共查询到20条相似文献，搜索用时 15 毫秒
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Abdalla SA Horiuchi H Furusawa S Matsuda H 《Veterinary immunology and immunopathology》2004,98(1-2):31-41
Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) were identified as signal transducers for the tumor necrosis factor receptor (TNFR) superfamily. In this study, we cloned and characterized two genes that encode chicken TNFR-II and TRAF5. The initial cDNA fragments were obtained by suppressive subtractive hybridization (SSH) of chicken spleen cells with or without lipopolysaccharide stimulation (Salmonella typhimurium SL1181 (RE-mutant)). The results showed that chicken TNFR-II is 1518 bp in length with an open reading frame (ORF) of 1386 bp having 31% homology with human TNFR-II. Expression analysis of chicken TNFR-II revealed that it is highly expressed in the spleen and bursa of Fabricius. The chicken cell lines IN24, MSB1 and 1104B express TNFR-II abundantly. The time course analysis of expression in spleen, bursa of Fabricius and IN24 cell line showed that TNFR-II is maximally expressed at 6 h after stimulation in bursa of Fabricius and after 8 h stimulation in the IN24 cell line. With regard to TRAF5, the complete sequence was 1936 bp in length with an ORF of 1671 bp that showed 71.3% homology with human TRAF5. Expression analysis showed that, among the tissues examined, TRAF5 was strongly expressed in spleen and bursa of Fabricius, while among the cell lines examined, it was maximally expressed in IN24. Thus, both genes were expressed in the same tissues and cell line among examined materials. These results suggest that chicken TNFR-II may interact with TRAF5 adaptor protein to complete its signal transduction pathway. 相似文献
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Lee SH Lillehoj HS Park MS Baldwin C Tompkins D Wagner B Del Cacho E Babu U Min W 《Comparative immunology, microbiology and infectious diseases》2011,34(3):3818-279
This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD80 (chCD80). A recombinant plasmid containing a chCD80/horse IgG4 fusion gene was constructed and expressed in CHO cells to produce recombinant chCD80/IgG4 protein. Chicken CD80 was purified from the chCD80/IgG4 fusion protein following enterokinase digestion, and used to immunize BALB/c mice, resulting in 158 hybridomas that produced mAbs against chCD80. Three mAbs with high binding specificity for recombinant chCD80/IgG4-transfected CHO cells were identified by flow cytometry, and one of these (#112) was selected for further characterization. Immunoprecipitation of CD80/IgG4-CHO cell extract, or lipopolysaccharide (LPS)-treated monocytes identified 35.0 kDa proteins. Immunohistochemical analysis revealed chCD80-expressing cells exclusively in the bursal follicles at the outer portion of the cortex, and throughout the red pulp and the outer boundary of the white pulp in the spleen. By immunofluorescence microscopy, chCD80 was observed on intestinal dendritic cells. LPS treatment of bursa or spleen monocytes for 24 or 48 h increased chCD80 expression. Finally, addition of chCD80 mAb to Con A-stimulated spleen cells inhibited the expression of major histocompatibility complex class II antigens and IL-2-driven proliferation of lymphoblast cells. In summary, these chCD80 mAbs will serve as valuable immunological reagents for basic and applied poultry immunology research. 相似文献
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法氏囊是鸟类特有的器官,是B淋巴细胞成熟的场所.本研究分析了鸡胚早期发育过程的中法氏囊形态变化及增殖细胞核抗原PCNA表达变化.结果表明鸡胚6.5 d(E6.5)时,法氏囊开始出现在鸡胚泄殖腔的最深处,从最开始E6.5的一些小的囊泡发育成E8的一个大腔,E8～E10,法氏囊继续发育.从E11开始,法氏囊开始出现一些小的皱褶,E11～E18,这些小的皱褶逐渐变得清晰,并贯穿整个法氏囊.在E9时,法氏囊轮廓形成,可以发现在表皮层和基膜层PCNA有着广泛的表达.统计分析表明表皮层的PCNA阳性率在100％左右,在整个发育早期几乎没有变化；而基层的阳性率随着胚胎的发育逐渐下降,在E9、E10、E11、E12、E14和E18表达率分别是97％、93％、71％、55％、56％和20％. 相似文献
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应用免疫组化超敏SP法对鸡不同生长阶段免疫器官中血管活性肠肽(VIP)的表达特点进行了研究,结果显示,胸腺中VIP阳性反应主要分布于被膜及髓质,而皮质内反应较弱;随日龄增加,VIP在被膜和髓质内一直维持较强的表达;实质中VIP强阳性淋巴细胞数量随日龄逐渐增多,至42 d开始减少.脾脏中VIP阳性反应主要出现于被膜及红髓,并在整个生长阶段维持较强的表达,红髓的脾索和脾窦间还有较多的巨噬细胞呈VIP强阳性反应;白髓中仅见动脉周围淋巴鞘中央动脉有VIP的表达,并随日龄的增加逐渐减弱.法氏囊中VIP的表达范围广泛,涉及黏膜、黏膜下层和肌层,尤以黏膜上皮细胞和法氏囊小结髓质的表达为典型;随着日龄增加,黏膜上皮细胞VIP表达逐渐减弱,囊小结髓质一直保持较强的VIP阳性着色,囊小结皮髓质交界处上皮细胞层内VIP强阳性细胞有逐渐增多的趋势.鸡免疫器官中VIP阳性反应物质的广泛分布,提示这3种器官是神经系统以外产生VIP的重要部位,法氏囊中的VIP可参与鸡免疫器官的功能调节,它可作为一种信号分子介导神经内分泌系统和免疫系统之间的相互作用. 相似文献
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为研究鸡冷诱导RNA结合蛋白(CIRP)基因的结构特征和组织定量分布情况,根据GenBank中原鸡CIRP基因(NM-001031347)的编码区序列自行设计引物,从健康雏鸡脾脏中扩增青脚麻羽鸡CIRP基因的编码区(CDS)并进行生物信息学分析;设计引物建立实时荧光定量RT-PCR,检测CIRP mRNA在雏鸡各组织器官中的分布及相对表达水平。结果显示,鸡CIRP基因CDS区全长573bp,编码190个氨基酸,其氨基端包含一个RNA结合域,羧基端富含RGG重复序列,推导的多肽链羧基端比鸭、斑胸草雀及哺乳动物要多18个氨基酸;CIRP基因在鸡各被检组织器官中均有表达,相对表达水平由低到高分别为法氏囊、肺脏、胰腺、胸腺、脾脏、睾丸、盲肠扁桃体、心脏、脑、肾脏、哈德氏腺、肝脏。本研究为进一步探讨鸡CIRP基因的遗传进化及其在病原感染中的作用提供了参考。 相似文献
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目的:研究血管活性肠肽(VIP)在痛风模型鸡免疫器官中的表达情况,从而探讨VIP在鸡痛风中的作用。方法:采用免疫组织化学(SABC)的方法,分别检测20只正常鸡和两组各20只由不同日粮所建立的痛风模型鸡的胸腺、脾脏、法氏囊中VIP的表达情况,并比较其差异。结果:高钙日粮组和高钙高蛋白日粮组模型鸡免疫器官中VIP的表达比正常组高(P<0.05),高钙日粮组与高钙高蛋白日粮组模型鸡免疫器官中VIP的表达无明显差异(P>0.05)。结论:高钙日粮组和高钙高蛋白日粮组模型鸡免疫器官中有较高VIP的表达,推测VIP作为一种信号肽介导了免疫系统和神经内分泌系统间的相互作用,并与鸡痛风的发生可能有密切关系。 相似文献
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Isolation and characterization of a reovirus from common eiders (Somateria mollissima) from Finland 总被引：3，自引：0，他引：3
Samples of brain, intestine, liver, lung, spleen, and bursa of Fabricius were collected from five common eider (Somateria mollissima) duckling carcasses during a die-off in the western Gulf of Finland (59 degrees 50'N, 23 degrees 15'E) in June 1996. No viral activity was observed in specific-pathogen-free chicken embryos inoculated with tissue suspensions, but samples of bursa of Fabricius from three birds were positive when inoculated into Muscovy duck (Cairina moschata) embryo fibroblasts. The isolates were characterized as nonenveloped RNA viruses and possessed several characteristics of the genus Orthoreovirus. Virus particles were icosahedral with a mean diameter of 72 nm and were stable at pH 3.0; their genome was separated into 10 segments by polyacrylamide gel electrophoresis. Mallard (Anas platyrhynchos) ducklings experimentally infected with the eider reovirus showed elevated serum activities of aspartate aminotransferase, creatine kinase, and lactate dehydrogenase enzymes and focal hemorrhages in the liver, spleen, and bursa of Fabricius. During 1997-99, the prevalence of neutralizing antibodies to the isolated virus ranged from 0 to 86% in 302 serum samples collected from incubating eider hens at three nesting areas along coastal Finland. The highest seroprevalence was found in Hanko in 1999, just weeks before reports of an uninvestigated mortality event resulting in the death of an estimated 98% of ducklings at that location. These findings raise the question of potential involvement of the virus in poor duckling survival and eider population declines observed in several breeding areas along coastal Finland since the mid-1980s. 相似文献
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建立并利用鸡白细胞介素18(Chicken interleukin-18,ChIL-18)荧光定量RT-PCR方法,对禽网状内皮组织增生症病毒(Reticuloendotheliosis virus,REV)感染肉鸡后主要免疫器官的IL-18 mRNA转录水平进行了初步研究.结果表明,与对照组相比,处理组脾脏、胸腺和法氏囊中IL 18的分泌水平在感染后7和14 d均显著升高(P＜0.05),在感染21、28、35和42d表达差异均有显著变化(P＜0.05).本研究为探讨REV感染的免疫抑制机理奠定了一定基础. 相似文献
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Dan WB Guan ZB Zhang C Li BC Zhang J Zhang SQ 《Veterinary immunology and immunopathology》2007,118(1-2):113-120
B-cell activating factor (BAFF), belonging to the TNF family, is critical for B cell survival and maturation. cDNA of goose BAFF (gBAFF) was amplified from goose spleen by RT-PCR. The open reading frame (ORF) of gBAFF encodes a protein of 288-amino acid. The gBAFF shows 98, 92, 44 and 55% amino acid sequence identity with duck (dBAFF), chicken (cBAFF), mouse (mBAFF) and human BAFF (hBAFF), respectively. RT-PCR results showed that gBAFF mRNA is expressed in thymus and more highly expressed in the bursa of Fabricius and spleen. Recombinant soluble gBAFF (gsBAFF) expressed in Escherichia coli has molecular weight of approximately 19kDa. In vitro, purified gsBAFF was able to promote bursa B cells survival/proliferation in goose, duck and chicken. Furthermore, recombinant dsBAFF and csBAFF have a positive effect on goose, duck and chicken bursa B cells survival/proliferation. These findings indicate that gBAFF plays an important role in the survival/proliferation of goose B cells and, owing to its high evolutionary conservation, functional cross-reactivity exists between chicken, duck and goose BAFF. 相似文献
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本研究旨在通过荧光定量PCR方法研究禽白血病J亚群病毒(ALV-J)在家禽体内各个器官的分布。根据ALV-J NX0101毒株基因5 258—5 510bp的碱基序列,设计了1对特异性引物,进行RT-PCR反应,扩增产物为253bp,将该产物连接T载体作为Real-time PCR反应的标准品,利用SYBR Green I染料进行Real-time PCR反应,建立了标准曲线,并进行反应灵敏性,特异性和重复性试验。然后用已建立的方法对人工感染ALV-J的SPF鸡心脏、肝脏、脾脏、肺脏、肾脏、胸腺、法氏囊、腺胃进行3次重复检测,将Ct值带入标准曲线公式得出各个组织的病毒拷贝数。试验结果表明:标准曲线线性关系R值均为0.991 4,检测极限约为81拷贝质粒DNA,比RT-PCR灵敏100倍以上;特异性分析表明只有ALV-J能检测到特异性的熔解度峰值;批内和批间重复性试验的变异系数均小于5%。通过比较数值以得出胸腺ALV-J基因含量是最高的,肺脏、脾脏、法氏囊含量也比较高,心脏拷贝数最低。自然感染发病鸡各器官ALV-J基因均高于人工感染ALV-J的基因含量。本研究所建立的ALV-J基因实时荧光定量RT-PCR方法灵敏度高、特异性强、检测周期短,初步探讨了禽白血病J亚群病毒在畜禽体内各个器官的病毒分布。这为以后禽白血病的诊断以及治疗提供了重要的技术支持。 相似文献
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The bursa of Fabricius is critical for the normal development of B lymphocytes in avian species. Productive colonization of bursal follicles by B cell precursors requires surface immunoglobulin expression. We have shown using retroviral gene transfer that expression of chimeric receptors containing the extracellular and transmembrane domains of murine CD8alpha and CD8beta fused to the cytoplasmic domains of chicken Igalpha and Igbeta can support productive bursal colonization in the chicken embryo in bursal cells lacking the expression of endogenous sIgM. We show here that chimeric receptor expression does not support continued bursal cell development after hatch. However intrabursal administration of anti-CD8 antibodies that ligate the CD8alpha:Igalpha chimeric receptor results in maintained numbers of bursal cells that express the chimeric receptor in the absence of endogenous sIgM. These results support a model in which sIgM receptor expression is required for productive bursal colonization in the chick embryo but sIgM receptor ligation is required to support later B cell development after hatch. 相似文献
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Lee SH Lillehoj HS Jang SI Baldwin C Tompkins D Wagner B Parcells M Del Cacho E Hong YH Min W Lillehoj EP 《Veterinary immunology and immunopathology》2011,144(3-4):396-404
This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD25 (chCD25), the alpha chain of the interleukin-2 (IL-2) receptor. A recombinant chimeric chCD25/IgG4 fusion protein was expressed in Chinese hamster ovary (CHO) cells and isolated from spent cell culture medium by protein G affinity chromatography. Purified chCD25 protein was used to immunize mice, from which 54 stable hybridomas secreting chCD25 mAbs were produced. Two mAbs, chCD25-32 and chCD25-54, with high binding affinity for chCD25-expressing CHO cells were selected for further characterization. By flow cytometry, both mAbs detected cells in the spleen, bursa of Fabricius, intestinal duodenum, and immunostained established chicken T cell, B cell, and macrophage cell lines. Both mAbs reacted with a 55 kDa protein on Western blots of lysates from concanavalin A (Con A)-stimulated spleen mononuclear cells. Intraperitoneal injection of chickens with bacterial lipopolysaccharide increased the percentage of chCD25(+) spleen cells by approximately 4-fold compared with untreated animals. In vitro stimulation of spleen cells with Con A increased the percentage of chCD25(+) cells by up to 50-fold compared with cells treated with medium alone. Finally, the chCD25-32 mAb suppressed IL-2-driven spleen cell proliferation and reduced IL-2-induced nitric oxide production. These mAbs may be useful for future investigation of chicken regulatory T cells. 相似文献
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Pigeon circovirus was identified by polymerase chain reaction (PCR) in young pigeons belonging to 12 different lofts. Viral DNA was extracted from formalin-fed, paraffin-imbedded tissues containing primarily bursa and occasionally liver and spleen with a commercial kit. PCR primers were selected from a published sequence for columbid circovirus and evaluated in a PCR assay. The histopathologic examination of various tissues revealed basophilic globular intracytoplasmic inclusions in the mononuclear cells of the bursa of Fabricius and occasionally in the spleen characteristic for a circovirus. Transmission electron microscopy of a few bursas of Fabricius revealed virus particles measuring 18-21 nm. All the samples were negative by PCR for psittacine beak and feather disease (PBFD) virus and chicken infectious anemia virus. The primers for both pigeon circovirus and PBFD virus did not react in PCR with the chicken anemia virus DNA. Most of the circovirus-infected pigeons had concurrent infections of Escherichia coli, Salmonella, Pasteurella, Aspergillus, candidiasis, nematodiasis, or capillariasis. 相似文献
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采用马齿苋多糖口腔灌注雏鸡,观察对雏难法氏囊免疫功能的影响。试验选取80只1日龄雏鸡随机分为4组,1个空白对照组及3个POP试验组,POP剂量分别为25,50和100mg/kg,每天口腔灌注1次,连续20d。结果显示,POP能显著提升外周血CD4+T淋巴细胞数量及CD4+/CD8+比值(P0.05),显著提高法氏囊指数(P0.05)及法氏囊淋巴细胞增殖指数(P0.05)。试验表明,马齿苋多糖可通过改变法氏囊免疫系统的动态变化,促进免疫器官发育,直接作用于免疫细胞,提高雏鸡的免疫能力。 相似文献
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Islam MR Johne R Raue R Todd D Müller H 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2002,49(7):332-337
Chicken anaemia virus (CAV) was detected in the bursa of Fabricius of a 4-week-old chicken obtained from an outbreak of acute infectious bursal disease in Bangladesh. Repeated attempts to grow this virus in MDCC-MSB1 cells were not successful. A full-length PCR amplicon of the genome of this strain, designated as BD-3 CAV, was cloned and sequenced. The complete nucleotide sequence and the deduced amino acid sequence were compared with those of 12 other CAV strains. The genetic analysis of the amino acid sequences of VP1 indicated the possible existence of genetic groups among CAV strains, as BD-3 CAV along with four other strains (CIA-1, L-028, Isolate 704 and TR-20) formed a distinct lineage. These strains have four signatory amino acids in VP1, such as 75I/T, 97L, 139Q and 144Q, out of which the latter two are located in a small hydrophilic peak. 相似文献
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本研究通过荧光定量PCR方法检测了TLR15、LY86和TRIM39 mRNA在不同日龄鸡法氏囊中的变化。结果显示,10日龄时,TLR15 mRNA的相对表达量显著高于18胚龄时的,10日龄和30日龄时的表达量基本相同;出壳后,LY86 mRNA的表达水平呈下降趋势,30日龄时的表达量极显著低于18胚龄时的;而TRIM39 mRNA在出壳后表达量逐渐增加,30日龄时的表达量极显著高于18胚龄时的。本研究结果表明,TLR15、LY86和TRIM39在鸡法氏囊发育过程中发挥着重要作用。 相似文献