猪蛔虫感染对小鼠肺部免疫病理反应的动态观察

目的:动态观察感染猪蛔虫后,小鼠肺组织的病理变化及肺泡灌洗液中相关细胞因子的变化,从而了解蛔虫感染对肺脏的侵害过程。方法:温箱孵育猪蛔虫受精卵至含蚴卵,用灌胃法感染小鼠,分别在感染后第0、7、14、45天处死小鼠,观察肺组织的病理改变,并通过ELISA方法检测肺泡灌洗液（bronchoalveolar lavage fluid,BALF）中IL-4、IFN-γ、IL-10和TGF-β₁含量的动态变化。结果:感染14天组,肺组织的病理改变最为严重:大量炎症细胞浸润,嗜酸性粒细胞增多,严重者支气管狭窄,肺泡塌陷;感染45天组,小鼠肺组织病理变化比前者显著减轻。BALF中IL-4含量在感染7天、14天后分别为（169.20±34.61pg/ml）,（381.33±57.39pg/ml）,均明显高于未感染组（38.03±6.09 pg/ml）;在感染45天后降低（98.49±25.33 pg/ml）,但仍高于未感染组。IL-10水平在感染后降低;但感染45天组,IL-10水平却有所增高（179.78±21.33 pg/ml）,并高于感染前。IFN-γ、TGF-β₁含量在感染后也有明显降低。结论:猪蛔虫感染初期,小鼠肺部损伤严重,小鼠BALF中促炎症细胞因子占主导地位;但在感染后期,BALF中促炎症细胞因子含量降低,炎症抑制因子IL-10有所增高,炎症缓和。     

猪蛔虫有三管型雌性生殖系统的解剖记录

1999年 1 0月 2 5日在学生解剖实验中 ,发现一例具有三管型雌性生殖系统的猪蛔虫 (Ａｓｃａｒｉｓｓｕ ｕｍ)。该猪蛔虫采自南充市冷冻厂 ,全长 1 82ｍｍ。外观发育良好 ,与一般雌性猪蛔虫无明显差别 ,体前端1 / 3处环状狭窄部分和位于该处腹面阴门 (雌性生殖孔 )均无异常。解剖发现它具有三管雌性生殖系统 ,其中两条直径分别为 1 0 8ｍｍ和 1 3 2ｍｍ的子宫汇合成一条短的阴道 ,并由阴门通往体外 ,第三条子宫在阴道中后部发出 ,其最大直径为 1 0 0ｍｍ。两子宫汇合成的阴道总长 6 0 4ｍｍ ,阴道中后部发出的第三条子宫至阴门距离为 3 2…     

登革病毒感染的免疫与免疫病理的研究进展

登革病毒感染是世界上热带与亚热带地区一个严重的公众卫生健康问题,本文综述了登革病毒感染后机体免疫的改变,并从抗体、Ｔ细胞激活及细胞因子过量释放方面探讨了登革出血热（ＤＨＦ）／登革休克综合征（ＤＳＳ）的发病机理。     

异型流感病毒感染小鼠肺细胞因子水平变化

为了制备能够抵御不同型别流感病毒感染的疫苗,揭示机体对异型流感病毒感染交叉免疫保护作用的主要机制,用流感病毒疫苗免疫小鼠后分别感染同型、异型流感病毒,另设使用免疫增强剂IL-2后感染异型流感病毒组,观察小鼠的一般状况和肺指数,并用ELISA方法测定肺匀浆中细胞因子IFN-γ、IL-2、IL-4及IL-10的含量。结果显示,异型免疫组和异型免疫加强组病毒感染后细胞因子IFN-γ含量明显高于感染前（P〈0.05）。研究表明,异型病毒感染后IFN-γ水平明显增高,此细胞因子可能在流感病毒异型间交叉保护免疫反应中起重要作用,其机制有待进一步研究确定。     

菌群失调小鼠感染鼠伤寒沙门菌后Th1和Th2细胞因子的动态研究

目的研究鼠伤寒沙门菌感染小鼠在菌群失调下Th细胞因子的动态变化,以探讨菌群失调对沙门菌感染的免疫机制。方法分别建立菌群失调、感染和空白对照的小鼠模型。各组动物在感染不同时点处死,观察小肠、肝脏和脾脏病理改变。采用流式细胞仪检测脾脏细胞中IFN-γ和IL-4表达,以此代表Th1和Th2细胞。结果菌群失调组病理损害最重,Th1明显增加,表达水平高,Th2变化不大,Th1/Th2比值升高。结论菌群失调后,能够加重小鼠感染鼠伤寒沙门菌引起的的Th1型反应,产生炎症性损伤。     

小鼠Ⅲ度烧伤模型的复制及皮肤病理形态改变的动态观察

目的利用近交系小鼠建立方法简便、效果稳定的Ⅲ度烧伤模型,对其皮肤病理改变进行动态观察,为烧伤实验治疗研究提供资料。方法小鼠实验前备毛,乙醚麻醉固定小鼠四肢,苏醒后于备毛部覆盖自制控制烧伤面积硬纸片,滴加95%酒精,点燃计时;烧伤早期和远期取皮肤进行病理检查,用Ki67抗体免疫组化观察细胞增殖。结果按本方法建立小鼠Ⅲ度烧伤模型,面积准确,深度一致,操作简便易行。小鼠烧伤皮肤病理改变与人类基本一致,但有其特点：小鼠Ⅲ度烧伤包括表皮至脂膜肌,烧伤后炎性渗出较晚,约需2 d;创面周围毛发生长活跃,倾向创面,Ki67抗体阳性细胞集中于毛囊底部。结论为利用小鼠烧伤模型进行实验治疗的研究提供了有实用价值的资料。     

光,电镜观察小鼠呼吸道合胞病毒性肺炎的免疫病理改变

Ｂａｌｂ／ｃ小鼠经鼻吸入呼吸道合胞病毒（ＲＳＶ）悬液感染成ＲＳＶ肺炎。于感染第５天后连续隔日取肺,光镜与透射电镜检查。感染第５～７天,肺组织病理改变最严重,多数小鼠表现为间质淋巴细胞（ＬＣ）套状浸润,肺泡隔增宽;少数小鼠出现间质内大量ＬＣ浸润与肺泡内大量单个核细胞渗出的两种病理改变。病毒包涵体出现于肺泡上皮细胞内,细胞受感染后发生肿胀、坏死。Ⅰ型细胞核周胞质内有核衣壳复制,表面病毒芽生形成长短不等的丝状体。第９天,肺泡隔增宽与间质ＬＣ浸润逐渐减轻。第１２天,病毒包涵体明显减少。     

益生菌对感染小鼠早期IFN-γIL-4的影响

目的研究口服益生菌对鼠伤寒沙门菌（STM）感染小鼠Th细胞因子的影响,以探讨益生菌抗沙门菌感染的免疫学机制。方法将95只Balb／c小鼠分为4组,分别为益生菌组（P）、益生菌对照组（Pc）、正常感染组（I）和对照组（C）。P组口服益生菌,I组口服生理盐水,均予等剂量STM口服感染,Pc组接种益生菌但不感染STM,C组不作任何处理。各组动物在11个不同时点处死,观察小肠、肝脏、脾脏病理改变,ELISA测量血清IFN-γ、IL-4表达。结果益生菌组器官组织的病理改变轻微,IFN-γ较正常感染组明显增加（Xp-1=66．52,P=0．001）,且其表达在感染最初的1h和后期的72h分别出现两个高峰;IL-4明显降低（Xp-1=-29．02,P〈0．001）,且较稳定。IFN-γ／IL4比值扩大（Xp-1=2．64,P〈0．001）。结论口服益生菌使小鼠保持有利于抗STM感染的Th1型反应,减轻了STM对机体的免疫损伤。     

流感病毒感染介导的免疫病理损伤研究进展

流感病毒感染(如暴发性流行或高致病性禽流感H5N1感染)可以造成广泛的病理损伤及严重的并发症,其肺部病理损伤以肺水肿及广泛的炎性渗出为特点,并伴有大量的中性粒细胞、巨噬细胞、淋巴细胞浸润及促炎因子和趋化因子的产生．组织学及病理学研究表明,过度的宿主应答反应是介导病理损伤的主要原因之一,而这些在流感病毒感染过程中介导组织损伤的免疫分子与细胞,在病毒的有效清除过程中同样至关重要．主要对甲型流感病毒感染过程中免疫系统的多种效应成分如何引发及加重病理性损伤等有害方面加以综述．为深入了解流感病毒感染防御机制及寻找并设计出既无害又能有效地治疗流感病毒感染的策略提供理论指导．     

感染和免疫反应对睡眠的影响

既往认为急性感染过程中睡眠结构发生的改变是由发热或神经中枢损伤引起的,现在则普遍认为这是微生物诱导的急性反应的一部分。感染可以激活免疫反应,进而导致一些免疫调节物质如白细胞介素-1β（IL-1β）和肿瘤坏死因子α（TNFα）等的表达,这些免疫调节物质通过复杂的分子网络直接参与睡眠调控。     

Identification of tubulin isoforms in different tissues of Ascaris suum using anti-tubulin monoclonal antibodies.

Three monoclonal antibodies specific to - and β-tubulin were used to examine the expression of tubulin isofoms in the intestine, reproductive tract and body wall muscle of A. suum. The tubulins were found to be different in their isoelectric points, number of isoforms and peptide maps with Western blot analysis of one-dimensional polyacrylamide gel confirming the presence of -, β₁- and β₂- tubulin. Commercial cross-reactive anti- and anti-β MAbs 356 and 357 recognized tubulin from A. suum tissues as well as from pig brain, whereas anti-A. suum β-tubulin specific MAb P3D6 recognized tubulin from the A. suum tissues only. Two-dimensional gel analysis showed different isoform patterns in different A. suum tissues with anti-A. suum β-tubulin MAb P3D6 and cross-reactive β-tubulin MAb 357 recognizing 2–4 β- tubulin isoforms and anti--tubulin MAb 356 recognizing 1–6 -tubulin isoforms. Different peptide maps of tubulin were observed in the three tissues, when subjected to limited proteolysis followed by SDS-PAGE. The data indicate that different tubulins are found in different tissues of adult A. suum.     

Quantifying the Early Immune Response and Adaptive Immune Response Kinetics in Mice Infected with Influenza A Virus

Seasonal and pandemic influenza A virus (IAV) continues to be a public health threat. However, we lack a detailed and quantitative understanding of the immune response kinetics to IAV infection and which biological parameters most strongly influence infection outcomes. To address these issues, we use modeling approaches combined with experimental data to quantitatively investigate the innate and adaptive immune responses to primary IAV infection. Mathematical models were developed to describe the dynamic interactions between target (epithelial) cells, influenza virus, cytotoxic T lymphocytes (CTLs), and virus-specific IgG and IgM. IAV and immune kinetic parameters were estimated by fitting models to a large data set obtained from primary H3N2 IAV infection of 340 mice. Prior to a detectable virus-specific immune response (before day 5), the estimated half-life of infected epithelial cells is ∼1.2 days, and the half-life of free infectious IAV is ∼4 h. During the adaptive immune response (after day 5), the average half-life of infected epithelial cells is ∼0.5 days, and the average half-life of free infectious virus is ∼1.8 min. During the adaptive phase, model fitting confirms that CD8⁺ CTLs are crucial for limiting infected cells, while virus-specific IgM regulates free IAV levels. This may imply that CD4 T cells and class-switched IgG antibodies are more relevant for generating IAV-specific memory and preventing future infection via a more rapid secondary immune response. Also, simulation studies were performed to understand the relative contributions of biological parameters to IAV clearance. This study provides a basis to better understand and predict influenza virus immunity.Current strategies for preventing or decreasing the severity of influenza infection focus on increasing virus-neutralizing antibody titers through vaccination, as experience indicates that this is the best way to prevent morbidity and mortality. Influenza A virus (IAV) undergoes mutations of the genes encoding the hemagglutinin (HA) and neuraminidase (NA) proteins that the neutralizing antibodies are directed against. When the variation is low (antigenic drift), prior vaccination often confers substantial heterologous immunity against a new seasonal IAV strain. In contrast, major genetic changes (antigenic shift) can result in pandemic IAV strains, since for novel strains, the humoral immune response is a primary response, and heterologous immunity is lacking. The emergence of such pandemic strains and the fact that young children are more vulnerable to influenza diseases highlight the need to better understand which viral and immune parameters determine the outcome of infection with viruses novel to the individual.Conventional experimental methods to measure influenza virus immunity have been limited to animal models and studies of adult human peripheral blood leukocytes. The advantages of using animal models include the ability to intensively sample multiple tissues and to utilize genetic and other interventions, such as blocking or depleting antibodies, to dissect the contribution of individual arms of the immune system. However, it is easy to question the relevance of these experiments to humans because of the many important biological differences between human and murine immune systems (29). In both the animal and human systems, we are limited to measuring those parameters and variables for which assays are available, most of them being ex vivo. Parameters such as cell-to-cell spread of the virus in vivo, trafficking of immune cells to the lung, and the in vivo interactions in an intact immune system are much more difficult or impossible to measure with contemporary techniques, particularly in humans. Computational approaches have the potential to offset some of these limitations and provide additional insight into the kinetics of the IAV infection and the associated immune response.Animal models of influenza virus infection in which different arms of the immune system have been suppressed suggest that some components of the adaptive immune system are required for complete viral clearance, often termed a sterilizing immune response. For example, abrogation of the CD4 T-cell response by cytotoxic antibody therapy or through knockout of major histocompatibility complex (MHC) class II slightly delays viral clearance but has little overall effect on the ability to control the infection (21, 54, 55). Elimination of the CD8 T-cell response typically results in delayed viral clearance (12, 20, 47), although animals with intact CD4 T-cell and B-cell compartments are able to control the infection in the absence of CD8 T cells. Presumably, this occurs through antibody-mediated mechanisms (54). Most animals depleted of both CD8 T cells and B cells are not able to clear the virus, which results in death (14, 32, 53). CD4⁺ T cells certainly contribute to the control of IAV infection, although cytotoxic CD4 T cells are not frequently observed unless cultured in vitro (8, 22, 45). Thus, it is generally accepted that CD8 T cells and/or antibodies are sufficient for timely and complete IAV clearance. Studies that strictly separate the relative roles of CD8 T cells and virus-specific antibodies are less satisfying. Animals depleted of both CD4 and CD8 T cells generally do not control the infection, despite substantial production of anti-IAV IgM antibodies (4, 23, 33, 34). However, adoptive transfer of IAV-specific IgM or IgG antibodies is protective (40, 51), suggesting that the timing and magnitude of the antibody response, i.e., the affinity, avidity, and antibody isotype, are important protective factors.While murine gene knockout or lymphocyte depletion studies are highly informative, they also have a number of limitations. Most importantly, the near-complete ablation of one component of the adaptive immune system often causes profound and unpredictable effects on many other immune components. Although the reported experimental measurements are highly quantitative, they often focus only on a limited number of time points or measurements and do not capture the complexity of the altered, or intact, immune response. In contrast, high-frequency experimental sampling, coupled with mathematical modeling techniques and new statistical approaches, can give insights into the complex biology of IAV infection and test the assumptions inherent in the model. We have learned from other systems, particularly HIV (19, 35, 37, 38, 56), that quantitative analysis of the biology can reveal important factors that are not intuitively obvious. For example, our current estimates for the rates of HIV production and the life span of productively infected cells in vivo were obtained via mathematical modeling (35).Mathematical models have long been used to investigate viral dynamics and immune responses to viral infections, including influenza A virus (3, 5, 7, 15, 16, 31, 36, 48). We recently described a complex differential equation model to simulate and predict the adaptive immune response to IAV infection (24). This model involves 15 equations and 48 parameters, and because of its complexity, many of the parameter values that could not be directly measured were unidentifiable. Thus, it is difficult to estimate all model parameters by fitting experimental data directly to this complex model, although the model can be used to perform simulation predictions (25). This issue can, however, be addressed by reducing the model into smaller submodels with smaller but identifiable sets of parameters, which can be estimated from experimental data. In this paper, we describe such an approach which focuses on IAV infection and the immune response solely within the lung.In the present report, we have fitted a model of primary murine influenza virus infection to data. In naïve subjects, our data suggested that there is no adaptive immune response (e.g., IAV-specific CD8⁺ T cells or antibodies) detectable in the spleen, lymph nodes, or lung until approximately 5 days after infection; therefore, we have divided the analysis into the following two phases: the initial preadaptive (innate) phase and the later adaptive phase. We use direct experimental data from infection of mice with the H3N2 influenza virus A/X31 strain (2, 24) to obtain key kinetic parameters. The model fitting has revealed that the duration of the infection depends on a small set of immune components, and even large fluctuations in other arms of the immune system or IAV behavior have surprisingly little impact on the outcome of the infection.     

增强DNA疫苗免疫应答的新进展

DNA疫苗为编码抗原蛋白的真核表达载体,注入体内后在原位表达所编码的抗原并诱导免疫应答,在预防感染、治疗自身免疫性疾病、过敏性疾病和肿瘤等疫病中有着很好的应用前景。但与灭活疫苗相比,其免疫效价还比较低。有多种策略能够增强或调节DNA疫苗诱导的免疫应答,其中,作为外源基因载体的质粒的组成及插入的有关基因均可直接或间接地影响免疫反应的效果,在构建DNA疫苗质粒时,加入细胞因子、融合信号、泛素等基因以及ISS序列,另外还可以通过设计一些对抗原提成细胞有影响的分子共注射,以及加入转移分子,都可以明显增强DNA疫苗的免疫效果,从而有利于研制更有效的DNA疫苗。     

扇贝糖胺聚糖对感染HSV-I小鼠免疫功能的影响研究

目的：研究扇贝裙边糖胺聚糖（glycosaminoglycan from Scallop Skirt,SS—GAG）对感染单纯疱疹病毒I型（herpes simplex virustype,HSV-I）小鼠免疫功能的影响。方法：通过在无菌条件下给予小鼠注射扇贝糖胺聚糖SS．GAG,连续11天,并在给药第三天给小鼠腹腔注射HSV—I病毒悬液建立小鼠感染模型,用MTT等方法观察SS—GAG对HSV—I感染小鼠腹腔巨噬细胞吞噬活性的影响、对脾脏指数、胸腺指数的影响以及对脾淋巴细胞转化能力等免疫指标的影响。结果：与病毒对照组相比,扇贝糖胺聚糖SS-GAG低剂量组、中剂量组、高剂量组均能显著增强HSV．I感染小鼠的腹腔巨噬细胞的吞噬活性和HSV—I感染小鼠的脾脏指数和胸腺指数（P〈0．01）,并且能促进其脾淋巴细胞转化增殖能力（P〈0．01）。结论：扇贝糖胺聚糖在体内有一定的抗I型单纯疱疹病毒作用。其抗病毒作用可能与增强机体免疫功能有关。     

Oxidative Stress in Lungs of Mice Infected with Influenza a Virus

As oxidative stress has been implicated in the pathogenesis of certain viral diseases we determined antioxidant and prooxidant parameters in lungs and bronchoalveolar lavage fluid (BALF) of mice infected with a lethal dose of influenza A/PR8/34 virus. Viral infection was characterized by massive infiltration of leukocytes, mainly polymorphonuclear leukocytes, into the alveolar space. The total number of BALF cells increased up to 8-fold (day 3 post-infection) and these cells appeared activated as judged by their increased rates of superoxide anion radical (O₂^-) generation upon stimulation. Maximal rates of radical generation by BALF cells during the early stages of infection were 15- or 70-fold higher than those of cells from control animals when expressed per cell or total BALF cells, respectively. At the terminal stages of infection the total capacity of BALF cells to release of declined to ≈ 35-fold the control values. Infection also resulted in increased in vivo formation of hydrogen peroxide (H²O²) within the lungs at a time that coincided with the maximal capacity of BALF cells to release O_2-.

Whereas pulmonary activities of glutathione peroxidase and reductase remained unaltered, levels of ascorbate in the cell-free BALF decreased significantly during the early stages of the infection and then returned to normal levels and above, late in infection. The oxidation state of the dehydroascorbic acid/ ascorbate couple increased concomitantly with the decrease in ascorbate concentrations early in infection and remained elevated throughout the infection. As assessed by the prevention of peroxyl radical-induced loss of phycoerythrin fluorescence, the total antioxidant capacity present in lung tissue homogenate from terminally ill animals was not diminished when compared to that prepared from lungs of control mice. We conclude that although early stages of influenza infection are associated with the presence of oxidative stress in the lung tissue and alveolar fluid lining the epithelial cells, this stress does not appear to overwhelm local antioxidant defenses. The results therefore do not support a direct causative role of oxidative tissue damage in the pathogenesis of influenza virus infection.     

具免疫时滞的HIV感染模型动力学性质分析

讨论了一类具免疫时滞的HIV感染模型.分析了未感染平衡点的全局渐近稳定性,给出了感染无免疫平衡点及感染免疫平衡点局部渐近稳定的充分条件.数值模拟结果表明,当易感细胞生成率的取值使得基本再生数满足平衡存在的条件且低于某一临界值时,时滞对平衡点的稳定性没有影响;若大于该临界值,随着时滞增大,稳定性开关发生,平衡点不稳定,出现一系列Hopf分支,最终表现为周期波动模式.     

乙型肝炎疫苗与卡介苗、百白破、脊髓灰质炎疫苗联合免疫的免疫应答观察

本文报道在对婴幼儿实施基础免疹(BCC、DPT和TOPV)的同时,接种国产血源乙肝疫苗的免疫应答。对八月龄的婴儿血清学检测结果表明:乙肝疫苗接种组1(10μg)、2(20μg)抗-HBs达到临界保护(P/N≥10.0)的分别为80.65％,78.49％,未检出HBV感染标志物;未接种乙肝疫苗组的HBV感染率边10.5％。血清学检测还表明乙肝疫苗与BCG、DPT和TOPV等生物制品联合免疫应答是好的,各抗原间无干扰作用,提示乙肝疫苗可列入扩大免疫规划,并将有效地控制乙肝病毒感染。