Th17及IL-17与实验性自身免疫性重症肌无力发病过程的相关性

目的探讨Th17及其相关因子IL-17与实验性自身免疫性重症肌无力(EAMG)发病过程的相关性。方法建立EAMG大鼠模型及CFA对照组,分别于两发病时相(早期发病高峰和晚期发病高峰),采用ELISA法检测血清以及淋巴细胞培养上清中IL-17的含量;流式细胞仪检测CD4+IL-17+淋巴细胞含量;3H增殖试验检测淋巴细胞的增殖能力;B-ELISPOT法检测B细胞的抗体分泌情况。结果与CFA组相比,EAMG组大鼠血清、淋巴细胞培养上清中IL-17的表达以及CD4+IL-17+淋巴细胞含量在早期发病时相差异均无统计学意义;而在晚期发病时相则均明显增多。与非刺激组相比,IL-17的刺激对CFA和EAMG组淋巴细胞的增殖能力及B细胞抗体分泌水平,在早期发病时相均无明显影响;而在晚期发病时相,EAMG组均明显升高。结论Th17及其相关因子IL-17参与大鼠EAMG的晚期发病时相,并促进疾病的发展。     

卡介苗诱导人外周血单个核细胞Th1活化

目的探讨卡介苗(BCG)对人外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)Th1活化的调节作用。方法经密度梯度离心法制备PBMCs悬液,用BCG进行体外刺激。WST1法测定BCG对PBMCs增殖能力的影响;ELISA法检测BCG对PBMCs分泌干扰素γ(interferonγ,IFNγ)及IL-4水平的影响;流式细胞术检测BCG对PBMCs Th1活化的影响。结果经BCG体外刺激后,PBMCs的增殖能力、IFNγ的分泌水平及CD4+IFNγ+细胞比例均显著增加(P0.05),IL-4的分泌水平显著下降(P0.05)。结论 BCG可显著诱导PBMCs发生Th1活化,为深入研究BCG的抗肿瘤作用机制奠定了基础。     

骨化三醇对实验性自身免疫性脑脊髓炎的治疗作用及相关机制

目的探讨骨化三醇(Calcitriol)对实验性自身免疫性脑脊髓炎(Experimental autoimmune encephalomyelitis,EAE)的治疗作用及相关机制。方法用含200μg髓鞘少突胶质细胞糖蛋白35-55肽段(Myelin oligodendrocyte glycoprotein 33-35,MOG33-35)、250μg结核菌素的50μl弗氏不完全佐剂(IFA)皮内免疫C57BL/6小鼠,并分别于免疫当天和第2天注射百日咳毒素,建立EAE实验性动物模型(EAE组);骨化三醇组从免疫当天起隔日腹腔注射骨化三醇100 ng进行治疗,观察两组小鼠临床评分的差异;于EAE发病高峰期处死小鼠,取脊髓及淋巴结,通过HE及LFB染色观察脊髓中炎细胞浸润及髓鞘脱失;采用流式细胞术检测两组淋巴细胞CD4+T细胞亚型的分布。结果与EAE组比较,骨化三醇组发病延缓且发病较轻,临床评分差异有统计学意义(P<0.05或P<0.01或P<0.001);骨化三醇组较EAE组小鼠脊髓白质炎性细胞浸润明显减少,脱髓鞘斑块明显减轻;骨化三醇组与EAE组相比,Th17细胞亚群明显受到抑制(P<0.05),而Th2和Treg细胞水平明显升高(P均<0.05)。结论骨化三醇可以延缓EAE发病,减轻临床症状及病理改变,并能够通过调节CD4+T细胞亚群平衡,即抑制Th17细胞,上调Th2和Treg细胞水平发挥对EAE的预防和治疗作用。     

二代测序法分析儿童肺炎支原体肺炎肺泡灌洗液中Th1/Th2的平衡

目的应用二代测序技术(next-generation sequencing,NGS)检测肺炎支原体肺炎(Mycoplasma pneumoniae pneumonia,MPP)患儿肺泡灌洗液(bronchoalveolar lavage fluid,BALF)中,Th1细胞因子IFNγ、Th2细胞因子IL-4的表达水平及免疫应答的平衡变化趋势。方法提取支气管异物、MPP轻症及重症患儿BALF样本,应用NGS技术中转录组测序技术进行深度测序,根据基因表达量检测IFNγ、IL-4表达水平,并计算IL-4/IFNγ比值;采用Real-time q PCR法对样本的IFNγ、IL-4、IL-4/IFNγ相关趋势进行验证。结果 MPP轻症患儿BALF中IL-4、IFNγ表达水平及IL-4/IFNγ比值均较支气管异物对照组升高,MPP重症患儿BALF中IL-4、IFNγ水平较轻症组和支气管异物对照组升高,IL-4/IFNγ比值较支气管异物对照组升高,较轻症组有所下降。结论在MPP患儿免疫反应中,MPP轻症及重症患儿BALF中两类细胞因子均有所升高,IL-4/IFNγ比值变化趋势表明Th1/Th2存在失衡现象,MPP轻症组、重症组与支气管异物对照组相比,平衡向Th2移动,因此,平衡以Th2反应占优势为主。     

淋巴细胞功能相关抗原-1基因敲除对实验性自身免疫性脑脊髓炎小鼠引流淋巴结T细胞活性的影响

目的探讨淋巴细胞功能相关抗原-1(lymphocyte function-associated antigen-1,LFA-1)基因敲除对实验性自身免疫性脑脊髓炎(experimental autoimmune encephalomyelitis,EAE)模型小鼠引流淋巴结T细胞活性的影响。方法采用髓鞘少突胶质细胞糖蛋白(myelin oligodendrocyte glycoprotein,MOG)35-55多肽诱导LFA-1a链CD11a基因敲除的C57BL/6小鼠及野生型C57BL/6小鼠,建立EAE小鼠模型,取MOG诱导后21 d EAE模型小鼠脊髓颈段切片,HE染色和LuxolFastBlue染色,观察小鼠脊髓组织学变化;于MOG诱导后7、14、21d处死小鼠,收集腹股沟引流淋巴结CD4+T淋巴细胞,加入终浓度为1μg/ml的MOG35-55多肽刺激48 h后,应用BrdU试剂盒检测细胞增殖情况;于MOG诱导后4、7、10、14、21 d处死小鼠,收集腹股沟引流淋巴结细胞,加入终浓度为1μg/ml的MOG35-55多肽刺激48 h后,进行细胞内细胞因子染色,观察分泌IFNγ和IL-17的T细胞比例。结果 MOG诱导的野生型小鼠脊髓出现明显淋巴细胞浸润及广泛的脱髓鞘变化,而LFA-1基因敲除小鼠无明显变化;在EAE疾病早期(7 d),LFA-1基因敲除可减少淋巴细胞向引流淋巴结聚集,降低MOG体外刺激T细胞增殖能力,分泌IFNγ及IL-17的T细胞比例也较野生型C57BL/6小鼠明显降低(P0.05);而在EAE疾病缓解期(21 d),LFA-1基因敲除小鼠淋巴细胞聚集及T细胞增殖能力与野生型C57BL/6小鼠相比无明显差异(P0.05),分泌IFNγ及IL-17的T细胞比例高于野生型C57BL/6小鼠(P0.05)。结论在EAE疾病发生早期,LFA-1可能起到了关键性的作用,LFA-1有可能成为自身免疫性疾病治疗的重要分子靶点。     

氢氧化铝佐剂对肠道病毒71型灭活疫苗诱导小鼠细胞免疫应答的影响

目的评价氢氧化铝佐剂对肠道病毒71型(Enterovirus 71,EV71)灭活疫苗诱导小鼠细胞免疫应答的影响。方法用含铝佐剂及无佐剂EV71灭活疫苗免疫BALB/c小鼠,并设生理盐水对照组。于加强免疫后7 d采血,进行小鼠脾单个核细胞(Mononuclear cell,MNC)的分离及CD8+T细胞的去除,采用酶联免疫斑点法(ELISPOT)检测经EV71疫苗原液或VP2-36刺激后的小鼠脾MNC分泌IFNγ水平;应用液相芯片技术(LUMINEX)检测经EV71疫苗原液刺激后的小鼠脾MNC分泌IL-2、IL-4、IL-5、IL-6和IL-10水平;采用体外微量中和试验法检测小鼠血清中和抗体效价。结果 BALB/c小鼠经2次免疫后,铝佐剂疫苗组小鼠脾MNC的IFNγSFC值显著高于无佐剂疫苗组(P均<0.05);铝佐剂疫苗组分泌IL-6和IL-10水平显著高于无佐剂疫苗组(P均<0.05);铝佐剂与无佐剂疫苗组小鼠血清EV71中和抗体阳转率均达到100%,铝佐剂疫苗组诱导的中和抗体效价显著高于无佐剂疫苗组(P<0.01)。结论 EV71灭活疫苗可诱导BALB/c小鼠产生特异性IFNγ、IL-2、IL-6和IL-10应答,铝佐剂可同时增强EV71灭活疫苗的Th1和Th2类免疫应答。     

附红细胞体感染小鼠CD4~+ T淋巴细胞相关因子的检测

目的动态检测小鼠感染附红细胞体后CD4+T淋巴细胞相关细胞因子的变化情况,探讨CD4+T淋巴细胞发挥的免疫学效应。方法将小鼠随机分为实验组(纯化的附红细胞体)和对照组(生理盐水),均经腹腔免疫接种,0.5 ml/只。分别于感染后第3、5、7、9 d,经小鼠尾尖采血,镜下观察附红细胞体形态并进行PCR鉴定。建模成功后,分别于感染后第3、5、7、9 d无菌取小鼠脾脏,采用RT-PCR法检测小鼠脾脏中IL-4、IL-17、IFNγ基因的转录水平。结果各时间点感染小鼠的红细胞均出现不同程度的变形,边缘被附红体附着,PCR扩增产物可见602 bp的特异条带。实验组小鼠脾脏IL-4、IL-17、IFNγ均有不同程度的表达,IL-17在感染在第3天上调,5 d达到高峰,7 d开始下降;IFNγ在感染第3天表达上调,5 d明显下降,7 d表达上升,9 d达到高峰;IL-4始终处于低表达状态。实验组小鼠脾脏IL-4、IL-17、IFNγ的转录水平均高于对照组(P<0.01),IL-17和IFNγ的表达呈相互抑制状态,IL-4呈被抑制状态。结论附红细胞体感染后,IL-17在早期发挥了促进炎症发生和抵抗感染的免疫学作用;IFNγ在感染后期发挥保护炎性反应,避免炎性反应过度发生的免疫学效应;IL-4在此感染过程中作用不明显。     

黄连上清胶囊联合抗敏牙膏治疗慢性牙周炎效果及对Th17/Treg、复发的影响

研究黄连上清胶囊联合高露洁专效抗敏牙膏治疗慢性牙周炎（CP）效果及对辅助性T细胞17/调节性T细胞（Th17/Treg）、复发的影响。选取CP患者134例，采用简单随机化法分为对照组（n=67）和观察组（n=67），分别给予高露洁专效抗敏牙膏、黄连上清胶囊+高露洁专效抗敏牙膏。观察两组治疗效果、不良反应、复发情况及治疗前后牙周袋深度（PD）、牙龈指数（GI）、龈沟出血指数（SBI）、吹气刺激敏感值（BST）、Th17/Treg细胞水平、牙龈卟啉单胞菌（PG）、乳酸杆菌（LB）。结果显示，治疗4周后，观察组治疗总有效率高于对照组（P<0.05）；治疗4周后观察组PD、PLI、GI、BST值低于对照组（P<0.05）；治疗4周后观察组Th17、Treg细胞及PG、LB水平低于对照组（P<0.05）；两组不良反应及复发率比较差异无统计学意义（P>0.05）。黄连上清胶囊联合高露洁专效抗敏牙膏有利于下调CP患者Th17/Treg细胞水平，阻止细菌定植，改善牙周症状，提高治疗效果，且安全性高。     

肠道病毒71型灭活疫苗的小鼠细胞免疫效果

目的评价肠道病毒71型(enterovirus 71,EV71)灭活疫苗诱导小鼠的细胞免疫效果。方法分别以不同剂量(1、2.5、5、10μg/ml,均含铝佐剂1 mg/ml)、不同剂型(含铝佐剂1 mg/ml或不含铝佐剂)、不同免疫剂次(单次免疫或加强免疫)EV71灭活疫苗经腹腔免疫小鼠,0.5 ml/只,均设铝佐剂对照组(仅注射铝佐剂1 mg/ml)。采用经典小鼠树突状细胞(dendritic cell,DC)培养方法制备正常小鼠DC,瑞氏染色法检测细胞形态,流式细胞术分析其表型及纯度;免疫磁珠分选法(magnetic activated cell sorting,MACS)分离小鼠脾脏CD4+、CD8+T淋巴细胞,流式细胞术检测细胞纯度。ELISPOT法检测各组免疫小鼠CD4+、CD8+T淋巴细胞分泌IL-2、IL-4、IFNγ的水平;细胞因子试剂盒检测小鼠淋巴细胞培养上清及小鼠血清中细胞因子的水平。结果 DC形态不规则,表面树枝状突起形态不同,细胞核较大且不规则;DC纯度为(81.39±9.24)%,可高水平表达MHⅡ(I-A/I-E)类分子,中度表达CD86和CD40。CD4+、CD8+T细胞纯度分别为95.27%和94.08%。随着疫苗免疫剂量的增加,CD4+T细胞分泌IL-2、IL-4、IFNγ及CD8+T细胞分泌IFNγ的SFC显著增加,5μg/ml疫苗组达最大值,且明显高于其他组(P均0.05);5μg/ml疫苗组淋巴细胞培养上清中IFNγ、IL-2、IL-4、IL-6、IL-10、IL-13、IL-5、TNF-α含量明显高于其他组(P0.05);1、2.5、5μg/ml疫苗组血清中IL-10含量明显高于铝佐剂对照组(P0.05),1μg/ml疫苗组明显低于2.5及5μg/ml疫苗组(P0.05)。含铝佐剂疫苗组CD4+T细胞分泌IL-2、IL-4、IFNγ及CD8+T细胞分泌IFNγ的SFC、淋巴细胞培养上清中IL-2、IFNγ、IL-4、IL-5、IL-6、IL-10、IL-13、TNF-α水平及血清中IL-10含量均明显高于无铝佐剂疫苗组(P均0.05)。加强免疫组CD4+T细胞分泌IL-2、IL-4、IFNγ及CD8+T细胞分泌IFNγ的SFC、淋巴细胞培养上清中IL-2、IFNγ、IL-4、IL-5、IL-6、IL-10、IL-13、TNF-α水平、血清中IL-10含量均明显高于单次免疫组(P均0.05)。结论 EV71灭活疫苗可诱导小鼠产生特异性细胞免疫反应,本实验为其进一步人体临床试验研究奠定了基础。     

转化生长因子-β在骨髓间充质干细胞治疗实验性自身免疫性重症肌无力机制中的作用

目的探讨转化生长因子-β(TGF-β)在骨髓间充质干细胞(BMSCs)治疗实验性自身免疫性重症肌无力(EAMG)机制中的作用。方法分离培养健康Lewis大鼠BMSCs,并进行大量体外扩增;以大鼠来源的乙酰胆碱受体(R-AChR)2次免疫Lewis大鼠,建立EAMG模型;第2次免疫的同时,经尾静脉移植BMSCs,1×107个/只,依据Lennon评分标准,进行体重测量和临床体征评定。并通过体外实验进一步探讨TGF-β在治疗EAMG过程中的具体机制。结果BMSCs移植明显缓解了EAMG的临床症状,临床评分及体重变化差异均有统计学意义,表明治疗有效;体外实验结果显示,BMSCs能通过TGF-β的分泌影响AChR特异性Th17/Treg细胞亚群的分布及其相关因子的分泌,以anti-TGF-β抗体封闭后,这种调节作用在一定程度上被抑制。结论BMSCs通过细胞因子TGF-β,能在一定程度上调节AChR特异性Th17/Treg细胞亚群的平衡,从而起到治疗EAMG的作用。     

Th17 Activation and Th17/Treg Imbalance in Prolonged Anterior Intraocular Inflammation after Ocular Alkali Burn

Ocular alkali burn (OAB) is a sight-threatening disease with refractory ocular inflammation causing various blinding complications. Th17 lymphocytes account for the pathogeneses of the autoimmune disease and chronic inflammation, but their role in prolonged anterior intraocular inflammation after OAB is still unknown. A rat OAB model was established for this purpose. Anterior intraocular inflammation was observed in both the acute and late phases of OAB, and histological examination confirmed the presence of inflammatory cell infiltration and fibrin exudation in the anterior segment. Luminex xMAP technology and qPCR were used to evaluate the intraocular levels of cytokines. The levels of IL-1β, IL-6, and TNF-α were significantly elevated during the acute phase. The expression of IL-17A gradually increased from day 7 onwards and remained at a relatively high level. Immunofluorescence was performed to identify Th17 cells. CD4 and IL-17A double positive cells were detected in the anterior chamber from days 7 to 28. Flow cytometry showed that the frequency of Th17 cells increased in both lymph nodes and spleen, while the frequency of Treg cells remained unchanged, resulting in an elevated Th17/Treg ratio. The present study suggests that Th17 activation and Th17/Treg imbalance account for prolonged anterior intraocular inflammation after OAB.     

Th17/Treg Cells Imbalance and GITRL Profile in Patients with Hashimoto’s Thyroiditis

Hashimoto’s thyroiditis (HT) is an organ-specific immune disease characterized by the presence of lymphocytic infiltration and serum autoantibodies. Previous studies have confirmed the critical role of Th17 cells in the pathopoiesis of HT patients. Additionally, regulatory T cells (Treg) display a dysregulatory function in autoimmune disease. The purpose of this study is to investigate the alteration of Th17 and Treg cells in HT patients and explore contributing factors. We found there was an increased ratio of Th17/Treg in HT patients and a positive correlation with autoantibodies (anti-TgAb). In addition, there was an increased level of GITRL, which has been demonstrated to be correlated with the increassement of Th17 cells in the serum and thyroid glands of HT patients; the upregulated serum level of GITRL has a positive correlation with the percentage of Th17 cells in HT patients. In summary, an increase in GITRL may impair the balance of Th17/Treg, and contribute to the pathopoiesis of Hashimoto’s thyroiditis.     

Imbalance between Th17 and Regulatory T-Cells in Sarcoidosis

Sarcoidosis is a systemic granulomatous disease, which is thought to result from an aberrant immune response. CD4⁺ T lymphocytes play an important role in the development of granulomas. Previously, the immunopathogenesis of sarcoidosis was focused on Th1/Th2 disturbances. The aim of this study was to evaluate the balance between newer CD4⁺ T lymphocytes, i.e., Treg and Th17 cells. In our studies, a decrease in Treg cells and an increase in Th17 cells were observed in the peripheral blood and BALF of sarcoidosis patients. A significant increase in the Th17/Treg cell ratio was observed in sarcoidosis patients. After treatment with prednisone, the expression of Foxp3 mRNA was elevated in the peripheral blood, and expression of (ROR)γt mRNA showed a downward trend. These findings suggest that sarcoidosis is associated with an imbalance between Th17 and Treg cells in peripheral blood and BALF. Therefore, targeting the cytokines that affect the Th17/Treg ratio could provide a new promising therapy for pulmonary sarcoidosis.     

Peritoneal Fluid from Patients with Ovarian Endometriosis Displays Immunosuppressive Potential and Stimulates Th2 Response

Endometriosis is a common gynaecological disorder characterized by the ectopic growth of endometrial tissue outside the uterine cavity. It is associated with chronic pelvic inflammation and autoimmune reactivity manifesting by autoantibody production and abrogated cellular immune responses. Endometriotic peritoneal fluid contains various infiltrating leucocyte populations and a bulk of proinflammatory and immunoregulatory cytokines. However, the nature and significance of the peritoneal milieu in women with endometriosis still remains obscure. Therefore, the aim of the present study was to investigate the immunoregulatory activity of the peritoneal fluid (PF) from women with endometriosis. The peritoneal fluid samples were collected during laparoscopic surgery from 30 women with and without endometriosis. Immunoregulatory cytokines (IL-2, IL-4, IL-6, IL-10, IL-17A, IFN-γ and TNF) and chemokines (CCL2, CCL5, CXCL8 and CXCL9) were evaluated in PF and culture supernatants generated by unstimulated and CD3/CD28/IL-2-stimulated CD4⁺ T cells cultured in the presence of PF. The effect of PF on the generation of Treg and Th17 cells in CD4⁺ T cell cultures, as well as the natural cytotoxic activity of peripheral blood mononuclear cells, was also investigated. Concentrations of IL-6, IL-10, CCL2, CXCL8 and CXCL9 were significantly upregulated in the PF from women with endometriosis when compared to control women, whereas concentrations of other cytokines and chemokines were unaffected. The culturing of unstimulated and CD3/CD28/IL-2-stimulated CD4⁺ T cells in the presence of endometriotic PF resulted in the downregulation of their IL-2, IFN-γ, IL-17A and TNF production as compared to culture medium alone. On the other side, endometriotic PF significantly stimulated the production of IL-4 and IL-10. Endometriotic PF also stimulated the release of CCL2 and CXCL8, whereas the production of CCL5 and CXCL9 was downregulated. Endometriotic PF stimulated the generation of Treg cells and had an inhibitory effect on the generation of Th17 cells in cultures of CD4⁺ T cells. It also inhibited the NK cell cytotoxic activity of the peripheral blood lymphocytes. These results strongly imply that the PF from patients with endometriosis has immunoregulatory/immunosuppressive activity and shifts the Th1/Th2 cytokine balance toward the Th2 response, which may account for deviation of local and systemic immune responses. However, a similar trend, albeit not a statistically significant one, was also observed in case of PF from women without endometriosis, thus suggesting that peritoneal milieu may in general display some immunoregulatory/immunosuppressive properties. It should be stressed, however, that our present observations were made on a relatively small number of PF samples and further studies are needed to reveal possible mechanism(s) responsible for this phenomenon.     

CD4+ T-Cell Plasticity in Non-Infectious Retinal Inflammatory Disease

Non-infectious uveitis (NIU) is a potentially sight-threatening disease. Effector CD4⁺ T cells, especially interferon-γ-(IFNγ) producing Th1 cells and interleukin-17-(IL-17) producing Th17 cells, are the major immunopathogenic cells, as demonstrated by adoptive transfer of disease in a model of experimental autoimmune uveitis (EAU). CD4⁺FoxP3⁺CD25⁺ regulatory T cells (Tregs) were known to suppress function of effector CD4⁺ T cells and contribute to resolution of disease. It has been recently reported that some CD4⁺ T-cell subsets demonstrate shared phenotypes with another CD4⁺ T-cell subset, offering the potential for dual function. For example, Th17/Th1 (co-expressing IFNγ and IL-17) cells and Th17/Treg (co-expressing IL-17 and FoxP3) cells have been identified in NIU and EAU. In this review, we have investigated the evidence as to whether these ‘plastic CD4⁺ T cells’ are functionally active in uveitis. We conclude that Th17/Th1 cells are generated locally, are resistant to the immunosuppressive effects of steroids, and contribute to early development of EAU. Th17/Treg cells produce IL-17, not IL-10, and act similar to Th17 cells. These cells were considered pathogenic in uveitis. Future studies are needed to better clarify their function, and in the future, these cell subsets may in need to be taken into consideration for designing treatment strategies for disease.     

Human CD4+CD45RA+ T Cells Behavior after In Vitro Activation: Modulatory Role of Vasoactive Intestinal Peptide

Naїve CD4⁺ T cells, which suffer different polarizing signals during T cell receptor activation, are responsible for an adequate immune response. In this study, we aimed to evaluate the behavior of human CD4⁺CD45RA⁺ T cells after in vitro activation by anti-CD3/CD28 bead stimulation for 14 days. We also wanted to check the role of the VIP system during this process. The metabolic biomarker Glut1 was increased, pointing to an increase in glucose requirement whereas Hif-1α expression was higher in resting than in activated cells. Expression of Th1 markers increased at the beginning of activation, whereas Th17-associated biomarkers augmented after that, showing a pathogenic Th17 profile with a possible plasticity to Th17/1. Foxp3 mRNA expression augmented from day 4, but no parallel increases were observed in IL-10, IL-2, or TGFβ mRNA expression, meaning that these potential differentiated Treg could not be functional. Both VIP receptors were located on the plasma membrane, and expression of VPAC₂ receptor increased significantly with respect to the VPAC₁ receptor from day 4 of CD4⁺CD45RA⁺ T activation, pointing to a shift in VPAC receptors. VIP decreased IFNγ and IL-23R expression during the activation, suggesting a feasible modulation of Th17/1 plasticity and Th17 stabilization through both VPAC receptors. These novel results show that, without polarizing conditions, CD4⁺CD45RA⁺ T cells differentiate mainly to a pathogenic Th17 subset and an unpaired Treg subset after several days of activation. Moreover, they confirm the important immunomodulatory role of VIP, also on naїve Th cells, stressing the importance of this neuropeptide on lymphocyte responses in different pathological or non-pathological situations.     

重组弓形虫磷酸甘油酸变位酶2联合白细胞介素-2或干扰素γ诱导小鼠的脾淋巴细胞免疫应答

目的观察重组弓形虫磷酸甘油酸变位酶2(Recombinant Toxoplasma gondii phosphoglycerate mutase 2,rTgPGAM2)联合白细胞介素-2(Interleukin-2,IL-2)或干扰素γ(Interferonγ,IFNγ)滴鼻免疫小鼠诱导的脾淋巴细胞免疫应答,探讨IL-2和IFNγ的佐剂效应。方法将48只BALB/c小鼠随机均分为6组:rTgPGAM2组(30μg)、IL-2组(500 IU)、IFNγ组(1 000 IU)、rTgPGAM2(30μg)+IL-2(500 IU)组和rTgPGAM2(30μg)+IFNγ(1 000 IU)组和对照组(20μl PBS),免疫途径均为滴鼻免疫,共免疫3次。末次免疫后第14天,颈椎脱臼处死小鼠,CCK-8法检测各组小鼠脾淋巴细胞增殖活性,ELISA法检测脾淋巴细胞培养上清中IL-2、IFNγ、IL-4和IL-10水平。结果经rTgPGAM2刺激后,与对照组比较,各免疫组小鼠脾淋巴细胞刺激指数(Stimulation index,SI)均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);经ConA刺激后,仅IL-2组SI显著高于对照组(P<0.01)。各免疫组小鼠脾淋巴细胞培养上清中IL-2和IFNγ含量均显著高于对照组(P<0.05或P<0.01),rTgPGAM2联合IL-2或IFNγ组显著高于rTgPGAM2组(P<0.01或P<0.05);rTgPGAM2组及其联合IL-2或IFNγ组IL-4含量显著高于对照组(P<0.05);而IL-10的含量,只有IFNγ组和rTgPGAM2+IFNγ组显著高于对照组(P<0.05)。结论 rTgPGAM2联合IL-2或IFNγ鼻内免疫小鼠诱导的脾淋巴细胞免疫应答优于rTgPGAM2单独免疫,表明IL-2和IFNγ具有良好的佐剂效应,IL-2的佐剂效应更佳。