低硒与Coxsackie B_(3m)病毒感染对BALB/c小鼠肝脏脂质过氧化物代谢的影响

用国产低硒食用酵母合成低硒饲料（硒０．０１３ｍｇ／ｋｇ，ＶＥ含量为２０ｍｇ／１００ｇ）喂养断奶后ＢＡＬＢ／Ｃ雄性小鼠，５周后腹腔接种嗜鼠心肌病毒ＣＶＢ３ｍ１０３ＴＣＤ５００．１ｍｌ，７天后处死建立低硒状态下病毒性心肌炎模型，测定肝脏组织中脂质过氧化物（ＬＰＯ）含量及全血中谷胱甘肽过氧化物酶（ＧＳＨ－Ｐｘ）活性。结果表明低硒感染ＣＶＢ３ｍ小鼠肝组织中ＬＰＯ含量明显高于常硒病毒感染组及常硒对照组（Ｐ＜０．０１）；低硒感染ＣＶＢ３ｍ病毒组小鼠全血ＧＳＨ－Ｐｘ活力也最低。结果提示：低硒因素加重病毒感染引起ＧＳＨ－Ｐｘ活力降低，ＬＰＯ堆积，降低机体的抗氧化能力     

弓形虫RH株ROP16I/III基因缺陷虫株感染BALB/c鼠的致病性研究

目的 旨在深入研究弓形虫棒状体效应分子ROP16与虫株毒力及致病性的关系。方法 采用弓形虫ROP16_I/III基因敲除RH株(RH_ΔROP16)攻击感染BALB/c小鼠,在感染后不同时间点与野生株感染鼠比较,动态观察感染动物发病症状、存活时间; HE染色观察脑组织、肺组织病理学差异,qRT-PCR检测脾细胞炎性及抑炎细胞因子表达水平。结果 两组小鼠的发病症状无明显差异;经HE染色显微镜下观察小鼠肺部及脑部病理学改变亦未见无明显差异;qRT-PCR检测并用Graph pad分析两组虫株感染小鼠的脾细胞Arg-1、IL-10、IL-12、TNF -α及IFN-γ等细胞因子表达水平。数据表明在感染相同时间ROP16缺陷株感染小鼠Arg-1的表达量明显低于野生型虫株(P< 0.05);而IL-10、IL-12、TNF-α和IFN -γ的表达水平无统计学差异(P> 0.05)。结论 I型弓形虫RH株的ROP16_I/III并非是决定急性感染期弓形虫毒力的唯一效应分子;ROP16_I/III可诱导宿主Arg-1高表达,提示与巨噬细胞内虫体的增殖有关。     

小鼠接种rHBs疫苗产生的特异性淋巴细胞增殖反应

目的　研究小鼠免疫接种基因重组乙型肝炎表面抗原疫苗(recombinant hepatitis B virus(HBV)surface vaccine,rHBs)产生的特异性淋巴细胞增殖反应。方法　40只BALB／c小鼠随机分为0.65、1.25、2.5、5 μg四组,腹腔分别接种0.65、1.25、2.5、5 μg的rHBs,一半4、鼠2周后加强免疫1次。分别在初次免疫后2周或加强免疫后2周分离小鼠脾T淋巴细胞;分别进行以下实验:实验组用rHBs(10 μg／m1)刺激脾T淋巴细胞,对照组用PBS代替rHBs刺激脾T淋巴细胞;3天后用3H掺入法检测脾T淋巴细胞特异性增殖反应,以3H掺入的同位素cpm值及刺激指数(Stimulation index,SI,实验组cpm值／对照组cpm值)表示。结果 只接受单次免疫的0.65,1.25,2.5,5 μg组小鼠脾T淋巴细胞特异性增殖反应SI分别为1.55、1.93、2.41、2.81;而接受加强免疫的0.65、1.25、2.5、5 μg组小鼠脾T淋巴细胞特异性增殖反应SI分别为1.61、2.05、3.74、3.62。结论 小鼠接种rHBs后产生特异性淋巴细胞增殖反应,加强免疫或增加接种剂量可以增强反应强度。     

Tularemia induces different biochemical responses in BALB/c mice and common voles

Background  

Both BALB/c mice and common voles (Microtus arvalis) are considered highly susceptible to tularemia. However, the common vole is reported to harbour Francisella tularensis in European habitats as well as to survive longer with chronic shedding of the bacterium. The purpose of the present study was to compare the response of these two rodents to a wild Francisella tularensis subsp. holarctica strain infection.     

The mucosal mast cell and IgA plasma cell responses to primary Trichinelia spiralis infection in BALB/c mice treated with ketotifen

The effect of ketotifen on inflammation in the intestine during T. spiralis infection in BALB/c mice was examined. The lack of enteropathy in treated mice was accompanied to affected the size of worms and the number of worms, but did not prevent the retardation of nematodes in the intestine. Fecundity of females in vitro and the number of muscle larvae were not statistically changed after treatment. Ketotifen reduced the number of mast cells and IgA plasma cells in the intestine. As inflammatory influx, epithelial damage and IgA response are related to the action of T cells, ketotifen affected T dependent reactions in the gut of T. spiralis infected mice.     

Proteoglycan-induced polyarthritis and spondylitis adoptively transferred to naive (nonimmunized) BALB/c mice

Mononuclear cells from BALB/c mice with progressive polyarthritis and spondylitis induced by injection of fetal human articular cartilage proteoglycan (PG) were used to transfer arthritis by intravenous injection into irradiated, nonimmunized syngeneic mice. Successful transfer of arthritis to BALB/c mice required the injection of lymphocytes from mice with arthritis, along with 50 m̈g of human fetal PG, or lymphocytes stimulated in vitro with either fetal human PG or with mouse cartilage PG. In addition, interleukin-2 or immune sera from animals with arthritis significantly reduced the time to onset of transferred disease. The onset of adoptively transferred arthritis, using cells and antigen, from the time of the first injection (38.2 ± 18.2 days, mean ± SD) was shortened if lymphocytes from mice with transferred arthritis were reinjected (retransferred) into other, irradiated syngeneic mice (6.1 ± 2.6 days). The appearance of autoreactive antibodies to mouse cartilage PG in the sera of mice with adoptively transferred arthritis (secondary or tertiary) preceded the appearance of the first clinical symptoms by a few days. The transfer of arthritis was blocked by pretreatment of donor (arthritic) lymphocytes with either anti-T cell or anti-B cell antibodies and complement. Exposure of mononuclear cells from mice with arthritis to PG, and its removal prior to transfer, also resulted in transfer of the arthritis. PG-induced arthritis was not transferred to nonirradiated mice, nor to irradiated mice injected with lymphocytes from animals with primary arthritis without chondroitinase ABC-digested fetal human PG. Arthritis never developed after injection of immune sera from mice with arthritis (without cells), nor when cells of nonarthritic animals were used with chondroitinase ABC-digested fetal human PG, with or without interleukin-2.     

白细胞介素-15重组体对HBsAg核酸疫苗的免疫佐剂作用

白细胞介素-15(IL-15)属螺旋结构细胞因子家族，它能促进T细胞、自然杀伤(NK)细胞和B细胞增殖及分化；具有诱导淋巴细胞产生γ干扰素(IFN-γ)及肿瘤坏死因子α的作用。IL-15与疫苗共同免疫时可刺激抗体产物增加，细胞毒性T淋巴细胞(CTL)活性加强。将IL-15表达质粒与HBsAg DNA疫苗共同免疫小鼠，以观察其对小鼠免疫应答的影响。     

致弱日本血吸虫尾蚴诱导BALB/c小鼠早期免疫应答动态变化研究

目的比较辐照致弱日本血吸虫尾蚴免疫小鼠与正常感染小鼠早期免疫活化程度及其动态变化差异。方法采用流式细胞术（FCM）和免疫组织化学（IHC）方法比较辐照致弱尾蚴免疫小鼠与正常感染小鼠早期脾组织和/或肺组织中树突状细胞（DC）表面分子CD11c、T细胞表面分子CD25表达差异及脾细胞和外周血中CD3＋CD25＋/CD3＋T细胞比例,分析T细胞免疫活化程度及其动态变化。结果在感染后7d,脾细胞中的CD3＋CD25＋/CD3＋T细胞比例致弱尾蚴免疫组为（19.52±3.65）%,明显低于正常感染组的（22.12±3.24）%;而在第14、21天,致弱尾蚴免疫组这一比例分别为（28.73±3.94）%和（26.43±0.40）%,均高于正常感染组的（13.68±3.64）%和（14.42±2.24）%。在攻击感染后7、14、21d,致弱尾蚴免疫组与正常感染组小鼠在肺组织中的CD11c＋DC表达率分别为（1.05±0.16）%和（0.96±0.15）%、（1.34±0.15）%和（1.09±0.17）%、（1.49±0.14）%和（0.97±0.16）%,脾组织中CD11c＋DC表达率分别为（2.05±0.26）%和（1.95±0.18）%、（2.24±0.25）%和（2.17±0.25）%、（2.18±0.26）%和（2.06±0.18）%。在攻击感染后7、14、21d,致弱尾蚴免疫组与正常感染组小鼠在肺组织中CD25＋T细胞表达率分别为（1.24±0.13）%和（1.17±0.16）%、（1.48±0.11）%和（1.25±0.13）%、（1.55±0.14）%和（0.97±0.12）%,脾组织中CD25＋T细胞表达率分别为（3.25±0.22）%和（2.93±0.20）%、（4.57±0.23）%和（3.69±0.24）%、（4.28±0.24）%和（3.86±0.26）%,紫外线辐照致弱尾蚴免疫小鼠较正常感染小鼠在攻击感染后第7、14、21天能在肺组织募集更多的CD11c＋DC并激活更多的CD25＋T细胞。结论致弱日本血吸虫尾蚴免疫小鼠在攻击感染后14、21d,小鼠体内T细胞激活程度和肺部DC活化程度均高于正常感染组,提示致弱尾蚴在肺部可募集更多抗原递呈细胞并使之活化。     

A comparison of mucosal inflammatory responses to Giardia muris in resistant B10 and susceptible BALB/c mice

In the first three weeks of primary Giardia muris infections B10 mice clear infection more rapidly than BALB/c mice. There is evidence that interferon-γ contributes to the relative resistance of B10 mice. The nature of the functional contribution of interferon-γ is unclear and does not relate to the secretory or serum antibody response. Mucosal inflammatory events in these strains have been studied. Apart from a small rise in both strains of goblet cell and mucosal mast cell numbers, associated with release of mast cell protease-1 in serum, no inflammatory infiltrate was observed at the time trophozoites were cleared from the intestinal lumen. Inhibition of mast cell products (5-hydroxytryptamine and histamine) by cyproheptadine enhanced the intensity of infection in both strains. The relative resistance of B10 mice could not be explained in terms of the mucosal inflammatory response.     

cDNA微阵列分析BALB/c小鼠肝脑组织中差异性表达基因的研究

目的分析BALB/c小鼠肝、脑组织差异基因表达谱。方法TRIZOL法提取总RNA,反转录cDNA,采用36K小鼠全基因组寡核苷酸芯片,运用荧光双交换方法分析BALB/c小鼠肝脑组织中mRNA转录本含量。经芯片扫描、图像采集后,运用MAS2.0软件对差异基因进行基因功能分析(GO)和KEGG通路分类。结果基因芯片结果表明与脑组织相比,在肝组织中共发现差异表达基因6 132个,其中包括上调基因2 974个,下调基因3 158个,GO分析上述差异表达基因主要涉及代谢、细胞信号转导、DNA结合与转录、细胞周期、细胞骨架、细胞粘附和发育等。KEGG信号通路分析显示:上调差异基因中71个通路的改变差异具有统计学意义(P0.05),下调差异基因中有80个通路的改变差异具有统计学意义(P0.05)。结论小鼠肝与脑组织基因表达谱的差异涉及多个基因表达调控途径,研究这些基因分子功能有助于深入了解各组织独特的功能表达系统。     

Effects of the antioxidant butylated hydroxytoluene (BHT) on mortality in BALB/c mice.

Butylated hydroxytoluene (BHT) was given in the feed to determine its effect on life span in genetically well-defined, barrier-derived BALB/c mice. Both sexes received 0.75% BHT for three different treatment periods: (A) 8 to 11 weeks of age; (B) for life, beginning at 11 weeks; (C) for life, beginning at 8 weeks of age. The control group (D) was untreated. All BHT treatment groups had mean survival times which exceeded that of controls. The order of survival was B greater than C greater than A greater than D (Males: 890, 832, 726, 684 days; Females: 875, 798, 759, 701 days). Most of the increases in mean survival time were related to a reduction in early deaths (350--600 days) in BHT-treated mice. The reason for the life-lengthening effect on BHT was not identified, but it may relate to alterations in specific disease incidences.     

泡球蚴感染对宿主肝细胞增殖影响的初步研究

目的 探讨泡球蚴感染对宿主肝细胞增殖和细胞周期的影响.方法 8～ 10周龄雌性BALB/c小鼠随机分为实验组和假手术组.分别于感染后2、8、30、60、90d采集小鼠肝脏标本.观察小鼠肝脏病理改变.检测泡球蚴感染不同阶段的增殖细胞核抗原(PCNA)和细胞周期蛋白(cyclins)表达的动态变化及其定位.组间数据比较采用t检验.结果 随着泡球蚴持续感染,实验组病灶与肝细胞交界区出现炎性细胞浸润、脂肪变性和纤维结缔组织增生等病理变化,感染早期(2、8、30 d)病灶周围肝细胞存在细胞核增大、双核或多核细胞增多等现象.感染30、60、90 d,实验组PCNA表达呈上升趋势,阳性率明显高于假手术组(7.01％±1.89％与1.03％±0.52％、8.41％±2.80％与0.93％±0.31％、13.40％±4.43％与1.07％ ±0.94％,t值分别为-6.817,-5.931,-6.102,P值均＜0.05).感染30d和60d时,实验组cyclin D1表达上升,阳性率明显高于假手术组(6.73％±2.52％与0.48％±0.43％、8.22％±3.09％与0.55％±0.34％,t值分别为-5.479,-5.504,P值均＜ 0.05);感染90d时,cyclin D1呈下降的趋势(从“+++”降至“+”).感染30、60、90d时,实验组cyclin A阳性率明显高于假手术组(7.75％±3.05％与0.69％±0.36％、3.42％±1.80％与1.14％±0.42％、3.03％±1.50％与0.69％±0.31％,t值分别为-5.131,-2.774,-3.415,P值均＜ 0.05).感染2、8、30d时,实验组cyclin B1表达呈上升趋势(从“+”升至“++”),感染60、90 d时,cyclin B1表达呈下降趋势(从“++”降至“+”).结论 体内泡球蚴感染促进宿主肝细胞的增殖和抗凋亡,并对其细胞周期产生影响.     

Differential responses of yellow Avy/A and agouti A/a (BALB/c X VY) F1 hybrid mice to the same diets: glucose tolerance, weight gain, and adipocyte cellularity

Differences in weight gain, efficiency of food utilization, glucose tolerance, insulin levels, and adipocyte cellularity were measured when three different diets were fed to lean agouti and obese yellow mice. Sets of adult and weanling agouti (A/a) and yellow (Avy/A) (BALB/c X VY) F1 hybrid mice were fed high-sucrose (HS), 10 percent fat, or regular lab chow (control). Some mice received the diets only after 12 weeks of eating lab chow (adult-fed); others ate the diets from the time of weaning. Since responses of both age groups were similar, only the data from the adult-fed groups are presented. The HS and 10 percent fat diets increased the body weight gain in both A/a and Avy/A adult mice more than the control diet; the HS diet was utilized more efficiently in the lean agouti females causing a slightly higher weight gain, whereas the 10 percent fat and HS diets were used with the same efficiency by the yellow mice. Short-term feeding (3-5 weeks) with the HS and 10 percent fat diets decreased the glucose tolerance of adult yellow but not of agouti mice. The pancreatic insulin response to a glucose load was higher in all mice fed the HS diet, whereas this response was blunted in those on the 10 percent fat diet. In agouti mice the HS and 10 percent diets increased the mean cell diameter of the parametrial adipocytes and deteriorated the glucose oxidation rate in response to insulin compared to the control diet. In the yellow littermates, on the other hand, the test diets decreased the mean cell diameter and also impaired insulin sensitivity of the adipocytes. The decrease of the mean adipocyte size was probably due to an increased number of small cells.     

Inflammatory cell infiltration and high antibody production in BALB/c mice caused by natural exposure to Lutzomyia longipalpis bites

Sand flies inject saliva into the mammalian host when probing for a blood meal. Understanding the initial vertebrate reactions against sand fly saliva is important for possible interventions because these insects transmit diseases to humans and other animals. Little is known of these reactions to New World sand flies. Repeated exposure of BALB/c mice to Lutzomyia longipalpis bites leads to local inflammatory cell infiltration comprised of neutrophils, macrophages, and eosinophils. Total IgG and IgG1 antibodies react predominantly with three major protein bands (45, 44, and 16 kD) of the insect saliva by Western blot. The injection of immune serum previously incubated with salivary gland homogenate induced an early infiltration with neutrophils and macrophages, suggesting the participation of immune complexes in triggering inflammation.     

Humoral immune responses induced by Kudoa sp. (Myxosporea: Multivalvulida) in BALB/c mice: oral administration, immunization and cross-reactions with Myxobolus aeglefini (Myxosporea: Bivalvulida)

The majority of Kudoa species infect the somatic muscle of fish, establishing cysts. Because there is no effective method to detect infected fish without destroying them, these parasitized fish reach the consumer. The elevated humoral responses detected previously by us in BALB/c mice immunized with Kudoa sp. pseudocyst extracts showed the possible immunopathological effects in man from the ingestion of Kudoa-infected fish. In this work, the high IgG1 and IgE levels induced by the oral administration of Kudoa pseudocysts to BALB/c mice confirmed the allergenic nature of some of their components. An alternative way of preparing the soluble extract by using a FastPrep' shaker indicated the inconvenience of using sonication to prepare the Kudoa sp. extract. IgG+M, IgG, IgG3 and IgA cross-reactions of Kudoa sp. with another myxosporean, Myxobolus aeglefini, were found.     

The presence of Syphacia obvelata in laboratory mice (BALB/c) — parasite antigens in immune response

Summary In conventional mice colonies, mouse pinworm, Syphacia obvelata is found very often. Several studies indicate that infection with this parasite can modulate the immune system of the host and can affect experimental final results. The aim of our study was to investigate the most immunogenic proteins of S. obvelata inducing both local and systemic immune response in naturally infected laboratory mice. Protein extracts of S. obvelata were analysed by Western blotting to examine their antigenic character. The antigens were probed with serum and mucosa of S. obvelata naturally infected mice. Surface and somatic antigens were recognized by serum and mucosal IgG, IgA and IgM antibodies. The most immunogenic and dominant proteins were observed. Proteins of Mw ∼ 70, 65 and 48 kDa showed the most evident reaction with serum and mucosa antibodies of infected animals. Surface and somatic antigens of nematode S. obvelata eliciting immune response in laboratory mice may be useful in development of a diagnostic test which could be applied for the infection control prior the experiments.     

Clonogenic mast cell progenitors and their excess numbers in chimeric BALB/c mice with inactivated GATA-1

In agar cultures of marrow cells from adult female BALB/c chimeric GATA-1(Plt13/+) mice, a high frequency of unusual dispersed colonies was noted. Analysis showed that these were colonies of mast cells and that mast cell colony-forming cells (progenitors) could be detected in clonal cultures of adult marrow, neonatal marrow, or fetal liver if the combined stimulus of stem cell factor and interleukin-3 was used. Mast cell progenitors were in active cell cycle and showed an extensive capacity for self-generation. Mast cell colonies both from control GATA-1(+/+) mice and GATA-1(Plt13/+) mice could generate growth factor-dependent cloned cell lines that grew for >18 months. Surprisingly, the majority of the excessive numbers of mast cell progenitors in chimeric GATA-1(Plt13/+) mice were transcribing the inactive Plt13 allele of GATA-1, suggesting that GATA-1 normally acts to restrict the emergence of committed mast cell progenitors. In sharp contrast, all eosinophil progenitors in these mice were transcribing the normal GATA-1 allele. No excess tissue mast cells were observed in GATA-1(Plt13/+) mice, suggesting that the excess mast cell progenitors in these mice might be generating mast cells with a defective in vivo proliferative or tissue homing capacity.