当归多糖对免疫系统作用的研究

当归多糖(AP)对动物机体具有明显的免疫作用。研究表明,AP通过活化巨噬细胞作用于各种免疫效应细胞,产生相应的细胞因子从而实现其免疫作用。AP对淋巴细胞也有较强的活化作用,能提高IL-2和免疫球蛋白IgG、IgM、补体C5的水平。     

The TLR Expression Pattern on Monocyte-Derived Macrophages for Lipopolysaccharid Stimulation of Calves

In this paper, toll-like receptor expression pattern in monocytes-derived macrophages by lipopolysaccharid （LPS） stimulation was examined. Jugular venous blood samples from 4 Japanese calves were obtained and the peripheral blood mononuclear cells （PBMC） were isolated. The PBMC were cultured for 7 d so as to collect monocytes-derived macrophages in Repcell. The PBMC were stimulated by LPS for 24 h and the mRNA expression pattern of TLR and cytokines in monocytes-derived macrophages （Mod-Mφ） was analyzed. Results showed that LPS stimulation of Mod-Mφ could increase the mRNA levels of the genes of TNF-α, IL-6, and IL-8. In addition, the mRNA levels of the genes of TNF-α and IL-6 in the group of LPS stimulation were most significantly （P 〈 0.01） higher than those in control group and the mRNA levels of TLR1, 3, 5, 8, and 10 were significantly （P 〈 0.05） decreased after LPS stimulation. There was no difference in the mRNA expressions of TLR2, 4, 6, and 7 between the groups of the control and LPS stimulation. Besides, expression of TLR9 was not found. It suggested that monocytes-derived macrophages could respond to LPS and they might take an important role in the innate immunity. The important function of the cells might contribute to better disease treatment.     

重组真菌免疫调节蛋白对巨噬细胞RAW 264.7激活应答的研究

真菌免疫调节蛋白(fugal immunomodulatory proteins,FIPs)是一类从高等大型真菌中分离的、具有生物活性的小分子蛋白质。在之前的研究中,通过基因改组技术,对来自赤芝(Ganoderma lucidum)和紫芝(G.sinense)的FIP进行了重组改造,获得了具有333bp的FIP突变体FIP-SN15。本研究通过原核表达系统获得了重组FIP-SN15(rFIP-SN15),并利用荧光定量PCR(quantitative real-time PCR,qRT-PCR)技术在转录水平上对rFIP-SN15诱导巨噬细胞TNF-α的功能及其机制进行研究。结果表明,rFIP-SN15在原核表达系统中的产量约为35.31mg/L;rFIP-SN15处理细胞后,TNF-α的转录水平显著上调(P0.05),并且Akt的转录水平上调极其显著(P0.01)。此外,ARG1和mTOR的mRNA水平上调显著(P0.05)。因此,rFIPSN15能够激活小鼠腹腔巨噬细胞RAW 264.7。本研究为进一步验证FIP-SN15的生物学功能提供了有益的参考。     

In infectious pancreatic necrosis virus carrier Atlantic salmon, Salmo salar L., post-smolts, almost all kidney macrophages ex vivo contain a low level of non-replicating virus

The level of infection by infectious pancreatic necrosis virus (IPNV) of kidney macrophages from 12 asymptomatic carrier Atlantic salmon post-smolts was studied. Kidney leucocytes were fractionated on 34/51% Percoll gradients, allowed to adhere to plastic wells overnight, washed to remove non-adherent cells and cultured for up to 7 days with or without renewal of medium on day 3. On day 1, supernatants were harvested, macrophages were counted, lysed and IPNV in the supernatants and lysates was titred in chinook salmon embryo (CHSE-214) cells. The multiplicity of infection ranged between 1:2.2 and 1:7.4 (virus:macrophages). On day 3, the titres of IPNV in macrophage lysates decreased and in wells where the medium was renewed on day 3, IPNV was no longer detectable on day 7. In the supernatants, one fish was positive for IPNV on day 1, four fish on day 3 but none were detectably positive on day 7. In parallel wells in which the medium was not renewed, on day 7 IPNV was detected in macrophage lysates of three fish and the supernatants were also IPNV positive in two of these fish. This suggests that virus might be shed from infected macrophages and then reinfect other macrophages. When macrophages were serially diluted in wells and cultured for 24 h, IPNV could be cultured from macrophage lysates of wells containing between two and 70 macrophages. These results indicate that a very high proportion of the adherent kidney macrophages must be infected with very few non-replicating virions.     

鲫鱼的细胞免疫应答研究

以嗜水气单胞菌（AhJ-1）灭活疫苗腹腔注射免疫鲫鱼（Crucian carp）,加强免疫后取血分离出淋巴细胞,进行巨噬细胞移动抑制试验,结果显示致敏淋巴细胞于体外培养时,在特异性抗原刺激下,产生了巨噬细胞移动抑制因子,抑制了巨噬细胞移动,移动抑制指数约为0.67,初步说明细胞免疫应答在鲫鱼的免疫应答中具有一定的地位。     

桔梗皂苷D对小鼠淋巴细胞和巨噬细胞免疫功能的影响

【目的】探讨桔梗皂苷D对小鼠淋巴细胞和巨噬细胞免疫功能的影响。【方法】无菌分离小鼠脾淋巴细胞和腹腔巨噬细胞,制备淋巴细胞和巨噬细胞悬液。分别培养这2类细胞,并分为空白对照组、阳性对照组(左旋咪唑处理细胞)及25,50,75和100μg/mL桔梗皂苷D组,采用MTT法检测桔梗皂苷D对淋巴细胞增殖和巨噬细胞吞噬功能的影响,采用ELISA法检测桔梗皂苷D对淋巴细胞IL-2、IL-4和巨噬细胞TNF-α、IL-12分泌的影响。【结果】桔梗皂苷D能够促进淋巴细胞增殖,增强巨噬细胞的吞噬功能,并可刺激淋巴细胞IL-2、IL-4和巨噬细胞TNF-α、IL-12的分泌,其中以50μg/mL组效果最佳。【结论】桔梗皂苷D能增强小鼠淋巴细胞和巨噬细胞的免疫调节活性。     

乳黄消口服液滋阴、扶正及祛邪功效的研究

为探讨乳黄消口服液滋阴、扶正及祛邪的功效,本研究观察乳黄消口服液对阴虚模型大鼠临床症状、饮食量、饮水量、体重、胸腺指数、脾脏指数、总三碘甲状腺原氨酸(T_3)、总甲状腺素(T_4)和促甲状腺激素(TSH)含量的影响,以及对小鼠巨噬细胞吞噬功能及对大肠杆菌、金黄色葡萄球菌、无乳链球菌和副乳房链球菌抑制作用的影响。结果显示,试验组大鼠在乳黄消口服液给药后精神正常,易抓取,但被毛光泽仍较差,有脱毛现象,大便正常。与模型组相比,试验组大鼠体重(P>0.05)、胸腺指数(P>0.05)、脾脏指数(P>0.05)、TSH含量(P<0.05)上升,饮食量(P<0.05)、T_3(P<0.05)、T_4含量(P>0.05)下降,但均与阴性对照组差异不显著(P>0.05);与模型组相比,试验组大鼠饮水量显著下降(P<0.05),但仍显著高于阴性对照组(P<0.05)。乳黄消口服液给药后高、中、低剂量组与阴性对照组间吞噬指数均差异显著(P<0.05),乳黄消口服液高、中剂量组与阴性对照组相比吞噬百分比差异显著(P<0.05);乳黄消口服液对大肠杆菌、金黄色葡萄球菌、无乳链球菌及副乳房链球菌的最小抑菌浓度(MIC)分别为12.50、1.56、0.20和0.39 mg/mL。综上所述,乳黄消口服液可改善大鼠阴虚临床症状,增强巨噬细胞吞噬功能,抑制奶牛乳房炎主要病原菌,具滋阴、扶正和祛邪的功效,可用于奶牛乳房炎的治疗。     

塑化剂DEHP暴露对小鼠巨噬细胞的免疫毒性作用

为了研究邻苯二甲酸二乙基己酯(DEHP)对巨噬细胞的免疫毒性,采用不同浓度剂量DEHP处理小鼠巨噬细胞株RAW264.7细胞和腹腔巨噬细胞,通过检测细胞吞噬、细胞因子分泌及活性氧水平变化探讨DEHP暴露对免疫系统的毒性作用。结果表明:高浓度(20、40、80 mg·L-1)DEHP处理48 h后对RAW264.7细胞可造成急性损伤。低浓度(1、5、10 mg·L-1)DEHP连续染毒处理10代后,RAW264.7细胞吞噬红细胞百分率及吞噬指数显著抑制(P0.05)。RAW264.7细胞分泌TNF-α、IL-12和IL-23等细胞因子能力明显降低,并表现出一定的剂量-效应关系。随着DEHP暴露浓度的增加,DEHP染毒造成RAW264.7细胞内活性氧产生,受损加剧。除此之外,DEHP暴露还能明显抑制小鼠腹腔巨噬细胞对红细胞的吞噬能力(P0.05)。以上结果表明,DEHP暴露对巨噬细胞具有免疫抑制作用,损伤了巨噬细胞正常免疫功能发挥。     

大黄鱼刺激隐核虫病组织中巨噬细胞移动抑制因子的检测分析

由刺激隐核虫(Cryptocaryon irritans)感染引发的\"白点病\"为大黄鱼(Larimichthys crocea)养殖业带来重大损失,但仍未找到安全、有效的治疗方法。本研究应用免疫组织化学方法,对健康大黄鱼和患刺激隐核虫病大黄鱼肠、脾、头肾与肝组织中巨噬细胞移动抑制因子(macrophage migration inhibitory factor,MIF)蛋白进行检测,研究刺激隐核虫病大黄鱼组织中MIF的表达情况及其对预后的影响。结果表明,MIF在健康大黄鱼肠、脾、头肾、肝各组织中均无表达;MIF在刺激隐核虫病大黄鱼各组织中高表达,表达强度从弱到强依次是肠、肝、脾、头肾,MIF表达阳性率分别为54%、80%、86%、90%。推测MIF在大黄鱼刺激隐核虫病发病过程中发挥作用。