Immunization of proteins from Toxascaris leonina adult worm inhibits allergic specific Th2 response

Recently, the influence of parasitic infections on the incidence of allergic diseases has become the focus of increased attention. In order to ascertain whether parasite-derived proteins could inhibit the allergic specific Th2 response, we applied excretory-secretory protein (Tl-ES) or total protein (Tl-TP) of the adult worm Toxascaris leonina to asthma model mice prior to or simultaneously with OVA challenge, after which we assessed the OVA-specific Th2 responses. The group subjected to immunization with Tl-ES and Tl-TP (immunized group) evidenced a thinning of the bronchial epithelial and muscle layer, a disruption and shedding of epithelial cells, a reduction in the number of goblet cells, and a reduction in mucus production as compared to the group treated with Tl-ES coupled with OVA challenge (challenge with OVA groups) and the OVA-induced asthma group. The administration of Tl-ES and Tl-TP, regardless of injection time, was shown to inhibit the recruitment of inflammatory cells into the airway, and in particular, macrophages, neutrophils, and lymphocytes were significantly reduced as the result of the parasite proteins. However, the total number of eosinophils was slightly reduced as the result of the administration of parasite proteins. Sensitization and OVA challenge was shown to accelerate the secretion of Th2 cytokines (IL-4 and IL-5) within the lung, but in the immunized groups, those levels were lower. The administration of Tl-TP and OVA challenge group also evidenced a significant reduction in IL-4 levels as compared to the OVA-challenged group. The concentrations of Th2 cytokines in the Tl-ES and OVA challenge group were more similar to those observed in the OVA-challenged group. The concentration of IL-10 and TGF-beta in the lung was decreased substantially in the OVA-only challenge group, but the Tl-TP immunized group exhibited significantly induced IL-10 cytokine. OVA-specific IgG2a, IgG1, and IgE levels in the immunized groups were significantly lower than those detected in the OVA-challenged group. In conclusion, parasite-derived protein is able to inhibit OVA-specific Th2 responses, and in particular, immunization with parasite proteins exerts a more profound protective effect than is seen with the treatment of allergic reactions. The results of our study are encouraging in terms of our further understanding of the molecular basis of immune evasion by nematodes.     

Characterization of the immune response to Mycoplasma bovis lung infection

To better understand the interaction between Mycoplasma bovis and its bovine host, we have characterized the immune response generated during an experimental lung infection with M. bovis. Proliferation ([3H]-thymidine blastogenesis) and Th1/Th2 cytokine production were used to monitor peripheral cellular immune responses. Flow cytometry analysis was used to determine T-cell subset activity by CD25 expression. Humoral immune response was monitored by the identification of antigen-specific IgG1 and IgG2 isotypes over time. Herein, we show that M. bovis antigen stimulates activation of CD4+ and CD8+ cells in vitro in a manner consistent with memory, and that gammadelta-T cells are activated by antigen in a manner consistent with innate immunity. In addition, the percentage of cells producing IFN-gamma during recall response is equal to that of IL-4 producing cells. Serological analysis shows M. bovis stimulates increased production of antigen-specific IgG1 while very little IgG2 is produced. We therefore submit that experimental lung infection of cattle with M. bovis results in a Th2-skewed immune response.     

L-茶氨酸对氧化应激断奶仔猪生长性能及血液生化指标的影响

本研究探讨了L-茶氨酸对氧化应激断奶仔猪生长性能、抗氧化能力及免疫功能的影响。试验采用2×2二因素设计,160头断奶仔猪（7.53 kg±0.51 kg）随机分为4个处理组：1组（日粮中不添加L-茶氨酸,腹腔注射5 mL生理盐水）;2组（日粮中不添加L-茶氨酸,腹腔注射8 mg/kg体重敌快死）;3组（日粮中添加1 000 mg/kg L-茶氨酸,腹腔注射5 mL生理盐水）;4组（日粮中添加1 000 mg/kg L-茶氨酸,腹腔注射8 mg/kg体重敌快死）。预试期7 d,正试期28 d。结果显示：①与1组相比,2组仔猪平均日增重（ADG）和平均日采食量（ADFI）显著降低,而料重比显著升高（P<0.05）;与2组相比,4组仔猪ADG显著增加（P<0.05）。②与1组相比,2组仔猪血清丙二醛（MDA）含量显著增加,而总抗氧化能力（T-AOC）和谷胱甘肽过氧化物酶（GSH-Px）活性显著下降（P<0.05）,3组仔猪血清MDA显著降低,而T-AOC显著增加（P<0.05）;与2组相比,4组仔猪血清MDA含量显著降低,而T-AOC和GSH-Px活性显著上升（P<0.05）。③与1组相比,2组仔猪血清IgA、IgG、IL-2、IL-4水平显著增加,C3和C4水平显著下降（P<0.05）,3组仔猪血清IgA、IgG和IL-4水平显著降低,而C3和C4水平显著增加（P<0.05）;与2组相比,4组仔猪血清IgA、IgG、IL-2、IL-4水平显著降低,而血清C3水平显著增加（P<0.05）。结果表明,注射敌快死可引发仔猪氧化应激,造成生长性能下降、体液免疫能力下降;日粮中添加1 000 mg/kg L-茶氨酸可缓解仔猪氧化损伤,提高氧化应激仔猪的生长性能、抗氧化能力和体液免疫能力。     

Correlation between the nature of immunity induced by different immunogens and the establishment of latent infection by wild-type pseudorabies virus

To assess the correlation between the nature of immunity induced by different types of immunogens and the establishment of latent infection by wild-type pseudorabies virus (PrV), we used a murine model immunized with different immunogens, the PrV modified live vaccine (MLV), inactivated vaccine (IAV), and commercial oil-adjuvant subunit vaccine (OSV), via either intranasal (i.n.) or intramuscular (i.m.) route. Both MLV and IAV induced a different nature of immunity biased to Th1- and Th2-type, respectively, as judged by the ratio of PrV-specific IgG isotypes (IgG2a/IgG1) and the profile of cytokine IL-2, IL-4, and IFN-gamma production. In contrast, the OSV induced a lower isotype IgG2a to IgG1 ratio and higher level of IL-2 production. The MLV (inducing Th1-type) provided more effective protection against a virulent wild-type PrV challenge than IAV and OSV (inducing Th2- and mixed type, respectively). In addition, the MLV impeded the establishment of a latent infection with wild-type PrV, and the decrease in the PrV latency load by immunization with the MLV appeared to be mediated by the immune T-cells. These results demonstrate the substantial role of the immune responses driven by preceding vaccination in modulating the establishment of PrV latency caused by the post-infection of a field virus.     

Immune response in mice and cattle after immunization with a Boophilus microplus DNA vaccine containing bm86 gene

Plasmid pBMC2 encoding antigen Bm86 from a Colombian strain of cattle tick Boophilus microplus, was used for DNA-mediated immunization of BALB/c mice, employing doses of 10 and 50microg, delivered by intradermic and intramuscular routes. Anti-Bm86 antibody levels were significantly higher compared to control mice treated with PBS. In the evaluation of immunoglobulin isotypes, significant levels of IgG2a and IgG2b were observed in mice immunized with 50microg of pBMC2. Measurement of interleukine (IL) levels (IL-4, IL-5, IL-12(p40)) and interferon-gamma (IFN-gamma) in the sera of mice immunized with pBMC2 indicated high levels of IL-4 and IL-5, although there were also significant levels of IFN-gamma. Mice immunized with pBMC2 showed antigen-specific stimulation of splenocytes according to the incorporation of bromodeoxyuridine and IFN-gamma secretion. In all trials, mice injected intramuscularly with 50microg of pBMC2 presented the highest immune response. Moreover, cattle immunized with this DNA vaccine showed antibody production significantly different to the negative control. In conclusion, these results suggest the potential of DNA immunization with pBMC2 to induce humoral and cellular immune responses against B. microplus.     

Cytokine responses in camels (Camelus bactrianus) vaccinated with Brucella abortus strain 19 vaccine

In the present study, we determined the levels of cytokines produced by camel (Camelus bactrianus) peripheral blood mononuclear cells (PBMCs) in response to live attenuated Brucella abortus (B. abortus) S19 vaccine. Seven camels were vaccinated with commercial B. abortus S19 vaccine, and their cytokine responses were determined using a real-time PCR assay. Cytokine responses to B. abortus S19 were examined at 6 hr, 48 hr and 1, 2 and 3 weeks post-vaccination. Serological tests were performed to further confirm these immune responses. The results revealed that IFN-gamma and IL-6 were upregulated during the first week post-vaccination. Low level expressions of IL-1alpha, IL-1beta, TNFalpha and IL-10 and no expression of IL-2 and IL-4 were observed compared with the control camels. The findings showed that B. abortus stimulates cell-mediated immunity by directly activating camel Th1 cells to secrete IFN-gamma. This quantification of cytokine expression in camels is essential for understanding of Camelidae disease development and protective immune responses. This is the first report of in vivo camel cytokine quantification after vaccination.     

Cytokine and antibody subclass responses in the intestinal lymph of sheep during repeated experimental infections with the nematode parasite Trichostrongylus colubriformis

The expression of interleukin (IL)-4, IL-5, IL-10, IL-13, TNF-alpha and IFN-gamma genes, and parasite-specific IgM, IgG1, IgG2, IgA and total IgE levels, were monitored daily in intestinal lymph of sheep infected repeatedly with the nematode parasite Trichostrongylus colubriformis. Host genotype had a significant influence on IL-13 gene activity, with resistant-line (R) sheep consistently expressing higher levels of mRNA than susceptible-line (S) sheep. Mean gene expression of IL-13, IL-4 and IFN-gamma did not differ significantly between the first and second nematode challenge. Field-primed R and S as well as field-primed R and na?ve S sheep had lower mean gene expression of IL-5 and IL-10, respectively, during the second when compared to primary challenge. Genes for IL-13 and IL-5 were transiently and strongly up-regulated after nematode infection, particularly in animals with previous exposure to nematodes. Genes for TNF-alpha and IFN-gamma were also transiently up-regulated, but to a lesser extent and more typically after primary challenge. Na?ve sheep of both genotypes produced relatively little antibody response after primary challenge. A second nematode challenge resulted in large increases in the lymphatic levels of all antibody sub-classes which were significant for adult antigen-specific IgA and larval antigen-specific IgG1. In na?ve S line sheep, the larval-specific IgA and IgG2 response appeared delayed when compared to the R line animals. Field-primed R and S line sheep had relatively high lymphatic IgG1 levels prior to experimental infection and these did not change significantly afterwards. These results demonstrate that during nematode infections, the intestinal micro-environment of sheep is transiently skewed towards Th2 cytokine dominance, although IFN-gamma gene expression continues. This response is accompanied by increases of nematode-specific IgG1, IgA, IgG2 and IgM, as well as of total IgE in lymph plasma.     

A role for balance of interferon-gamma and interleukin-4 production in protective immunity against Neospora caninum infection

A suitable balance in the production of Th1/Th2-type cytokines has a crucial role in the control of microbial infections. We investigated cytokine production patterns and effects during Neospora caninum infection, based on two mouse models and an in vitro system. In the acute infection of N. caninum, BALB/c-background IFN-gamma-deficient mice that were sensitive to the N. caninum infection showed high levels of IL-10 production, whereas significant levels of interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) production were observed in resistant wild type mice. BALB/c mice vaccinated with recombinant vaccinia virus expressing N. caninum surface protein NcSRS2 resisted parasite spread throughout the body, low levels of IFN-gamma production and high levels of IL-4 production were observed compared to unvaccinated animals. The treatment of N. caninum-infected cells with IFN-gamma or IL-10 decreased the host-cell viability in an in vitro system using mouse macrophage J774A.1 cells. On the other hand, IL-4, but not IL-10 administration, increased the viability of N. caninum-infected and IFN-gamma-treated cells. In the light of the balance of Th1/Th2-type cytokine production, an IFN-gamma/IL-4 balance may have a crucial role for the control of cellular responses against the parasite invasion.     

L-茶氨酸对黄羽肉鸡生产性能和免疫功能的影响

本试验旨在探讨L-茶氨酸对黄羽肉鸡生产性能和免疫功能的影响。选用1日龄黄羽肉鸡690只,随机分为6个处理,每个处理5个重复,每个重复23只。6个处理分别为对照组(饲喂无抗生素基础饲粮)、抗生素组[饲喂在基础饲粮中添加10%硫酸黏杆菌素的饲粮,前期(1～3周)添加200 mg/kg,后期(4～7周)添加150 mg/kg]、4个不同梯度(100、200、400和800mg/kg)L-茶氨酸试验组,试验期为49 d。试验检测了黄羽肉鸡的生产性能、免疫器官指数、肠道分泌性免疫球蛋白A(sIgA)含量及血清溶菌酶活性、抗体水平和免疫细胞因子含量等指标。结果表明:在试验前期,100 mg/kg L-茶氨酸组黄羽肉鸡的体增重显著低于其他各组(P<0.05),100、200和800 mg/kg L-茶氨酸组黄羽肉鸡的法氏囊指数显著低于对照组(P<0.05),各L-茶氨酸组黄羽肉鸡的肠道sIgA含量和血清干扰素-γ(IFN-γ)含量显著高于对照组(P<0.05)。在试验后期,与对照组及抗生素组比较,L-茶氨酸对黄羽肉鸡生产性能、免疫器官指数、肠道sIgA的含量影响不显著(P>0.05);100、200和400 mg/kg L-茶氨酸组黄羽肉鸡血清白细胞介素-2(IL-2)的含量显著高于对照组(P<0.05),200和400 mg/kg L-茶氨酸组黄羽肉鸡血清IFN-γ的含量显著高于对照组(P<0.05)。在整个试验期,各组黄羽肉鸡血清溶菌酶的活性和白细胞介素-4的含量差异不显著(P>0.05);一免后第6天,100和200 mg/kg L-茶氨酸组黄羽肉鸡血清新城疫抗体滴度显著低于对照组和抗生素组(P<0.05),一免后第14天,100、200和400 mg/kg L-茶氨酸组黄羽肉鸡抗牛血清白蛋白(BSA)抗体水平显著高于对照组(P<0.05),其他各阶段各组间的抗BSA抗体水平和新城疫抗体滴度均差异不显著(P>0.05)。由此可知,饲粮添加L-茶氨酸对黄羽肉鸡生长后期生产性能、免疫器官指数均无显著影响,说明其对黄羽肉鸡后期生长是安全的;同时L-茶氨酸可提高黄羽肉鸡生长前期肠道sIgA的含量、后期血清IL-2的含量及全期血清IFN-γ的含量,从而增强其机体免疫功能。     

Adjuvant regulation of cytokine profile and antibody isotype of immune responses to Mycoplasma agalactiae in mice

Traditionally, adjuvants have been administered with antigens to enhance immunity. We studied the effect of several adjuvants such as Freund's complete adjuvant (FCA), Freund's incomplete adjuvant (FIA), lipopolysaccharide (LPS), homopolymers of polyinosinic-polycytidylic acid (poly I:C) and polyadenylic-polyuridylic acid (poly A:U), lithium chloride (LiCl), saponin Quil A and calcium phosphate gel (CaHPO(4)) on the immune response of mice to formalin-inactivated Mycoplasma agalactiae. The specific antibody or cytokine producing splenocytes were detected by ELISAspot and immunocytochemistry, respectively. Depending on the adjuvant given, the number of M. agalactiae-specific antibody producing cells was increased 2.5-6-fold. IgG was the major class of M. agalactiae-specific antibodies followed by IgM, IgA and IgE. Among IgG isotypes, FCA, FIA, Quil A and CaHPO(4) induced an IgG1 response with substantial increase of the IgG2a, IgG2b and IgG3 isotypes while poly I:C shifted the response toward an IgG2a/IgG3 production. Finally, poly A:U induced an IgG2b response while LPS and LiCl augmented the IgG3/IgG1/IgG2a secretion. FCA augmented IL-4, IL-5 and IL-10 production suggesting a strong Th2 response, while IFN-gamma and IL-12 remained low; poly I:C enhanced IFN-gamma, IL-12 and TNF-alpha eliciting a Th1 response; poly A:U resulted in a IL-10, IL-5, IL-6 and IL-12 secretion; and LPS enhanced the IL-10, IL-6 and TNF-alpha production. Our data show that adjuvants augment M. agalactiae-specific antibody production and lead to B cell isotype-switching via the appropriate cytokine milieu. Certain adjuvants, such as poly I:C, therefore, appear as promising immune enhancers for vaccination against M. agalactiae infections.     

Cytokine and immunoglobulin isotype profiles during CP7_E2alf vaccination against a challenge with the highly virulent Koslov strain of classical swine fever virus

CP7_E2alf is a promising marker vaccine candidate against classical swine fever (CSF). To better understand the mechanisms of protection, cytokine and isotype-specific antibody profiles were investigated in CP7_E2alf vaccinated pigs before and after challenge with the highly virulent CSFV strain “Koslov” at 14 days or 6 months post-vaccination. The interference of vaccination with CSFV pathogeny-related cytokine responses, previously described following a moderately virulent challenge, was confirmed. However, the levels of additional cytokines, TNF-α and IL-6, were significantly attenuated by vaccination following highly virulent challenge. This vaccine interference with cytokine response was not dependent on the immunization route or the consequence of competition between vaccine and challenge strain. Interestingly, IFN-γ enhancement and persistent high IgG2 levels suggested an important role of cell-mediated immunity in long-term protection against CSFV induced by CP7_E2alf vaccination. IgA production also revealed a stimulation of mucosal immunity, especially after oral administration of the vaccine.     

Antigen-specific antibody isotypes, lymphocyte subsets and cytokine profiles in cattle naturally infested by Hypoderma sp. (Diptera: Oestridae)

Antigen-specific antibody responses, T cell subsets and cytokine profiles were studied in 7 heifers naturally infested by Hypoderma sp. in Northwestern Spain. Immunoglobulin G (IgG) levels increased significantly at the end of the endogenous life cycle of the parasite (Mr). Similarly, IgG1 subclass increased considerably when first instars (L1) started their migration towards the back (Nv-Ja), whereas IgG2 increased earlier, coinciding with the arrival of L1 to the resting sites (Jn-Jl). Both subclasses decreased significantly when L3 began to leave the host. IgM levels and CD4 and CD8 profiles hardly oscillated throughout the life cycle of the parasite into the host. The CD4/CD8 ratio showed helper T cell predominance. Serum interferon-γ (IFN-γ) concentrations decreased from October to the end of the study. Interleukin 4 (IL-4) concentrations decreased in January and increased in February and May. There were a significant positive relationship between IL-4 and IgG2 subclass and a negative correlation between IFN-γ, IgG and IgG1 and also between IgM and CD2 and CD8 counts. These results suggest that in the early phases of natural primoinfestations by Hypoderma there is a slight predominance of a Th1 response, characterized by high IgG2 and IFN-γ levels, which is followed by a Th2 response with a clear predominance of IgG1 and IL-4.     

Analysis of Immune Response Induced by Recombinant Lactobacillus reuteri Expressing Cap Protein of Porcine Circovirus Type 2 in Mice

The purpose of this study was to construct a recombinant Lactobacillus reuteri (L. reuteri) expressing the cap protein of porcine circovirus type 2 (PCV2) and evaluate its effect on immune response in mice. The cap protein gene of PCV2b strain isolated and stored in the laboratory was amplified by PCR. A recombinant strain pPG-T7 g10-PPT-cap / L. reuteri expressing the cap protein was constructed using L. reuteri of pig origin as the host strain and explored the immune effect of BALB/c mice with recombinant bacteria orogastrically. Indirect ELISA was used to determine the level of antigen-specific IgG antibodies in the serum of mice after immunization, the levels of antigen-specific sIgA antibodies in stool, nasal wash, reproductive tract wash, and intestinal mucus, and the levels of various cytokines in mouse serum; MTT method was used to detect mouse spleen lymphocyte proliferation levels; flow cytometry (FCM) was used to detect the levels of CD4⁺ T cells and CD8⁺ T cells in mouse spleen lymphocytes; fluorescence quantitative PCR was used to detect the viral load of organs in challenged mice after immunization. The results showed that the serum levels of IgG antibodies in the mice of the oral immune recombinant strain group (OIG) were significantly higher than those in the control group (P<0.01); the levels of sIgA antibodies in the stool, nasal wash, reproductive tract wash, and intestinal mucus of the mice in OIG were significantly higher than those in the control group (P<0.01); Compared with the control group, the levels of cytokines in the serum of OIG were as follows: The levels of IFN-γ, IL-2, IL-4, IL-12 increased, the levels of IL-10 decreased, and the levels of IFN-α did not change significantly; Incubation of PCV2 and mouse spleen lymphocytes in vitro showed that the proliferation stimulating index of spleen lymphocytes in OIG was significantly higher than that of the control group (P<0.01); FCM results showed that CD4⁺ T cells and CD8⁺ T cells were higher than those of the control group; the results of fluorescent quantitative PCR showed that compared with the control group, the viral load in the OIG was significantly lower than that of the control group. In summary, the recombinant L. reuteri expressing the PCV2 cap protein were successfully constructed, and the constructed recombinant L. reuteri can stimulate mice to produce humoral and cellular immune responses after oragastrical immunization, and can exert a certain immune protection effect.     

表达猪圆环病毒2型cap蛋白的重组罗伊氏乳酸杆菌在小鼠诱导的免疫应答分析

旨在构建表达猪圆环病毒2型（PCV2）cap蛋白的重组罗伊氏乳酸杆菌（Lactobacillus reuteri,L.reuteri）,并评价其在小鼠体内诱导的免疫应答效果。利用PCR扩增实验室分离保存的PCV2b型毒株的cap蛋白基因,以猪源L.reuteri为宿主菌,构建表达cap蛋白的重组菌株pPG-T7 g10-PPT-cap/L.reuteri,通过口服免疫BALB/c小鼠。采用间接ELISA方法测定免疫后小鼠血清中抗原特异性IgG抗体水平,粪便、鼻腔洗液、生殖道洗液、肠黏液中抗原特异性sIgA抗体水平,小鼠血清中各细胞因子水平;MTT法检测小鼠脾淋巴细胞增殖水平;流式细胞技术检测小鼠脾淋巴细胞中CD4⁺T细胞、CD8⁺T细胞的水平;荧光定量PCR检测免疫后攻毒的小鼠体内器官的病毒载量。结果显示,口服免疫重组乳酸菌组小鼠血清IgG抗体水平显著高于对照组（P<0.01）;小鼠粪便、鼻腔洗液、生殖道洗液、肠黏液中sIgA抗体水平显著高于对照组（P<0.01）;小鼠血清中细胞因子水平和对照组相比,IFN-γ、IL-2、IL-4、IL-12水平升高,IL-10水平降低,IFN-α无显著变化;体外孵育PCV2和小鼠脾淋巴细胞结果表明,重组乳酸菌组小鼠脾淋巴细胞增殖刺激指数显著高于对照组（P<0.01）;流式细胞技术检测结果显示,口服免疫重组乳酸菌组小鼠脾细胞中CD4⁺T细胞、CD8⁺T细胞含量高于对照组;荧光定量PCR结果显示,相比于对照组,口服免疫重组乳酸菌组小鼠体内的病毒载量明显低于对照组。综上所述,本研究成功构建了表达PCV2 cap蛋白的重组罗伊氏乳酸杆菌,经口服途径免疫动物,构建的重组乳酸杆菌能够刺激小鼠产生体液免疫和细胞免疫应答,且具有一定的免疫保护效果。     

旋毛虫肌幼虫期胞外囊泡对小鼠免疫保护作用

本试验通过差速超速离心法获得旋毛虫肌幼虫期胞外囊泡(Trichinella spiralis muscle larvae extracellular vesicles,Ts-ML-EVs),经透射电镜观察、纳米颗粒追踪分析、流式细胞术和Western blot鉴定.选择6～8周龄的健康雌性BALB/c小鼠,随机分为4组...     

Disease outcome and cytokine responses in cats immunized with an avirulent feline infectious peritonitis virus (FIPV)-UCD1 and challenge-exposed with virulent FIPV-UCD8

Eight cats were immunized with an avirulent strain of feline infectious peritonitis virus (FIPV)-UCD1, then challenge-exposed to a highly virulent cat passaged strain (FIPV-UCD8). Th1 and Th2 cytokine profiles in the peripheral blood mononuclear cells (PBMCs) were measured throughout in the experiment. No clinical signs of FIP were evident in the experimental cats after immunization. After challenge, the immunized cats demonstrated one of four clinical outcomes: (1) classical effusive FIP; (2) accelerated FIP; (3) non-effusive FIP, or (4) resistance to challenge. Only minor cytokine changes were observed following immunization, however, several cytokine changes occurred following challenge-exposure. The most noteworthy changes were in tumor necrosis factor-alpha (TNF-alpha) and interferon gamma (IFN-gamma) levels. Our preliminary findings suggest that immunity against FIP is associated with TNF-alpha and IFN-gamma response imbalance, with high TNF-alpha/low IFN-gamma mRNA responses favouring disease and low TNF-alpha/high IFN-gamma mRNA responses being indicative of immunity.     

Antibody responses in pregnancy-induced transmammary transmission of Ancylostoma caninum hookworm larvae.

Third stage larvae of the Ancylostoma caninum hookworm nematode have the capacity to infect a dog, abort the normal maturation pathway to become blood-feeding intestinal worms, and instead distribute throughout the body in a developmentally arrested state that is relatively resilient to most chemotherapeutic agents. During pregnancy, a percentage of the arrested larvae reactivate and transmit via the mammary glands to infect the nursing puppies with resulting iron-deficiency anemia and potential mortality. To determine if the suppression of parasite-specific antibody responses during pregnancy facilitates the reactivation and transmammary transfer of hookworm larvae, a murine model of A. caninum infection was used to compare the infected versus uninfected animals that were either bred or not bred. Initial comparisons of genetically divergent BALB/c versus C57BL/6 mice showed that both the strains mounted strong Th2 biased IgG1and IgE antibody responses to A. caninum infection. Using the BALB/c strain for the breeding analyses, it was confirmed that larval transfer to the mouse pups only occurred during the post-partum lactational period. In the dams, levels of total and antigen-specific IgG1 and total IgE were highly correlated with parasite burden. During most phases of pregnancy and lactation, infected dams had lower total IgG1, IgG2a and IgE levels as compared to unbred mice at comparable times post-infection; this downward modulation of antibody responses supports the established dogma of a generalized immunosuppression associated with pregnancy. However, at parturition and post-partum lactation, antigen-specific IgG1 levels measured at 1 : 5000 serum dilutions were comparable between bred and unbred mice, and antigen-specific IgG2a levels at 1 : 100 serum dilutions were also not significantly different except for a marginal reduction in the bred mice at the lactational timepoint. The comparable anti-A. caninum IgG1 levels between bred and unbred mice, and low correlation between IgG2a levels and larval burden suggest that parasite-specific antibody responses do not play a major role in the pregnancy-associated transmammary transmission of A. caninum larvae. This conclusion does not rule out the possibility that underlying fluxes in the levels of specific cytokines associated with pregnancy and infection may be involved in the process of larval reactivation and transmission.     

Control of immunoglobulin isotype production by porcine B-cells cultured with cytokines

Cytokines regulate immunoglobulin (Ig) isotype production following the Th1/Th2 paradigm, derived from studies of inbred mice. In pigs, it is not known which, if any, Ig isotypes may reflect a Th1/Th2 response. To evaluate this, purified porcine CD21(+) B-cells were co-cultured with Staphylococcus aureus Cowan strain 1 or Escherichia coli lipopolysaccharide as B-cell mitogens together with recombinant human IL-2, and recombinant porcine (rp) interferon (IFN)-gamma, IL-12 or IL-10. While the mitogens increased B-cell proliferation, cytokines had no additional effect. A quantitative competitive enzyme-immuno assay was used to measure concentrations of porcine IgM, IgG(1) and IgG(2) in B-cell culture supernatants. In vitro, porcine B-cells produced IgG(2), 106 +/- 17.3 microg/ml; IgG(1) 107 +/- 38.3 microg/ml and IgM 25.6 +/- 8.45 microg/ml. In some individuals, Th1 cytokines such as rpIFN-gamma and IL-12, enhanced IgG(2) in the face of low concentrations of IgG(1). Furthermore, individual responses, in some cases, tended to be diametrically opposed, reminiscent of previously documented categorical immune responses in pigs such that some individuals produced high concentrations of IgG(1) in response to the various doses of rp cytokines, while others produced lower concentrations. Pigs may generate a high IgG(1):IgG(2) ratio in response to rpIL-10, and possibly to other Th2-associated cytokines. However, B-cell response to rp cytokines in vitro exhibits marked variation by pig, a feature that is likely a function of highly variable individual genotypes and their interaction with complex environments.     

Immunoglobulin isotypes of lactating Holstein cows classified as high, average, and low type-1 or -2 immune responders

Infectious diseases are detrimental to the health and economy of the livestock industry. Observations of cattle resistant to natural infections have implied the feasibility of breeding livestock for disease resistance. Studies of pigs selected for antibody (AMIR)- and cell (CMIR)-mediated immune responses have demonstrated increased immune responsiveness suggesting enhanced protection by both type 2 and type 1 responses, respectively. Additionally, natural or artificial infections of cattle suggest that the production of particular immunoglobulin (Ig) M, IgG1 and IgG2 isotypes are important for protecting against pathogens. In fact, IgG1/IgG2 ratios are often used to establish whether type 1 (CMIR) or type 2 (AMIR) responses predominate following immunization or infection. The objectives of this study were therefore; (1) to evaluate the Ig isotype bias responses to Candida albicans and hen-egg white lysozyme (HEWL) in cows classified as high responders (HR), average responders (AR) or low responders (LR) based on AMIR or CMIR; (2) to determine if ranking based on IFN-γ (a type 1 cytokine) and DTH responses were analogous in terms of ranking; and (3) to estimate IFN-γ, Ig isotypes, and DTH correlations. Antibody responses to HEWL and DTH to C. albicans were detected such that cows were phenotypically classified as HR, AR and LR for AMIR or CMIR with significant differences (p ≤ 0.05) among classified groups. C. albicans-induced IFN-γ allowed classification of cows, some of which had the same ranking as that of DTH response. The lowest IgG1/IgG2 ratio was to the C. albicans purified antigen (candin), but no differences were observed in anti-HEWL or anti-candin IgG1/IgG2 ratios between classified groups. Anti-HEWL IgG1 and IgG2 responses at day 21 post-immunization were negatively and significantly correlated with DTH to candin at 24h. There were no significant correlations between anti-HEWL or anti-candin IgG1 or IgG2 responses with IFN-γ. Based on Ig isotype bias, IFN-γ and DTH responses, it was concluded that immunization with C. albicans can be used to classify CMIR responder cows based on DTH read-out.