双歧杆菌LTA、EPS 对小鼠巨噬细胞活力的影响

脂磷壁酸（LTA）和胞外多糖（EPS）是双歧杆菌的细胞壁表面物质，在抗肿瘤和提高免疫力方面已成为研究热点。据此，本研究采用MTT法、荧光显微镜形态观察及激光扫描共聚焦分子生物学观察法检测LTA和EPS对小鼠腹腔巨噬细胞的免疫激活能力，并通过脂多糖（LPS）作用组做对照，进一步探讨LTA和EPS提高免疫活性机理及影响程度。     

LBP对LPS激活巨噬细胞内p38信号通路的影响 

目的:探讨脂多糖结合蛋白(LBP)对脂多糖(LPS)激活肺泡巨噬细胞内p38信号通路的调节作用。方法:经硫酸铵盐析、Bio-Rex70阳离子交换层析和MonoQ阴离子交换层析, 从大鼠急性期血清中分离纯化LBP。分别用0.01mg/L和1mg/L的LPS刺激肺泡巨噬细胞, 并加入不同浓度LBP(0mg/L、0.01mg/L、0.1mg/L、1mg/L和10mg/L), 观察肺泡巨噬细胞中p38蛋白激酶的磷酸化程度。结果:纯化的大鼠LBP在SDS-PAGE的60kD处呈现单一条带, 并可增强LPS与单核细胞的结合。当LPS浓度为0.01mg/L时, 1mg/L以下的LBP可明显增敏LPS对肺泡巨噬细胞内p38信号通路的激活, 并且这种增敏作用随LBP浓度的增加而增强。但LBP为10mg/L时, LBP对LPS的增敏作用反而有所减弱;当LPS为1mg/L时, LBP对LPS激活肺泡巨噬细胞内p38信号通路无调节作用。结论:LBP对低浓度LPS(0.01mg/L)激活肺泡巨噬细胞内p38信号通路有明显的调节作用;而高浓度LPS(1mg/L)不需LBP的增敏作用, 可能通过LBP非依赖途径直接激活p38信号通路。     

p38蛋白激酶对大鼠肺泡巨噬细胞活化机制的调控

目的:探讨p38蛋白激酶对LPS诱导大鼠肺泡巨噬细胞激活机制的作用。方法:提取细胞核蛋白,采用Western印迹分析p38蛋白激酶水平。用放射免疫法检测细胞上清TNF-α、IL-8的含量。结果:LPS显著增加肺泡巨噬细胞TNF-α、IL-8的合成,呈剂量依赖性诱导p38的活化。特异性p38蛋白激酶抑制剂SB203580能显著降低LPS诱导的肺泡巨噬细胞核蛋白p38的含量及细胞上清TNF-α、IL-8水平。结论:LPS刺激肺泡巨噬细胞释放炎性细胞因子TNF-α、IL-8受p38蛋白激酶调控。     

IL-6阳性细胞在小鼠胸腺内的定位

目的：检测IL－6阳性细胞在小鼠胸腺的分布。方法：采用免疫酶细胞化学，免疫荧光双染及免疫电镜技术方法原位显示小鼠胸腺不同部位IL－6阳性神经的表达情况。结果：光镜免疫酶组织化学确定了IL－6阳性细胞的分布范围，即IL－6在胸腺皮质和髓质的某些特定的细胞表达；进一步采用免疫荧光双染色及免疫电镜技术对IL－6阳性细胞进行了解析，确定了小鼠胸腺内IL－6阳性细胞主要是胸腺实质中不同形态的巨噬细胞，IL－6不存在于胸腺上皮细胞及胸腺细胞。结论：巨噬细胞是小鼠胸腺IL－6的主要来源，参与胸腺细胞发育微环境的构成。     

巨噬细胞炎症蛋白1-α、解整合素样-金蛋白酶8、12 和CD68蛋白在颌骨巨细胞病变及长骨骨巨细胞瘤中的表达

目的检测巨噬细胞炎症蛋白1-α(MIP-1α)、解整合素样-金属蛋白酶(ADAM)8、12和CD68在颌骨巨细胞病变和长骨骨巨细胞瘤中的表达,探讨其在这两种巨细胞病变中的多核巨细胞发生中的作用.方法采用免疫组织化学SP法检测24例颌骨中心性巨细胞病变,24例长骨骨巨细胞瘤石蜡组织中的MIP-1α、ADAM8、ADAM12和CD68蛋白的表达.结果 MIP-1α在24例颌骨中心性巨细胞病变和24例长骨骨巨细胞瘤中的血管、类骨质和新骨形成区中强表达,而在多核巨细胞和圆形单核细胞中为阴性;ADAM8、ADAM12和CD68在颌骨中心性巨细胞肉芽肿和长骨骨巨细胞瘤中几乎所有多核巨细胞和部分圆形单核基质细胞均为阳性.结论颌骨中心性巨细胞病变和长骨骨巨细胞瘤中的多核巨细胞可能由CD68+的圆形单核基质细胞融合而成,这些圆形单核细胞可能是由骨微环境中的趋化因子,募集外周血中循环的单核细胞到病变局部分化而成的具有破骨细胞前体细胞性质的细胞,继而在某些融合蛋白作用下发生融合,成熟为多核破骨样巨细胞.     

Distinct Pattern of Human Vδ/51 γδ/5 T Cells Recognizing MICA

γδ T cells represent one unique recognition pattern, the limited recognition, which distinguishes from the specific recognition for αβ T cells and pattern recognition for macrophages. Vδ1 γδT cell is the major subset of human γδT cells, which predominates in mucosal tissue including the intestinal epithelia. Presently, a few antigens that human Vδ1TCR can recognize have been identified. Among them, MHC class I chain-related molecules A （MICA） have been studied most intensively. Besides Vδ1TCR, MICA is also the ligand of NKG2D, a C-type lectin-like activating immunoreceptor. In human, only Vδ1 cells can simultaneously express both types of receptors of MICA while NK cells, αβ T cells and other subsets of γδT cells likewise express NKG2D. Although the precise mechanisms are still enigmatic, this distinct pattern of Vδ1 cells recognizing MICA predicts unique biological significance of Vδ1 cells in immune defense. Recent years, some progresses have been made in this issue. In this review we summarize the related reports and put forward some novel views based on our group＇s studies.     

Kinetics of the phenotype and function of murine peritoneal macrophages following acute inflammation

This study was undertaken to have a better understand for the process and the underlying mechanisms to limitmacrophage activation and population of activated macrophages.A comprehensive kinetics of cytokineproduction was performed in murine peritoneal macrophages recovered from Balb/c mice at various timeduring the course of an intraperitoneal injection with thioglycollate (TG).The expression of cell surfacemolecules such as MHC-Ⅰ,MHC-Ⅱ,B7-1 and B7-2 of these macrophages were also determined by flowcytometry.The present findings of our research suggested that the population of activated macrophages and theactivation of macrophages (including cytokines production and expression of cell surface functional molecules)were strictly controlled during inflammation process.This is one of the important mechanisms to retain the hosthomeostasis.Cellular & Molecular Immunology.2004;1(1):57-62.     

巨噬细胞激活因子的分离纯化

由Ｔ细胞株（Ｍｏｌｔ ４）产生的巨噬细胞激活因子（ＭＡＦ）用阴离子交换和凝胶过滤色谱纯化。纯化的ＭＡＦ进行ＰＡＧＥ出现一条带，进行ＳＤＳ－ＰＡＧＥ证实其为单亚基，分子量为６００００；ＩＥＰ表明其等电点在ｐＨ３．５～４．５之间。     

rhGM-CSF作为成人乙肝疫苗无(弱)应答者免疫佐剂初探

Objective To investigate the effect of recombinant human granulocyte macrophage colony stimulating factor (rhGM-CSF) as adjuvant on immune response in adults of non-and hyporesponders to hepatitis B vaccine. Methods Those who were once immunized with recombined yeast gene hepatitis B vaccine more than one standard scheme in two years and negative for hepatitis B markers were randomly sorted as group A and group B. 33 adults of group A were given hepatitis B vaccine 10 μg each time. The immune procedure was O, 1 and 6month. 34 adults of group B were given rhGM-CSF 300 μg for the first day, then 10 μg each time for routine immune. The blood samples were collected before the first injection and in 1, 2 and 8 months (T1, T2, TS)following the first injection to test Anti-HBs. Results Anti-HBs positive conversion rates of group A and B at T8 was 39.39% and 64.71% respectively(P=0.038). Anti-HBs levels of group B at TI, T2, T8 were(113.85±198.56) mIU/ml, (312.40±349.44) mIU/ml, (427.74±411. 58) mIU/ml (P=0.001). There was significant difference between group A and B in T8 Anti-HBs levels(P=0.010). Conclusion Better immune response was found in the group of rhGM-CSF with hepatitis B vaccine. So rhGM-CSF can induce the immune respond to hepatitis B vaccine.     

小白鼠腘淋巴结淋巴窦的超微结构

我们观察了幼年小白鼠腘淋巴结淋巴窦的超微结构,窦壁由腔面向外一般出3层构成:(1)一层连续的内皮;(2)由一薄层细胞间质构成的基膜;(3)一层外膜网状细胞及其扁平的突起。内皮细胞核扁圆,异染色质细小,核仁不明显,胞质极薄,含粗面内质网极少,而有大量的吞饮小泡。细胞邻接处相互重叠或嵌合,有20nm宽的间隙相隔。有的地方可见不发达的细胞连接。当巨噬细胞或淋巴细胞穿越内皮时,可出现临时性间隙,内皮边缘与穿越细胞相贴。基膜由电子透明的无定形基质及细的胶原原纤维构成。外膜网状细胞的核异染色质较多,核仁明显,胞质丰富,含许多粗面内质网而很少吞饮小泡。外膜网状细胞之间常见0.5μm宽的间隙。在窦腔内有巨噬细胞、淋巴细胞及星状的网状细胞等。由于小白鼠腘淋巴结的淋巴窦小,故窦内星形的网状细胞很少,其超微结构特点,与外膜网状细胞及淋巴组织内的网状细胞相似,在突起的切面上,常见包有小束的胶原原纤维。巨噬细胞较多,形态不规则,常贴附于内皮细胞表面,核椭圆,常有凹陷,胞质丰富,含许多溶酶体。在取材前1h,于足垫注射中国墨汁的小鼠,巨噬细胞吞噬了大量粗的墨汁颗粒。本文认为,淋巴窦壁可能具有一定的屏障作用。