NY-ESO-1/HSP65融合蛋白腹腔免疫小鼠的免疫效果

目的  观察NY-ESO-1/热休克蛋白65（heat shock protein 65,HSP65）融合蛋白（NY-H）免疫BALB/c小鼠诱导的体液和细胞免疫应答及其抗肿瘤作用。 方法  将BALB/c小鼠按简单随机方法分组,分别用 NY-H、NY-ESO-1与HSP65混合（NY+H）、NY-ESO-1、HSP65和PBS腹腔免疫4次,间隔2周。每次免疫后第14天,对小鼠尾静脉采血,检测血清抗体滴度。末次免疫后第7天,取小鼠脾细胞,检测细胞因子、淋巴细胞增殖水平、CD4+和CD8+ T细胞的百分比及对肿瘤细胞的特异性杀伤作用,采用t检验或2检验对结果进行比较。结果 NY-H组小鼠在第1次免疫后就产生了较高水平的抗体（吸光度值为0.841±0.059）,与NY+H组（0.333±0.077）和NY-ESO-1组（0.275±0.183）相比差异有统计学意义（t=12.753,P<0.01;t=7.204,P<0.01）。NY-H组小鼠的IL-2和IFN-γ分泌能力均高于其他各组,并且CD4+、CD8+ T细胞数量大幅增高,细胞增殖率（38.00%±3.84%）明显高于NY+H组（12.90%±0.65%）和NY-ESO-1组（10.90%±1.44%）（x=830.1,P<0.01;x²=994.0,P<0.01)。小鼠免疫NY-H后,脾淋巴细胞能特异性杀伤表达NY-ESO-1的小鼠B16肿瘤细胞。 结论  NY-H能诱导小鼠产生较强的体液和细胞免疫应答,且具有良好的抗肿瘤作用。     

川芎嗪联用环孢素A抗皮肤移植排斥反应的作用机制研究

目的：探讨川芎嗪（TMPZ）联用亚剂量环孢素A（CsA）对同种异体小鼠皮肤移植的抗排斥反应作用。方法：将BALB/c小鼠作为供体,C57BL/6小鼠作为受体,行背-背皮肤移植术。随机分为模型组、TMPZ组（50 mg·kg^-1）、CsA组（10 mg·kg^-1）、TMPZ+亚剂量CsA组（TMPZ 50 mg·kg^-1+CsA 5 mg·kg^-1）、假手术组,腹腔注射给药10 d。观察受体鼠移植皮片的存活情况,酶联免疫吸附法（ELISA）检测血浆中白介素-2（IL-2）、IL-4、IL-10、γ-干扰素（IFN-γ）含量,流式细胞术分析脾脏CD4⁺、CD25⁺T淋巴细胞亚群的比例。结果：各给药组小鼠移植皮片的存活天数均有不同程度的延长,与模型组相比差异均有显著性（P<0.01）。TMPZ+CsA组的移植皮片存活天数比TMPZ组或CsA组均有显著延长（P<0.05或P<0.01）;与模型组相比,TMPZ+CsA组以及TMPZ单用组均可明显减少血浆Th1类细胞因子（IL-2、IFN-γ）的表达（P<0.01）,明显增高Th2类细胞因子（IL-4、IL-10）的表达（P<0.01）。与模型组相比,各给药组均可明显增高脾脏CD4⁺ CD25⁺T淋巴细胞的比例（P<0.05或P<0.01）。结论：TMPZ具有抑制同种异体小鼠皮肤移植排斥反应的效果,与CsA联合应用可能具有协同作用,其作用机制可能与影响Th1/Th2细胞因子、刺激CD4⁺、CD25⁺T淋巴细胞的表达等有关。     

TAK-242通过调控JAK2/STAT3通路抑制小鼠心肌缺血再灌注损伤炎症反应

目的：探讨Toll样受体4拮抗剂TAK-242抑制小鼠心肌缺血/再灌注损伤（ischemia/reperfusion,I/R）炎症反应的分子机制。方法：选用48只雄性C57BL/6小鼠随机分为4组：假手术组（sham）、模型组（I_30min /R_24h）、给药组[I/R+TAK-242（3 mg·kg^-1）]、干预组[I/R+TAK-242+AG490（15 mg·kg^-1）]。再灌注24 h后心脏超声检测小鼠心功能,氯化三苯基四氮唑（TTC）染色法测定心肌梗死面积,HE染色观察心肌病理改变,WB检测心肌JAK2/STAT3磷酸化水平,ELISA检测血清IL-6、TNF-α、IL-10和高迁移率族蛋白B1（HMGB1）浓度。结果：与sham组比较,I/R组小鼠左心室收缩期直径（LVIDs）延长（P<0.01）,左心室射血分数（LVEF）和左心室短轴缩短分数（LVFS）显著降低（P<0.001或P<0.01）,心梗面积明显增加并出现心肌炎性浸润,心肌p-JAK2/p-STAT3表达明显升高（P<0.01或P<0.05）,血清IL-6、IL-10、TNF-α和HMGB1水平显著升高（P<0.001或P<0.01）。与I/R组比较,TAK-242给药组小鼠LVIDs缩短（P<0.05）,LVEF和LVFS显著升高（P<0.01或P<0.05）,心梗面积缩小（P<0.01）,心肌炎症浸润减轻,心肌p-JAK2/p-STAT3表达降低（P<0.01或P<0.05）,血清IL-6和TNF-α水平明显下降（P<0.001或P<0.01）,而IL-10和HMGB1浓度进一步升高（P<0.01）。与TAK-242给药组比较,AG490干预可显著加强TAK-242治疗作用,包括心肌收缩功能增强,心梗面积缩小及炎性浸润程度减轻,心肌p-JAK2/p-STAT3表达降低（P<0.05）,血清IL-6、TNF-α浓度下降而IL-10、HMGB1浓度升高（P<0.01或P<0.05）。结论： Toll样受体4拮抗剂TAK-242抑制小鼠I/R炎症反应与JAK2/STAT3信号通路失活有关。     

金茵清热口服液对甲型H1N1流感病毒致细胞病变效应的影响

目的：研究金茵清热口服液（试药）抑制甲型H1N1流感病毒的作用及其对喉癌上皮细胞（Hep-2细胞）免疫机能的影响。方法：甲型H1N1流感病毒感染狗肾细胞（MDCK细胞）和Hep-2细胞后,给予不同浓度的试药。显微镜观察细胞病变效应（cytopathic effect,CPE）,MTT法检测细胞活性,荧光定量PCR（qRT-PCR）检测试药作用于Hep-2细胞后细胞因子（TNF-α,IFN-1α,IFN-1β,IL-1α和IL-6）mRNA的水平。结果：实验细胞感染病毒后用试药处理,当试药质量浓度为3 mg·mL^-1时,MDCK细胞存活率提高了61.6%;试药质量浓度为4 mg·mL^-1时,Hep-2细胞存活率提高了40.6%,显著高于药物空白组细胞的存活率（P<0.05）。同时,试药显著上调Hep-2细胞的TNF-α（2.39倍）和IFN-1α（1.98倍）水平,下调炎症因子IL-1α（90%）和IL-6（64%）水平（P<0.05）,而IFN-1β水平的差异无统计学意义（P>0.05）。结论：金茵清热口服液可以显著提高染毒细胞的存活率,并能显著上调Hep-2细胞抗病毒免疫分子,同时抑制炎症反应,推测金茵清热口服液调节细胞免疫应答可能是其抗流感病毒作用机制之一。     

重组毕赤酵母HBsAg诱导小鼠细胞免疫应答的研究

目的   研究毕赤酵母表达的重组HBsAg(Pichia-rHBsAg)在小鼠中诱导T细胞免疫应答的能力,评价疫苗的免疫原性.   方法  以rHBsAg免疫雌性BALB/c小鼠,制备小鼠脾淋巴细胞,并在体外以抗原/特异性多肽刺激.采用ELISA法测定抗原特异性T淋巴细胞分泌的细胞因子,酶联免疫斑点法(ELISPOT)测定CTL频数.   结果 由毕赤酵母表达的rHBsAg可在小鼠中诱导Th1和Th2型免疫应答;加铝佐剂的rHBsAg诱导IFN-γ的水平及CTL克隆明显高于无佐剂抗原;Pichia-rHBsAg刺激小鼠细胞免疫应答的水平强于酵母-rHBsAg,与中国仓鼠卵巢细胞-rHBsAg相仿.   结论   Pichia-rHBsAg可在BALB/c小鼠中诱导较强的细胞免疫应答.     

贝伐珠单抗注射液在改善银屑病小鼠模型中的作用及机制

目的：研究贝伐珠单抗注射液（安维汀）在改善银屑病小鼠模型中所发挥的作用及作用机制。方法：通过对银屑病小鼠模型的构建,安维汀药物处理,小鼠病情等级评定,细胞因子检测,mRNA定量,蛋白免疫印迹的生物学方法,研究安维汀用药后,银屑病小鼠病情,体内细胞因子和血管生成因子mRNA含量及蛋白含量变化。结果：小鼠病情级别评定结果显示,安维汀能减轻银屑病的症状;mRNA定量结果和免疫蛋白印迹结果均显示,安维汀可以改变银屑病小鼠体内炎症细胞因子含量和血管生成因子含量,从而缓解银屑病的症状。结论：安维汀通过影响Th1类细胞因子IFN-γ,Th17类细胞因子IL-17A,Th2类细胞因子IL-4及Treg细胞因子IL-10及血管生成相关因子的含量,抑制炎症和血管的生成,从而改善银屑病症状。     

初诊2型糖尿病短期胰岛素泵强化治疗对血清炎症因子及胰岛功能的影响

目的：观察短期胰岛素泵强化治疗对初诊2型糖尿病患者（T2DM）血清C反应蛋白（CRP）、肿瘤坏死因子-α（TNF-α）、白介素-6（IL-6）水平及胰岛β细胞功能的影响。方法：初诊T2DM患者64例,应用胰岛素泵强化治疗14 d,比较治疗前后血清CRP、TNF-α、IL-6、胰岛素分泌指数（HOMA-IS）、胰岛素抵抗指数（HOMA-IR）的变化,并与38例健康对照组人群进行比较。结果：T2DM患者治疗前血清CRP、TNF-α、IL-6水平较正常对照组明显升高（P<0.01）;强化治疗后较治疗前血清CRP、TNF-α、IL-6水平明显降低（P<0.01）,HOMA-IS明显升高（P<0.01）,而HOMA-IR则明显降低（P<0.01）。结论：初诊T2DM患者存在慢性炎症反应,胰岛素具有抗炎作用,胰岛素强化治疗可改善患者糖代谢,减轻胰岛素抵抗,从而降低炎症因子的表达。     

异型南五味子醇提物对D-半乳糖胺/脂多糖致小鼠急性肝损伤的保护作用

目的：研究异型南五味子对D-半乳糖胺/脂多糖诱导小鼠肝损伤的保护作用。方法：ICR小鼠按体质量随机分为 6组,即正常组、模型组、联苯双酯（200 mg·kg^-1）组,异型南五味子醇提物低、中、高剂量（200、400、800 mg·kg^-1）组。每天给药一次,连续给药14 d,末次给药1 h后,除正常组外其余各组用600 mg·kg^-1 D-半乳糖胺和40 μg·kg^-1脂多糖腹腔注射以复制小鼠急性肝损伤模型,末次给药12 h后,摘眼球取血,取材。用生化法测定血清中ALT、AST水平,用酶联免疫法测定各组小鼠肝匀浆中SOD、MDA、MPO及GPx-2含量,测定血清及肝组织中 TNF-α、IL-6、IL-10炎性因子表达水平,并于光镜下检查肝组织病理变化。结果：与模型组比较,异型南五味子200、400、800 mg·kg^-1剂量均能显著降低血清中ALT、AST水平（P<0.05或P<0.01）,明显降低肝匀浆中MDA、MPO含量,显著升高SOD、GPx-2水平（P<0.05或P<0.01）,并能降低肝脏及血清中TNF-α、IL-6含量,升高IL-10的表达水平（P<0.05或P<0.01）。病理观察显示异型南五味子组能明显减轻肝脏灶性坏死、水肿,减少肝脏炎性细胞浸润。结论：异型南五味子对腹腔注射600 mg·kg^-1D-半乳糖胺和40 μg·kg^-1脂多糖诱导的小鼠急性肝损伤具有较好的保护作用。     

参麦注射液联合双抗治疗对老年非ST段抬高型急性冠脉综合征患者血清MMP-9、TNF-α及IL-6水平的影响

目的：探讨参麦注射液联合双抗治疗对老年非ST段抬高型急性冠脉综合征患者血清MMP-9、TNF-α及IL-6水平的影响。方法：研究对象为80例老年非ST段抬高型急性冠脉综合征患者,均接受常规治疗,对照组给予阿司匹林及氯吡格雷双抗治疗,研究组在此基础上再予参麦注射液静点,连续治疗4 d。检测治疗前、后血清炎性因子及心肌损伤标志物,胸痛发作情况及出血情况,血小板聚集率及主要心血管发生率,比较临床疗效。结果：与治疗前比较,两组血清血清肿瘤坏死因子α（tumor necrosis factor,TNF-α）、白介素-6（interleukin-6,IL-6）及C反应蛋白（C-reactionprotein,CRP）水平均显著降低（P<0.01）,肌钙蛋白T（cardiac troponin T,cTnT）、肌酸激酶同工酶（isoenzyme of creatine kinase,CK-MB）水平均明显升高（P<0.05）,基质金属蛋白酶-9（matrix metallo preteinases-9,MMP-9）水平明显降低（P<0.05）,胸痛发作频率显著降低（P<0.01）,持续时间显著缩短（P<0.01）;与对照组比较,研究组血清TNF-α、IL-6及CRP水平较低（P<0.01）,cTnT、CKMB水平较低,MMP-9水平较低（P<0.05）,胸痛发作频率较低（P<0.01）,持续时间较短（P<0.01）,血小板聚集率较低（P<0.01）,主要心血管发生率较低（P<0.01）,治疗有效率较高（P<0.01）。结论：参麦注射液联合双抗治疗老年非ST段抬高型急性冠脉综合征有显著疗效,能明显改善胸痛,且能降低血清MMP-9、TNF-α及IL-6水平。     

三叶青藤正丁醇部位对DBA/1小鼠胶原性关节炎的作用及机制

目的：观察三叶青藤正丁醇部位（nbuIR）对胶原性关节炎小鼠关节病变及血清炎症因子的抑制作用。方法：用DBA/1小鼠建立胶原性关节炎（CIA）动物模型,将造模成功小鼠随机分为模型对照组、甲氨蝶呤组、nbuIR低、中、高剂量组,另设正常对照组。灌胃给药,给药期间每周进行一次关节炎指数评分。干预结束后,ELISA法检测各组小鼠血清IL-23和IL-17水平,H-E染色观察各组小鼠踝关节组织病理学改变。结果：与模型对照组比较,nbuIR高剂量组血清IL-23和IL-17水平明显降低（P<0.01）,滑膜细胞增生与炎症细胞浸润程度明显减轻。结论：nbuIR能改善CIA小鼠局部关节肿胀,下调血清IL-23和IL-17水平,减少滑膜炎症程度,其机制可能与调节IL-23/IL-17轴有关。     

Ganoderma tsugae in vivo modulates Th1/Th2 and macrophage responses in an allergic murine model

We have reported that Ganoderma tsugae supplementation alleviates bronchoalveolar inflammation in an airway sensitization and challenge model with female BALB/c mice. However, the effects of G. tsugae supplementation in vivo on serum antibody levels, splenocyte and peritoneal microphage immune responses have not yet been determined. In this study, serum antibody levels, cytokines and splenocyte chemical mediators and peritoneal macrophage cultures from ovalbumin (OVA)-sensitized and -challenged mice were examined after continuously consuming G. tsugae supplementation diets for 5 weeks. The results showed that OVA sensitization and challenge significantly (P < 0.05) decreased the spontaneous production of IL-2 (Th1) cytokine, but significantly (P < 0.05) increased spontaneous and OVA-stimulated IL-4 (Th2) production in splenocyte cultures from experimental mice. OVA administration significantly decreased both spontaneous and LPS/IFN-γ-stimulated IL-1β and IL-6 levels in peritoneal macrophage cultures from experimental mice. However, dietary supplementation with G. tsugae significantly increased spontaneous IL-2 level, but slightly decreased spontaneous IL-4 level in cultured splenocyte supernatants in the experimental groups. G. tsugae supplementation enhanced pro-inflammatory cytokines IL-1β and IL-6 production in cultured peritoneal macrophages. However, the nitric oxide level from cultured peritoneal macrophages and serum OVA-specific IgE and IgG_2a antibody levels was not significantly affected. These results suggest that OVA sensitization and challenge induced a Th2-skewed splenocyte response and decreased peritoneal macrophage cytokine secretion. G. tsugae supplementation in vivo modulated the Th1/Th2 balance and enhanced macrophage immune responses. However, the supplementation diet could not fully reverse the Th2-skewed responses to level of Th1-skewed responses.     

Mechanism of TCDD-induced suppression of antibody production: effect on T cell-derived cytokine production in the primary immune reaction of mice.

2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is known to suppress antigen-specific antibody production in humoral immune reactions, but the precise mechanism remains unclear. Since T cell activation and subsequent production of type 2 helper T (Th2) cell-derived cytokines are required for antigen-specific antibody production in humoral immunity, we examined the effects of TCDD on splenic T cell numbers, T cell growth factor IL-2 production, and Th2 cell-derived cytokine production. C57BL/6N mice were orally given TCDD (20 microg/kg) or vehicle, and immediately intraperitoneally immunized with ovalbumin (OVA) adsorbed to alum, and cellular changes in the spleen and cytokine production by spleen cells were investigated from Day 1 to Day 14. In vehicle-control mice the numbers of splenic CD3(+) T cells increased from Day 7 onward, but no increase was observed in the TCDD-exposed mice. When spleen cells from control mice were cultured and restimulated with OVA ex vivo, a significant amount of IL-2 was found from Day 1, but it decreased on Day 7, whereas TCDD exposure promptly suppressed the increase on Day 4. TCDD exposure significantly suppressed the production of Th2 cell-derived cytokines IL-4, IL-5, and IL-6, which were prominently increased from Day 4 onward in control mice. The dose-dependent study showed that IL-5 production was significantly suppressed in a dose-dependent manner starting at 1 microg/kg TCDD. Moreover, separation and reconstitution studies showed that the TCDD-induced suppression of IL-5 production was due to the impaired function of T cells rather than that of antigen-presenting cells. The results of this study suggest that TCDD-induced suppression of T cell activation and Th2-type cytokine production is involved in the impairment of antigen-specific antibody production.     

Immunological adjuvant effect of Boswellia serrata (BOS 2000) on specific antibody and cellular response to ovalbumin in mice

In this study, the biopolymeric fraction BOS 2000 from Boswellia serrata was evaluated for its potential ability as adjuvants on the immune responses to ovalbumin (OVA) in mice. Balb/c mice were immunized subcutaneously with OVA 100 μg alone or with OVA 100 μg dissolved in saline containing alum (200 μg) or BOS 2000 (10, 20, 40 and 80 μg) on Days 1 and 15. Two weeks later, OVA specific antibodies in serum; concanavalin A (Con A), OVA stimulated splenocyte proliferation, CD4/CD8/CD80/CD86 analysis in spleen cells and its estimation of cytokines (IL-2 and IFN gamma) from cell culture supernatant were measured. OVA specific IgG, IgG1 and IgG2a antibody levels in serum were significantly enhanced by BOS 2000 (80 μg) compared with OVA control group. Moreover, the adjuvant effect of BOS 2000 (80 μg) on the OVA-specific IgG, IgG1, and IgG2a antibody responses to OVA in mice were more significant than those of alum. BOS 2000 significantly enhanced the Con A and OVA induced splenocyte proliferation in the OVA immunized mice especially at a dose of 80 μg (p<0.001). However, no significant differences were observed among the OVA group and OVA/alum group. At a dose of 80 μg (p<0.001), there was a significant increase in the CD4/CD8 and CD80/CD86 analysis in spleen cells and cytokine (IL-2 and IFN-gamma) profile in the spleen cell culture supernatant was observed. In conclusion, BOS 2000 seems to be a promising balanced Th1 and Th2 directing immunological adjuvants which can enhance the immunogenicity of vaccine.     

Immunomodulatory activity of novel adjuvant formulations based on Montanide ISA oil-based adjuvants and peptidoglycan monomer

The aim of the study was to directly compare the potential of Montanide ISA720 and ISA206 oil-based adjuvant formulations on the induction of Th1/Th2-type of immune response, and to compare their effect to Complete Freund's adjuvant (CFA), a well known Th1 inducer. IgG isotype profiles (IgG1/IgG2a ratios) and specific cytokine secretion (IFN-gamma and IL-4) as specific markers of Th1/Th2-type of immune response were monitored in experimentally immunised mice using ovalbumin (OVA) as an antigen. Specifically, we wanted to evaluate whether the incorporation of immunostimulating peptidoglycan monomer (PGM) into two oil-based adjuvants (ISA720(PGM) and ISA206(PGM)) influences their capability on Th1/Th2-type of immune response switching. The experiments were carried out using two genetically different inbred strains of mice, i.e. CBA and NIH/OlaHsd mice, respectively. We found significant differences in immune responses related to the genetic background of the two mice strains used in the study. In both mice strains, ISA720 formulations had similar effect to the positive control, CFA, and induced the switch towards Th1-type of immune response specific for OVA. However, ISA206 formulations were less effective in inducing the switch towards Th1 in CBA mice, while in NIH/OlaHsd mice promoted the switch towards Th2-type of immune response.     

Diesel exhaust, carbon black, and silica particles display distinct Th1/Th2 modulating activity

Certain particulate air pollutants may play an important role in the increasing prevalence of respiratory allergy by stimulating T helper 2 cell (Th2)-mediated immune responses to common antigens. The study described here examined different particles, diesel exhaust particles (DEP), carbon black particles (CBP), and silica particles (SIP) for their immunomodulating capacity in both primary and secondary immune responses in female BALB/C mice. The primary response was studied after subcutaneous injection of 1 mg of particle together with 10 microgram of reporter antigen TNP-OVA (2,4,6-trinitrophenyl coupled to ovalbumin) into the hind paw. Interferon-gamma (IFN-gamma) and interleukin 4 (IL-4) production was assessed in the popliteal lymph node (PLN) at Day 2 and Day 5 after injection by flow cytometry and ELISA. The number of IL-4-containing CD4(+) T cells increased between Day 2 and Day 5 in DEP- and CBP-exposed mice, in contrast to SIP-treated animals. IL-4 production by cultured PLN cells was also significantly increased for DEP- and CBP-treated animals. The secondary response was studied in different organs after an intranasal challenge with TNP-OVA (50 microgram), which was given 4 weeks after the initial subcutaneous injection. Five days after challenge the number of antibody-forming cells (AFCs) was assessed in peribronchial lymph nodes (PBLN), spleen, bone marrow, and PLN, and antibody levels were determined in weekly obtained blood samples. It appeared that all particles acted as adjuvant, but the different particles stimulated distinct types of immune responses to TNP-OVA. DEP-treated animals show high IgG1 and IgE levels in serum and high IgG1 and IgE-forming AFC numbers in PBLN, bone marrow, and spleen. CBP-treated animals show even higher IgG1 and IgE levels and AFC numbers, and in addition display IgG2a production. SIP-injected animals display predominantly IgG2a responses. It is concluded that DEP are able to skew the immune response toward the T helper 2 (Th2) side, whereas SIP stimulate a Th1 response and CBP have a mixed activity, stimulating both Th1 and Th2 responses in this model.     

The ex vivo study of synergistic effects of polycyclic aromatic hydrocarbon, benzo(a)pyrene with ovalbumin on systemic immune responses by oral route

The present study was undertaken in order to examine whether oral administration of soluble antigen together with one of polycyclic aromatic hydrocarbons (PAHs) which is present in diesel exhaust particles (DEPs) called benzo(a)pyrene (BP), induced the systemic immune response in mice or not. Mice were orally given 1mg of ovalbumin (OA), a common food allergen, every 3 days over a period of 15 days. The results showed that oral administration of OA plus BP produced anti-OA IgE antibodies in serum, whereas either OA or BP alone failed to show the antigen-specific IgE antibody production. Production of anti-OA IgE antibody, which is dependent on Th2 CD4(+) T cells, was seen in mice fed with combined OA and BP was significantly higher than that of other groups. The anti-OA antibody production was associated with marked secretion of the Th1 cytokines, IFN-gamma and IL-12p70 as well as the Th2 cytokines IL-4, and IL-10. These results suggest that BP may act as a mucosal adjuvant in the gut enhancing systemic Th1 and Th2 immune responses and might play a role in oral immunization and food allergy.     

Diosgenin, a steroidal sapogenin, enhances antigen-specific IgG2a and interferon-gamma expression in ovalbumin-sensitized BALB/c mice

The effect of diosgenin, the most abundant sapogenin in Chinese yam, on humoral immunity was investigated. Ovalbumin (OVA)-sensitized and challenged BALB/c mice were administered daily with diosgenin for 34 days. The production of OVA-specific serum IgG2a was significantly enhanced by diosgenin treatment, whereas total IgE and OVA-specific IgG1, IgG2a and IgM were unaffected. In parallel with the enhancement of IgG2a, OVA-induced IFN-gamma secretion and mRNA expression were markedly elevated in splenocytes of diosgenin-treated mice, whereas IL-4 expression was unaltered. Furthermore, the expression of T-bet, but not of GATA-3, in splenocytes was up-regulated by diosgenin administration. However, diosgenin treatment did not modulate IL-4 mRNA expression and inflammatory cell infiltration in the lung of OVA-sensitized and challenged mice. Collectively, these data suggest that diosgenin regulates the systemic immune response towards the Th1 direction in response to OVA sensitization. The present study provides evidence to show that intake of diosgenin modulates certain aspects of acquired immunity, including the enhancement of antigen-specific IgG2a and IFN-gamma expression, which may be mediated through the up-regulation of Th1 differentiation.     

Haemolytic activities and adjuvant effect of Anemone raddeana saponins (ARS) on the immune responses to ovalbumin in mice

In this study, saponins (ARS) extracted from the rhizoma of Anemone raddeana were evaluated for their haemolytic activities and its potential ability as adjuvant on the cellular and humoral immune responses of ICR mice against ovalbumin. The haemolytic activity of ARS was determined using 0.5% rabbit red blood cell. ARS showed a slight haemolytic effect, with its haemolytic percents being 16.50 and 3.56% at the concentrations of 500 and 250 microg/ml, respectively. ICR mice were immunized subcutaneously with OVA 100 microg alone or with OVA 100 mug dissolved in saline containing Alum (200 microg), QuilA (10 and 20 microg) or ARS (50, 100 or 200 microg) on Days 1 and 15. Two weeks later (Day 28), concanavalin A (Con A)-, lipopolysaccharide (LPS)- and OVA-stimulated splenocyte proliferation and OVA-specific antibodies in serum were measured. ARS significantly enhanced the Con A-, LPS-, and OVA-induced splenocyte proliferation in the OVA-immunized mice especially at a dose of 100 microg (P<0.01 or P<0.05). The OVA-specific IgG, IgG1 and IgG2b antibody levels in serum were also significantly enhanced by ARS compared with OVA control group (P<0.01 or P<0.05). Moreover, no significant differences (P>0.05) were observed between enhancing effect of ARS and QuilA on the OVA-specific IgG2b antibody responses to OVA in mice. The results suggest that ARS showed a slight haemolytic effect and enhanced significantly a specific antibody and cellular response against OVA in mice.     

Co-encapsulation of an antigen and CpG oligonucleotides into PLGA microparticles by TROMS technology.

It seems well established that CpG oligonucleotide Th1-biased adjuvant activity can be improved when closely associated with a variety of antigens in, for example, microparticles. In this context, we prepared 1-mum near non-charged poly(lactic-co-glycolic) acid (PLGA) 502 and PLGA 756 microparticles that loaded with high-efficiency antigen (50% ovalbumin (OVA), approximately) into their matrix and CpG-chitosan complexes (near to 20%) onto their surface maintaining OVA and CpG integrity intact. In the intradermal immunization studies, whereas OVA microencapsulated into PLGA 756 alone induced a strong humoral immune response assisted by a very clear Th1 bias (IgG2a/IgG1=0.88) that was decreased by CpG co-delivery (IgG2a/IgG1=0.55), the co-encapsulation of CpG with OVA in PLGA 502 particles significantly improved the antibody response and isotype shifting (IgG2a/IgG1=0.73) in comparison with mice immunized with OVA-loaded PLGA 502 (IgG2a/IgG1=0). This improvement was not correlated with the cellular immune response where the effect of co-encapsulated CpG was rather negative (2030 and 335pg/mL IFN-gamma for OVA PLGA 502 and OVA CpG PLGA 502, respectively). These results underscore the critical role of polymer nature and microparticle characteristics to show the benefits of co-encapsulating CpG motifs in close proximity with an antigen.