A recombinant BCG vaccine generates a Th1-like response and inhibits IgE synthesis in BALB/c mice

The tubercle vaccine, bacille Calmette-Guérin (BCG), is a strong inducer of T-helper type 1 (Th1) responsiveness, and it has been suggested that recombinant BCG (rBCG), which produces and secretes antigens, may be used to prevent allergic diseases. The effects of rBCG vaccination on allergic responses in a murine model were examined in this study. A BCG-Escherichia coli shuttle vector was developed with the promoter and signal sequence of the alpha-antigen of Mycobacterium bovis, and the vector was tested using E. coli beta-galactosidase as the model antigen and allergen. This vector enabled the expression of the E. coli beta-galactosidase gene in BCG, which was detected in its protein extract by immunoblotting analysis. Vaccination of mice with a single dose of 106 recombinant BCG generated a beta-galactosidase-specific antibody response. The splenocytes of vaccinated mice compared with controls produced significantly higher amounts of interferon-gamma (IFN-gamma) (P<0. 01) and interleukin-2 (IL-2) (P<0.05) and lower amounts of IL-5 (P<0. 01). Mice vaccinated with rBCG had significantly less (P<0.01) serum IgE compared with controls. These results together demonstrate that rBCG secreting antigens or allergens may be utilized for the induction of a Th1-like response and the down-regulation of IgE antibody response.     

Effect of Angiostrongylus costaricensis extract on eosinophilic pulmonary response in BALB/c mice

Epidemiological and experimental studies have demonstrated an association between parasitic infections and the allergic diseases. A protective effect in asthma was shown in animals infected with helminths. The aim of this study was to determine the effect of Angiostrongylus costaricensis extract on inflammatory lung response to ovalbumin (OVA) in mice. Four BALB/c mice received A. costaricensis extract by intraperitoneal (i.p.) injection on the first day. Mice were immunised against OVA by i.p. injection on day (D) 5 and D12 and received a daily intranasal OVA challenge (40 μl) between the D19 and D21. On D23, we performed a bronchoalveolar lavage (BAL) on the mice. Four BALB/c mice (control group) were immunised against OVA using the same protocol, but did not receive parasite extract. Total cell counts (TCC) and differential cell counts were performed in BAL fluid samples. Eosinophil cell counts in BAL fluid were lower in the group that received A. costaricensis extract when compared with the control group (0.04×10⁶ cells/ml and 0.01×10⁶ cells/ml, respectively; p=0.04). TCC were not different between the groups studied. A. costaricensis extract in mice decreases eosinophilic response to OVA in BAL fluid.     

A Kudoa septempunctata antigen induces production of IgE in BALB/c mice

Parasitology Research - Kudoa septempunctata, a myxosporean parasite, is the causative agent of a foodborne illness associated with consumption of raw Paralichthys olivaceus (olive flounder)....     

Induction of anti-DNA IgG and IgE antibodies in BALB/c mice.

Immunization of BALB/c mice with denatured DNA (dnDNA)-methylated bovine serum albumin (MBSA) complex along with aluminium hydroxide gel as adjuvant, resulted in the induction of anti-DNA antibodies of both IgG and IgE isotypes demonstrable by avidin-biotin micro enzyme-linked immunosorbent assay (ELISA) and solid phase radioimmunoassay (SPRIA), respectively. In contrast to the high levels of IgG2a and IgG2b anti-DNA antibodies observed in SLE-prone autoimmune mice, more than 90% of the anti-DNA antibodies of IgG isotype were found to be of IgG1 subclass. Specificity of both IgG and IgE antibodies which recognized activated DNA, dnDNA and double-stranded DNA but not RNA was established by competitive ELISA and SPRIA inhibition assays. These antibodies cross-reacted with cibacron blue and chondroitin sulfate but not with various other proteoglycans, nucleosides and nucleotides. Passive cutaneous anaphylaxis reaction in rats showed that these antibodies are capable of inducing in vivo degranulation of mast cells in a dose-dependent manner. These studies lend support to the concept that IgE antibodies directed against DNA may mediate mast cell degranulation and thus contribute to immediate-type hypersensitivity phenomena including hives seen in patients with systemic lupus erythematosus and to the localization of IgE-nucleic acid complexes.     

Individual variations of the immune response in BALB/c mice

The nature and the causes of variations of the immune response to thyroid hormones are analyzed in BALB/c mice. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 1, pp. 80–82, January, 1995 (Presented by Yu. A. Romanov, Member of the Russian Academy of Medical Sciences)     

A malaria 'mitogen'-induced depressed immune response to meningococcal polysaccharide vaccine in BALB/c mice

Uninfected female BALB/c mice were given a 4-daily intraperitoneal injection, of supernatants obtained from 24-h cultures of Plasmodium berghei infected and control mouse red blood cells, for 20 days. Each mouse was subsequently injected subcutaneously with 10 mg meningococcal (groups A and C combined) polysaccharide vaccine. Mean meningococcal haemagglutinating antibody titres obtained in mice pretreated with control supernatants were consistently higher, than those obtained in mice pretreated with supernatants from malaria-infected red blood cell cultures, over a period of 14 days. The results suggest that a malaria 'mitogen' may be involved in the pathogenesis of the immunosuppression characteristic of this infection.     

Suppression of phosphorylcholine-specific IgE antibody formation in BALB/c mice by isologous anti-T 15 antiserum

In order to study the regulation of IgE antibody formation, isologous anti-idiotypic antisera against the phosphoryl choline (PC)-specific BALB/c myeloma proteins T 15 and M 167 were passively administered to BALB/c in the course of an anti-PC IgE response. Isologous anti-T 15 antiserum had a long-lasting suppressive effect on the formation of IgE antibodies with PC specificity, whereas administration of anti-M 167 antiserum had no or only little effect, similar to that of normal BALB/c serum. This indicates that anti-PC IgE antibodies consist mainly of the T 15 idiotype or of cross-reacting idiotypes, and that IgE response is accessible to regulation with anti-idiotypic antibodies. This murine model may permit the study of regulation of an IgE response largely restricted to few defined idiotypes characterized as tumor proteins.     

Association of matrix metalloproteinase-9 in eosinophilic meningitis of BALB/c mice caused by Angiostrongylus cantonensis

Induction of gelatinase in eosinophilic meningitis of BALB/c-strain mice was caused by Angiostrongylus cantonensis. Time-course studies showed that the molecular weight of 94-kDa gelatinase was detected at day 10 post-inoculation (PI), and reached a high intensity from days 15 to 25 PI. The 94-kDa gelatinase activity was clearly inhibited by EDTA and 1,10-phenanthroline, but not by leupeptin and phenylmethanesulphonyl fluoride. When immunoblots were performed using specific antiserums against the 94-kDa gelatinase B (matrix metalloproteinase-9; MMP-9) with cerebrospinal fluid (CSF), the 94-kDa immunopositive band was MMP-9. Immunohistochemistry studies demonstrated MMP-9 localisation within eosinophils and macrophages. The increased MMP-9 activity was closely associated with the rapid rise of CSF eosinophils, and the inflammatory reaction of the subarachnoid space. In contrast to changes in MMP-9, MMP-2 activity was constitutive and unaffected in this parasitic meningitis. These results show that MMP-9 was associated with eosinophilic meningitis, and that the enzyme may be a useful marker for angiostrongyliasis meningitis.     

卵清白蛋白特异性T细胞免疫诱导BALB/c小鼠的调节性免疫应答

目的：研究T细胞免疫后正常小鼠的调节性免疫应答，方法：应用体外扩增的卵清白蛋白（OVA）特异的T细胞克隆免疫BALB／c小鼠，3H－TdR掺入法分析细胞增殖，3H－TdR标记靶细胞检测杀伤T细胞的杀伤效应，间接免疫荧光法分析血清中抗T细胞抗体水平。结果：T细胞免疫后能诱导BALB/c小鼠产生调节性T细胞的增殖反应，对靶细胞的杀伤效应以及针对于活化的T细胞的体液免疫应答，并进一步降低机体对OVA抗原的应答，结论：T细胞免疫能诱导正常机体的调节性免疫应答。     

T细胞免疫BALB/c小鼠诱导调节性体液免疫应答的研究

目的 :研究T细胞免疫后正常小鼠的调节性体液免疫应答。方法 :用经γ射线照射的体外扩增的活化OVA特异T细胞克隆免疫BALB c小鼠 ,FACS法分析血清中抗T细胞抗体水平 ,免疫共沉淀法分析靶抗原。结果 :T细胞免疫能诱导BALB c小鼠产生抑制T细胞增殖的抗T细胞抗体 ,这种抗体不是多克隆活化的结果 ,其靶抗原可能是T细胞上 93、87、5 5和 4 5kD的蛋白。结论 :T细胞免疫能诱导正常小鼠产生调节性体液免疫应答     

Echinococcus multilocularis: immunity response to purified alkaline phosphatase in BALB/c mice

The study of purified alkaline phosphatase and crude extract antigen immunogenicity from Echinococcus multilocularis was carried out on BALB/c mice. The animals were immunized, then infected with E. multilocularis metacestode. The immune response against purified alkaline phosphatase was studied. Flow cytometry analysis of the CD4+ and CD8+ lymphocyte populations showed a predominance of CD4+ populations in infected immunized mice. The specific humoral response to purified alkaline phosphatase was analyzed by enzyme-linked immunosorbent assay method. We noted a stimulation of an immunoglobulin IgG response. The isotypic profile showed a prevalence of IgG1 and IgG3 in immunized infected mice compared to IgG2a and IgG2b. In addition, analysis of the profiles of the in vitro secreted cytokines, after stimulation of the splenocytes from immunized mice, was performed. The cytokine profile was a mix of Th1/Th2 types in the infected and uninfected immunized mice. The results of this study suggest a humoral mixed Th1/Th2 response, with a high predominance of Th2 response. A similar study was conducted in mice immunized with crude total antigen. The comparison of the immune response showed an important immune response in mice immunized with purified alkaline phosphatase compared to mice immunized with the crude total antigen.     

Effect of cocaine on the immune response and host resistance in BALB/c mice

This study focuses on the effect of varying regimens of cocaine administration on three parameters of the immune response: antibody production, resistance to infection by Streptococcus pneumoniae following immunization, and resistance to tumors. The effect of cocaine on antibody production of female and male BALB/c mice was investigated to both a T-independent (pneumococcal polysaccharide type III [SSS-III]) and a T-dependent antigen (the 2,4-dinitrophenyl ligand [DNP]). It was found that high doses of cocaine injected 3 times/day prior to SSS-III resulted in a small rise in antibody levels in male mice. Low doses given for 4 days prior to or subsequent to SSS-III injection had no effect on the antibody response nor on the susceptibility to infection by live S. pneumoniae. High dosages of cocaine administered 3-5 times/day had no effect on the anti-DNP immune response of male mice but resulted in an almost 2-fold increase of anti-DNP plaque-forming cells in female mice.     

幽门螺杆菌感染小鼠及其抗原免疫小鼠后体液免疫应答的比较

目的：比较BALB／c小鼠感染H.pylori后以及经H.pylori UreB抗原口服免疫后的体液免疫应答的差异。方法：80只BALB／c小鼠分为感染组和免疫组，感染组灌喂H.pylori小鼠适应株；免疫组灌喂重组H.pylori UreB抗原和佐剂LTB。在试验的0，2，6和10周时每组分别取10只小鼠收集唾液及血液标本，用ELISA法检测抗UreBIgG和IgA抗体，同时采集胃组织作H.pylori感染检测。结果：感染组小鼠在灌喂H.pylori后2，6，10周，感染率均为100％，其血清中抗UreB IgG增高明显，但血清及唾液中均未见IgA的明显升高，；免疫组小鼠在初次免疫后第6，10周后，血清及唾液中抗UreB,IgG和IgA抗体的均显著升高。结论：H.yplori感染BALB／ c小鼠不能诱导其产生明显的特异性黏膜免疫应答，而重组UreB可作为良好的免疫原诱导其产生分泌型IgA抗体。     

动物源Ⅰ型胶原蛋白可引起BALB/c小鼠细胞免疫反应和组织免疫毒性

背景：天然纯胶原被认为具有较低的免疫原性和较好的生物相容性。目的：体外评估动物源Ⅰ型胶原蛋白的免疫原性。方法：将牛跟腱经免疫原性清除后,提取制成Ⅰ型胶原蛋白,HPLC测定胶原蛋白纯度,荧光染色定量胶原蛋白中残留DNA。将50只BALB/c小鼠随机均分为5组,分别皮下注射生理盐水(阴性对照)、小牛来源Ⅰ型胶原蛋白标准品(阳性对照)、33.4,66.8,133.4 mg/kg的动物源Ⅰ型胶原蛋白,1次/d,连续注射12 d后,检测注射胶原蛋白后小鼠淋巴细胞的增殖、细胞分型及NK细胞杀伤功能;连续注射3周后,取脾脏、肝脏及肺组织进行病理组织学检查。结果与结论：相比Ⅰ型胶原蛋白标准品,纯化后的牛源性Ⅰ型胶原蛋白纯度可达到99%以上,而残留DNA低于1 mg/L,远远低于目前常规脱细胞基质中DNA的残留水平50-100 µg/g(干质量)。注射12 d后,各组均未发生淋巴细胞增殖、NK细胞杀伤功能及淋巴细胞亚群的比例的变化。注射3周后,66.8,133.4 mg/kg动物源胶原蛋白组小鼠脾小动脉周围淋巴鞘面积明显增厚变大,有可能引发偶发性的肝损伤和肺损伤,而脾小体生发中心面积未发生明显变化。表明连续皮下注射动物源胶原蛋白可引发BALB/c小鼠发生较低水平的脾淋巴细胞免疫应答,可能引起偶发性肝、肺损伤。中国组织工程研究杂志出版内容重点：组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松;组织工程     

创伤弧菌感染BALB/c小鼠诱导适应性免疫应答的研究

为探讨创伤弧菌感染BALB/c小鼠后引起的适应性免疫应答类型及规律,我们首先通过半数致死量实验筛选创伤弧菌MO6-24/O腹腔注射感染菌量,随后通过流式细胞术检测创伤弧菌感染后小鼠脾脏Th细胞亚群应答情况,通过ELISA实验检测血清中IFN-γ浓度,采用real-time PCR检测创伤弧菌感染小鼠腹腔巨噬细胞后Th1细胞调控因子的表达。最后,采用流式细胞术动态检测Th1细胞亚群应答规律。结果显示,创伤弧菌MO6-24/O腹腔注射感染小鼠的半数致死量是2.5×105cfu,平均存活时间是16h,我们后续实验选择感染菌量为5×104cfu/只。创伤弧菌感染后能够诱导Th1型细胞应答,并且感染后血清中IFN-γ浓度显著增加(P0.05),并且创伤弧菌体外感染小鼠腹腔巨噬细胞后能够引起IL-12/IL-23p40、IL-12p35以及IFN-γmRNA表达显著增高(P0.05)。最后,流式结果显示,创伤弧菌感染后,脾脏Th1细胞的应答逐渐增强,在感染后3天达到高峰,随后逐渐下降到正常水平。因此,Th1型应答是创伤弧菌诱导的获得性免疫应答的主要类型。     

Immunomodulatory activity of diethylcarbamazine on humoral, cellular cytokine response and respiratory burst in BALB/c mice

Diethylcarbamazine (DEC) is an anthelmintic piperazine derivative drug with putative immunomodulating properties, including increased platelet and granulocyte adhesion to parasites and enhanced production of cytokines. To further analyse these properties in a well-established animal model, we evaluated the effect of DEC on antibody, cellular cytokine response and respiratory burst in BALB/c mice. Animals were challenged with a thymus-dependent (tetanus toxoid, (TT)) and with a thymus-independent (lipopolysaccharide, (LPS)) antigen and treated with DEC for seven days with two different doses (50?mg/day and 500?mg/day). Serum was assessed for antibody production at 0, 4, 7, 14, 21 and 28 days after stimulation and at 0, 24 and 48?h for IL-2, IFN-γ, IL-10 and IL-12 release. Respiratory burst of neutrophils and monocytes from peripheral blood was measured by flow cytometry. We found low-dose treatment with DEC enhanced cytokine production vs. TT and antibody production vs. LPS, whereas a higher dose enhanced significantly the respiratory burst of both polymorphonuclear leukocytes and monocytes, with a significant higher effect on the former. Our results suggest a stimulating, dose-dependent immunomodulatory effect of DEC with a higher effect on the phagocytic cells.     

A comparative analysis of radiation- and virus-induced leukemias in BALB/c mice

Endogenous murine leukemia virus (MuLV) proviral copies were analyzed in thymomas induced in normal BALB/c (Fv-1b) and in Fv-1n congenic mice by X-irradiation. Both strains of mice developed leukemia with similar kinetics, indicating that N-tropism of endogenous MuLV was not a rate-limiting factor in development of disease. Southern blot analysis, using a probe specific for ecotropic virus and for ecotropic-specific sequences retained in pathogenic, env-recombinant viruses, showed that the majority of radiation leukemias lacked newly acquired, clonally integrated, proviruses. This was in contrast to virus-induced leukemias, which routinely exhibited several new proviral integration sites. When an internal proviral DNA restriction fragment was monitored, some radiation leukemias showed evidence of nonclonal infection, accounting for more frequent isolation of infectious virus from such leukemias. Differences in expression of T-cell surface antigens were found in X-ray-induced and virus-induced leukemias. All radiation leukemias were TL positive, whereas virus-induced leukemias were primarily negative for TL. Some differences were also found in Lyt-1 and Lyt-2 expression. The data as a whole suggest that, in the majority of cases, radiation leukemogenesis is not initiated by a viral route--that is, the sort of viral mechanism for which exogenous infection by known pathogenic MuLV is the paradigm.     

Neonatal stimulation, maternal behavior, and accelerated maturation in BALB/c mice

An experiment was conducted to examine the effects of neonatal handling and hypothermia on infant physical maturation and growth and on maternal behavior in BALB/c mice. Stringent methodological and statistical controls were employed for experimenter bias and litter effects. No evidence appeared to support claims that neonatal stimulation leads to accelerated physical maturation. Alterations in maternal behavior patterns were found only immediately following hypothermia of pups. Maternal behavior did not correlate highly with offspring maturational rates, but maternal weight and age and pup birth weight were highly predictive of infant physical maturation.