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犬瘟热病毒的分离鉴定、致病特点和单克隆抗体的研究 总被引：5，自引：0，他引：5
犬瘟热是由犬瘟热病毒（ Ｃａｎｉｎｅ Ｄｉｓｔｅｍ ｐｅｒ Ｖｉｒｕｓ, Ｃ Ｄ Ｖ）引起的犬的一种高度接触传染性疾病。近年来,随着我国军、警犬、实验用犬和宠物犬饲养量的大幅度增加,以及异地交流的增多,犬瘟热在我国犬群中的发病率和致死率均有升高的趋势,而且临床表现也与以往有所不同。分析原因,除母源抗体干扰、疫苗效价低及使用不当外,犬群中是否出现了 Ｃ Ｄ Ｖ 新毒株亟待研究证实。本研究在从国内病犬分离到 Ｃ Ｄ Ｖ 的基础上,进行了 Ｃ Ｄ Ｖ 理化特性、致病特点、回归动物试验以及研制成功分泌抗 Ｃ Ｄ Ｖ 单克隆抗体,并进行了初步应用,取得了较为满意的研究结果。在研究中应用免疫组化方法进行病原检查,证实可很好地解决以往病理观察中无法进行病毒性疾病的特异性诊断的问题。１）采用免疫组化间接 Ｂ Ａ 法,对临诊疑似犬瘟热病犬脏器组织进行了 Ｃ Ｄ Ｖ 抗原定位检查及系统病理学观察。结果表明：发病犬大脑、小脑、心、肺、肝、脾、肠、肾、肾上腺、淋巴结、膀胱等组织细胞及血管内皮、支气管上皮细胞胞浆内均可见抗原阳性反应。同时,抗原阳性反应组织、器官均可见不同程度的病理变化。２）从病死犬肺、肝和淋巴结分离到 ２株 Ｃ Ｄ Ｖ,即 Ｃ Ｄ Ｖ９３０３９ 和 相似文献
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肿瘤是人类及犬主要死亡原因之一，在有的地区死亡病例的三分之一为恶性肿瘤所致。肿瘤抗原比较弱，有的肿瘤可能很难检测到肿瘤特异性抗原，所以研究难度比较大。虽然，有些研究者利用肿瘤抗原研究制备出特异性单克隆抗体，但研究成果不太理想。利用免疫耐受法，选择性地杀死非肿 瘤抗原刺激活化的淋巴细胞。经过多次处理后，使小鼠失去对正常组织的反应能力，从而使抗原性较弱的肿瘤抗原产生阳性反应，出现有反应性的 Ｂ淋巴细胞。经过与 Ｓ Ｐ２／０ 细胞融合和筛选，成功地得到犬肿瘤特异性单克隆抗体。 相似文献
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摘要根据免疫组织化学染色结果发现在犬瘟热急性病例的淋巴结皮质犬瘟热抗原呈强阳性，且密度很高。而在犬的肺脏虽然表现出严重的肺炎症状，但是犬瘟热病毒抗原阳性反应弱，仅在支气管上皮和少量的巨噬细胞胞浆中观察到． 相似文献
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《中国兽医杂志》2020,(2)
为探讨河北省部分地区屠宰猪戊型肝炎病毒(HEV)的感染情况,采集了55例屠宰猪肝脏,采用免疫组织化学方法对肝脏样品的戊型肝炎病毒抗原进行检测,同时应用H.E.染色和Mallory三色染色法对其病理形态学变化进行观察。结果显示,55例肝脏样品中有52例呈抗原阳性反应,阳性率为94.55%。光学显微镜观察可见,病毒抗原在肝细胞核、胞浆和胞膜上均有分布,但主要呈现胞浆弥漫型和胞核型。组织病理学观察结果显示,在HEV-抗原免疫组化阳性反应肝脏样品的H.E.染色切片中,均有不同程度的病理变化,主要表现为肝细胞变性散在单个核固缩性坏死,淋巴细胞等炎性细胞浸润,汇管区胆管增生,肝细胞萎缩,并常见有纤维结缔组织增生。Mallory三色染色结果显示,有29.1%的肝脏样品中有纤维结缔组织增生。 相似文献
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应用免疫组化ＡＢＣ法研究了催产素和加压素在犬泡状绦虫的定位和分布。在犬泡状绦虫中枢神经系统的脑神经节、２条主要纵神经索和环神经内观察到大量阳性纤维和少量阳性细胞。周围神经系统可见阳性神经纤维在皮层和实质形成神经网。从纵神经索和环神经发出的阳性纤维向生殖器官和皮层的肌肉分布。顶突的小钩之间的肌肉存在阳性反应物，小钩周围有较明显的阳性反应环。生殖器官呈现着色深浅不同的阳性反应，以卵囊—梅氏腺复合体最为明显。结果表明，催产素和加压素的免疫反应物具有相同的分布，它们可能参与虫体的神经内分泌调节。 相似文献
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临床中犬瘟热多分为急性和亚急性,3月龄犬多发本病,急性病例和亚急性病例的临床症状和病毒复制的脏器与慢性病例有很大区别。临床上急性病例的病变主要集中在肺脏和淋巴组织,组织病理学可观察到淋巴结炎和多核巨细胞性肺炎。根据免疫组织化学染色结果发现,在犬瘟热急性病例的淋巴结皮质中犬瘟热抗原呈强阳性,且密度很高。而病犬虽然表现出严重的肺炎症状,但在犬的肺脏中犬瘟热病毒抗原阳性反应弱,仅在支气管上皮和少量的巨噬细胞胞浆中观察到。犬瘟热病毒首先在犬的淋巴组织中进行复制、增殖,破坏被膜下淋巴窦和淋巴小节中的T淋巴细胞和B淋巴细胞,影响淋巴组织输出到外周血液循环中的淋巴细胞数量。本研究结果表明,犬瘟热虽然临床上以肺脏病变最严重,但在急性和亚急性病例,犬瘟热病毒主要在淋巴组织的T淋巴细胞、B淋巴细胞和巨噬细胞中增殖,故应以淋巴组织作为犬瘟热病毒的主要分离脏器。 相似文献
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３０ 只２ 月龄健康兔，随机分为 ３组，活体结扎圆小囊，分别注入辣根过氧化物酶（ｈｏｒｓｅｒａｄｉｓｈ ｐｅｒｏｘｉｄａｓｅ， Ｈ Ｒ Ｐ）、兔巴氏杆菌等抗原及生理盐水（对照组）后，于不同时间剖杀取材，观察圆小囊的细胞化学变化，并对其上皮内淋巴细胞及杯状细胞数量变化进行了统计分析。结果表明：注入 Ｈ Ｒ Ｐ 及兔巴氏杆菌后，均可见到圆顶表面贴附一层 Ｐ Ａ Ｓ阳性反应物，并且在圆顶上皮被包裹的细胞中可见少数阳性细 胞，小囊内淋巴组织中阳性反应物增多。本研究首次采用 Ｇｏｍ ｏｒｌ技术，对圆小囊的碱性磷酸酶的分布进行了观察，发现对照组及 Ｈ Ｒ Ｐ组圆小囊粘膜上皮碱性磷酸酶活性均较弱，圆顶上皮呈阴性。硝酸银反应进一步证实了在动物的肠道淋巴组织中存在嗜银细胞，注入示踪物后可见圆小囊内嗜银细胞显著增加，弥散于淋巴组织内，在滤泡处呈片状分布。同时，圆小囊圆顶两侧粘膜上皮内淋巴细胞及杯状细胞的数量在总体上呈增加趋势，但 Ｈ Ｒ Ｐ 组较巴氏杆菌组更为明显。注入 Ｈ Ｒ Ｐ及兔巴氏杆菌后所引起的圆小囊的组织细胞化学的不同结果提示，圆小囊的主要功能不是吸收营养物质，它对上述两种不同抗原的刺激有不同的反应机制并且圆小囊内嗜银细胞的 数量变化与兔肠道粘 相似文献
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10.
采用免疫组化ＢＡ法进行了犬传染性肝炎病毒（ＩＣＨＶ）在ＭＤＣＫ细胞上定位的研究。结果表明，接毒后１０小时可见细小阳性反应颗粒出现于肿胀的病变细胞核中，ＨＥ染色所见包涵体形态与临发病犬肝实质细胞包涵体形态相似，提示可能是ＩＣＨＶ感染细胞所引的特征性变化。同时测定的病毒血凝滴度结果表明，包涵体的形成与病毒增殖有一定关系，８４～９６小时收毒较为适宜。 相似文献
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对实验感染犬瘟热病毒的病犬进行了系统的病理学观察，并用酶标ＳＰＡ法对病犬脏器组织中ＣＤＶ抗原进行了定位检查。结果表明，淋巴系统各器官组织是ＣＤＶ急性感染早期首先侵犯的靶器官。脏器组织的病理改变与ＣＤＶ抗原检出呈正相关。脏器组织中包涵体的检出与形态结构具有一定的特征性和示病意义，但采用免疫组化方法检查ＣＤＶ抗原，更具优越性。作者认为，ＣＤＶ９３０３９株和ＣＤＶ９３０４１株是致病力很强的泛嗜性ＣＤＶ。 相似文献
12.
为了调查患犬瘟热病犬淋巴组织中T、B细胞变化的特点及淋巴细胞减少的发病机制,试验通过免疫组织化学的方法观察了T细胞(用CD3和CD45RO检测T细胞)、B细胞(用IgG、IgM抗血清检测B细胞)和犬瘟热病毒(抗犬瘟热病毒抗体)在病犬淋巴组织中的分布。结果表明:在淋巴组织中的淋巴细胞、淋巴小结中树突状细胞和巨噬细胞中均检出了抗病毒阳性反应细胞。在骨髓组织的前髓细胞中也发现抗病毒阳性反应细胞和嗜酸性胞浆内及核内包涵体的存在。与对照组相比,CD3和CD45RO阳性细胞主要存在于T细胞的分布域;但CD3和CD45RO阳性T细胞的数量较少。位于淋巴组织中的巨噬细胞有的被CD45RO染成阳性。在B细胞分布的区域中,IgG、IgM阳性细胞的数量明显减少;一些位于淋巴组织的浆细胞也被IgG或IgM染成阳性。在淋巴组织中淋巴细胞减少的顺序为:IgG阳性细胞减少最明显,其次为IgM和CD45RO阳性细胞,再次为CD3阳性细胞。依据试验结果,作者认为病犬淋巴组织中淋巴细胞减少主要是由B细胞缺乏所引起的;淋巴细胞的增殖能力减弱是引起淋巴组织中淋巴细胞减少的重要原因。 相似文献
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Bauder B Suchy A Gabler C Weissenböck H 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2000,47(10):775-784
Tissue samples of cats and dogs with panleukopenia and parvovirus enteritis, respectively, were examined for the presence of viral antigen-positive cells and apoptotic cells by immunohistochemistry and by TUNEL assay (Terminal Transferase-Mediated dUTP Nick End Labelling). Compared to control animals, infected cats and dogs generally had more TUNEL-positive cells. Cell types positive for parvovirus antigen, for example digestive tract epithelial and mesenchymal cells, and lymphocytes and macrophages in lymphoid tissues were also positive for TUNEL signals. Occasionally, TUNEL signal and viral antigen were present in the same tissue areas, suggesting a direct viral trigger of apoptosis. More frequently, however, there was no complete overlap of antigen and TUNEL-positive areas. The results of this study indicate that apoptotic cell death contributes significantly to the widespread tissue damage of parvovirus infection in cats and dogs. 相似文献
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Phenotypical characterization of T and B cell areas in lymphoid tissues of dogs with spontaneous distemper 总被引：5，自引：0，他引：5
CD3, CD4, CD5, and CD8 antigen expression of T cells and IgG expression of B cells and canine distemper virus (CDV) antigen distribution were immunohistochemically examined in lymphoid tissues (lymph node, spleen, thymus, and tonsil) of control dogs and animals with spontaneous canine distemper. In addition, CNS tissue of all animals was studied for neuropathological changes and CDV antigen distribution. Based on the degree of depletion distemper dogs were classified into two groups. Group I represented animals with moderate to marked lymphoid depletion, while group II dogs displayed mild or no depletion. CDV antigen was mainly found in lymphocytes and macrophages of group I dogs, whereas CDV expression was most prominent in dendritic cells of group II animals. In group I dogs, a marked loss of CD3, CD4, CD5, CD8, and IgG expression was noticed, hereby loss of CD4+ cells was more prominent than depletion of CD8+ cells. In the lymphoid tissues of group II animals, a significant increase in the number of T and B cells was observed compared to group I dogs. The number of CD3+, CD4+, and CD8+ cells in group II dogs was similar to the findings in controls, however, CD5 and IgG expression was mildly reduced in T and B cell areas, respectively. Additionally, in groups I and II dogs, CD3+ and CD5- T cells were detected in T cell areas. Whether this cell population represents a cell type with autoimmune reactive potential remains to be determined. Surprisingly in group II animals, viral antigen was found predominantly in dendritic cells indicating a change in the cell tropism of CDV during chronic infection and a possible mechanism of viral persistence. The two patterns of lymphoid depletions correlated to two different types of canine distemper encephalitis (CDE). Group I dogs displayed acute non-inflammatory CDE, whereas group II dogs suffered from chronic inflammatory demyelinating CDE, indicating a pathogenic relationship between lymphocytic depletion and inflammatory brain lesions in distemper. 相似文献
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The cerebella of 12 dogs infected with canine distemper virus (CDV) and those of three normal dogs were examined. The avidin-biotin-peroxidase complex technique was used to detect alphaB-crystallin (alphaB-c) immunoreactivity and immunolocalisation of the CDV antigen. CDV antigens, immunopositive astrocytes, oligodendrocytes and granular neurons were seen in both the white and grey matter of the infected dogs. In the controls, alphaB-c immunopositive glial cells were seen in the white matter and around the Purkinje cells. In dogs with distemper, alphaB-c immunoreactivity was not observed in some of the glial cells around the Purkinje cells. A significant negative correlation of P < 0.01 level was found between areas of severe demyelination and the number of alphaB-c immunopositive cells in dogs infected with CDV. Such correlation was not observed between mild and moderate demyelinating areas and alphaB-c immunostaining. The alphaB-crystallin/ total number of cells ratio was found to be significant in severely affected demyelinating areas (P < 0.05). These data indicate that there was a relationship between the degrees of CDV associated with demyelination and the level of alphaB-c expression in the glial cells. 相似文献
16.
为了进一步调查脑组织的髓鞘脱失与神经胶质细胞等成份的关系,用12只犬瘟热自然病例通过病理组织学和免疫组织化学染色法进行了本试验.结果表明:髓鞘脱失部位的脑组织伴有明显的血液循环障碍,即淤血、水肿、血栓形成和弥漫性血管内凝血;少突胶质细胞发生代谢紊乱和凋亡;用抗犬瘟热病毒(CDV)抗体染色,星状胶质细胞呈现强阳性反应;用抗GFAP染色,纤维性星状胶质细胞在脱髓区呈较强阳性反应,用TUNEL染色可检出发生凋亡的星状胶质细胞;一些室管膜细胞也被CDV感染,许多含有包涵体币口凋亡的室管膜细胞在脑室壁被发现;少数神经元变性和皱缩,其核发生浓缩.据此认为,脑组织的髓鞘脱失主要与血液循环障碍和少突胶质细胞的代谢紊乱及凋亡有关;脑组织的髓鞘脱失是许多病因共同作用的结果,并非是一种病因所致. 相似文献
17.
Rhalem A Sahibi H Guessous-Idrissi N Lasri S Natami A Riyad M Berrag B 《Veterinary parasitology》1999,81(3):173-184
Cell-mediated and humoral immune response in naturally and experimentally infected dogs was studied using crude and pure antigens. Both types of infections induced severe signs of visceral disease, but the symptoms observed in natural infections were more pronounced than in experimental infections. In addition, asymptomatic infections were not observed in experimentally infected animals. Disease evolution in laboratory infections was rapid and an increase in antibody titer to crude parasite antigen was correlated with the appearance and aggravation of clinical symptoms. Peripheral blood lymphocyte proliferation to crude antigen and pure gp63 was observed early following experimental infection, but was abolished once the infected dogs began to exhibit clinical signs. A similar pattern was observed in naturally infected dogs. Serum from all patent dogs showed high antibody titers to rK39 in enzyme-linked immunosorbent assays (ELISA), and reacted by western blotting with several antigens, 12 to 120 KDa, including gp63 and gp70. In the case of asymptomatic dogs. antibody titers to crude antigen were low and only a few antigens were identified by western blotting. None of the pure proteins examined, gp63, gp70, and rK39 were recognized by western blotting or ELISA. However, asymptomatic dogs exhibited specific lymphocyte proliferation to both crude antigen and the potential vaccine candidate gp63. 相似文献
18.
Kumagai K Yamaguchi R Uchida K Tateyama S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2004,66(2):175-181
The relationship between the canine distemper virus (CDV) infection and apoptosis in the canine lymphoid tissues was investigated using immunostaining for single stranded DNA (ssDNA), TdT-mediated dUTP-biotin nick end-labeling (TUNEL) method, and electron microscopy. Twenty-six lymphoid tissues from 8 spontaneously CDV-infected dogs and 1 non-infected dog were used, and lesions were classified into 4 groups according to frequency of the CDV-antigen. Histologically, the degree of lymphoid depletion tended to depend on amount of CDV antigen. The numbers of ssDNA- and TUNEL-labeling cells were significantly high in the lymphoid tissues with abundant viral antigen. However, ssDNA- and TUNEL-positive lymphocytes were also frequently found even in the lymphoid tissues where there was only a small amount of CDV-antigen in sinus histiocytes. The incidence and distribution of apoptotic cells in the CDV-antigens-negative lymphoid tissues from infected dogs were equal to those from a non-infected dog. Double labeling immunostaining using a ssDNA and a CDV nucleocapsid protein (CDV-NP) antibody revealed that there were ssDNA positive but CDV-NP negative cells besides those stained doubly positive. Ultrastructurally, lymphocytes in the CDV-infected lymphoid tissues frequently had characteristic morphological features of apoptosis such as apoptotic bodies. All these results suggest that CDV leads to lymphocytic apoptosis directly or indirectly, resulting in severe lymphoid depletion and immunosuppression in acute or subacute phase of CDV infection. 相似文献
19.
Pathogenesis of canine parvovirus-2 in dogs: histopathology and antigen identification in tissues 总被引：2，自引：0，他引：2
The pathogenesis of canine parvovirus-2 was studied in orally inoculated conventional dogs using histopathological and peroxidase anti-peroxidase staining techniques. Lymphoid necrosis and depletion of lymphocytes from lymphoid tissues were most notable on days 5 and 6 after exposure. Lymphocyte hyperplasia occurred following day 7. Epithelial cell changes in segments of the small intestine were more severe on days 6 to 9 after exposure in areas associated with Peyer's patches and in the upper segments of the small intestine. The lymphocyte was the primary infected cell. Virus infected cryptal epithelial cells were not detected until 24 hours after the identification of infected cells in lymphoid tissues on day 4 after exposure. The majority of virus infected epithelial cells were found in crypts intimately associated with or adjacent to Peyer's patches in the upper segments of the small intestine. 相似文献
20.
Leandro C Santos-Gomes GM Campino L Romão P Cortes S Rolão N Gomes-Pereira S Riça Capela MJ Abranches P 《Veterinary immunology and immunopathology》2001,79(3-4):273-284
In the present study, we have followed up Leishmania infantum infection in dogs: (1) naturally infected; (2) experimentally infected with amastigotes; and (3) experimentally infected with culture promastigotes. The main objective was to evaluate the differences of the humoral and cellular immune responses of each group. Sera from 12 beagle dogs were analysed for total anti-leishmanial antibodies and IgG1 and IgG2 subclasses by enzyme-linked immunosorbent assay (ELISA). Lymphoproliferation to L. infantum antigen was also performed. All naturally infected animals were symptomatic with a marked humoral response. Dogs inoculated with amastigotes were asymptomotic and presented lower antibody titres than naturally infected. Dogs inoculated with culture promastigotes were asymptomotic with no significant humoral response. Strong proliferative responses to Leishmania antigen was observed in dogs inoculated with promastigotes. In our experimental model, IgG1 antibody levels presented a similar pattern in all infected animals, and IgG2 reactivity was high in naturally infected dogs. 相似文献