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     

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。     

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     

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)     

T cells regulate the IgM antibody response of BALB/c mice to dextran B1355.

The IgM antibody response of BALB/c mice to bacterial (Leuconostoc) dextran B1355 is influenced in a positive and negative manner by regulatory CD4+ and CD8+ T cells, respectively. Treatment with concanavalin A (ConA) at the time of immunization or 2 days later caused suppression and enhancement of the antibody response, respectively. Priming of mice with a sub-immunogenic dose of dextran resulted in profound suppression upon subsequent immunization 3 days later. None of these effects were demonstrable in athymic mice. Transfer of T cells from mice primed 18 h previously with a subimmunogenic dose of dextran suppressed the antibody response in immunized recipients; such suppression was abolished by the treatment of transferred cells with anti Thy 1.2 or anti Lyt 2.2 (CD8) antibody in the presence of complement. By contrast, the transfer of T cells from mice, which had been given an immunogenic dose of dextran 4 days previously, increased the antibody response in immunized recipients; such enhancement was abolished by treating transferred cells with anti Thy 1.2 or anti L3T4 (CD4) antibody in the presence of complement. These findings indicate that the immune response to dextran B1355 is regulated by CD4+ T-amplifier cells (Ta cells) and by CD8+ T-suppressor cells (Ts cells) which are activated during the course of a normal antibody response.     

Regulation of an anti-polysaccharide immune response in BALB/c mice through a tight T and B lymphocyte idiotypic connection

The isotype expression in the J558 idiotype-associated humoral immune response against α(1 → 3)-dextran in BALB/c mice is controlled by idiotype-specific T cells which silence in situ B lymphocytes primed and committed to an IgG response. This leads to a restriction of the type II thymus-independent response to the sole production of IgM antibodies. The availability of the T cell receptor (TcR) α and β sequences for such a regulatory T cell clone allows the investigation of the degree of heterogeneity of the TcR usage of these T cells. It is found that all α(1 → 3)-dextran-primed BALB/c mice use a very similar, possibly identical TcR. This suggests a tight, possibly genetically programmed, interaction between the J558 idiotype-bearing dextran-specific B cells and their idiotype-specific regulatory T cell counterparts.     

Maternal immunization modulates the primary immune response to 2-phenyl-oxazolone in BALB/c mice

The development of the antibody repertoire in newborn mice is greatly influenced by idiotypic network interactions. It has been demonstrated that anti-idiotypic antibodies either directly injected or transferred from the mother may alter the repertoire for life. For an elucidation of the underlying mechanisms we have analyzed the primary immune response to 2-phenyl-5-oxazolone (phOx) coupled to chicken serum albumin (CSA) in BALB/c mice after complete disappearance of maternal antibodies which originated from different stages of affinity maturation. Depending on the serum titers of the mothers after primary (1° mo), secondary (2° mo) or tertiary (3° mo) immunization, maternal anti-phOx IgG persisted in F1 mice for up to 9 months. In addition, F1 mice born to 2° mo developed – even without immunization – an anti-phOx IgM titer which reached levels similar to an antigen-induced primary response. An enhancement of the early primary anti-phOx as well as anti-CSA response was seen in F1 mice born from 1° mo, whereas the response was delayed when born to 2° mo and 3° mo. The antibody titers in the latter group of mice remained at a lower level for 3 months. In contrast, mice of the F2 generation which received a smaller amount of the same collection of maternal antibodies as F1 mice from 3° mo exhibited a quite different primary response: (i) They showed an earlier onset in their anti-CSA response. (ii) Whereas normally a plateau in antibody titer was reached by the 4th weak after immunization, in 55 % of the F2 mice a prolonged increase of the anti-phOx and anti-CSA antibody titers was observed. At 12 weeks after antigenic challenge, titers reached plateau levels of 6 × 10⁵ which were never before seen in a primary phOx or CSA response. Thus, depending on its own immunological experience, the maternal immune system induces a state of memory in the offspring which results in a faster and/or enhanced immune response in the F1 and F3 generations.     

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。     

The impact of aluminium in acid-suppressing drugs on the immune response of BALB/c mice

BACKGROUND: Recently we have shown that anti-acid drugs lead to an enhanced risk of food allergy. This may be due to hindered peptic digestion, caused by an elevation of the gastric pH. Additionally, it is known that aluminium-linked antigens lead to an increased probability of sensitization. OBJECTIVE: Our aim in this study was to show whether sucralfate promotes sensitization not only by preventing peptic digestion but also by acting as a T-helper type 2 (Th2) adjuvant. METHODS: To avoid the effect of sucralfate on the gastric pH and to show only the adjuvant effect, BALB/c mice were immunized on the parenteral route with codfish extract plus sucralfate, and control groups with aluminium hydroxide (alum) (Th2 adjuvant) or monophosphoryl lipid A (MPL) (Th1 adjuvant). Antigen-specific antibodies and cytokine levels were determined. The in vivo effect was investigated by intradermal skin tests. RESULTS: Codfish-specific high IgG1 and IgE antibody levels as well as elevated IL-4 and IL-5 levels in alum- and MPL-treated mice, but more importantly also in sucralfate-treated mice, indicated a Th2 shift. Positive skin tests confirmed this Th2 response. CONCLUSIONS: Our data show that parenterally applied sucralfate is able to induce a Th2 response probably due to the aluminium content. This indicates that orally applied sucralfate may lead to an enhanced risk of food allergy not only by inhibiting peptic digestion but also by acting as a Th2 adjuvant.     

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.     

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.     

The early immune response in the liver of BALB/c mice infected with S. typhimurium

Gram-negative bacteria acquired through gastrointestinal infection can be a serious cause for the development of septic shock especially in immunosuppressed patients. Thus, the aim of this study was to examine the early events of the immune reaction against S. typhimurium. Bacteria were injected into mice at different concentrations. Four animals from each group were killed at five different points of time. Liver cytokine mRNA expression was determined by semiquantitative rt-PCR and liver histology was examined. Serum cytokine levels of interferon (IFN)-gamma, tumor necrosis factor (TNF)-alpha, interleukin (IL)-6, IL-4 and IL-10 were determined. intravenous (i.v.) infection with 109 bacteria led to lethal septic shock within 24 h. A delayed production of IFN-gamma, TNF-alpha, IL-18 and IL-10 and milder histological alterations in the liver were observed in these animals. The highest expression of cytokines in the liver and the strongest histological alterations were seen after infection with 107 bacteria. Here, an increased mRNA expression of all proinflammatory cytokines began 1 h after infection. Animals infected with 1 x 102 bacteria had the highest detectable serum levels of IL-6 and IL-10. These data indicate that the immediate events in the immune reaction within the liver after infection with S. typhimurium are associated with the outcome of the subsequent sepsis.     

Interleukin-13 induces T helper type 2 immune responses in OVA-immunized BALB/c mice bearing a T cell lymphoma

T lymphocytes play a crucial role in the regulation of immune responses against the tumour cells. Tumour progression results in dysfunction and inhibition of T cells, which ultimately leads to impairment in the antitumour immune response. The impaired antitumour immune response in the host is represented by the decreased number of T cells and their incomplete and improper function. The immunosuppressive network in tumour-bearing host mediated by tumour cells also leads to the inequities of T cell subsets and imbalance of Th1/Th2 dichotomy. Therefore, in the present study, we sought to investigate the role of tumour progression in the development of T cell phenotype and the involvement of interleukin-13 thereof selecting Dalton's lymphoma (DL) as a tumour model. It was observed that a significant increase in the number of CD4(+) T cell population, whereas a significant decline in the CD8(+) T cells among lymphoid cell population of OVA-immunized DL-bearing BALB/c mice occurs. Similar observation was found following the administration of IL-13 to the normal healthy mice. It was further confirmed that expansion in Th2 type cells among CD4(+) T cell population occurs following the progression of tumour and administration of IL-13 to normal healthy mice by an yet to define mechanism. Therefore, it can be concluded that IL-13 has immense role in polarizing the immune responses by inducing the differentiation of Th2 type of cells.     

Immunogenicity of the antigenic determinants of influenza virus type B hemagglutinin

The immunogenic activity of antigenic determinants of influenza type B viruses in primary and booster immunizations was studied. In primary immunization, the highest immunogenicity was found with the type- and strain-specific antigenic determinants whereas the antigenic determinants of group specificity were less immunogenic. After booster immunizations the animal sera had antibody to all 3 antigenic determinants.     

IL-2 preS DNA疫苗免疫正常小鼠和HBV转基因鼠的免疫应答研究

目的 探讨IL—2 preS DNA疫苗作为预防和治疗性疫苗的可行性及作用机理。方法 应用基因重组技术，构建人白细胞介素2(hIL-2)和前表面抗原(preS)的真核表达载体，将此重组载体用基因枪分别注射正常的BALB／c小鼠和HBV转基因小鼠，通过ELISA方法检测BALB／c小鼠和HBV转基因鼠的抗-preS2、HBsAg及抗-HBs抗体水平；荧光定量PCR方法检测HBV转基因鼠血清中HBV DNA拷贝数，并用免疫病理HE染色观察小鼠肝组织炎症活动度，同时检测肝功能指标。结果 ①基因枪注射真核表达质粒免疫正常小鼠后，100％小鼠能在第4、6周检测到抗preS1抗体，持续时间长达10周。②用IL-2 preS真核表达质粒基因枪肌肉注射方式优于正常肌肉注射和皮下注射的方式，且所需质粒量(10μg／只)仅为后者(100μg／只)的1／10。③在第4周高峰期检测IgG亚类，是诱导以TH1(IgG2a)细胞免疫为主的反应。④基因枪注射真核表达质粒(1μg／只)免疫转基因小鼠后，80％的小鼠产生了抗体，HBV DNA量下降，其中20％的小鼠HBsAg转阴。⑤HE肝组织染色显示：肝组织有明显的炎细胞浸润、肝细胞肿大、颗粒样变性、转氨酶升高。结论 IL-2 preS DNA疫苗能刺激小鼠机体产生体液和细胞免疫，可部分打破小鼠机体的免疫耐受，为新型乙肝疫苗和抗HBV持续性感染的特异性免疫治疗剂的设计和构建提供理论与实践依据。     

The immune response of BALB/c mice to phosphorylcholine is restricted to a limited number of VH- and VL-isotypes

To study the diversity of the anti-phosphorylcholine (PC)§ repertoire in BALB/c mice, hybridomas were produced by cell fusion of immune spleen cells of normal or T 15 idiotypically suppressed mice with a myeloma cell line. Idiotypic suppression was achieved by injection of anti-PCBMP T 15 idiotypic serum into newborn mice. Thirteen homogeneous PC-specific hybridoma-derived antibodies of classes IgM and IgG were isolated and analysed by partial N-terminal amino acid sequence determination of the variable (V) regions of their heavy (H) and light (L) chains. Eleven hybridoma antibodies displayed V_H-regions of isotype V_H−4 like T 15 and all other PCBMPs, whereas two belonged to a new V_Hisotype, designated V_H−12, not previously encountered in any PCBMP or mouse V_H-region known so far. The sequences of the variable regions of the L-chains expressed the V _K−8-, V_K−22- and V_K−24-isotypes described in PCBMPs M 603, M 167 and T 15 respectively. One hybridoma antibody expressed a L-chain of the V_K−15 -isotype, which probably originated from the immunoglobulin of the parental myeloma cell line used for cell fusion. The data suggest that a limited number of V_H- and V_L-isotypes are being used to generate the PC-specific repertoire in BALB/c mice and that the majority of the hybridoma antibodies contain V_H−V_L association pairs known from PCBMPs.     

BALB/c小鼠急、慢性弓形虫感染时Th细胞应答的基因表达谱分析

目的:分析弓形虫感染BALB/c小鼠后诱导Th细胞应答的基因表达谱特征.方法:以刚地弓形虫速殖子感染BALB/c小鼠,分别于感染后第6天和第12天各处死小鼠2只,制备小鼠脾细胞悬液并混合,提取细胞总RNA.采用基因芯片技术检测小鼠Th细胞应答各相关基因的表达情况,并对部分表达变化的基因应用Real time-PCR予以验证.结果:BALB/c小鼠感染弓形虫后,基因芯片检测共有20种基因表达差异超出2倍以上.感染后第12天比第6天,8种基因(Gata-3、Ccr3、Ccr4、Bcl6、Nfatc1、CD80、Fos、CD69)表达上调,12种基因(T-bet、CIITA、Irf1、Mapk9、Nfatc3、Fasl、Tyk2、Lat、Mapk10、Socs3、Socs6、Yy1)表达下调.总体呈现Th2型免疫应答基因上调,Th1型免疫应答基因呈下调趋势.Gata-3和T-bet基因的Real time-PCR检测结果与基因芯片检测一致.结论:从基因表达谱分析来看,BALB/c小鼠感染弓形虫后在感染急性期以Th1细胞应答为主,度过急性期后以Th2细胞应答为主.     

BALB/c小鼠滴鼻免疫灭活SARS病毒对全身性免疫的影响

目的　了解灭活SARS病毒滴鼻免疫小鼠对全身性免疫的影响。方法　将 5周龄雌性BALB c小鼠随机分为两组 ,每组 10只 ,免疫组小鼠每只免疫 5 0 μg的灭活病毒悬液 ,对照组注射等体积PBS。分别在免疫的 8d和 14d处死小鼠 ,分离脾细胞和外周血淋巴细胞 ,流式细胞仪观察其淋巴细胞表型 ;间接ELISA法测定其血清中抗SARS病毒特异性抗体IgM和IgG。结果　第 8天脾脏的CD4 T细胞的构成比增加 (P =0 .0 2 7)。外周血淋巴细胞表型在第 8天即开始发生改变 ,在第 14天变化明显 ,与对照组差异有显著性 ,CD4 ,CD8 ,CD3 T细胞构成比和T B比值显著降低 (P <0 .0 5、0 .0 1、0 .0 1、0 .0 1) ,同时CD4 5R B2 2 0 细胞 (B细胞 )构成比显著增加 (P =0 .0 0 0 )。但是此时血清中特异性抗体未能被检测到。结论　在未应用佐剂的情况下 ,小鼠滴鼻接种灭活SARS病毒虽然可以导致诱导机体免疫细胞表型的变化 ,却未能诱导抗体的产生。