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1.
Sensitization to allergens of Japanese cedar pollen is known to cause canine atopic dermatitis as approximately 10% of atopic dogs in Japan were positive to the pollen allergen. Among the two major allergens of Japanese cedar pollen, since Cry j 1 is more important than Cry j 2 as an antigen to increase IgE in atopic dogs sensitized to Japanese cedar pollen, Cry j 1 can be a target for immunotherapy. In our study, efficacy of DNA vaccination with a plasmid containing the gene of a major allergen of Japanese cedar (Cryptomeria japnonica, CJ) pollen, Cry j 1, was examined using a dog model experimentally sensitized to CJ pollen allergen. Cry j 1 DNA plasmid and a vector plasmid (pCAGGS) were injected into six dogs and three dogs, respectively, five times with an interval of 1.5 month. After the treatment with Cry j 1 DNA plasmid, production of IgE against Cry j 1 decreased in four of the six dogs in the treatment group, whereas it increased in the three dogs of the control group. The reactivity to the pollen allergen in intradermal testing and provocation testing were obviously reduced in the treatment group, but not in the control group. The number of mast cells in alveolar area of the lung in the treatment group was smaller than that in the control group. Cry j 1 DNA plasmid was also injected into three atopic dogs sensitive to Cry j 1, resulting in improvement of clinical signs in the pollination season. These findings indicated that Cry j 1 DNA plasmid could regulate mast cell-mediated reaction against Cry j 1, which could be an alternative and effective treatment for CJ pollinosis.  相似文献   

2.
Using both in vivo and in vitro tests, dogs with atopic dermatitis were examined for sensitization with Japanese cedar (Cryptomeria japonica, CJ) pollen allergen. Ten dogs with clinical manifestation of atopic dermatitis were shown to be sensitized to CJ pollen based on the results of intradermal skin test and serum antigen-specific IgE test. In vitro lymphocyte stimulation test showed blastogenic response after stimulation with crude antigen of CJ pollen in all of the 5 cases examined. The peripheral leukocytes showed increased histamine release after stimulation with crude antigen of CJ pollen in 2 cases examined. These data indicate that a proportion of dogs with atopic dermatitis is sensitized to CJ pollen in a cell-mediated manner and show immediate phase reaction of type I hypersensitivity.  相似文献   

3.
Japanese cedar (Cryptomeria japonica, CJ) pollen has been known to cause atopic dermatitis in dogs in Japan. However, since the mechanism of the CJ antigen recognition is not well understood in dogs, it is difficult to develop effective immunotherapy for atopic dermatitis caused by sensitization to CJ pollen. In order to aim at development of a peptide immunotherapy, we tried to identify T-cell epitopes of a major allergen of CJ pollen, Cry j 1, in dogs sensitive to CJ pollen allergen. Peripheral blood mononuclear cells (PBMCs) obtained from 22 dogs experimentally sensitized to CJ pollen allergen and 5 atopic dogs sensitive to CJ pollen allergen were used for mapping of T-cell epitopes of Cry j 1 using 35 kinds of synthesized overlapping peptides of Cry j 1. Reactive peptides were identified based on the results of blastogenic responses of PBMCs against the peptides when the stimulation indices were beyond 2.0. Three reactive peptides were identical in a relatively high population of experimental dogs, which were Nos. 8 (p71-90) (41%), 10 (p91-110) (50%), and 11 (p101-120) (41%). It was considered that these synthesized peptides should contain T-cell epitopes of Cry j 1 in the dogs. However, there were no reactive peptides identical among the five atopic dogs spontaneously sensitive to CJ pollen. The population of dogs experimentally sensitized to CJ pollen antigen will be used in order to investigate effects of a peptide immunotherapy using the reactive peptides. The results in atopic dogs sensitive to CJ pollen antigen will also provide useful information on necessity to develop a tailor-made immunotherapy using reactive peptides in each dog.  相似文献   

4.
The dog is the main reservoir of Leishmania infantum, the parasite responsible for visceral leishmaniasis in Mediterranean countries. The infection in dogs shows different clinical presentations, from subclinical/asymptomatic to a fully developed disease, depending on the host's immune responses. The Th1/Th2 dichotomy is not clear in the different forms of canine leishmaniasis, since the data available from studies of immunity response in canine leishmaniasis are scarce and fragmented. The present work describes the cytokine expression in peripheral blood mononuclear cells (PBMC) obtained from asymptomatic dogs experimentally infected with L. infantum that present a cellular protective immune response. The results obtained from freshly isolated PBMC showed expressions of TNF-alpha, IL-2, IFN-gamma, IL-10 and IL-18 mRNA, similar to those from non-infected dogs. However, there was almost no expression of IL-4 mRNA detected in the asymptomatic infected dogs compared to the control dogs. Unspecific stimulation with ConA promoted the expression in a greater or lower degree of all the cytokines studied. In vitro stimulation of PBMC with soluble leishmanial antigen (SLA) promoted the expression of IL-2, IFN-gamma, TNF-alpha, IL-18, IL-4, IL-6 and IL-10 mRNA, with the two first being specifically induced. Although both Th1 and Th2 cytokines are produced, cell mediated immunity observed in these L. infantum-infected asymptomatic dogs depended on the preferential expression of Th1 cytokines.  相似文献   

5.
The natural occurrence of Japanese cedar (Cryptomeria japonica) pollinosis has been reported in dogs with atopic dermatitis. However, the reactivity to Japanese cypress (Chamaecyparis obtusa) pollen allergens in these dogs has not been reported. The present study was designed to investigate the reactivity to Japanese cypress pollen allergens in dogs sensitized to Japanese cedar pollen allergens. In 19 dogs with specific IgE to C. japonica pollen allergen, we measured the specific IgE to C. obtusa pollen allergen and examined the reactivity to the allergen by intradermal test. Of the 19 dogs, 18 had specific IgE to crude and purified major allergens (Cha o 1) of C. obtusa pollen. Most of the dogs showed a positive reaction to C. obtusa pollen allergens in the intradermal test. Allergenic cross-reactivity between Cha o 1 and Cry j 1 (a major allergen in C. japonica pollen) was observed by the ELISA inhibition method. Dogs sensitized to Japanese cedar pollen allergens demonstrate reactivity to Japanese cypress pollen allergens.  相似文献   

6.
CD80, CD86, CD28 and Cytotoxic T lymphocyte antigen-4 (CTLA-4) are well-known co-stimulatory molecules that form the major co-stimulatory pathway essential for full activation of T cells. To investigate their role in pathogenesis of immune-mediated diseases, 12 dogs were sensitized experimentally to Japanese cedar pollen antigen (CPAg) as models of allergic diseases in dogs. After sensitization, lymphocyte stimulation test (LST) was carried out to evaluate reactivity to CPAg, and semi-quantitative real-time RT-PCR analysis of CPAg-stimulated peripheral blood mononuclear cells (PBMCs) to evaluate the expression of co-stimulatory molecules. As a result, CPAg-specific enhancements of CD80 expression were detected in all sensitized dogs. Furthermore, two different kinetics of its enhancements according to the blastgenic responses to CPAg were also observed. Expression of CD28, CTLA-4 and CD86 were suppressed following CPAg-stimulation. The result of the present study indicated the potential role of the CD28-CD80 co-stimulation pathway in pathogenesis of allergic diseases in dogs.  相似文献   

7.
A cat showing seasonal allergic symptoms of rhinitis was examined for reactivities to Japanese cedar (Cryptomeria japonica, CJ) pollen allergen by intradermal skin test (IDST), Prausnitz-Kustner (P-K) test, and lymphocyte blastogenic response. In IDST for 26 common allergens. the cat showed a positive reaction to CJ pollen allergen. P-K test using CJ pollen allergen also showed a positive reaction, indicating the presence of serum IgE specific to CJ pollen. In the lymphocyte blastogenic response, the stimulation index in the presence of CJ pollen allergen was 2.4. These data suggested that the seasonal rhinitis observed in the cat was caused by the sensitization to CJ pollen allergen.  相似文献   

8.
Three dogs were examined because of episodes of recurrent pruritic dermatitis in the spring, the season of Japanese cedar (Cryptomeria japonica, CJ) pollination in Japan. The dogs were shown to be sensitive to CJ pollen allergen using intradermal testing and antigen-specific IgE measurement. Fluorometric enzyme-linked immunosorbant assay (ELISA) showed increased concentrations of IgE specific to Cry j 1 and a negative result for Cry j 2 in the three dogs. The concentrations of IgE specific to Cry j 1 during the season of CJ pollination were higher than the concentrations found during the off-season in all the dogs, and the variation in the concentrations correlated with the variation in clinical signs. Peripheral blood mononuclear cells showed apparent proliferative responses to crude CJ pollen antigen and Cry j 1 during CJ pollination season. These findings indicated that Cry j 1 was the major allergen recognized by IgE and lymphocytes and resulted in the development of type I hypersensitivity to CJ pollen allergen in these atopic dogs.  相似文献   

9.
BACKGROUND: Flea allergy dermatitis (FAD) is a common skin disease in dogs and can be induced experimentally. It often coexists with other allergic conditions. So far no studies have investigated the quantitative production of cytokine mRNA in skin biopsies and peripheral blood mononuclear cells (PBMC) in flea allergic dogs. OBJECTIVE: The aim of our study was to improve the understanding of the immunopathogenesis of allergic dermatitis as a response to fleabites. MATERIAL AND METHODS: Allergic and non-allergic dogs were exposed to fleas. Before and after 4 days of flea exposure mRNA was isolated from biopsies and PBMC. Production of chymase, tryptase, IL-4, IL-5, IL-13, TNF-alpha and IFN-gamma mRNA was measured by real-time RT-PCR. The inflammatory infiltrate in the skin was scored semi-quantitatively. The number of eosinophils, mast cells (MC) and IgE+ cells/mm2 was evaluated to complete the picture. RESULTS: FAD was associated with a higher number of MC before flea exposure and with a significant increase of eosinophils after flea exposure as compared to non-allergic dogs. The number of IgE+ cells was higher in allergic dogs before and after flea exposure. In allergic dogs mRNA for most cytokines and proteases tested was higher before flea exposure than after flea exposure. After exposure to fleas an increased mRNA production was only observed in non-allergic dogs. In vitro stimulation with flea antigen resulted in a decreased expression of most cytokines in allergic dogs before flea exposure. In contrast, in PBMC, only increased levels of IL-4 and IL-5 mRNA were observed in allergic dogs before flea exposure. However, after flea exposure and additional stimulation with flea antigen the production of mRNA for all cytokines tested was significantly increased in allergic dogs. CONCLUSION: We demonstrated that the response in biopsies and PBMC is different and that FAD is associated with a TH2 response.  相似文献   

10.
The effects of the calcineurin inhibitors cyclosporine A (CsA) and FK506 on the mRNA expressions of various cytokines were evaluated in dogs to determine whether the effects of CsA and FK506 in dogs were similar to those in humans. The mRNA expression levels of the cytokines IL-2, IL-4, IFN-gamma and TNF-alpha were measured in PHA-stimulated canine PBMC using real-time RT-PCR after incubation with CsA or FK506 for 5 hr. Both reagents inhibited IL-2, IL-4 and IFN-gamma mRNA expressions in a dose-dependent manner. However, CsA hardly inhibited the mRNA expression of TNF-alpha. These findings are important for assessing the indications of CsA treatment in dogs.  相似文献   

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13.
Japanese cedar pollinosis is a type I allergic disease mediated by immunoglobulin E (IgE) antibodies to Japanese cedar (Cryptomeria japonica) pollen antigen (CPAg). By using 22 dogs consisting of 20 dogs aged 3 months and 2 dogs aged 3 years, immunization was performed by subcutaneous injections of CPAg with aluminum hydroxide gel. Variable levels of CPAg-specific IgE antibody response were detected in 21 of the 22 immunized dogs two weeks after the second immunization. This study provided an experimental sensitization system with CPAg in dogs, which will be useful for further immunological studies on Japanese cedar pollinosis.  相似文献   

14.
Using RT-PCR and semi-quantitative PCR, mRNA expression for canine interferon (IFN)-gamma, interleukin (IL)-2, IL-4, IL-5, IL-10, tumor necrosis factor (TNF)-alpha and transforming growth factor (TGF)-beta in peripheral blood mononuclear cells (PBMCs) was examined in dogs with or without demodicosis. mRNA expression for IFN-gamma as well as TNF-alpha in dogs with demodicosis (localized (LD) and generalized (GD)) was slightly lower than those in dogs without demodicosis (healthy controls). Expression of IL-5 mRNA in dogs with demodicosis was higher than that in control dogs, but there were no significant differences in IL-4 and IL-10 mRNA expression levels among the three groups. On the other hand, expression levels of TGF-beta mRNA in dogs with GD were higher than those in control dogs and dogs with LD. The expression levels of IL-5 and TGF-beta mRNA decreased in all three dogs with GD which showed resolution of the clinical signs. Taken together, these results suggest that the Th2-like response in PBMCs from dogs with demodicosis is up-regulated, and that subsequent increased expression of IL-5 and TGF-beta mRNA in dogs with GD is reversible after treatment. Therefore, these cytokines, particularly IL-5, might be a useful clinical index of the clinical course in demodicosis. Also, increased TGF-beta mRNA expression might be a key factor for revealing the difference in the mechanism of onset between LD and GD.  相似文献   

15.
Cyclosporine A (CsA) has been widely used for suppression of transplant rejection and controlling pruritus in allergic dermatitis in humans, dogs and cats. CsA is known to suppress the expression of inflammatory cytokines, including IL-2, IL-4, IFN-gamma and TNF-alpha in humans, dogs and experimental mice. However, little is known about the immunomodulating effect of CsA in cats. The aim of this study was to evaluate the effects of CsA on the expression of inflammatory cytokines in feline peripheral blood mononuclear cells (PBMC). Real-time PCR analyses with Concanavalin A (ConA)-stimulated PBMC obtained from 5 cats revealed that the expression of mRNAs for IL-2, IL-4, IFN- gamma and TNF-alpha was inhibited by CsA in a dose-dependent manner. Moreover, an enzyme-linked immunospot (ELISPOT) assay, which is capable of detecting IL-2 secreting cells as single spots, revealed that the frequency of IL-2 secreting cells in ConA-stimulated feline PBMC was significantly reduced in the presence of CsA. These results might provide an explanation for the mechanisms of action of CsA in the suppression of transplant rejection and the control of pruritus in cats.  相似文献   

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18.
试验旨在探究UBC13蛋白对支气管哮喘小鼠体内Th2、Th17细胞极化的影响。18只BALB/c小鼠随机均分为3组:PBS组、哮喘模型(OVA)组和UBC13蛋白(UBC13)组,OVA组和UBC13组采用卵清蛋白(OVA)和脂多糖(LPS)进行致敏和雾化建立哮喘模型,其中UBC13组于每次雾化前0.5 h腹腔注射100 μg UBC13蛋白。末次雾化激发24 h后处死小鼠,采集样品,通过HE染色观察肺脏切片病理学变化、PAS染色观察气道黏液的分泌,ELISA法检测血清IgE浓度和肺支气管肺泡灌洗液(BALF)上清中IL-4、IL-5和IL-17A的水平,实时荧光定量PCR法检测肺脏Th2型相关因子IL-4、IL-5、IL-13、IFN-γ mRNA和Th17型细胞因子IL-1β、IL-6、IL-17A、IL-23 mRNA的相对表达量。结果显示,试验成功建立了哮喘模型,与PBS组相比,OVA组IgE浓度极显著上升(P < 0.01),肺脏病理切片炎性细胞浸润和中性黏液分泌现象明显,肺脏中IL-1β、IL-4、IL-6、IL-13和IL-17A mRNA相对表达量均极显著升高(P < 0.01),IL-5和IL-23 mRNA相对表达量显著升高(P < 0.05),IFN-γ mRNA相对表达量极显著下降(P < 0.01),BALF中IL-4、IL-5和IL-17A的水平极显著上升(P < 0.01);与OVA组相比,UBC13组IgE浓度显著降低(P < 0.05),病理切片炎性细胞浸润现象减轻、黏液分泌减少,肺脏IL-1β、IL-4、IL-17A、IL-13和IL-23 mRNA相对表达量均显著下降(P < 0.05),IFN-γ mRNA相对表达量显著升高(P<0.05);IL-6 mRNA相对表达量极显著下降(P < 0.01),BALF中IL-5的水平极显著下降(P < 0.01),IL-4的水平显著下降(P < 0.05),IL-17A的水平无显著差异(P > 0.05)。由此可见,UBC13蛋白能下调Th2和Th17型细胞因子的表达量,影响哮喘模型Th2和Th17细胞的极化。  相似文献   

19.
The kinetics of cytokine expression was evaluated in whole blood from high-IgE beagles previously sensitized to house dust mites (HDM) and known to develop clinical signs compatible with atopic dermatitis (AD) upon allergen exposure. Six high-IgE beagles were environmentally challenged daily for 3 h on three consecutive days with a HDM solution. Clinical signs were evaluated before, during, and after the conclusion of the challenge (days 0, 2, 4 and 17) and expression analyses of Th2 (IL-4 and IL-13) and regulatory (IL-10 and TGF-beta) cytokine mRNA were undertaken on blood samples at each time point using real-time polymerase chain reaction. Multiple comparison used to detect significant differences in clinical scores and expression levels of cytokine mRNA revealed that the clinical scores on days 2 and 4 were higher than those on days 0 and 17 (P < 0.05) but no temporal differences in the expression levels of IL-4 and IL-13 mRNA. Expression of TGF-beta mRNA was, however, significantly lower on day 4 (P < 0.05) and the expression of IL-10 mRNA on days 4 and 17 was significantly lower than those on days 0 and 2 (P < 0.05). The results indicate that allergen challenge decreases mRNA expression of regulatory cytokines in whole blood without enhanced mRNA expression of Th2 cytokines and suggest aberrant regulatory T-cell function in the immunopathogenesis of AD in high-IgE beagles.  相似文献   

20.
Recent studies suggest that asymptomatic dogs infected with canine visceral leishmaniasis (CVL) develop a Th1 immunological profile whilst oligosymptomatic and symptomatic CVL-infected animals present a Th2 profile. In the present study, an RT-PCR method has been standardised and employed to evaluate the frequency and the semi-quantitative level of expression of the cytokines IL-4, IL-10, IL-12, INF-gamma and TNF-alpha in splenocytes of 30 dogs naturally infected with Leishmania chagasi and of 7 non-infected dogs (NID). An increase in the level of expression of IL-12 (p=0.059) was detected in all CVL-infected dogs compared with NID. In dogs exhibiting high parasitism, the frequency of expression of IL-10 was higher (p=0.011) than in animals presenting low parasitism or medium parasitism (MP) and in NID animals, whilst the level of expression of IL-10 was higher (p=0.0094) than in animals exhibiting MP and in the NID group. Positive correlations between the levels of expression of IL-10 with respect to the progression of the disease (IL-10: r=0.3510; p=0.0337) and the levels of expression of IL-10 and INF-gamma increase in parasitism (IL-10: r=0.3428; p=0.0438 and INF-gamma: r=0.4690; p=0.0045) were observed. Such data suggest that CVL is marked by a balanced production of Th1 and Th2 cytokines, with a predominant accumulation of IL-10 as a consequence of an increase in parasitic load and progression of the disease, and INF-gamma was related with the increase in parasitic load.  相似文献   

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