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Real-time evaluation of cytokine and protease expression in flea-allergic and nonallergic dogs
C.Brachelente K.Wuersch M.Doherr M.Reist J. E.Peel M.Welle 《Veterinary dermatology》2004,15(S1):31-31
Atopic dermatitis (AD) is very common in dogs, but its pathogenesis is not yet fully understood. It has been suggested that a Th2-dominant status may be associated with the occurrence of canine AD. IL-12 is thought to be important for the differentiation of Th1 cells. The IL-12 receptor β2 (IL-12Rβ2) gene is considered to play a critical role in signal transduction and is attracting attention as one of the causative genes of AD in humans. The purpose of this study was to investigate the relationship between IL-12Rβ2 gene expression and canine AD. The canine IL-12Rβ2 gene was cloned by RT-PCR and its nucleotide sequences were determined. Canine IL-12Rβ2 showed 76.8% homology at the amino acid level with human IL-12Rβ2, and its structural motifs were well conserved. cDNA with a 91 bp deletion including the transmembrane region was also cloned, which consequently produced a frame shift and an early stop codon. The deletion region corresponded to exon 14 of the human IL-12Rβ2 gene on chromosome 1. The expression of deleted canine IL-12Rβ2 mRNA in phytohemagglutinin-stimulated peripheral blood mononuclear cells was examined in seven healthy dogs and 11 AD dogs. Both deleted and intact mRNAs were expressed at constant ratios in healthy and AD dogs. The results indicate that the deletion of the transmembrane region is not associated with the occurrence of AD, and that the expression of the deleted mRNA may be constitutive and produced by alternative splicing.
Funding: Self-funded. 相似文献
Funding: Self-funded. 相似文献
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This study cloned canine interleukin-12 receptor beta2 (IL-12Rbeta2). Its nucleotide sequences were determined. Canine IL-12Rbeta2 showed 85.4% homology at the nucleotide level and 76.8% homology at the amino acid level with human IL-12Rbeta2. Its structural motifs were well conserved. We also cloned cDNA with a 91-bp deletion including the transmembrane region, which produced a frame shift and an early stop codon. Examination of the expression of deleted canine IL-12Rbeta2 mRNA revealed that both deleted and intact mRNAs were expressed at a constant ratio in all the dogs. Results suggested that expression of the deleted mRNA was constitutive and produced by alternative splicing. 相似文献
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T. Iwasaki Y. Kagawa S. J. Park N. Kanazawa Y. Momoi H. Tanikawa 《Veterinary dermatology》2004,15(Z1):5-5
Atopic dermatitis (AD) is thought to be caused by immunologic abnormalities expressed as a Th1/Th2 cytokine imbalance in both humans and dogs. Several studies have focused on the therapeutic effects of IFNγ in human AD with successful results; however, the mechanism of action of IFNγ is not fully understood. We investigated the effect of recombinant canine interferon gamma (rCaIFNγ) on 10 dogs with AD and evaluated the ratio of IL‐4 mRNA to IFNγ mRNA in peripheral blood mononuclear cells, serum total IgE levels, and histological changes in skin. After six injections of rCaIFNγ over a span of 2 weeks, seven of the 10 dogs showed improvement, and six of these seven dogs exhibited decreased IL‐4:IFNγ mRNA ratios. Two of the three cases that did not improve had increased IL‐4:IFNγ mRNA ratios. Total serum IgE levels were significantly decreased in nine of 10 cases. The number of IgE‐positive cells detected by immunostaining and the number of mast cells in skin biopsy samples were decreased. A reduction of epidermal cell layers was demonstrated by histopathology after treatment. These results demonstrated that rCaIFNγ may be a novel safe and effective therapeutic option for the treatment of canine AD, and the mechanism of action of rCaIFNγ may be related to the modulation of Th2 cytokines to Th1 cytokines with the reduction of serum IgE production. Funding: Self‐funded. 相似文献
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Andrea J. Gonzales William R. Humphrey James E. Messamore Timothy J. Fleck Gregory J. Fici John A. Shelly Janet F. Teel Gary F. Bammert Steven A. Dunham Troy E. Fuller Robert B. McCall 《Veterinary dermatology》2013,24(1):48-e12
Background – Interleukin‐31 (IL‐31) is a member of the gp130/interleukin‐6 cytokine family that is produced by cell types such as T helper 2 lymphocytes and cutaneous lymphocyte antigen positive skin homing T cells. When overexpressed in transgenic mice, IL‐31 induces severe pruritus, alopecia and skin lesions. In humans, IL‐31 serum levels correlate with the severity of atopic dermatitis in adults and children. Hypothesis/Objective – To determine the role of IL‐31 in canine pruritus and naturally occurring canine atopic dermatitis (AD). Animals – Purpose‐bred beagle dogs were used for laboratory studies. Serum samples were obtained from laboratory animals, nondiseased client‐owned dogs and client‐owned dogs diagnosed with naturally occurring AD. Methods – Purpose‐bred beagle dogs were administered canine interleukin‐31 (cIL‐31) via several routes (intravenous, subcutaneous or intradermal), and pruritic behaviour was observed/quantified via video monitoring. Quantitative immunoassay techniques were employed to measure serum levels of cIL‐31 in dogs. Results – Injection of cIL‐31 into laboratory beagle dogs caused transient episodes of pruritic behaviour regardless of the route of administration. When evaluated over a 2 h period, dogs receiving cIL‐31 exhibited a significant increase in pruritic behaviour compared with dogs that received placebo. In addition, cIL‐31 levels were detectable in 57% of dogs with naturally occurring AD (≥13 pg/mL) but were below limits of quantification (<13 pg/mL) in normal, nondiseased laboratory or client‐owned animals. Conclusions – Canine IL‐31 induced pruritic behaviours in dogs. Canine IL‐31 was detected in the majority of dogs with naturally occurring AD, suggesting that this cytokine may play an important role in pruritic allergic skin conditions, such as atopic dermatitis, in this species. 相似文献
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Maeda S Okayama T Omori K Masuda K Sakaguchi M Ohno K Tsujimoto H 《Veterinary immunology and immunopathology》2002,90(3-4):145-154
CC chemokine receptor 4 (CCR4) is a G protein-coupled seven transmembrane receptor that is selectively expressed on Th2 cells and plays an important role in the trafficking of Th2 cells into inflammatory sites. In this study, a full-length canine CCR4 cDNA was cloned and characterized in order to examine the potential role of CCR4 in allergic responses that produce skin lesions in canine atopic dermatitis (AD). The canine CCR4 cDNA reported in this study contained an open reading frame of 1083 nucleotides encoding 360 amino acids. The predicted amino acid sequence of canine CCR4 showed 91.9, 85.3 and 84.5% similarity with those of the human, mouse and guinea pig counterparts, respectively. Expression of CCR4 mRNA was detected in various tissues including thymus, spleen, heart, small intestine and lymph node. Furthermore, it was found that CCR4 mRNA was preferentially expressed in lesional skin of dogs with AD, together with the mRNA of thymus and activation-regulated chemokine (TARC), which is a ligand for CCR4. The present study demonstrates that CCR4 contributes strongly to the immunopathogenesis of canine AD. 相似文献
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Sarah J. Deitschel DVM Marie E. Kerl DVM DACVIM DACVECC Chee‐Hoon Chang DVM Amy E. DeClue DVM MS DACVIM 《Journal of Veterinary Emergency and Critical Care》2010,20(5):494-502
Objective – To determine whether older dogs will have a more pronounced pro‐inflammatory response and blunted anti‐inflammatory response to pathogen‐associated molecular patterns (PAMPs) compared with younger dogs. Design – Prospective. Setting – University teaching hospital. Animals – Thirty‐eight privately owned sexually altered dogs of various ages. Interventions – Blood was collected for HCT, WBC count, plasma biochemical analysis, and whole blood culture. Whole blood was stimulated with lipopolysaccharide (LPS) or, lipoteichoic acid or, peptidoglycan or, addition of phosphate‐buffered saline. Tumor necrosis factor (TNF), interleukin (IL)‐6, and IL‐10 production from whole blood were compared among young, middle aged, and geriatric dogs. Measurements and Main Results – LPS, lipoteichoic acid, and peptidoglycan stimulated significant TNF, IL‐6, and IL‐10 production from canine whole blood compared with phosphate‐buffered saline. Whole blood from geriatric dogs had a blunted IL‐10 response to LPS stimulation and middle‐aged dogs had increased LPS‐induced TNF production compared with the other groups. Conclusion – PAMPs from gram‐positive and gram‐negative bacteria stimulate TNF, IL‐6, and IL‐10 production from canine whole blood. The inflammatory mediator response to PAMPs from gram‐negative bacteria alters with age and may be one factor contributing to mortality in geriatric dogs with sepsis. 相似文献
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AP Herman T Misztal A Herman Dorota Tomaszewska – Zaremba 《Reproduction in domestic animals》2010,45(6):e426-e433
This study was designed to determine the effect of intravenous lipopolysaccharide (LPS) administration on the secretion of interleukin (IL)‐1β and IL‐1 receptors (IL‐1Rs) gene expression in the hypothalamus of anoestrous ewes. Gene expression of IL‐1β and its receptors was assayed by the real‐time polymerase chain reaction. The expression of IL‐1β in the hypothalamus was detected using Western blot. Our results showed that IL‐1β mRNA is transcribed in the ovine hypothalamus. Lipopolysaccharide increased (p ≤ 0.01) the IL‐1β gene expression in the pre‐optic area 2.4‐fold, the anterior hypothalamus (AHA) 3.4‐fold, the medial basal hypothalamus 3.7‐fold and the medial eminence 3.9‐fold. The pro‐form and mature form of IL‐1β protein were found in the hypothalamus after endotoxin injection. In general, the endotoxin also increased more than two times (p ≤ 0.01) the expression of IL‐1 receptor type I (IL‐1R1) and type II (IL‐1R2) genes in the hypothalamus, except the AHA, where the number of IL‐1R2 mRNA was extremely low and not sufficient to the quantitative analysis. These results demonstrate that the peripheral immune/inflammatory challenge increases the IL‐1β expression in the hypothalamus. This endogenous IL‐1β seems to be involved in the modulation of processes which are regulated at the hypothalamic level. One of these processes could be a reproduction. 相似文献
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Jolanta Klukowska‐Rötzler Ludovic Chervet Eliane J. Müller Petra Roosje Eliane Marti Jozef Janda 《Veterinary dermatology》2013,24(1):54-e14
Background – In humans, thymic stromal lymphopoietin (TSLP) plays a central role in the development of allergic inflammation, such as atopic dermatitis (AD), but it is unknown whether it is involved in the pathogenesis of canine AD (CAD). Hypothesis/Objectives – Our aim was to characterize canine TSLP and to assess its expression in CAD. Methods – Canine TSLP was identified based on sequence homology with human TSLP and the complementary DNA (cDNA) cloned by RT‐PCR. Real‐time quantitative RT‐PCR was established to assess the expression of canine TSLP in cultured canine keratinocytes and in skin biopsy specimens from lesional and nonlesional skin of 12 dogs with CAD and eight healthy control dogs. Results – Partial canine TSLP cDNA was cloned and characterized. It contained four exons that shared 70 and 73% nucleotide identity with human and equine TSLP, respectively, encoding the signal peptide and full‐length secreted protein. We found significantly increased TSLP expression in lesional and nonlesional skin of dogs with CAD compared with healthy control dogs (P < 0.05), whereas no difference was measured between lesional and nonlesional samples. In cultured primary canine keratinocytes, we found increased TSLP expression after stimulation with house dust mite allergen extract or Toll‐like receptor ligands lipopolysaccharide and poly I:C. Conclusions and clinical importance – Increased TSLP expression in the skin of dogs with CAD supports an involvement of TSLP in the pathogenesis of CAD similar to that in humans. Further studies should elucidate the function and therapeutic potential of TSLP in CAD. 相似文献
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Efficacy and safety of electrochemotherapy combined with peritumoral IL‐12 gene electrotransfer of canine mast cell tumours 
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下载免费PDF全文 M. Cemazar J. Ambrozic Avgustin D. Pavlin G. Sersa A. Poli A. Krhac Levacic N. Tesic U. Lampreht Tratar M. Rak N. Tozon 《Veterinary and comparative oncology》2017,15(2):641-654
Electrochemotherapy combined with peritumoral interleukin‐12 (IL‐12) gene electrotransfer was used for treatment of mast cell tumours in 18 client‐owned dogs. Local tumour control, recurrence rate, as well as safety of combined therapy were evaluated. One month after the therapy, no side effects were recorded and good local tumour control was observed with high complete responses rate which even increased during the observation period to 72%. IL‐12 gene electrotransfer resulted in 78% of patients with detectable serum IFN‐γ and/or IL‐12 levels. In the treated tumours vascular changes as well as minimal T‐lymphocytes infiltration was observed. After 1 week, the plasmid DNA was not detected intra‐ or peritumorally and no horizontal gene transfer was observed. In summary, our study demonstrates high antitumour efficacy of electrochemotherapy combined with IL‐12 electrotransfer, which also prevented recurrences or distant metastases, as well as its safety and feasibility in treatment of canine mast cell tumours. 相似文献
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Osteosarcoma (OSA) is the most common bone tumour in humans and companion animals, and has a poor long‐term prognosis. The identification of new markers and targeted therapies may help increase long‐term survival of these patients. Previous studies have demonstrated that interleukin‐11 receptor alpha (IL‐11Rα) is expressed in human and murine OSA but not in normal bone. The current study demonstrated via western analysis, immunoflourescence and immunohistochemistry that IL‐11Rα was expressed in primary canine OSA tissues as well as in a number of canine OSA cell lines, but not in normal canine bone. Cytotoxin‐conjugated antibodies targeting IL‐11Rα‐mediated canine OSA cytotoxicity. Thus, canine OSA may be a valuable model for the evaluation of IL‐11Rα directed therapies. 相似文献
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C. Pucheu‐Haston D. Shuster T. Olivry P. Brianceau P. Lockwood T. Mcclanahan R. De Waal Malefyt J. Mattson B. Hammerberg 《Veterinary dermatology》2004,15(Z1):38-38
IgE‐mediated late‐phase reactions can be induced in the skin of normal and atopic dogs by intradermal injections of anti‐IgE antibody. The histology of these reactions is very similar to that of naturally occurring atopic dermatitis. To characterize the cellular, cytokine and chemokine responses in the skin of placebo‐ and prednisolone‐treated dogs, normal beagles received either placebo or 0.5 mg/kg prednisolone twice daily for three days prior to intradermal injection of polyclonal rabbit anti‐canine IgE. Eight‐millimetre punch biopsy skin samples were taken before injection and at the injection sites after 6, 24 and 48 h. Histological and immunohistochemical examination revealed a rapid cellular influx. Eosinophil and neutrophil numbers increased from <1 to 61.4 ± 14.1, and from 7 to 62.2 ± 10.8 cells/mm2, respectively, within 6 h after injection, and remained moderately elevated 48 h later. The numbers of CD1c+, CD3+ and CD4+ mononuclear cells were also increased by 6 h. Taqman analysis demonstrated 2.5‐ to 72‐fold increases in mRNA expression for IL‐13, IL‐5, MCP (CCL2), RANTES (CCL5) and TARC (CCL17). Levels of mRNA for IL‐2, IL‐4, IL‐6, and IFNγ remained negligible. Prednisolone administration suppressed the influx of neutrophils and eosinophils, and the expression of IL‐13, CCL2, CCL5 and CCL17 (33, 97, 58, 86, 73 and 90%, respectively), as well as the influx of CD1c+ and CD3+ cells. These data document the cytokine and chemokine response to anti‐IgE injection and demonstrate the anti‐inflammatory effect of prednisolone. Funding: Schering‐Plough Animal Health. 相似文献
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The purpose of this study was to evaluate a combination of immunostimulatory bacterial DNA sequences and allergen‐specific immunotherapy for the treatment of canine atopic dermatitis. Seven dogs with nonseasonal atopic dermatitis diagnosed by history, clinical signs and exclusion of differential diagnoses were included. All dogs had been on allergen‐specific immunotherapy for at least 12 months with incomplete responses, were on additional antipruritic therapy and showed residual pruritus. Pruritus was marked by the owner on a visual analogue scale, lesions were determined by a clinician using the Canine Atopic Dermatitis Extent and Severity Index (CADESI), and concurrent medications were recorded before entering the study and after 14 weeks of treatment. Peripheral blood mononuclear cells were isolated and cultured; canine cytokine message for IFNγ, IL‐4, TNF and IL‐10 was quantitated using RT‐PCR. A mixture of allergen extract and liposome‐DNA complexes was injected intradermally at the beginning of the study and after 2, 4, 6, 10 and 14 weeks. CADESI, pruritus and medication scores, and cytokine messages at the beginning and end of the study were compared with a paired t‐test. There were significant improvements in pruritus scores (P = 0.0277). Reductions in medication scores and CADESI were not statistically significant. IL‐4 production decreased significantly (P = 0.0428); decreases in other cytokines were not significant. Although the number of dogs in this pilot study was small, the results warrant further investigation of a combination of immunostimulatory bacterial DNA sequences and allergen‐specific immunotherapy for the treatment of canine atopic dermatitis. Funding: Self‐funded. 相似文献
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β‐carotene is one of the most abundant carotenoids, has potential anti‐inflammatory effect, it has been reported that β‐carotene could suppress LPS‐induced inflammatory responses by inhibiting nuclear factor kappa B (NF‐κB) translocation, but the more detailed molecular mechanisms underlying the anti‐inflammatory action of β‐carotene remain to be fully understood. In this study, we investigated the influence of β‐carotene on the activation of JAK2/STAT3, MAPK, and NF‐κB signaling pathway induced by LPS in RAW264.7 cells and peritoneal macrophages. Cells were treated with different concentrations of β‐carotene for 3 hr after LPS treatment for 24 hr. The mRNA expression and the release of IL‐1β, IL‐6, and TNF‐α were evaluated by RT‐PCR and ELISA, and the level of signaling proteins of JAK2/STAT3, MAPK, and NF‐κB signaling pathway were detected by Western blot. The results showed that β‐carotene significantly suppressed (p < 0.05) LPS‐induced release of IL‐1β, IL‐6, and TNF‐α and their mRNA expression. LPS‐induced JAK2/STAT3, IκB/NF‐κB p65, JNK/p38 MAPK signal activation were significantly attenuated (p < 0.05) by β‐carotene in a dose‐dependent manner. In conclusion, β‐carotene could attenuate LPS‐induced inflammation via inhibition of the NF‐κB, JAK2/STAT3, and JNK/p38 MAPK signaling pathways in macrophages. 相似文献
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J. M. Jankovsky K. M. Newkirk M. R. Ilha S. J. Newman 《Veterinary and comparative oncology》2013,11(1):63-69
Gliomas are among the most common primary neural tumours of dogs. Cyclooxygenase‐2 (COX‐2) and c‐kit overexpression are associated with increased aggressiveness of gliomas and decreased survival in human beings. COX‐2 is the inducible form of cyclooxygenase, which catalyzes prostaglandin formation and may increase tumour proliferation and angiogenesis. C‐kit is a tyrosine kinase receptor involved in normal cell physiology; c‐kit is upregulated in some canine tumours. In this retrospective study, 20 canine gliomas were identified: 11 (55%) oligodendrogliomas, including 1 anaplastic variant; 1 (5%) oligoastrocytoma; and 8 (40%) astrocytomas, of which 2 were glioblastoma multiforme. None of the gliomas expressed COX‐2. None of the gliomas were immunoreactive for c‐kit, although all three high‐grade tumours had intramural vascular expression. Consequently, COX‐2 inhibitors would likely be ineffective against canine gliomas. C‐kit inhibitors may have an anti‐angiogenic effect in high‐grade gliomas, but would likely be ineffective in low‐ and medium‐grade tumours. 相似文献