Ghrelin对雌激素诱导小鼠胸腺萎缩过程中细胞因子及凋亡相关蛋白基因表达的影响

应用光镜和实时定量PCR技术研究了Ghrelin对雌激素诱导小鼠胸腺萎缩过程中胸腺形态学以及部分细胞因子和凋亡相关蛋白基因表达的影响。结果显示,注射Ghrelin后,雌激素诱导的小鼠萎缩胸腺在形态学上基本恢复到正常水平,胸腺中IL-6、TGF-[31、Caspase3、FADDmRNA含量显著降低,Caspase9、FasLmRNA含量略为下降,而IL-7、Bcl-2mRNA含量有所上升。结果表明,Ghrelin可能通过促进胸腺上皮细胞的增殖以及阻断Caspase级联程序和Fas／FasL凋亡信号通路抑制胸腺细胞的凋亡,从而逆转雌激素诱导的小鼠胸腺萎缩。     

高致病性猪繁殖与呼吸综合征病毒依赖Caspase途径诱导仔猪胸腺细胞凋亡

为研究高致病性猪繁殖与呼吸综合征病毒(HP-PRRSV)Hu N4株诱导断奶仔猪胸腺细胞凋亡的分子机制,本实验通过免疫荧光技术、流式细胞术、荧光定量PCR和western blot等方法检测HP-PRRSV感染仔猪胸腺内细胞凋亡相关因子Caspase-8、Caspase-9和Caspase-3表达量的变化。结果显示:HP-PRRSV感染仔猪后,胸腺内表达凋亡相关蛋白Caspase-8、Caspase-9和Caspase-3的细胞比例显著升高,并且凋亡相关蛋白Caspase-8、Caspase-9和Caspase-3表达量也明显升高。以上结果表明:HP-PRRSV感染仔猪引起胸腺细胞凋亡是通过Caspase依赖性通路介导。本研究为进一步了解HP-PRRSV感染机制提供了实验依据。     

Ghrelin对雌激素诱导雄性小鼠胸腺萎缩的逆转及对相关细胞因子基因表达的影响

SPF级6周龄Balb/c雄性小鼠60只,随机分为3组,每组20只,适应饲养1周后进行动物试验.雌激素+Gh-relin试验组:前2周隔日腹腔注射苯甲酸雌二醇注射液(含苯甲酸雌二醇0.1 mg)0.05 mL/只,后2周按每日腹腔注射Ghrelin(含Ghrelin 60 μg)0.1mL/只;雌激素+生理盐水对照组:前2周隔日腹腔注射苯甲酸雌二醇注射液(含苯甲酸雌二醇0.1mg)0.05mL/只,后2周按每日腹腔注射0.9％生理盐水0.1 mL/只;生理盐水对照组:前2周按0.05 mL/只隔日腹腔注射0.9％生理盐水,后2周按0.1 mL/只每日腹腔注射0.9％生理盐水.动物试验结束后,统计小鼠胸腺指数,应用光镜观察胸腺形态学变化,采用实时荧光定量PCR法检测胸腺中12种细胞因子mRNA表达量的变化.结果显示,注射Ghrelin后,雌二醇诱导的小鼠胸腺萎缩在形态学上基本恢复到正常水平,胸腺中白细胞介素1(IL-1)、IL-2、IL-4、IL-6、IL-12、干扰素(1FN-γ)、白血病抑制因子(LIF)、抑瘤素(OSM)、干细胞因子(SCF)、胸腺体液因子(THF)、肿瘤坏死因子(TNF-α) mRNA含量显著降低(P＜0.05),而IL-7 mRNA含量稍微升高(P＞0.05).结果表明,Ghrelin对雌激素诱导的小鼠胸腺萎缩具有逆转作用,其机制可能是:一方面通过抑制IL一6、OSM、LIF、SCF等细胞因子的表达,从而促进胸腺细胞的增殖;另一方面通过抑制IL-1、IL-2、IL-4、IL-12、THF等细胞因子的表达,从而减少TNF-α和IFN-γ的分泌,进而抑制胸腺细胞的凋亡.     

C_(17)-吡唑琳基甾体衍生物诱导Hela细胞凋亡研究

通过电镜、AO/EB染色检测细胞形态学变化,应用流式细胞仪检测细胞凋亡率,通过检测Caspase活性及相关凋亡蛋白的变化确定C17-吡唑琳基甾体衍生物1诱导细胞凋亡的机制。经化合物1处理后的Hela细胞出现典型的凋亡特征,细胞凋亡率与处理时间呈正相关,同时Caspase-3与Caspase-9活性也逐渐增加,而促凋亡蛋白Bax表达量上升,抑凋亡蛋白分子Bcl-2表达量显著下降。说明C17-吡唑琳基甾体衍生物1通过线粒体通路诱导Hela细胞凋亡。     

死亡受体通路在黄曲霉毒素B_1致雏鸡胸腺细胞凋亡中的作用

旨在探究摄食黄曲霉毒素(AF)后,死亡受体通路活化对雏鸡胸腺细胞过度凋亡的信号调控机制。将90只1日龄健康雏鸡随机分为两组,分别饲喂对照日粮和AFB_1日粮(在对照日粮中添加0.6mg·kg~(-1) AFB_1)。于7、14和21日龄时,检测雏鸡胸腺的脏器指数、T淋巴细胞亚群、细胞凋亡率及死亡受体通路上相关基因的mRNA相对转录量。结果显示,与对照组比较,AFB_1组雏鸡胸腺的脏器指数降低,CD3~+、CD3~+CD4~+和CD3~+CD8~+T淋巴细胞的百分率减少,胸腺细胞凋亡率升高,死亡受体通路上RIP1、Caspase-8、Caspase-9、Caspase-3、Fas、FasL、ASK1、IKIP和JNK的mRNA相对转录量升高,Bcl-2、NF-κB_1和Bid的mRNA相对转录量降低。本试验中,雏鸡胸腺细胞凋亡上调的过程中,死亡受体通路上的三条下游信号途径均有参与:(1)通过Fas-FasL-FADD-Caspase-8-Caspase-3途径直接诱导凋亡;(2)通过Fas-ASK1-JNK-Bcl-2途径,并在线粒体的介导下,启动下游调控因子Caspase-9和Caspase-3诱导细胞凋亡;(3)通过TNF-α-RIP1-IKIP-NF-κB_1-Caspase-3信号途径诱导凋亡。     

鼠源重组UBC13蛋白对脂多糖诱导的小鼠急性炎症的影响

试验旨在探讨鼠源重组UBC13蛋白对脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性炎症的影响。将24只SPF雌性小鼠随机分成4组:PBS组,LPS模型组,重组UBC13蛋白高、低剂量组(分别为100和25μg/只),每组6只。LPS模型组与各蛋白剂量组腹腔注射20 mg/kg LPS,PBS组腹腔注射等体积PBS;注射结束1 h后,各蛋白组按相应剂量背部皮下多点注射重组UBC13蛋白,PBS组与LPS模型组注射等体积PBS。给予蛋白24 h后处死小鼠。收集小鼠肺脏、脾脏、胸腺及肝脏组织,计算脏器指数,HE染色观察组织病理学变化,实时荧光定量PCR检测肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA的相对表达量,以及肺脏中iNOS mRNA的相对表达量,综合评价鼠源重组UBC13蛋白对LPS诱导小鼠急性炎症的影响。结果显示,与PBS组相比,LPS模型组小鼠肺脏、脾脏及肝脏指数均显著或极显著升高(P0.05;P0.01),且肺脏、脾脏和肝脏组织均出现病理变化。实时荧光定量PCR结果显示,与PBS组相比,LPS模型组肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA相对表达量均极显著升高(P0.01),肺脏中iNOS mRNA相对表达量也极显著升高(P0.01);与LPS模型组相比,UBC13蛋白高剂量组肺脏、脾脏和肝脏中病理变化明显改善,肺脏、肝脏、脾脏中IL-1β、TNF-α、IL-6及肺脏中iNOS mRNA表达量均极显著降低(P0.01);胸腺中TNF-αmRNA表达量显著降低(P0.05),IL-6 mRNA和IL-1β表达量极显著降低(P0.01)。表明鼠源重组UBC13蛋白可下调炎性因子的表达,从而改善LPS诱导的小鼠急性炎症反应。     

Ghrelin对雌激素诱导小鼠胸腺细胞凋亡的抑制作用

6周龄BALB/c雌性小鼠80只,分为4组：雌激素＋Ghrelin组、雌激素＋生理盐水组、生理盐水组、空白对照组。雌激素＋Ghrelin组首先隔日腹腔注射苯甲酸雌二醇（0.1mg/只）2周,然后采用相同方法注射Ghrelin（120μg/只）2周;雌激素＋生理盐水组首先注射苯甲酸雌二醇2周,然后注射生理盐水2周;生理盐水组注射生理盐水4周;空白对照组不注射任何试剂。测量各组小鼠的胸腺指数,同时应用光镜、电镜和流式细胞术检测胸腺的显微结构、超微结构和胸腺细胞凋亡率。结果雌激素＋Ghrelin组的胸腺指数极显著大于雌激素＋生理盐水组,胸腺细胞凋亡率极显著小于雌激素＋生理盐水组（P〈0.01）,而二者与生理盐水组和空白对照组均差异不显著（P〉0.05）;雌激素＋Ghrelin组的胸腺显微结构和超微结构基本恢复正常,并与生理盐水组和空白对照组相似。结论得出Ghrelin可以通过抑制胸腺细胞凋亡和促进胸腺细胞增殖,从而逆转雌激素诱导的小鼠胸腺萎缩。     

鼠源重组UBC13蛋白对脂多糖诱导的小鼠急性炎症的影响

试验旨在探讨鼠源重组UBC13蛋白对脂多糖(lipopolysaccharide,LPS)诱导的小鼠急性炎症的影响。将24只SPF雌性小鼠随机分成4组:PBS组,LPS模型组,重组UBC13蛋白高、低剂量组(分别为100和25μg/只),每组6只。LPS模型组与各蛋白剂量组腹腔注射20 mg/kg LPS,PBS组腹腔注射等体积PBS;注射结束1 h后,各蛋白组按相应剂量背部皮下多点注射重组UBC13蛋白,PBS组与LPS模型组注射等体积PBS。给予蛋白24 h后处死小鼠。收集小鼠肺脏、脾脏、胸腺及肝脏组织,计算脏器指数,HE染色观察组织病理学变化,实时荧光定量PCR检测肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA的相对表达量,以及肺脏中iNOS mRNA的相对表达量,综合评价鼠源重组UBC13蛋白对LPS诱导小鼠急性炎症的影响。结果显示,与PBS组相比,LPS模型组小鼠肺脏、脾脏及肝脏指数均显著或极显著升高(P<0.05;P<0.01),且肺脏、脾脏和肝脏组织均出现病理变化。实时荧光定量PCR结果显示,与PBS组相比,LPS模型组肺脏、脾脏、胸腺和肝脏中IL-1β、TNF-α、IL-6 mRNA相对表达量均极显著升高(P<0.01),肺脏中iNOS mRNA相对表达量也极显著升高(P<0.01);与LPS模型组相比,UBC13蛋白高剂量组肺脏、脾脏和肝脏中病理变化明显改善,肺脏、肝脏、脾脏中IL-1β、TNF-α、IL-6及肺脏中iNOS mRNA表达量均极显著降低(P<0.01);胸腺中TNF-αmRNA表达量显著降低(P<0.05),IL-6 mRNA和IL-1β表达量极显著降低(P<0.01)。表明鼠源重组UBC13蛋白可下调炎性因子的表达,从而改善LPS诱导的小鼠急性炎症反应。     

TNF-α对仔猪睾丸支持细胞中Fas/FasL表达的调节作用

本研究旨在探索TNF-α对睾丸支持细胞(SCs)中Fas/FasL表达的影响及机制。用不同质量浓度的TNF-α(0.0,0.1,1.0,10.0,25.0,50.0,100.0,200.0μg/L)处理SCs不同时间(0,6,12,24,36,48,72h)后,采用CCK-8细胞增殖检测试剂盒检测细胞的增殖;通过流式细胞术检测细胞周期进程和细胞凋亡;用Western blot和荧光定量PCR技术检测Fas/FasL、MMP-2/MMP-9和TIMP-2/TIMP-1蛋白水平及其mRNA丰度;利用ELISA试剂盒检测IL-1β、IL-6、IGF-1、TGF-β1等细胞因子水平。结果显示,当TNF-α质量浓度为50.0μg/L、作用时间为36h时,细胞的增殖活力最低;这一条件显著促进了Fas/FasL、MMP-2/MMP-9蛋白及mRNA的表达(P0.05),极显著降低了TIMP-1和TIMP-2mRNA与蛋白表达(P0.01),且极显著促进了细胞促炎性因子IL-1β和IL-6分泌(P0.01),显著抑制了促生长因子IGF-1和TGF-β1分泌(P0.05)。结果表明:TNF-α以剂量和时间依赖方式诱导SCs中Fas/FasL mRNA及蛋白表达,从而诱发SCs凋亡,且Fas/FasL表达受到MMP-2/MMP-9、TIMP-2/TIMP-1以及细胞因子水平的影响。     

受体相互作用蛋白1(RIP1)对BCG诱导小鼠巨噬细胞凋亡的调控作用

旨在通过构建受体相互作用蛋白1(RIP1)腺病毒干扰载体,研究其对BCG诱导的RAW264.7细胞凋亡相关指标的影响,以探讨其在BCG诱导RAW264.7凋亡过程中的调控作用。笔者构建RIP1腺病毒干扰载体,并转染感染BCG的小鼠RAW264.7细胞系,利用流式细胞仪检测各处理细胞凋亡率、细胞线粒体膜电位、细胞活性氧水平及细胞周期等指标,并用Western blot检测凋亡相关蛋白的表达水平。结果显示:BCG感染显著上调了RIP1的蛋白表达水平并提高了小鼠巨噬细胞RAW264.7的凋亡率,当RIP1被干扰后,BCG感染后的RAW264.7细胞凋亡率和活性氧水平显著降低,而促凋亡蛋白Bax表达量显著下调,线粒体膜电位和抑凋亡蛋白表达量上调。同时,BCG感染后细胞周期滞留于G_1期。BCG感染可有效上调RIP1表达量并诱导RAW264.7细胞凋亡。RIP1通过下调BCG感染后RAW264.7细胞的线粒体膜电位,上调活性氧含量并提高凋亡相关蛋白Bax/Bcl-2比值,使细胞周期阻滞于G_1期从而参与诱导细胞凋亡。     

冷应激环境下补充Vc对雄性小鼠免疫器官和呼吸器官组织结构的影响

选用清洁级雄性昆明小鼠30只，随机分均为冷应激组、冷应激＋Vc组和对照组，冷应激和冷应激＋Vc组每天在0℃、下给2h的冷刺激，冷应激＋Vc纽补充300μg／g的Vc，试验期1周。结果，在冷应激环境中，小鼠早晚体质量均表现明显下降趋势，胸腺、脾脏和肺脏的重量均显著降低（P〈0．05），器官指数有所下降，但差异不显著（P〉0．05）；胸腺小体数量增加，胸腺细胞数量明显减少，动脉周围淋巴鞘变薄，肺泡腔变大，肺泡壁变薄，肺泡间毛细血管减少。补充Vc后，小鼠早晚体质量下降趋势减缓，肺脏重量和器官指数显著增加（P〈0．05），胸腺和脾脏重量及器官指数也有所增加，但差异不显著（P〉0．05）。胸腺小体体积减小，淋巴细胞数量有所增加，脾小结数量以及体积都有所增加，动脉周围淋巴鞘增厚，肺泡壁明显增厚，肺泡间结缔组织增加，毛细血管增多。结果表明补充Vc可以改善冷应激对免疫器官和呼吸器官造成的不良影响。     

Effects of cytokines from thymocytes and thymic stromal cells on chicken intrathymic T cell development

We have studied the ability of thymic stromal cells (TSC) and thymocytes to produce cytokines and the involvement of cytokines in intrathymic T cell development. When thymocytes were co-cultured with thymic stromal cells in absence of direct contact and mitogenic stimulation, induction of thymocyte proliferation was observed. Supernatants of cultured stromal cells (TSC-CS) promoted a high proliferative response on CD3- thymocytes but had little effect on CD3+ thymocytes. These results indicate that stromal cells have produced a cytokine which can induce immature thymocyte proliferation. Moreover, stromal cells express the MRNA for stem cell factor (SCF) and c-kit (the receptor for SCF) was detected on CD3- thymocytes but not on CD3+ thymocytes. Since SCF can enhance the proliferation of immature thymocytes in synergy with IL-7 in mammals, there is a possibility that chicken stromal cells may produce a IL-7-like factor. Thymocytes have clearly expressed interferon (IFN)-gamma. In contrast, thymic stromal cells showed no detectable expression of IFN-gamma. CD3+ thymocytes express IFN-gamma MRNA more strongly than CD3 thymocytes, suggesting that IFN-gamma from thymocytes may operate on stromal cells and then may indirectly induce clonal elimination of CD3+ cells on stromal cells. The expression of these cytokines and receptors by thymic stromal cells and thymocyte subpopulations suggests that these cytokines participate in paracrine interactions between these cell populations during thymocyte differentiation.     

硒缺乏通过TLR4/MyD88/NF-κB信号通路介导鸡胸腺细胞凋亡

本试验旨在探讨硒缺乏是否通过激活Toll样受体信号通路诱导鸡胸腺细胞凋亡。复制硒缺乏模型,将200只1日龄健康的肉鸡随机分为对照组（C组）和缺硒组（L组）,对照组饲喂硒含量0.2 mg·kg^-1的正常日粮,缺硒组饲喂硒含量0.004 mg·kg^-1的缺硒日粮。在15、25、35、45、55日龄时,每组分别选取15只鸡,观察胸腺组织病理形态变化和超微结构变化;检测胸腺组织中TLR4信号转导通路与细胞凋亡相关因子的表达。又建立了MDCC-MSB1（MSB1）细胞硒缺乏模型,并在此基础上设立6个分组：对照（C group）组、缺硒（L group）组、缺硒+转染空质粒（L+pCMV-HA-N）组、缺硒+siRNA阴性对照（L+NCsiRNA）组和缺硒+过表达TLR4（L+pCMV-HA-TLR4）组、缺硒+干扰TLR4（L+siChTLR4）组,低硒处理5 d后,检测细胞活力、细胞凋亡情况、TLR4信号转导通路与细胞凋亡相关因子的表达。结果显示：1）与C组相比,L组皮质髓质的淋巴细胞数量减少、细胞排列紊乱、皮质髓质充血、核碎裂,广泛的局灶性坏死。L组鸡胸腺组织淋巴细胞间出现裂隙,体积缩小,细胞碎裂,核染色质边集,线粒体肿胀,嵴断裂。2）与15日龄C组相比,L组鸡胸腺中TLR4、MyD88、NF-κB、Caspase-3、Caspase-9、Bax mRNA和蛋白表达水平在15~55日龄中均显著上调,Bcl-2 mRNA和蛋白表达水平呈相反趋势。3）与C组相比,L组MSB1细胞活力显著下降、细胞凋亡数量明显增加、TLR4、MyD88、NF-κB、Caspase-3、Caspase-9、Bax mRNA和蛋白的表达均显著增加、Bcl-2 mRNA和蛋白表达水平极显著降低;与L组相比,L+siChTLR4组细胞活力明显增强、细胞凋亡数量明显减少、相关因子mRNA和蛋白表达显著下降,而L+pCMV-HA-TLR4组细胞活力明显降低、细胞凋亡数量明显增加、相关因子mRNA和蛋白表达均明显增加。综上所述,硒缺乏通过TLR4/MyD88/NF-κB信号通路介导鸡胸腺细胞凋亡,并进一步诱导鸡胸腺损伤。     

Regulation of early chicken thymocyte proliferation by transforming growth factor-beta from thymic stromal cells and thymocytes

We examined expression of TGF-betas in chicken thymic stromal cells and thymocytes and roles of TGF-betas in thymocyte development within the thymus. Thymic stromal cells expressed TGF-beta 2 and 3 genes but not TGF-beta 4 gene. Thymocytes showed expressions of TGF-beta 2, 3 and 4 genes and each TGF-beta gene was expressed more strongly in CD3- than CD3+ thymocytes. When anti-TGF-beta antibody was added with supernatants of stromal cells into thymocyte culture, only proliferative activity of CD3- thymocytes was enhanced and the cells in S and G2/M compartments of cell cycle increased. These results suggest that TGF-beta which is expressed in the thymus may regulate the ability of immature thymocytes to progress through the cell cycle and to differentiate to CD3+ thymocytes.     

Study on the Effect of Ricin on Human Peripheral Blood B Lymphocyte and the Expression of Related Genes

This study was amied to learn the toxic effects of ricin on human peripheral blood B lymphocytes（IM-9）and the expression of related genes.The extracted ricin of different concentrations were added to culture cells for 6 and 12 h to make sure the median inhibitory concentration,then culture cells at the median inhibitory concentration for 2,6,10 and 12 h,at each time point detect the expression of immune related genes CD40,IL-1β,TNF-α and apoptosis related genes Bcl-2,Bax,Caspase-3.The results showed that the inhibitory effect of ricin on IM-9 cells was increased with the increase of concentration and time.The expression of TNF-α and CD40 genes increased significantly in experimental group than control group at 6 h（P<0.05).10 and 12 h reached extremely significant level（P<0.01);There was no significance of IL-1β gene expression between experimental and control groups（P>0.05).About apoptosis gene,the Bax gene expression decreased extremely significantly in experimental group at 10 h（P<0.05）and decreased significantly at 12 h（P<0.05);Bcl-2 gene had reached significant levels of four time periods（P<0.05), and 2,12 h were extremely significant（P<0.01);The expression of Caspase-3 reduced at 6 h,the other time points increased significantly（P<0.05),at 2 h reached extremely significant level（P<0.01).It showed that the gene expression of IM-9 cells could be significantly affected by ricin,TNF-α and CD40 genes were two potential genes to evaluate the toxicity by IM-9 cells.     

蓖麻蛋白对人外周血B淋巴细胞的作用及相关基因表达的研究

试验旨在研究蓖麻蛋白对人外周血B淋巴细胞（IM-9）毒性作用及对相关基因表达的影响。将提取的蓖麻蛋白按不同浓度添加到培养基培养6、12 h,研究剂量－时间效应,确定半数抑制浓度,然后以半数抑制浓度培养细胞2、6、10、12 h,在每个时间段检测免疫相关基因CD40、IL-1β、TNF-α和凋亡相关基因Bcl-2、Bax、Caspase-3的表达情况。结果表明,蓖麻蛋白对IM-9细胞的毒性作用随浓度和时间的增加而增强。取接近半数抑制浓度20 ng/mL作用IM-9细胞2、6、10、12 h,免疫相关基因中,TNF-α和CD40基因在6 h试验组表达量与对照组相比显著升高（P<0.05）,10、12 h达到极显著水平（P<0.01）;IL-1β基因表达量试验组、对照组间差异不显著（P>0.05）。凋亡基因中,与对照比相比,试验组Bax基因表达量10 h极显著降低（P<0.01）,12 h显著降低（P<0.05）;Bcl-2基因表达量4个时间段均达到显著水平（P<0.05）,2、12 h达到极显著水平（P<0.01）;Caspase-3基因除6 h表达量降低外,其他时间段表达量均显著升高（P<0.05）,2 h为极显著水平（P<0.01）。表明蓖麻蛋白能显著影响IM-9细胞的基因表达,TNF-α和CD40基因是两个潜在用IM-9细胞评价蛋白毒性的检测基因。     

Effect of calf thymus extract and zinc supplementation on the cellular response of mice exposed to restraint stress

The studies were carried out on Balb/c mice exposed to restraint stress twice for 12 h at 24 h intervals. Prior to stress exposure, the mice were treated with calf thymus extract (TFX - Jelfa) i.p. at a dose of 10 mg/kg, ten times at 24 h intervals. TFX was used per se or with zinc ions interaction, by adding zinc ions (as sulfate salt) to drinking water at a dose of 72 microg/mouse per day. The results obtained show that restraint stress dramatically decreased the total number of thymocytes and splenocytes which is also accompanied by decreasing weight ratio of the thymus and spleen. The decreasing number of thymic and spleen cells corresponded to a diminishing percentage of immature, double-positive CD4+CD8+ thymocytes, mature single-positive CD4+ thymic cells and CD4+, CD8+ and CD19+ splenocytes. Changes in the number of thymic cells affect their activity, which is expressed as a decreased proliferative response of thymocytes stimulated in vitro with concanavalin A (Con A) and phytohaemagglutinin (PHA). Besides, exposure to the restraint stress decreased interleukin-1 (IL-1) production by murine intraperitoneal macrophages stimulated in vitro with lipopolisacharide (LPS) from E. coli. Previous treatment with TFX counteracted restraint stress-induced immunosuppression, which is expressed as partial normalisation of the total number of thymic and spleen cells, accelerated regeneration of these two lymphatic organs, shortned suppressive action of restraint stress on the percentage of immature CD4+CD8+ thymocytes and CD4+ splenocytes and in total normalisation of the CD4+ thymocytes and CD8+ splenocytes. TFX administered prior to restraint stress not only counteracted the suppresive effects of stress on the proliferative activity of thymic cells stimulated in vitro with Con A and PHA, but also augmented the proliferative response of these cells to two mitogens. The immunorestorative effect of TFX was augmented by zinc supplementation.