阿奇霉素对哮喘致敏大鼠气道炎症及Th1/Th2失衡的调节作用

目的:探讨阿奇霉素对哮喘(OVA)致敏大鼠气道炎症及Th1/Th2失衡的调节作用。方法:SD大鼠40只,随机分为生理盐水组、哮喘模型组、地塞米松组以及阿奇霉素组,每组10只。利用卵白蛋白(Ovalbumin,OVA)/Al(OH)3致敏与OVA雾化吸入激发建立大鼠过敏性气道炎症模型,收集肺泡灌洗液(BALF)进行白细胞分类计数。采用ELISA法测定肺泡灌洗液中IL-2、IL-4、TNF-α与ET-1的表达情况。光镜观察肺组织病理结构变化。结果:OVA模型大鼠肺泡灌洗液中的中性粒细胞、淋巴细胞以及嗜酸性粒细胞含量明显增加;HE染色观察肺组织病理结构出现明显的支气管上皮脱落、杯状细胞增生,支气管周围嗜酸性粒细胞明显浸润现象;BALF中IL-2、IL-4、TNF-α与ET-1的表达均明显高于生理盐水对照组(P<0.05)。阿奇霉素则显著降低肺泡灌洗液中中性粒细胞、淋巴细胞以及嗜酸性粒细胞含量;明显改善支气管上皮脱落、杯状细胞增生,支气管周围嗜酸性粒细胞浸润现象;BALF中IL-2、IL-4、TNF-α与ET-1的表达也明显低于OVA模型大鼠(P<0.05)。结论:阿奇霉素通过调节Th1/Th2失衡对过敏性哮喘的气道炎症具有明显的治疗作用。     

麦芪益肺合剂对哮喘小鼠Th1/Th2平衡的影响及安全性研究

目的:通过观察麦芪益肺合剂对系统性呼吸道变态反应性模型小鼠外周血T淋巴细胞的变化,探讨其对辅助性T淋巴细胞1/辅助性T淋巴细胞2(Th1/Th2)平衡的影响及安全性。方法:1%卵蛋白(OVA)刺激复制哮喘模型小鼠,随机分为正常、哮喘模型和麦芪益肺合剂组,采用流式细胞仪测定小鼠外周血(摘除眼球取血)细胞内白细胞介素4(IL-4)及干扰素(IFN-γ)的表达。按最大给药量对小鼠灌胃,对照组给予等量生理盐水,观察小鼠症状及体质量改变。结果:模型小鼠表现出明显非特异性炎症反应征象,而麦芪益肺合剂组小鼠无明显炎症反应征象。与正常组比较,哮喘模型组小鼠IL-4明显升高(P<0.01);与哮喘模型组比较,麦芪益肺合剂组小鼠Th1细胞表达明显增强(P<0.01)。以最大给药剂量灌胃,麦芪益肺合剂组与正常组小鼠症状及体质量无明显改变(P>0.05)。结果:麦芪益肺合剂对小鼠系统性呼吸道变态反应性模型在激发期有干预作用,调节Th1/Th2平恒是其药效作用机制之一。     

支气管哮喘患者外周血Th1、Th2与Th17细胞表达水平及临床意义

目的 探讨外周血Th1、Th2与Th17细胞在支气管哮喘患者中的表达水平及临床意义.方法 选取2015年1月-2016年2月诊治的支气管哮喘40例,均为急性发作期,治疗1周后为缓解期,选取同期健康人40例作为对照组.检测支气管哮喘急性发作期、缓解期和对照组外周血Th1、Th2与Th17细胞水平,观察外周血中干扰素(IFN)-白介素(IL)-4及IL-17的表达情况.结果 支气管哮喘组急性发作期和缓解期Th1、Th1/Th2、IFN-γ的表达水平低于对照组,且急性发作期低于缓解期(P＜0.05);支气管哮喘组急性发作期和缓解期Th2、Th17、IL-4、IL-17表达水平高于对照组,且急性发作期高于缓解期(P＜0.05).结论 支气管哮喘患者外周血Th1、Th2与Th17细胞表达水平失衡,导致IFN-γ、IL-4及IL-17异常分泌,可能参与支气管哮喘的发病.     

支气管哮喘患儿Th1/Th2细胞免疫平衡变化研究

目的探讨支气管哮喘患儿Th1/Th2细胞免疫平衡变化及其机制。方法选择急性发作期支气管哮喘患儿30名作为发作组,选择30名缓解期支气管哮喘患儿作为缓解组,选择30名健康小儿作为对照组,采用流式细胞仪检测各组静脉血Th1、Th2表达情况,采用酶联免疫吸附法检测三组患儿血清IL-4、IL-10、IFN-γ水平,比较各组患者Th1/Th2及血清IL-4、IL-10、IFN-γ表达变化水平。结果发作组静脉血Th1/Th2低于对照组及缓解组,血清IL-4、IL-10水平高于对照组及缓解组,IFN-γ水平低于对照组及缓解组。缓解组静脉血Th1/Th2低于对照组,血清IL-4、IL-10水平高于对照组,血清IFN-γ水平低于对照组。结论支气管哮喘患儿存在Th1/Th2免疫失衡,其可能是小儿支气管哮喘的发病机制之一。     

Dexamethasone alleviate allergic airway inflammation in mice by inhibiting the activation of NLRP3 inflammasome

Dexamethasone (DEX) is the mainstay treatment for asthma, which is a common chronic airway inflammation disease. However, the mechanism of DEX resolute symptoms of asthma is not completely clear. Here, we aimed to analyze the effect of DEX on airway inflammation in OVA-induced mice and whether this effect is related to the inhibition of the activation of NLRP3 inflammasome. Female (C57BL/6) mice were used to establish the allergic airway inflammation model by inhalation OVA. The number of inflammatory cells in the bronchi alveolar lavage fluid (BALF) was counted by Swiss-Giemsa staining, and the contents of IL-1β, IL-18, IL-5 and IL-17 were detected by ELISA. The degree of inflammatory cells infiltration and mucous cells proliferation in lung tissue were separately observed by H&E and PAS staining. The proteins expression of NLRP3, pro-caspase-1, caspase-1, IL-1β, IL-6 and IL-17 in lung tissue were detected by Western blotting. We found that DEX significantly inhibited OVA-induced inflammatory cells infiltration, airway mucus secretion and goblet cell proliferation in mice. The total and classified numbers of inflammatory cells and the levels of IL-1β, IL-18, IL-5 and IL-17 in the BALF of the experimental group were significantly lower than those of the model group after DEX treatment. DEX also significantly inhibited the activity of NLRP3 inflammasome and reduced the protein contents of Pro-Caspase-1, Caspase-1, Capase-1/Pro-Caspase-1, IL-1β, IL-6 and IL-17 in lung tissues. Our study suggested that DEX alleviates allergic airway inflammation by inhibiting the activity of NLRP3 inflammasome and the levels of IL-1β and IL-18.     

Zerumbone enhances the Th1 response and ameliorates ovalbumin-induced Th2 responses and airway inflammation in mice

Zerumbone is a sesquiterpene compound isolated from the rhizome of wild ginger, Zingiber zerumbet Smith. The rhizomes of the plant are used as a spice and traditional medicine. Zerumbone was shown to possess anticarcinogenic, anti-inflammatory, and antioxidant properties. However, the antiallergic activity and the underlying mechanism of zerumbone have not been reported. Herein, we investigated the immunomodulatory effects of zerumbone on antigen-presenting dendritic cells (DCs) in vitro and its potential therapeutic effects against ovalbumin (OVA)-induced T helper 2 (Th2)-mediated asthma in mice. In the presence of zerumbone, lipopolysaccharide-activated bone marrow-derived DCs enhanced T cell proliferation and Th1 cell polarization in an allogeneic mixed lymphocyte reaction. In animal experiments, mice were sensitized and challenged with OVA, and were orally treated with different doses of zerumbone after sensitization. Circulating titers of OVA-specific antibodies, airway hyperresponsiveness to methacholine, histological changes in lung tissues, the cell composition and cytokine levels in bronchoalveolar lavage fluid, and cytokine profiles of spleen cells were assessed. Compared to OVA-induced hallmarks of asthma, oral administration of zerumbone induced lower OVA-specific immunoglobulin E (IgE) and higher IgG_2a antibody production, attenuated airway hyperresponsiveness, prevented eosinophilic pulmonary infiltration, and ameliorated mucus hypersecretion. Zerumbone treatment also reduced the production of eotaxin, keratinocyte-derived chemokine (KC), interleukin (IL)-4, IL-5, IL-10, and IL-13, and promoted Th1 cytokine interferon (IFN)-γ production in asthmatic mice. Taken together, these results suggest that zerumbone exhibits an antiallergic effect via modulation of Th1/Th2 cytokines in an asthmatic mouse model.     

卡介苗对哮喘患者血Th1／Th2平衡和IgE的影响

目的：探讨口服卡介苗（BCG）对哮喘患者外周血Th1／Th2平衡、嗜酸性粒细胞（EOS）和IgE水平的影响。方法：对40例急性发作期的哮喘患者（治疗组）在常规治疗的基础上加用BCG口服（50mg／支）,100mg用5％碳酸氢钠溶液250ml稀释后饭前口服,每周1次,共16周。分别在治疗前后采用双抗体夹心ELISA法检测外周血培养上清单个核细胞（PBMC）中白介素4（IL4）和 γ-干扰素（IFN-γ）水平,同时测定外周血IgE和嗜酸性粒细胞（EOS）计数,并进行肺功能的测定。并以同期入选的健康体检者30例作对照（对照组）。结果：治疗组患者治疗前外周血IL4、IgE水平和EOS细胞数明显高于对照组,而IFN-γ水平明显低于对照组（P〈0．05或P〈0．01）,经BCG口服治疗16周后患者外周血IL-4、IgE水平和EOS细胞数均明显降低,而IFN-γ水平明显上升（P均〈0．05）;同时患者肺功能指标FVC、FEV,和PEF均明显改善（均P〈0．05）。结论：哮喘患者存在外周血Th1／Th2平衡失调。口服BCG治疗哮喘的临床疗效确切,作用机制可能通过纠正外周血Th1／Th2细胞因子平衡,抑制EOS聚集,使B细胞分泌IgE减少,从而改善患者肺功能,达到治疗哮喘的目的。     

Th1／Th2／Th17 T／reg 对鉴别儿童支气管哮喘与单纯性肺炎的意义

目的：检测Th1（IFN-γ＋）、调节T细胞（CD2＋5Foxp3＋）、辅助T 细胞17（Th17）及细胞因子IL-2、IL -4、IL-17和IL-21在儿童支气管哮喘以及肺炎患儿外周血中的表达,探讨其在儿童支气管哮喘发病中的作用以及与肺炎鉴别的意义。方法收集哮喘患儿32例,肺炎患儿37例作为研究对象,选取同期体检的22例健康儿童为对照组。分离外周血单个核细胞,采用流式细胞术检测CD 4＋IFN-γ＋、CD 4＋CD 2＋5Foxp 3＋、CD4＋Th17＋细胞百分率。采用酶联免疫吸咐（ ELISA）法检测3组IL-2、IL-4、IL-17、IL-21表达水平。结果哮喘组患儿外周血CD4＋CD2＋5 Foxp3＋细胞占CD4＋T细胞的比率明显低于肺炎组及对照组;哮喘组患儿者外周血CD4＋IFN-γ＋细胞比例明显低于肺炎组和对照组;哮喘组患儿者外周血CD4＋Th17＋细胞所占比例高于肺炎组及对照组,差异均有统计学意义（ P ＜0*．05）。哮喘组患儿外周血IL-2的浓度明显低于对照组（ P ＜00．5）,肺炎组患儿外周血IL-2浓度明显低于对照组（ P ＜0．05）,但与哮喘组比较差异无统计学意义（ P ＞0．05）。哮喘组患儿外周血 IL4-、IL-17和IL-21的浓度明显低于肺炎组和对照组,差异有统计学意义（ P ＜0．05）。结论 Th1／T2h ／Th17／Treg功能失调参与了儿童支气管哮喘的发病机制,临床上可以通过检测Th1／Th2／Th17／Treg相关细胞因子的变化评估儿童支气管哮喘病情并辅助与肺炎的鉴别。     

Airway inflammation and remodeling of cigarette smoking exposure ovalbumin-induced asthma is alleviated by CpG oligodeoxynucleotides via affecting dendritic cell-mediated Th17 polarization

Cigarette smoking (CS) is common in asthma, aggravating inflammatory reactions. However, the current treatment strategies for asthma are still not effective enough, and novel therapeutic approaches are required for CS-induced asthmatic disorders. We here investigated the ability of CpG oligodeoxynucleotides (CpG-ODNs) to inhibit airway inflammation and remodeling in ovalbumin (OVA)-associated asthma in mice exposed to chronic CS, revealing potential mechanistic insights. Lung tissue specimens were histologically analyzed. Th1/Th2/Th17 associated cytokines in serum, bronchoalveolar lavage fluid (BALF), and lung specimens were quantitated by ELISA, qRT-PCR and immunoblot. Parameters of bone marrow-derived dendritic cells (BMDCs) functions were evaluated as well. The results showed that BALB/c mice after CS and OVA treatments developed an asthmatic phenotype with airway inflammation involving both eosinophils and neutrophils, goblet cell metaplasia, airway remodeling, and elevated OVA-specific serum IgE, serum IL-17A, and BALF Th17/Th2 associated cytokines. CpG-ODNs and budesonide were found to synergistically inhibit inflammatory cell recruitment in the lung, airway remodeling, IgE synthesis, and Th17/Th2 associated cytokines. Mechanistically, CpG-ODNs and budesonide acted synergistically on BMDCs via downregulation of TSLP receptor (TSLPR) and IL-23 production, and subsequently contributed to dampen Th17/Th2 polarization in CS-associated asthma. In conclusion, combined administration of CpG-ODNs and budesonide, in a synergistic manner, inhibits airway inflammation, and tissue remodeling mediated by BMDCs by regulating IL-23 secretion and blocking TSLP signaling, which subsequently contribute to alleviate Th17/Th2 imbalance in CS-associated asthma.     

环境真菌链格孢霉对小鼠Th1/Th2平衡及气道反应性的影响

目的 探讨环境真菌链格孢霉（Alt,Alternaria）对小鼠Th1/Th2平衡及气道反应性的影响.方法 通过将小鼠暴露于链格孢霉提取物建立小鼠模型,以鸡卵蛋白（OVA）为阳性对照,探索真菌链格孢霉对Th1/Th2平衡及哮喘特征性指标的影响.结果 与OVA相似,Alt暴露引起了气道炎症细胞总数（P＜0.01）、嗜酸性细胞数增加（P＜0.01）;诱导了气道敏化和Th2反应并引起肺部和支气管的病理学变化.Alt与OVA间存在协同作用.结论 实验结果表明,环境真菌可通过干扰气道Th1/Th2平衡继而导致炎症,最终引起哮喘的发生.     

Breast regression protein-39 (BRP-39) promotes dendritic cell maturation in vitro and enhances Th2 inflammation in murine model of asthma

Aim:

To determine the roles of breast regression protein-39 (BRP-39) in regulating dendritic cell maturation and in pathology of acute asthma.

Methods:

Mouse bone marrow-derived dendritic cells (BMDCs) were prepared, and infected with adenovirus over-expressing BRP-39. Ovalbumin (OVA)-induced murine model of acute asthma was made in female BALB/c mice by sensitizing and challenging with chicken OVA and Imject Alum. The transfected BMDCs were adoptively transferred into OVA-treated mice via intravenous injection. Airway hyperresponsiveness (AHR), inflammation and pulmonary histopathology were characterized.

Results:

The expression of BRP-39 mRNA and protein was significantly increased in lung tissues of OVA-treated mice. The BMDCs infected with adenovirus BRP-39 exhibited greater maturation and higher activity in vitro. Adoptive transfer of the cells into OVA-treated mice significantly augmented OVA-induced AHR and eosinophilic inflammation. Meanwhile, BRP-39 further enhanced the production of OVA-induced Th2 cytokines IL-4, IL-5 and IL-13, but significantly attenuated OVA-induced IFN-γ production in bronchoalveolar lavage fluid.

Conclusion:

In OVA-induced murine model of acute asthma, BRP-39 is over-expressed in lung tissue and augments Th2 inflammatory response and AHR. BRP-39 promotes dendritic cell maturation in vitro. Therefore, BRP-39 may be a potential therapeutic target of asthma.     

翘芩清肺剂对哮喘大鼠Th1/Th2细胞因子失衡的影响

目的 观察翘芩清肺剂对哮喘模型大鼠Th1/Th2失衡的免疫调节作用及机制。方法 以10%卵蛋白（OVA）在实验第1天、第8天对大鼠进行腹腔注射。第15天后用1%OVA雾化吸入,每天1次,连续2周;同时,分别用生理盐水、地塞米松、氨茶碱及翘芩清肺剂干预致哮喘模型大鼠,每天1次,连续4周,观察各组大鼠哮喘潜伏期、喘息程度、肺组织、支气管病理学变化;测定肺泡灌洗液（BALF）中IL-4、IL-5、IFN-γ水平变化,并与阳性药地塞米松及氨茶碱组对照。结果 翘芩清肺剂能有效延长OVA致哮喘大鼠模型的潜伏期、减轻喘息程度、改善升高的IL-4水平,降低病鼠的IL-5、INF-γ水平及IL-4/IFN-γ的比值,结果均有显著差异（P〈0.05或P〈0.01）。病理组织学检查模型组大鼠支气管黏膜上皮细胞及杯状细胞增生,肺泡内有分泌物及炎性细胞浸润,管腔狭窄;各用药物组病理改变均显著减轻。结论 翘芩清肺剂可能通过调节哮喘时失衡的Th1/Th2水平,有效改善哮喘状态,减轻哮喘发作。     

射干麻黄药对对支气管哮喘小鼠模型气道炎症及外周血Th1/Th2的影响

目的:观察不同比例射干麻黄药对对支气管哮喘小鼠模型气道中炎症细胞及外周血Th1/Th2平衡的影响.方法:将80只BALB/c小鼠随机分为A正常对照组、B哮喘模型组、C地塞米松组、D麻黄单药组、E射干单药组及不同比例射干麻黄药对组F、G、H),每组10只.采用鸡卵蛋白(OVA)致敏和激发建立小鼠哮喘模型,给予不同药物对小鼠进行干预.干预完成后取支气管肺泡灌洗液(BALF)测定白细胞总数及嗜酸性粒细胞计数,采用ELISA检测小鼠血清中IL-4、IL-5、IL-13及IFNγ的水平,HE染色观察小鼠肺组织病理炎症等改变.结果:射干麻黄药对组可以显著减轻哮喘小鼠的哮喘症状,降低外周血中IL-4、IL-5、IL-13水平、升高IFN-γ水平,并能降低BALF中白细胞总数及嗜酸粒细胞水平.结论:射干麻黄药对可以显著减轻哮喘小鼠气道炎症,抑制炎症介质的释放,纠正Th1/Th2失衡,从而达到治疗哮喘的作用.     

p38蛋白激酶抑制剂SB203580对支气管哮喘小鼠气道炎症和Th2类细胞因子的影响

目的观察特异性p38蛋白激酶(p38 MAPK)抑制剂SB203580对哮喘小鼠气道炎症和Th2类细胞因子的影响。方法BALB/c小鼠30只随机分成3组,即正常对照组、哮喘模型组和SB203580干预组。通过原位分子杂交和酶联免疫吸附法(ELISA)检测肺组织IL-4、IL-5 mRNA和支气管肺泡灌洗液(BALF)中白细胞介素(IL-4、IL-5)含量的变化,并观察BALF中炎症细胞和肺组织病理学改变。结果哮喘模型组小鼠BALF中炎症细胞计数和IL-4、IL-5含量以及肺组织IL-4、IL-5mRNA的表达较正常对照组明显升高,差异具有显著性(P<0.01);SB203580干预组小鼠上述指标较哮喘模型组小鼠明显降低,差异亦具有显著性(P<0.01),肺组织病理学改变明显减轻。结论SB203580能降低气道炎症细胞的聚集和炎症介质的表达。抑制p38 MAPK的活性可能成为哮喘治疗的新途径。     

当归对阴虚哮喘BALB/c小鼠Th2优势免疫应答的影响

目的 探讨当归对阴虚哮喘BALB/c小鼠Th2细胞优势免疫应答的影响。方法 采用注射卵清白蛋白(ovalbumin,OVA)致敏、吸入OVA激发的方法来复制BALB/c小鼠哮喘模型,实验后期给予甲状腺素以复制阴虚哮喘模型,通过观测哮喘行为学、血清及支气管肺泡灌洗液(broncho-alveolar lavage fluid,BALF)中IL-4、IL-13的水平、BALF及肺组织中炎性细胞的含量、肺组织中GATA-3蛋白表达水平,探讨当归的平喘作用及对Th2细胞优势免疫应答的影响。结果 当归可明显减少阴虚哮喘小鼠哮喘发作的次数及其发作时的症状,改善肺组织病理学而减少肺组织及BALF中炎性细胞的数量,降低血清及BALF中IL-4、IL-13的水平,抑制肺组织中GATA-3蛋白的表达(P<0.05或0.01);同时,当归与地塞米松配伍后对嗜酸粒细胞、IL-4及GATA-3的抑制作用具有一定的协同作用(P<0.05或0.01)。结论 当归具有平喘作用,抑制细胞因子IL-4、IL-13及转录因子GATA-3的表达而缓解Th2优势免疫应答可能是其作用机制之一。     

Effect of Xiaofenghuoxue decotion on Th1 and Th2 cytokine production in BALF in allergen-induced asthma guinea pigs

Objective To investigate the effect of Xiaofenghuoxue decotion(XFHX)on the production of Th1 and Th2 cytokine in bronchoalveolar lavage fluid(BALF)in asthma guinea pigs sensitized and challenged with ovalbumin.Methods The animals were randomly divided into 5 groups:the normal control group,the asthma group,the XFHX high dose group,the XFHX low dose group and the budesonide group.2 weeks after sensitization with a mixture of 10% OVA and 10% Al(OH)3,the drugs were given to the animals for 7 days,and then the animals were challenged with aerosol of 1% ovalbumin,the latency time for the onset of respiratory abnormalities was examined,BALF was collected 16 h after.Leukocyte in BALF was counted.The level of IFN-γ reduced by Th1 cells and IL-4 and IL-5 produced by Th2 cells were determined by EILSA.NO contents were also examined.Results The latency time for the onset of respiratory abnormalities and leukocyte infiltration in both XFHX high dose group and low dose group significantly decreased compared with the asthma group.The level of IFN-γ in both XFHX high dose group and low dose group significantly increased compared with the asthma group,the level of IL-4 and IL-5 in both XFHX high dose group and low dose group significantly decreased compared with the asthma group.The production of NO was inhibited in both XFHX high dose group and low dose group compared with the asthma group.Conclusions These results suggest that the anti-asthma effect of Xiaofenghuoxue decotion is associated with NO regulated reversal of Th1/Th2 cytokine balance in allergen-induced asthma guinea pigs.     

Immunomodulatory effects of gyokuheifusan on INF-gamma/IL-4 (Th1/Th2) balance in ovalbumin (OVA)-induced asthma model mice

Asthma, a common, chronic lung disease in industrialized countries, is characterized by the production of large quantities of IgE antibody by B cells and a decrease of the IFN-gamma/IL-4 (Th1/Th2) ratio. Gyokuheifusan (GHS) is a classical formulation of traditional Chinese medicine (TCM) that is usually prescribed to prevent or treat respiratory tract diseases, such as respiratory infection and bronchial asthma. In order to evaluate the possible effectiveness of GHS on bronchial asthma, its immunomodulatory activity was examined in ovalbumin (OVA)-induced asthma model mice. All mice, except those in the normal group, were sensitized by intraperitoneal (IP) administration of OVA emulsified with Al(OH)(3), and a second immunization was given 6 d later. After a further 13, 17 and 21d, mice were challenged with inhalation of aerosolized OVA solution, except for the normal group, which received mock sensitization using saline-Al(OH)(3) emulsion and were challenged with an aerosol of saline without OVA. Allergen-specific IgE and total IgE in plasma were both significantly increased in the disease-control group. These increases were markedly blocked by GHS treatment. IFN-gamma released by splenocytes was significantly increased after co-culture with OVA for 24 h, 48 h, and 72 h. GHS treatment further elevated the IFN-gamma content compared with the disease-control group. The production of IL-4 was significantly increased when splenocytes were simulated with OVA for 72 h, but this increase was blocked by GHS treatment, so that GHS returned the decreased IFN-gamma/IL-4 (Th1/Th2) ratio of the disease-control group to the normal range. These results indicate that GHS may inhibit the development and severity of asthma.     

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.