脾虚湿蕴证溃疡性结肠炎模型构建及T淋巴细胞变化的研究

目的 构建脾虚湿蕴证溃疡性结肠炎小鼠模型及探明T淋巴细胞在该模型中的特征。方法 C57BL/6小鼠，随机分为Control组、FXY组、3%DSS组、FXY+3%DSS组和FXY+2%DSS组。采用DSS联合番泻叶(FXY)建立脾虚湿蕴证溃疡性结肠炎小鼠模型，造模结束后，HE染色观察结肠组织结构变化；ELISA法检测结肠组织炎性细胞因子的浓度，流式细胞术测定CD4/CD8 T淋巴细胞、Th1/Th2、Th17/Treg细胞水平。结果 FXY组小鼠的临床症状以腹泻为主，3%DSS组的小鼠以便血为主，而FXY+3%DSS组和FXY+2%DSS组小鼠表现出腹泻和便血同行，但FXY+3%DSS组小鼠生存率低至50%。与Control组相比，3%DSS组、FXY+3%DSS组和FXY+2%DSS组体重、结肠长度及抗炎性细胞因子IL-10、TGF-β1浓度均显著降低，而DAI、结肠重量、促炎性细胞因子TNF-α、IL-1β、IL-6浓度均显著上升；FXY+3%DSS组和FXY+2%DSS组肠系膜淋巴结中的CD4、Th1、Th17细胞百分比显著上升，而CD8、Th2、Treg细胞显著下降。结论 番泻...     

乳杆菌对牛乳β-乳球蛋白致敏小鼠淋巴细胞分泌Th1/Th2型细胞因子和抗体的体外影响

为了分析乳杆菌对致敏小鼠脾淋巴细胞分泌Th1/Th2细胞因子及抗体的体外影响,用牛乳β-乳球蛋白腹腔注射BALB/c小鼠建立过敏症模型,造模成功后,分离致敏小鼠的脾淋巴细胞并与4种活/死乳杆菌(107 CFU/mL)体外共同孵育,ELISA法检测细胞上清液中细胞因子(IL-12、IFN-γ和IL-4)和抗体(总IgE、β-Lg特异性IgE和总IgG)含量。4种活/死乳杆菌均可体外调节致敏小鼠脾淋巴细胞分泌细胞因子和抗体的水平,特别是热致死的发酵乳杆菌和嗜酸乳杆菌可提高淋巴细胞IL-12和IFN-γ的分泌,抑制IL-4的分泌,使其IFN-γ/IL-4比值(代表Th1/Th2细胞平衡)高于活菌,与空白对照组比较差异显著(P<0.05)。同时,这两株热致死菌还可显著下调细胞上清液中总IgE、特异性IgE和总IgG抗体的浓度(P<0.05)。试验结果表明乳杆菌可提高牛乳β-乳球蛋白致敏小鼠脾淋巴细胞的IFN-γ/IL-4比值,进而纠正Th2占优势的Th1/Th2失衡,下调抗体分泌量,且具有菌株特异性。     

幽门螺杆菌感染对DSS结肠炎小鼠Th17/Treg细胞亚群的影响

目的探讨幽门螺杆菌(H.pylori)感染对葡聚糖硫酸钠(DSS)诱导的小鼠溃疡性结肠炎(UC)的影响。方法 80只雌性BALB/c小鼠随机分为空白对照组(CON组,n=20)和实验组(n=60),实验组制作DSS诱导UC模型,然后分组建立成为结肠炎组(CLi组)、H.pylori持续感染的结肠炎组(Hp组)和H.pylori根除的结肠炎组(Hp-Era组),每组20小鼠。实验过程中观察小鼠一般情况和疾病活动指数(DAI);在第25天、45天时处死动物,分离外周血和结肠组织中单个核细胞,流式细胞仪检测其Th17/Treg细胞亚群变化。结果 CLi组、Hp组、Hp-Era组小鼠DAI评分显著高于CON组(均P0.05)。Hp-Era组小鼠体质量下降较Hp组更明显(P=0.012)。CLi组、Hp组、Hp-Era组小鼠结肠组织及外周血单个核细胞中Foxp3、IL-17A、RoRγT表达多显著高于CON组(大多数P值小于0.05)。与CLi组(实验第25天、45天)相比,Hp组小鼠结肠组织及外周血Foxp3表达显著升高(均P0.05),IL-17A、RoRγT表达显著降低(均P0.05)。与Hp-Era组(实验第45天)相比,Hp组小鼠结肠组织Foxp3表达显著升高(P0.05),结肠组织及外周血IL-17A、RoRγT表达显著降低(均P0.05)。结论 H.pylori感染使DSS结肠炎小鼠UC的Th17亚群比例降低、Foxp3~+Treg细胞比例增加,这种变化似乎对DSS诱导的结肠炎具有保护作用。     

乳杆菌肽聚糖对β-乳球蛋白致敏小鼠脾细胞Th1/Th2及Treg/Th17失衡的体外影响

【目的】观察嗜酸乳杆菌完整肽聚糖(Whole peptidoglycan,WPG)对致敏脾淋巴细胞Th1/Th2及Treg/Th17平衡的体外调节作用。【方法】通过腹腔注射β-乳球蛋白(β-Lg)建立BALB/c小鼠牛乳过敏模型。造模成功后,分离致敏小鼠的脾淋巴细胞并分别与不同剂量的WPG共同孵育,酶联免疫法检测细胞上清液中抗体(总IgE和特异性IgE),Th1/Th2及Treg/Th17相关细胞因子(IFN-γ,IL-4,TGF-β,IL-17)水平,流式细胞术检测脾淋巴细胞中CD3+、CD4+和CD8+的百分含量,荧光定量PCR法检测过敏小鼠脾细胞中Th1/Th2及Treg/Th17相关转录因子T-bet、GATA-3、Foxp3和RORγt mRNA的表达量。【结果】WPG体外刺激致敏脾细胞后可显著抑制IgE的产生,上调CD3+、CD4+细胞数及CD4+/CD8+比值,下调Th2型因子(IL-4,GATA-3mRNA)和Th17型因子(IL-17,RORγt mRNA)的表达,且与过敏组相比,中、高剂量WPG的作用效果显著(P0.05);另外,WPG体外作用还上调了Th1型因子(IFN-γ,T-bet mRNA)及Treg型因子(TGF-β,Foxp3 mRNA)的表达,且具有剂量依赖性。【结论】嗜酸乳杆菌WPG体外刺激可有效纠正致敏脾淋巴细胞的Th1/Th2及Treg/Th17失衡。     

粪菌移植对小鼠实验性结肠炎TLR4信号通路及肠黏膜屏障的影响

目的探讨粪菌移植(FMT)对溃疡性结肠炎(UC)小鼠肠黏膜屏障的影响及可能机制。方法小鼠饮用2.0%葡聚糖硫酸钠(DSS)溶液构建小鼠UC模型;50只成年雄性C57BL/6J小鼠,随机留取10只取粪便(这10只不参与后续的实验),其余40只称重、编号,随机分为空白对照组(Con组)、DSS模型对照组(Model组)、美沙拉嗪组(Model+5-ASA组)和粪菌液组(Model+FMT组),每组10只,Con组和Model组均给予0.9%NaCl溶液灌肠,给药组分别给予美沙拉嗪、粪便滤液灌肠;评估疾病活动指数(DAI)、各组结肠组织病理情况,用透射电镜检测各组小鼠的结肠黏膜上皮细胞结构的变化情况,ELISA检测各组血清内毒素、炎症因子TNF-α水平变化,免疫组化法检测结肠组织Toll样受体4(TLR4)及核因子-κB(NF-κB)的表达变化,Western blot检测各组ZO-1蛋白表达。结果与Model组相比,粪菌移植明显改善小鼠的DAI指数和结肠组织的病理损伤,结肠上皮细胞间隙增宽程度减轻,腺上皮细胞间连接较紧密,结肠黏膜上皮细胞微绒毛完整,排列整齐,内毒素、TNF-α的含量明显下降,TLR4及NF-κB在结肠组织的表达明显下降,ZO-1蛋白表达明显升高,促进结肠黏膜屏障的修复,差异具有统计学意义(t=7.9543,P<0.0001;t=3.7641,P=0.0010;t=4.5899,P=0.0020;t=13.2886,P<0.0001;t=4.9750,P=0.0010;t=6.9388,P<0.0001;t=8.3744,P<0.0001)。结论FMT可减少内毒素及炎症因子的产生,改善结肠炎症,TLR4-NF-κB信号通路可能是FMT修复结肠黏膜屏障功能的机制之一。     

一株猪肠道Lactobacillus animalis LGM对结肠炎小鼠Th细胞分化转录因子基因表达的影响

【目的】从体外和体内研究Lactobacillus animalis LGM对Th细胞分化转录因子T-bet、GATA-3、ROR-γt和Foxp3的调节作用,以及探究L. animalis LGM对小鼠结肠炎的影响。【方法】本试验采用改良型的Hungate滚管技术从猪结肠内容物中分离一株L. animalis LGM,根据其16S rRNA序列进行鉴定。收集L. animalis LGM培养液上清,与细菌脂多糖(LPS,2μg/mL)同时孵育Caco-2细胞24 h,体外研究L. animalis LGM对Caco-2细胞内Th细胞分化转录因子(T-bet,GATA3,ROR-γt和Foxp3) mRNA表达的影响;配制L. animalis LGM细菌悬液,研究L. animalis LGM灌胃对DSS诱导结肠炎小鼠症状及结肠Th细胞分化转录因子和细胞因子(IFN-γ,IL-4,IL-17和IL-10) mRNA表达的影响,表达结果采用荧光定量PCR法检测。【结果】与对照组相比,L.animalisLGM培养液上清显著上调Caco-2细胞内ROR-γt与Foxp3 mRNA表达(P0.05),显著下调GATA3、IL-4、IL-17和TGF-βmRNA表达(P0.05)。L. animalis LGM灌胃显著上调小鼠结肠内ROR-γt和Foxp3的表达(P0.05),显著降低了促炎因子IL-4和IL-17的表达(P0.05),阻止了小鼠结肠长度缩短(P0.05)。【结论】猪肠道分离L. animalis LGM表现出对Th细胞分化转录因子的选择性调节,显著上调Caco-2细胞及结肠炎小鼠ROR-γt与Foxp3mRNA表达。降低DSS诱导结肠炎小鼠炎症水平,对DSS诱导结肠炎起保护作用,有助于维护肠道环境稳态。     

Th17及Th1相关细胞因子与支气管哮喘的关系研究

目的:分析外周血Th17、Th1及相关细胞因子表达水平和支气管哮喘(bronchial asthma,BA)发生、发展的相关性研究。方法:回顾选取我院收治的BA病例57份,称作BA组,另选取呼吸系统正常的病例55例为对照组,检测两组入选者的外周血IL-2、TNF-α、Th17、IL-6、Th1指标表达差异,并进行多因素回归分析。结果:BA组Th17(0.62±1.67)%、Th1(1.45±0.48)%及Th1/Th17(2.33±1.28)均显著低于对照组(P均0.05);BA组TNF-α(27.46±8.12)pg/mL、IL-6(11.69±2.14)pg/mL表达量显著高于对照组,IL-2(2.58±3.89)pg/mL、IFN-γ(3.74±6.15)pg/mL含量均显著低于对照组(P均0.05);经Logistic回归分析,TNF-α、IL-6、IFN-γ、Th1/Th17、IL-2均和BA有密切相关性(P均0.05)。结论:IL-2、IFN-γ、Th1/Th17、TNF-α、IL-6表达水平均与BA有密切关联,可能是参与BA发病的主要原因,及早进行Th17、Th1及相关细胞因子检查有助于明确病情。     

宫颈癌组织中Th1/Th2类细胞因子的漂移研究

目的:观察宫颈癌组织中Th1/Th2类细胞因子的漂移情况。方法:选以IL-2和IFN-γ代表Th1类细胞因子,IL-4和IL-6代表Th2类细胞因子,通过逆转录聚合酶链反应(RT-PCR)检测25例宫颈癌癌组织中Th1/Th2类细胞因子mRNA的表达。结果:IIIB期宫颈癌组织中,Th1型细胞因子的表达显著低于IB期、IIA期、IIB期,Th2型细胞因子的表达显著高于IB期、IIA期、IIB期,差异均有统计学意义(P0.05)。Ⅰ期和Ⅱ期宫颈癌以Th1型细胞因子表达为主。25例宫颈癌组织中,13例呈典型的Th1类细胞因子的强势表达,7例为Th2型,5例为Th0型,随着宫颈癌分期的增高,由Th1向Th2漂移(P0.05)。结论:IB期、IIA期、IIB期宫颈癌患者组织中细胞因子呈Th1状态,IIIA期呈Th0状态,IIIB期呈Th2状态,随着宫颈癌分期的增高,由Th1向Th2漂移。     

木犀草素调节SIRT3/AMPK/mTOR信号通路对溃疡性结肠炎小鼠Th17/Treg免疫平衡的影响CSCD

探究木犀草素对溃疡性结肠炎(ulcerative colitis,UC)小鼠模型中辅助性T细胞17(Th17)/调节性T细胞(Treg)免疫平衡的影响,并分析其潜在机制。60只BALB/c雄性小鼠随机分为正常组、模型组、阳性对照组、木犀草素组、木犀草素+3-TYP组,每组12只。采用葡聚糖硫酸钠(DSS)诱导建立UC模型,观察并记录小鼠体重变化、大便稠度和大便潜血情况,计算疾病活动指数(DAI);HE染色观察结肠组织病理学变化;流式细胞术检测脾脏Th17、Treg细胞比例,计算Th17/Treg比值;ELISA检测结肠组织中IL-6、IL-10、IL-17和IL-23含量;RT-qPCR检测结肠组织RORγt和Foxp3的mRNA表达;Western blot检测结肠组织SIRT3、AMPK、p-AMPK、mTOR、p-mTOR蛋白表达。与正常组相比,模型组小鼠DAI评分、组织病理学评分、脾脏Th17/Treg比值、结肠组织IL-6、IL-17、IL-23、RORγt mRNA水平和p-mTOR/mTOR比值升高,IL-10、Foxp3 mRNA和SIRT3蛋白水平以及p-AMPK/AMPK比值降低(P<0.05)。经药物干预后,小鼠DAI评分、组织病理学评分、脾脏Th17/Treg比值、结肠组织IL-6、IL-17、IL-23、RORγt mRNA水平和p-mTOR/mTOR比值降低,IL-10、Foxp3 mRNA和SIRT3蛋白水平以及p-AMPK/AMPK比值升高(P<0.05);3-TYP可减弱木犀草素对UC小鼠Th17/Treg细胞分化平衡的影响(P<0.05)。木犀草素可改善DSS诱导的UC小鼠模型中Th17/Treg失衡,其机制可能与激活SIRT3/AMPK/mTOR通路有关。     

菌群失调小鼠感染鼠伤寒沙门菌后Th1和Th2细胞因子的动态研究

目的研究鼠伤寒沙门菌感染小鼠在菌群失调下Th细胞因子的动态变化,以探讨菌群失调对沙门菌感染的免疫机制。方法分别建立菌群失调、感染和空白对照的小鼠模型。各组动物在感染不同时点处死,观察小肠、肝脏和脾脏病理改变。采用流式细胞仪检测脾脏细胞中IFN-γ和IL-4表达,以此代表Th1和Th2细胞。结果菌群失调组病理损害最重,Th1明显增加,表达水平高,Th2变化不大,Th1/Th2比值升高。结论菌群失调后,能够加重小鼠感染鼠伤寒沙门菌引起的的Th1型反应,产生炎症性损伤。     

Th1/Th2 differentiation and cross-regulation

The T helper (Th) phenotypes, Th1/Th2, are acquired upon interaction of a naive T helper cell and an antigen presenting cell (APC). Naive T helper cells may differentiate into either phenotype, and the actual outcome is determined by the density and avidity of the antigenic determinants presented by the APC, and the APCs inherent costimulatory properties. Until recently it was thought that differentiation is further affected by cytokines. However, Murphy et al. (1996, J. Exp. Med. 183, 901) have demonstrated that the experimental results, formerly interpreted as Th1/Th2 differentiation, in effect comprise an observation of two consecutive processes. (i) An interaction between naive T cells and APC creates a mixture of mature cells irreversibly committed to Th1 or Th2 phenotype. (ii) Subsequent addition of regulatory cytokines, promotes expansion of one phenotype while suppressing the other. The consequent shift in the per culture production of marker cytokines mimics the appearance of a cellular phenotype switch. We present and analyse a mathematical model that extrapolates these experimental facts into systemic behavior during an immune response. Despite the fact that differentiation produces cells of Th1 and Th2 phenotypes with the same receptor specificity, our results indicate that competition for antigenic stimulation, mediated by the APCs, combines with cytokine mediated cross-suppression between phenotypes to yield a response that is eventually dominated by T helper cells that are uniform in both receptor specificity (clonotype) and in cytokine secretion phenotype.     

In vitro Th1 cytokine-independent Th2 suppressive effects of bifidobacteria

A comparison between 17 strains of lactic acid bacteria and 15 strains of bifidobacteria indicated that bifidobacteria induced significantly lower levels of interleukin-12 (IL-12) in murine splenic cells. The present study aims to evaluate the effect and mechanism of Bifidobacterium longum BB536, a probiotic strain, in suppressing antigen-induced Th2 immune response in vitro. BB536 suppressed immunoglobulin (Ig) E and IL-4 production by ovalbumin-sensitized splenic cells, but induction of Th1-inducing cytokine production, such as IL-12 and gamma interferon (IFN-gamma) tended to be lower compared with lactic acid bacteria. Neutralization with antibodies to IL-12, IFN-gamma, IL-10 and transforming growth factor beta indicated negative involvement of Th1-inducing cytokines and regulatory cytokines in the suppression of Th2 immune response by BB536, especially when treated at higher doses of BB536 (>10 microg cells/ml). Furthermore, BB536 induced the maturation of immature bone marrow-derived dendritic cells (BM-DCs), and suppressed antigen-induced IL-4 production mediated by BM-DCs. These results suggested that BB536 suppressed Th2 immune responses, partially independent of Th1-inducing cytokines and independent of regulatory cytokines, mediated by antigen-presenting cells such as dendritic cells.     

CD2 facilitates differentiation of CD4 Th cells without affecting Th1/Th2 polarization.

The role of CD2 in murine CD4 helper T cell differentiation and polarization was examined using TCR-Cyt-5CC7-I transgenic recombination activating gene-2-/- H-2(a) mice on CD2+/+ or CD2-/- backgrounds. In the absence of CD2, thymic development was abnormal as judged by reduction in the steady state number of total, double-positive, and CD4 single-positive (SP) thymocytes, as well as a defect in their restorative dynamics after peptide-induced negative selection in vivo. In addition, in CD2-/- animals, lymph node CD4 SP T cells manifest a 10- to 100-fold attenuated activation response to cytochrome c (CytC) agonist peptides as judged by induction of CD25 and CD69 cell surface expression or [(3)H]TdR incorporation; differences in the magnitude of responsiveness and requisite molar peptide concentrations were even greater for altered peptide ligands. Although the presence or absence of CD2 did not impact the final Th1 or Th2 polarization outcome, CD2 expression reduced the CytC peptide concentration threshold necessary to facilitate both Th1 and Th2 differentiation. In vivo administration of CytC peptide to CD2-/- animals yielded an impaired CD4 SP T cell effector/memory phenotype compared with similarly treated CD2+/+ mice. Analysis of TCR-Cyt-5CC7-I human CD2 double-transgenic mice similarly failed to reveal a preferential Th1 vs Th2 polarization. Collectively, these results indicate that CD2 is important for the efficient development of CD4 SP thymocytes and TCR-dependent activation of mature CD4 lymph node T cells, but does not direct a particular helper T cell subset polarity.     

Elevated Frequencies of Circulating Th22 Cell in Addition to Th17 Cell and Th17/Th1 Cell in Patients with Acute Coronary Syndrome

Background

Atherosclerosis is a chronic inflammatory disease mediated by immune cells. Th22 cells are CD4⁺ T cells that secret IL-22 but not IL-17 or IFN-γ and are implicated in the pathogenesis of inflammatory disease. The roles of Th22 cells in the pathophysiologic procedures of acute coronary syndrome (ACS) remain unclear. The purpose of this study is to investigate the profile of Th22, Th17 and Th17/Th1 cells in ACS patients, including unstable angina (UA) and acute myocardial infarction (AMI) patients.

Design and Methods

In this study, 26 AMI patients, 16 UA patients, 16 stable angina (SA) patients and 16 healthy controls were included. The frequencies of Th22, Th17 and Th17/Th1 cells in AMI, UA, SA patients and healthy controls were examined by flow cytometry. Plasma levels of IL-22, IL-17 and IFN-γ were measured by enzyme-linked immunosorbent assay (ELISA).

Results

Th22, Th17 and Th17/Th1 cells were significantly increased in AMI and UA patients compared with SA patients and healthy controls. Moreover, plasma IL-22 level was significantly elevated in AMI and UA patients. In addition, Th22 cells correlated positively with IL-22 as well as Th17 cells in AMI and UA patients.

Conclusion

Our findings showed increased frequencies of both Th22 and Th17 cells in ACS patients, which suggest that Th22 and Th17 cells may play a potential role in plaque destabilization and the development of ACS.     

The Th1/Th2/Th17 cytokine profile of HIV-infected individuals: A multivariate cytokinomics approach

HIV infection causes the dysregulation of cytokine production. A cytokinomics approach employing cytometric bead array (CBA) technology, flow cytometry and multivariate analysis was applied to the investigation of HIV-induced T helper cell type 1 (Th1), Th2 and Th17 cytokine changes in the sera of treatment naive individuals. Stepwise linear discriminant analysis (LDA) and logistic regression identified interleukin (IL)-6 to be discriminatory for HIV infection with 74.6% and 71.2% of the cases correctly classified. Analysis of variance (ANOVA) confirmed IL-6 and IL-10 concentrations to be significantly (p = 0.001 and p = 0.025) different between the groups. A scatter plot of the log IL-6 and IL-10 concentrations for the groups largely overlapped, with improved differentiation where patients were advancing to the acquired immunodeficiency syndrome (AIDS). IL-17A levels were higher than other cytokines but did not significantly distinguish the groups suggesting that the HIV? and HIV+ individuals had similar immune profiles. This possibility was supported by other clinical indicators. Taken together, the measured cytokines (IL-6, 10 and 17) have potential prognostic value.     

微卡对支气管哮喘Th1/Th2类细胞因子失衡的调节作用

目的 探讨微卡对哮喘ThⅠ/Th2类细胞因子失衡的调节作用.方法 选取确诊的轻中度哮喘40例,所有患者深部肌肉注射微卡22.5μg,每2周1次,共8周,并在治疗前、治疗后1月、2月分别抽取静脉血3 ml检测IFN-γ和IL-4水平（ELISA法）.结果 微卡治疗后1月即可纠正失衡的IFN-γ/IL-4,其中以治疗后2月作用较明显,且未见明显的药物不良反应.结论 微卡通过调节失衡的Th1/Th2平衡而达到抗气道炎症作用,可作为哮喘的防治药物.     

Age-related differentiations of Th1/Th2 cytokines in newborn infants

OBJECTIVE: To evaluate age-related differentiation of immune response in newborns by measuring serum concentrations of interleukin-2 (IL-2), interleukin-4 (IL-4) and interferon-gamma (IFN-gamma) during the perinatal period. SUBJECTS AND METHODS: Fifty-seven healthy term neonates, their mothers and 25 healthy adults (controls) age-matched to the mothers were included in the study. Cytokine concentrations were measured in the umbilical cord (UC), and in first-day (1N) and fifth-day (5N) neonatal samples, compared with those in maternal serum (MS) and control serum samples. RESULTS: Serum IL-2 concentrations in the UC were markedly elevated compared with those in MS and controls (p < 0.0001), decreasing significantly thereafter up to 5N (p < 0.001). IL-4 serum concentrations did not differ significantly between the UC, 1N and 5N samples; they were, however, markedly elevated compared with those in MS (p < 0.001, p < 0.0007 and p < 0.0001, respectively) and controls (p < 0.05, p < 0.01 and p < 0.006, respectively). IFN-gamma serum concentrations were significantly lower in the UC compared with those in controls (p < 0.04), increasing significantly up to 5N (p < 0.03). Both IFN-gamma/IL-2 and IFN-gamma/IL-4 ratios increased significantly in 5N, compared with those in the UC (p < 0.001 and p < 0.03). CONCLUSION: Our findings indicate a differential cytokine balance at birth with enhanced expression of IL-2 and IL-4 against IFN-gamma. However, a regularization of immune response seems to proceed quickly during the early neonatal life.     

Proteomic profiling of surface proteins on Th1 and Th2 cells

We utilized mass spectrometry to profile cell surface protein differential expression on primary human T helper (Th1 and Th2) cells with the stable isotope labeling by amino acids in cell culture (SILAC) approach. Proteomic and microarray analyses were done concurrently and results were compared for 38 different genes. Although microarray studies displayed wide variability between donors for mRNA expression, these two approaches were shown to be corroborative for most gene products with the exception of a small subset of uncorrelated protein and message levels. The greatest differing Th1 to Th2 ratios were observed for BST2 (bone marrow stromal protein 2) and TRIM (T cell receptor interacting molecule). Both showed greater Th1 expression by proteomic methods, even though mRNA levels were approximately equal for both. To validate this method, we compared protein expression levels of a recently cloned molecule, B and T cell lymphocyte attenuator (BTLA), on Th1 and Th2 cell populations and showed greater protein expression on Th1 cells, which agrees with a previous analysis of higher BTLA mRNA expression in Th1 cells.(1).