheral nerve tissue was infiltrated with dendritic cells, CD4(+), and CD8(+) T cells. Finally, CD4(+) T cells isolated from affected animals induced the disease in NOD.SCID mice. Thus, the B7-2-deficient NOD mouse constitutes the first model of a spontaneous autoimmune disease of the peripheral nervous system, which has many similarities to the human disease, chronic inflammatory demyelinating polyneuropathy (CIDP). This model demonstrates that NOD mice have "cryptic" autoimmune defects that can polarize toward the nervous tissue after the selective disruption of CD28/B7-2 costimulatory pathway.     

B cell-deficient NOD.H-2h4 mice have CD4+CD25+ T regulatory cells that inhibit the development of spontaneous autoimmune thyroiditis

Wild-type (WT) NOD.H-2h4 mice develop spontaneous autoimmune thyroiditis (SAT) when given 0.05% NaI in their drinking water, whereas B cell-deficient NOD.H-2h4 mice are SAT resistant. To test the hypothesis that resistance of B cell-deficient mice to SAT was due to the activity of regulatory CD4+CD25+ T (T reg) cells activated if autoantigen was initially presented on non-B cells, CD25+ T reg cells were transiently depleted in vivo using anti-CD25. B cell-deficient NOD.H-2h4 mice given three weekly injections of anti-CD25 developed SAT 8 wk after NaI water. Thyroid lesions were similar to those in WT mice except there were no B cells in thyroid infiltrates. WT and B cell-deficient mice had similar numbers of CD4+CD25+Foxp3+ cells. Mice with transgenic nitrophenyl-specific B cells unable to secrete immunoglobulin were also resistant to SAT, and transient depletion of T reg cells resulted in severe SAT with both T and B cells in thyroid infiltrates. T reg cells that inhibit SAT were eliminated by day 3 thymectomy, indicating they belong to the subset of naturally occurring T reg cells. However, T reg cell depletion did not increase SAT severity in WT mice, suggesting that T reg cells may be nonfunctional when effector T cells are activated; i.e., by autoantigen-presenting B cells.     

Effects of vitamin D on thyroid autoimmunity markers in Hashimoto’s thyroiditis: systematic review and meta-analysis

ObjectiveTo perform a meta-analysis of randomized controlled trials to evaluate the efficacy of vitamin D supplementation on thyroid autoimmunity markers in Hashimoto’s thyroiditis (HT).MethodsThis meta-analysis included randomized controlled clinical trials identified by a systematic search of electronic databases (PubMed®, MEDLINE®, EMBASE, The Cochrane Library, China National Knowledge Infrastructure) from inception to August 2020. All studies included patients with HT that received vitamin D supplementation irrespective of the doses administered or the duration of treatment. The primary and secondary outcome measures were thyroid peroxidase antibody (TPOAb) and/or thyroglobulin antibody (TGAb) titres.ResultsEight studies (n = 652) were included. There was significant heterogeneity between the studies. Using a random-effect model, vitamin D supplementation reduced TPOAb titre (standardized mean difference [SMD]: –1.11; 95% confidence interval [CI]: 1–1.92, –0.29) and TGAb titre (SMD: –1.12; 95% CI: –1.96, –0.28). A subgroup analysis demonstrated that vitamin D supplementation for >3 months resulted in a decrease in TPOAb titre (SMD: –1.66, 95% CI: –2.91, –0.41) but treatment ≤3 months was ineffective. Treatment with vitamin D₃ decreased TPOAb titre (SMD: –1.48; 95% CI: –2.53, –0.42) whereas vitamin D did not.ConclusionThese data suggest that vitamin D reduces autoantibody titre in patients with HT.     

Vitamin D receptor gene polymorphisms are associated with risk of Hashimoto's thyroiditis in Chinese patients in Taiwan

The etiology of the autoimmune thyroid, Hashimoto's thyroiditis (HT) is not very clear. However, genetic susceptibility is thought to play a critical role. The vitamin D receptor (VDR)-related endocrine system has been demonstrated to be able to carry out modulation of the immune response. Here, we investigated whether single nucleotide polymorphisms (SNPs) of VDR are associated with HT patients. VDR SNP was detected by polymerase chain reaction (PCR)-based restriction analysis in 109 patients with HT and 90 normal controls. Significant differences were found in the genotype distribution of VDR SNP between Hashimoto's thyroiditis patients and controls (P=0.0458). Allelic frequency of the VDR gene distinguished HT patients from controls (P=0.0089). The results revealed a significant difference between HT patients and normal controls in VDR SNP and a statistic correlation between VDR-FokI polymorphisms and HT formation. It could be concluded that patients who carry the C/C homozygote of the VDR-FokI gene polymorphism in exon 2 may have a higher risk of developing HT in Chinese patients in Taiwan.     

NOD鼠体内CD_4~+NKG2D~+T细胞与CD_8~+T细胞间关系

目的探讨NOD鼠体内CD_4~+NKG2D~+T细胞与CD_8~+T细胞间关系。方法动态监测NOD鼠外周血中CD_4~+NKG2D~+T及CD8+NKG2D~+T细胞在不同周龄频率。利用IGRP206-214特异性CTL清除的NOD鼠,对其外周血中CD_4~+NKG2D+/CD_4~+T及CD8+NKG2D+/CD_8~+T细胞频率进行分析。将磁珠分选所得CD_4~+NKG2D~+T细胞分别与经CFSE标记的纯CD_4~+NKG2D-T细胞、CD_8~+T细胞共培养4 d,检测体系中CD_4~+NKG2D-T细胞、CD_8~+T细胞CFSE的荧光强度变化。结果 NOD鼠外周血CD_4~+NKG2D+/CD_4~+T及CD8+NKG2D+/CD_8~+T细胞频率均随其1型糖尿病疾病进程发展逐渐升高,且二者之间呈正相关。IGRP206-214特异性CTL清除的NOD鼠,外周血中CD_4~+NKG2D+/CD_4~+T细胞频率随CD8+NKG2D+/CD_8~+T细胞频率降低显著下降。与单独培养的CD_4~+NKG2D-T细胞相比,加入CD_4~+NKG2D~+T细胞的培养体系中,CD_4~+NKG2D-T细胞增殖加快。但CD_4~+NKG2D~+T细胞对CD_8~+T增殖作用不明显。结论 NOD鼠体内存在着一群与1型糖尿病疾病进程正相关的CD_4~+NKG2D~+T细胞,且该群细胞极有可能是通过直接作用于CD_4~+NKG2D-T细胞而间接对CD_8~+T细胞产生作用。     

康复预处理对实验性自身免疫性脑脊髓炎小鼠IL-23/IL-17轴的影响

目的：探讨康复预处理对实验性自身免疫性脑脊髓炎（EAE）小鼠白介素23（IL-23）／白介素17（IL-17）轴的影响。方法：对C57BL／6小鼠进行为期8周的康复预处理（跑台训练）,复制C57BI．／6小鼠EAE模型,建立模型组和预处理加模型组（预加模组）,并设立空白对照组,观察各组小鼠临床评分,用ELISA法检测各组小鼠血清IL-23和1L--17的表达。结果：预加模组小鼠临床评分明显低于模型组（1．44土0．56、2．75±1．13,P〈o．05）。模型组IL-23及IL-17的表达均较空白对照组高（P〈0．05）,预加模组较模型组低（P〈0．05）,预加模组和空白对照组比较差异无统计学意义。结论：康复预处理能够延迟EAE发生,这一作用可以通过抑制IL-23／IL-17轴而实现。     

维生素D3对小鼠肝脏病理学及肝纤维化指标的影响

目的探讨维生素D3(VitD3)对四氯化碳(CCl4)诱导雌性BALB/C小鼠急性肝损伤(acute liver injury)中肝纤维化指标的影响及其病理学改变。方法将25只SPF级雌性BALB/C小鼠随机分为正常对照组、模型组及VitD3高、中、低剂量干预组。VitD3高、中、低剂量组分别每日给予VitD3 15.0μg、7.5μg、l.0μg腹腔注射,正常对照组与模型组腹腔注射等量生理盐水,预处理2周。VitD3预处理1周后,模型组与VitD3高、中、低剂量组每只小鼠隔日腹腔注射0.1%CCl4橄榄油溶液0.2mL造模,正常对照组与模型组同步隔日腹腔注射生理盐水,CCl4给药7d后,全部小鼠经麻醉眼眶采血,分离肝脏并称质量,光镜下观察肝组织HE、Masson及网状纤维染色的病理变化;常规生化方法检测血清丙氨酸氨基转移酶(ALT)、天门冬氨酸氨基转移酶(AST);放射免疫法检测血清透明质酸(HA)、Ⅲ型前胶原N端肽(PⅢNP)、Ⅳ型胶原(CⅣ)、层粘连蛋白(LN)。结果与正常对照组比较,模型组BALB/C小鼠肝脏指数和血清ALT、AST、HA、LN、PⅢNP、CⅣ皆显著升高,差异有统计学意义(P0.01)。与模型组比较,VitD3高、中剂量组的肝脏指数、ALT、AST、HA、LN、PⅢNP、CⅣ均显著降低,差异有统计学意义(P0.01)。结论 VitD3能有效减轻CCl4诱导急性肝损伤小鼠肝组织的炎性活动度和纤维化程度,提示VitD3具有一定的抗炎、抗纤维化作用。     

小鼠H-2半相合非清髓同种异基因造血干细胞移植模型的建立及其相关研究

为了探讨小鼠主要组织相容性抗原 (H 2 )半相合非清髓移植中预处理方案的可行性、移植后的造血重建、嵌合体水平及GVHD的发生情况 ,以CB6F1小鼠为受鼠 ,分为 3组 ,移植前 5天开始给予预处理 ,A组给予清髓(10 .5Gy)预处理方案 ,B组给予全身照射 (2Gy) Ara C Cy ,C组为全身照射 (2Gy) Ara C Cy Flu ,对所有受鼠均未进行GVHD防治。所有受鼠在第 0天经尾静脉注射C5 7BL 6小鼠混合细胞悬液 (2× 10 7骨髓细胞 1×10 7脾细胞 ) ,然后观察其造血恢复、植入及移植物抗宿主病 (GVHD)的情况。结果表明 :A组植入结果始终为完全供者型嵌合体 ,B和C组则为混合型或完全供者型嵌合体 ,其中B组混合嵌合体保持在 80 %以下 ,且在移植 5 0天以后有下降趋势 ,而C组嵌合体水平保持在 80 %以上 ,其嵌合接近或为完全供者型 ,移植 5 0天以后仍保持稳定。由于没有给予GVHD防治措施 ,各组均发生不同程度的GVHD ,其中A组受鼠GVHD的发生率和死亡率明显高于B和C两组 (P≤ 0 .0 1) ,但B和C两组之间无显著差异。结论 :以氟达拉宾为主的非清髓预处理方案 ,能够在受者体内形成稳定且持久的植入 ,并通过在受者体内形成混合性嵌合体 ,诱导供体对受体的特异免疫耐受 ,减少或避免GVHD的发生。