Cetacean-reconstituted severe combined immunodeficient (SCID) mice respond to vaccination with canine distemper vaccine

Morbillivirus infections have been responsible for mass mortalities in several species of marine mammals. Nevertheless, relatively little is known on the pathogenesis of the disease and the immune response to the agent, especially in cetaceans, hindering the treatment of individuals and the development of appropriate vaccines, given the difficulty of performing experimental work in marine mammals. The reconstitution of severe combined immunodeficient (SCID) mice, which do not have the ability to reject grafts, with lymphocytes from different species has been used with increasing success as a surrogate species model to study the immune system. We injected NOD/SCID mice with lymphocytes from different species of cetaceans and further vaccinated those mice with a commercial canine distemper virus (CDV) vaccine to develop a practical model to study cetacean immune response to a morbillivirus. Reconstitution was detected in 10/20 mice reconstituted with harbor porpoise spleen, 6/10 mice reconstituted with harbor porpoise lymph node cells, 8/10 mice reconstituted with fresh beluga PBMCs and none of the mice reconstituted with neonate bottlenose dolphin spleen or thymus cells when assessed 42-63 days after reconstitution. While a humoral immune response was detected in none of the reconstituted mice, a cell-mediated immune response to the CDV vaccine was detected in 6/15 (40%) and 2/18 (11%) of the SCID mice after reconstitution with cetacean immune cells after a single or booster vaccination, respectively, for a combined total of 8/33 (24%). This represents the first demonstration of successful reconstitution of SCID mice with marine mammal cells, and to the authors' knowledge, the first direct demonstration of a primary antigen-specific cell-mediated immune response in reconstituted SCID mice. This model will be useful for further research on the physiology of the marine mammal immune system and its response to infectious agents and vaccines, with possible important outcomes in conservation issues.     

T cell-mediated immunity to Cowdria ruminantium in mice: the protective role of Lyt-2+ T cells

The inability of athymic nude mice to make a drug-aided recovery from infection with either the Kümm or the Welgevonden stocks of Cowdria ruminantium and their inability to mount an immune response, suggest that immunity in heartwater is cell-mediated. The adoptive transfer of immunity with the spleen cells of mice immune to the Welgevonden stock is supportive evidence. Immune spleen cells depleted of Lyt-2+ T cells are unable to confer resistance to challenge to recipient mice, whereas the depletion of L3T4+ T cells had no effect on the protection conferred by immune spleen cells. This is conclusive evidence that immunity in heartwater is largely cell-mediated. Immune serum, C. ruminantium and complement incubated in the presence of mouse peritoneal macrophages, inhibits the infectivity of the heartwater agent, but not in the absence of macrophages. The decreased resistance to challenge of immune mice treated with gloxazone adds further support to the concept that in heartwater persistence of C. ruminantium in the host is associated with immunity.     

Heligmosomoides polygyrus and Trypanosoma congolense infections in mice: effect of immunisation by abbreviated larval infection.

Concurrent African trypanosome and gastrointestinal helminth infections are prevalent in sub-humid savannah where they are endemic. However, acquired resistance in animals varies with their responder status and exposure. As a guide to study in the definitive hosts, the effects of Trypanosoma congolense infection on the development and maintenance of homologous Heligmosomoides polygyrus resistance were investigated in outbred TO mice. These mice were immunised by abbreviation of larval infection. Immune or naive mice were either infected with 500 infective larvae (L3) of H. polygyrus and/or 10(4) bloodstream forms of T. congolense or were not infected. The outcome of infection was monitored by routine parasitological and immunological techniques for 30 days after the day of the T. congolense infection. Significantly more immune mice concurrently infected with both parasites survived than did immune mice in which H. polygyrus was superimposed on a 10-day-old T. congolense infection. Although all the mice in this latter group died before the end of the experiment, larval immunisation prolonged their survival, relative to similarly treated naive mice. The antibody titres to H. polygyrus in the sera of immune mice challenged with H. polygyrus alone were significantly higher than those of immune mice concurrently infected with both parasites but the levels of protection obtained were comparable. It is concluded that T. congolense may not completely block the strong acquired resistance induced by abbreviated H. polygyrus larval infection in TO mice but is capable of interfering with protective responses, especially if the trypanosome infection occurs prior to H. polygyrus challenge infection.     

Regulating effects of porcine interleukin-6 gene and CpG motifs on immune responses to porcine trivalent vaccines in mice

In order to develop novel immunoadjuvants to boost immune response of conventional vaccines, experiments were conducted to investigate the regulating effects of porcine interleukin-6 gene and CpG motifs as the molecular adjuvants on immune responses of mice that were co-inoculated with trivalent vaccines against Swine fever, the Pasteurellosis and Erysipelas suis. Synthetic oligodeoxynuleotides containing CpG motifs were ligated into pUC18, forming recombinant pUC18-CpG plasmid. Eukaryotic plasmid expressing porcine interleukin-6 (VPIL-6) were also constructed as molecular adjuvants in an attempt to enhance levels of immune responses of mice co-administered with the trivalent vaccines in this paper. The cellular and humoral immune responses of mice were systematically analysed, and the experimental results were observed that the number of white blood cells, monocytes, granuloytes and lymphocytes significantly increased, respectively, in the mice immunized with VPIL-6, compared with those of the control; the IgG content and titre of specific antibodies to the trivalent vaccine mounted remarkably in the sera from the VPIL-6 vaccinated mice; the proliferation of lymphocytes and induced IL-2 activities were significantly increased in the vaccinated groups. The above-mentioned immune responses of mice co-inoculated with pUC18-CpG plasmid were significantly stronger than those of co-inoculated with pUC18 plasmid, suggesting that the immunostimulatory effect of oligodeoxynuleotides CpG is closely connected with the number of CpG motifs. These results suggest that the porcine IL-6 gene and CpG motifs could be employed as effective immunoadjuvants to elevate immunity to conventional vaccines.     

三种多糖对小鼠免疫功能的影响

为探讨黄芪多糖、酵母胞壁多糖和虫草菌丝体多糖对动物免疫功能的影响,本试验分为对照组和黄芪多糖、酵母胞壁多糖、虫草菌丝体多糖,3种多糖等量复配4个试验组。每组选用昆明种小鼠40只,对照组小鼠每只灌胃1mL生理盐水,黄芪多糖、酵母胞壁多糖、虫草菌丝体多糖和3种多糖等量复配组小鼠每只灌胃量为140mg多糖/kg体重,每隔10d取样1次,分别检测增重率、免疫器官指数、血细胞数和外周血IgG含量。结果表明,黄芪多糖组、酵母胞壁多糖组和虫草菌丝体多糖组均可显著提高小鼠增重率、免疫器官指数、血细胞数目及外周血IgG含量(P<0.05),复合多糖组则与单多糖组相比差异显著(P<0.05)。说明3种多糖组及其混合多糖组均能提高小鼠免疫力,且最好的为混合多糖组。     

Humoral immune response to non-viable Mycoplasma pulmonis in mice enhanced by dextran sulfate

The enhancing effect of dextran sulfate on the humoral immune response to nonviable Mycoplasma pulmonis in mice was evaluated by means of the indirect haemagglutination test. The serum antibody titres in mice immunized subcutaneously with a mixture of non-viable M. pulmonis and dextran sulfate were greater and persisted longer than those in mice immunized with non-viable M. pulmonis alone. DEAE-dextran also enhanced the humoral immune response to non-viable M. pulmonis in mice.     

不同地理株伊氏锥虫灭活苗交叉免疫保护及免疫方法研究

用伊氏锥虫湖北株和广西株制备纯虫灭活苗和含血灭活苗,经小鼠１次、２次免疫,再经强毒伊氏锥虫湖北株、广西株和江苏株交叉攻击。结果湖北株纯虫灭活苗２次免疫鼠,对同株（湖北株）获得３０／３０保护,对异株即广西株和江苏株分别获０／１０和７／１０保护,该苗一次免疫鼠对同株仅获得５／１０保护。广西株纯虫灭活苗２次免疫鼠对同株（广西株）获２０／２０保护,对异株即湖北株和江苏株分别获得０／１０和４／１０保护。含血湖北株伊氏锥虫灭活苗对同株无保护作用,１３只免疫小鼠全部死亡,对江苏株（异株）为９／１０死亡。对照小鼠全部发病死亡。交叉免疫保护试验结果说明,纯虫灭活苗经两次免疫后对伺株虫体产生坚强的免疫保护作用,免疫保护率达１００％,不同地理株伊氏锥虫因抗原变异现象出现不同的免疫保护作用。两次免疫接种小鼠其免疫保护效果比一次免疫的效果好。疫苗中如含有大量非特异物质,虫苗的免疫保护作用将完全丧失。稳定试验表明,灭活苗在室温下保存１个月,在４℃、－２０℃下保存６个月,免疫效果不变。     

家兔与BALB/c小鼠对猪瘟病毒E2蛋白靶向化树突状细胞疫苗敏感性的比较

为比较家兔与BALB/c小鼠对猪瘟病毒E2蛋白靶向化树突状细胞疫苗的敏感性,本研究首先根据小鼠与家兔体重比,分别免疫糖基化E2蛋白与E2蛋白（家兔2 mg/只,小鼠20 μg/只）,在不同时间采血制备血清,通过ELISA方法测定血清中IgG水平。结果显示,接种后BALB/c小鼠出现了轻度的免疫应答,而家兔产生了强而持久的免疫保护。本研究对BALB/c小鼠的接种量进行了进一步摸索,结果表明,50与100 μg/只免疫效果明显优于20 μg/只;其中50 μg/只E2蛋白与糖基化E2蛋白免疫后,血清IgG水平较高且稳定;而100 μg/只糖基化E2蛋白免疫后,免疫初期,糖基化E2蛋白组产生了免疫抑制,至21 d抗体水平逐渐升高且达极高水平;但100 μg/只E2蛋白免疫后,整个免疫期内抗体水平均较低,只有轻微波动。以上结果表明,家兔对糖基化E2蛋白与E2蛋白的敏感性均显著高于BALB/c小鼠,且糖基化E2蛋白免疫效果明显好于E2蛋白,BALB/c小鼠的糖基化E2蛋白最佳免疫剂量为50 μg/只。     

阿胶泡腾颗粒对小鼠免疫功能的影响

研究了阿胶泡腾颗粒对小鼠免疫功能的影响.通过免疫器官重量法,考察了阿胶泡腾颗粒对小鼠中枢免疫器官和外周免疫器官发育的影响;通过碳廓清试验,考察了阿胶泡腾颗粒对巨噬细胞吞噬功能的影响;通过迟发型变态反应试验,考察了阿胶泡腾颗粒对小鼠细胞免疫功能的影响.结果表明,阿胶泡腾颗粒能增加免疫抑制小鼠的脾脏、胸腺指数,与空白对照组...     

Immunity to Babesia in mice. I. Adoptive transfer of immunity to Babesia rodhaini with immune spleen cells and the effect of irradiation on the protection of immune mice

Immunisation of Balb/c mice against Babesia rodhaini by an amicarbalide-controlled infection resulted in a solid immunity which lasted for 216 days. With spleen cells of immune mice protection could be transferred both to naive mice pretreated with cyclophosphamide. Treatment of naive mice with cyclophosphamide (300 mg/kg) five days before a lethal B. rodhaini inoculation resulted in over 50% survival. This protective effect of cyclophosphamide is explained by its inhibiting effect on suppressor T-cells. The protection against B. rodhaini challenge infection afforded to immune Balb/c mice was completely resistant to a sublethal irradiation of 400 rad. Since B-lymphocyte function in antibody production is suppressed by this dose, the role of antibodies in the effector phase of the immunity appears to be of minor if any importance. A considerable degree of protection was still preserved after irradiation of immune animals with 875 rad. Sensitivity to this irradiation dose of all immunocompetent cells except macrophages and a small fraction of T-lymphocytes indicates the involvement of these cell types in the effector phase of the specific immunity. Highly radioresistant macrophages are therefore considered to play the major role but T-lymphocytes are also required for complete protection.     

Recovery of age-dependent immunological deterioration in old mice by thyroxine treatment

On the basis that multiple interactions exist between thyroid hormones and immune system, and ageing is accompanied by changes in thyroid hormone secretion, it seems possible that thyroid hormones may be involved in the age-related immune dysfunction. The present study was conducted to evaluate in vivo and in vitro effects of thyroxine (T(4)) treatment on both cell-mediated and humoral immune responses of aged mice. In a trial to improve age-associated immune dysfunction, T(4) (0.2, 1.0 and 5.0 microg) was subcutaneously supplemented to BALB/c mice (over 18 months old) for 30 consecutive days. The present results showed that exogenous treatment of aged mice with T(4) was associated with a marked increase in serum T(4) level, and the total number of peripheral blood leukocytes as well as the total cellularity of thymus, spleen, peripheral lymph nodes (PLNs), mesenteric lymph nodes (MLNs) and bone marrow (BM). T(4) treatment also caused a significant increase in the total and differential numbers of peritoneal exudate cells (PECs), while it caused a slight increase in macrophages' phagocytic activity of PEC. Moreover, T(4) treatment elicited a statistically significant increase in both plaque-forming cell and rosette-forming cell responses. In vitro results showed that the addition of T(4) at concentrations of 0.001, 0.005 and 0.025 microg/well substantially potentiated the ability of splenocytes from aged mice to proliferate in the presence of concanavalin-A mitogen. Histological examination of thymuses from T(4)-treated aged mice revealed that the cortex was preferentially enlarged and repopulated with immature thymocytes. The present study postulates that thyroid hormones may be involved in the observed decrease in the immune responsiveness during ageing, and that T(4) treatment to aged mice is able to restore the age-related decline of the immune efficiency.     

蚯蚓肽对小鼠非特异性免疫功能的影响

通过用不同剂量的蚯蚓肽(EP)对健康和免疫抑制小鼠灌胃15 d,观察EP对小鼠淋巴细胞增殖反应,巨噬细胞细胞毒效应以及巨噬细胞和脾细胞产生NO的影响,以探讨EP对小鼠非特异性免疫功能的影响。试验发现0.1,0.5 mg/mL组明显提高淋巴细胞增殖率,增强巨噬细胞细胞毒效应,提高巨噬细胞和脾细胞分泌NO的水平(P<0.0l),0.5 mg/mL明显提高免疫抑制小鼠的免疫功能(P<0.01)。结果表明:一定剂量下,EP具有调节免疫功能、拮抗环磷酰胺引起的免疫抑制的作用。     

Assessment in mice of vapA-DNA vaccination against Rhodococcus equi infection

There is a need to produce a vaccine against Rhodococcus equi pneumonia in foals in which immunity against infection is largely based on a type 1, cell-mediated, immune response. The VapA protein of the virulence plasmid of R. equi is highly immunogenic. To assess the potential of vapA-DNA to produce immunity, C57BL/6 and BALB/c mice were immunized with a DNA vaccine constructed from vapA incorporated into pcDNA3.1. The plasmid construct expressed VapA in a COS-7 cell line. Intramuscular immunization of mice resulted in enhanced clearance of R. equi from the liver of intravenously challenged mice compared to non-immunized controls. This effect was more marked when pORF-IL-12, a plasmid expressing murine IL12, was included with the vaccine. Antibody developed to VapA, with an IgG2a response being more marked in mice immunized with pcDNA-vapA than in non-immunized or in mice immunized with the mixed vapA and IL-12 plasmid constructs. In conclusion, this study has shown for the first time that DNA immunization with vapA enhances the immune responses of mice against R. equi infection, that the IgG subisotype response is consistent with a type 1-based immune response, and that this can be enhanced by injection of the IL-12 gene.     

Effects of mild stress on the immune response against pseudorabies virus in mice

Stress is a recognised problem in intensive pig husbandry, which might lead to changes in immune reactivity. To study the effect of stress on the development of an anti-viral immune response, we used a murine model in which mice were immunized with an attenuated strain of pseudorabies virus (PRV). The effect of two stress treatments, both relevant to intensive pig husbandry, on the development of the specific immune response against PRV was investigated. The stress treatments consisted of restraint, social isolation, and transport and they differed in predictability. The specific immune response against PRV, which developed in the draining lymph nodes, was measured by a lymphocyte proliferation assay and cytokine production assays. Our results showed that the unpredictable stress treatment had no effect on the development of the immune response against PRV in mice, whereas the predictable stress treatment actually hastened the immune response.     

纳米级小肽螯合铜对小白鼠生长性能和免疫器官的影响

此试验的目的是探讨以硫酸铜、碱式氯化铜、蛋氨酸铜、纳米铜和纳米小肽螯合铜等不同铜源供给形式对小白鼠生长性能和免疫器官的影响。将120只18日龄小白鼠随机分为6个处理组,分别为Ⅰ（小肽＋硫酸铜组）、Ⅱ（硫酸铜组）、Ⅲ（碱式氯化铜组）、Ⅳ（蛋氨酸铜）、Ⅴ（纳米铜组）和Ⅵ（纳米小肽螯合铜）等,每个处理组设4个重复,每个重复5只小白鼠。饲喂试验分0～5d、6～12d、13～19d、20～26d和27～53d五个阶段。其中0～5d为预试验期,后四个阶段为试验期。预试期和试验期各处理组采用相同的基础日粮。试验期Ⅰ～Ⅵ组饲粮铜的含量（以铜计）在6～12d、15～19d、20～26d和27～55d四个阶段分别为2．5mg、5mg、10mg及15mg。结果表明,纳米小肽螯合铜能有效促进小鼠增重。采食不同铜源日粮对小鼠免疫器官指数的影响随饲养日龄增大而减小,并随铜的供给量由低剂量变化到正常剂量而差异减小。     

Mice immune responses to Brucella abortus heat shock proteins. Use of baculovirus recombinant-expressing whole insect cells,purified Brucella abortus recombinant proteins,and a vaccinia virus recombinant as immunogens

Brucella abortus resists the microbicidal mechanisms of macrophages, and the expression of its heat shock proteins (HSPs) such as GroEL, GroES and HtrA may play a role in this resistance. Bacterial HSPs can be very immunogenic, inducing protective immunity in various types of bacterial infections. However, the significance of immune responses directed against B. abortus HSPs in the protection against brucellosis is currently unresolved. To elucidate the role of these proteins in protection against Brucella challenge, individual, divalent or trivalent baculovirus (BV) recombinants of B. abortus GroEL, GroES and/or HtrA were injected into BALB/c mice either as protein-expressing whole cells or as purified proteins. The preparations were given to mice in combination with Freund's or Ribi adjuvant, respectively. In addition, some mice were primed with a vaccinia virus-GroEL recombinant, followed by inoculation with purified GroEL-Ribi adjuvant combination. Antibodies were observed against B. abortus GroEL and HtrA, but not against GroES. Cellular immune response was demonstrated by observing significant IFN-gamma release by lymphocytes of mice immunized with the purified HtrA-Ribi adjuvant combination. However, none of the mice inoculated with individual, divalent or trivalent HSP-expressing cells combined with complete Freund's adjuvant or inoculated with purified B. abortus HSPs combined with Ribi adjuvant, were protected against challenge with B. abortus virulent strain 2308. Priming with vaccinia virus-GroEL recombinant and boosting with GroEL-Ribi combination did not induce protective immunity. Based on the results obtained, we suggest that although humoral and cell-mediated immune responses are induced, but protective immune response is not induced by B. abortus HSPs.     

猪源植物乳杆菌的诱变选育及其对小鼠免疫性能的影响

To get a Lactobacillus plantarum strain with good acid resistance, cholate tolerance, and in vitro adhesion rate and study its effects on immune function of mice. After screening Lactobacillus plantarum by acid resistance and cholate tolerance after UV mutagenesis, we got BL-15 strain, which was cultured, freeze-dried, dissolved and administered to BALB/c mice orally (gastric perfusion) by three times. Blood, spleen and excrement were collected to detect changes in immune function index.One Lactobacillus plantarum strain BL-15 was selected from 17 mutant strains, with higher acid resistance, cholate tolerance and in vitro adhesion rate. The experimental data of animal tests indicated that IL-2 level and spleen index of mice at early growth stage had been increased. Furthermore, the mice treated with BL-15 as the immunity enhancer revealed higher level of SIgA in intestinal contents compared with control (P<0.05). The results showed that Lactobacillus plantarum strain BL-15 had been proved to be able to elevate immune response and enhance immune function.     

蒲公英多糖对小鼠免疫器官的影响

从蒲公英中提取多糖,观察其对小鼠免疫机能的影响。30只昆明种小鼠随机分成试验组和对照组,每组15只。试验持续10 d,于试验第3天给试验组小鼠灌服制备的蒲公英多糖溶液,对照组小鼠则以生理盐水灌胃。试验结束后脱颈处死小鼠,剖检,取胸腺和脾脏分别称重,并取适当组织制备切片镜检。结果显示,蒲公英多糖能显著提高脾脏指数和胸腺指数,改善器官内部组织结构,促进小鼠免疫器官的生长发育,有利于提高小鼠的免疫功能。     

表达H1N1、H3N2猪流感病毒HA基因重组质粒在小鼠的免疫效力

应用已构建的真核表达质粒pCI-H1-HA、pCAGGS-H1-HA、pCI-H3-HA和pCAGGS-H3-HA作为DNA疫苗，利用BALB／c小鼠进行免疫保护试验，通过测定不同免疫期HI抗体滴度、分析攻毒后BALB／c小鼠体重变化及肺脏病毒含量，评价DNA疫苗的免疫效力。结果表明：构建的DNA疫苗均可诱导小鼠产生免疫力；BALB／c小鼠体重变化统计学分析显示，免疫组与对照组差异极显著（P〈0．01），pCAGGS表达载体构建的DNA疫苗免疫效果优于pCI表达载体构建的DNA疫苗（P〈0．05）。     

In vitro effect of T-2 mycotoxin on the immune response of mice

The in vitro biologic effects of T-2 mycotoxin on the immune response of mice was undertaken. Twenty nanograms of toxin abrogated the immune response to the T-dependent antigen sheep RBC, whereas a partial response was observed when 2 ng was used. Analysis of cell culture viabilities indicated that cell death occurred with toxin doses that coincided with the diminished immune responses. A similar decreased response was observed against the T-independent antigen, TNP-lipopolysaccharide, indicating toxic effects on both B and T lymphocyte populations. Delay of toxin administration as much as 116 hours of the 120-hour incubation period still resulted in a substantially diminished immune response, indicating the toxin acts on both the afferent and efferent immune systems. Equal effects were observed for mice of the b, d, and k haplotype, indicating no apparent strain variability in sensitivity to T-2 mycotoxin effects. These results indicated that T-2 mycotoxin can modulate the immune response, and that this modulation is attributable to direct toxic effects on the cells of the immune system.