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1.
用犬瘟热病毒（ＣＤＶ）２种不同毒株感染非洲绿猴肾细胞（Ｖｅｒｏ）,用流式细胞术分析了病毒诱导的感染细胞的凋亡。结果发现,２种毒株感染引起的细胞病变类型不同,分为圆缩型与合胞体型;２种毒株均可诱导感染细胞凋亡,但毒株间的凋亡细胞化例存在差异,即产生合胞体型细胞病变的毒株凋亡出现较晚,凋亡细胞比例显著低于产生圆缩型细胞为的毒株（Ｐ〈０．０１）。结果提示,不同毒株导致细胞死亡的机制可能不同。 相似文献
2.
根据对培养细胞致细胞病变的能力将牛病毒性腹泻病毒2型毒株(BVDV-2)分为致细胞病变型和非致细胞病变型。BVDV-2的致细胞病变机制还不是很清楚。试验通过显微镜检查和微阵列分析检测由BVDV-2感染造成的牛肾细胞发病机理。结果发现,BVDV-2可激活内质网应激信号通路,该通路对感染细胞的凋亡起作用。在感染BVDV-2期间用RT-PCR监测内质网应激标记基因的表达,结果表明,用BVDV-2致细胞病变毒株感染牛肾细胞可诱导葡萄糖调节蛋白78的表达。感染BVDV-2也可诱导DNA损伤诱导转录物3的表达并抑制外源凝集素半乳糖苷结合溶解物1的水平。结果表明,BVDV-2致细胞病变毒株可诱导内质网应激反应从而导致细胞凋亡。 相似文献
3.
《中国预防兽医学报》2016,(3)
为研究猪流行性腹泻病毒(PEDV)诱导体外细胞产生病变及细胞凋亡的动态变化,本研究应用western blot、间接免疫荧光、细胞活性计数(CCK-8)等方法检测PEDV感染Vero E6细胞后不同时间点细胞内病毒含量、细胞活性、凋亡小体的形成,以及相关凋亡因子的变化。结果表明,感染后24 h细胞内病毒含量开始明显升高,48 h时达到峰值。同时在病毒感染24 h后出现明显的细胞病变,感染48 h后出现典型的凋亡特征,细胞聚堆,细胞核内染色质固缩,形成凋亡小体。CCK-8分析检测结果显示,与对照组相比感染病毒细胞活性明显降低。另外,western blot结果表明促凋亡因子蛋白水解酶Caspase-3在病毒感染过程中被活化。该研究结果进一步证实了PEDV感染可以诱导细胞凋亡,其中Caspase-3的激活在病毒诱导细胞凋亡的过程中发挥重要作用。 相似文献
4.
试验旨在对不同毒力水貂阿留申病毒（Aleutian mink disease virus,AMDV）在猫肾细胞（feline kidney cell,CRFK）中的增殖规律及其诱导细胞凋亡情况进行比较研究。将标准毒株AMDV-G及分离到的野毒株AMDV-DL124、AMDV-DL125、AMDV-QD2、AMDV-QD3、AMDV-ZJ3接种CRFK细胞,应用间接免疫荧光、实时荧光定量PCR、TCID50测定技术研究病毒在细胞中的复制及表达情况,同时检测病毒诱导的细胞凋亡情况。间接免疫荧光结果显示,5株野毒株荧光着色趋势差异不大,均在感染后12 h出现荧光,随感染时间延长荧光增多,AMDV-G荧光出现时间比野毒株晚,但病毒感染后72 h几乎所有细胞均出现荧光;实时荧光定量PCR结果显示,基因组复制趋势大致相同,AMDV-DL125感染后3 h复制开始,AMDV-G感染后24 h复制才开始并呈快速增长趋势,但感染后72 h均达到峰值。TCID50检测结果表明,0~12 h为病毒感染潜伏期,AMDV-G感染后60 h达到峰值,野毒株均在感染后72 h达到峰值,但是6株病毒均能在感染后48~72 h维持较高的感染滴度,其后随细胞崩解而降低。SPSS 23.0统计软件分析凋亡检测结果显示,与对照组相比,野毒株感染细胞后2~12 h诱导细胞凋亡差异显著（P<0.05）,AMDV-G诱导细胞凋亡差异明显低于野毒株,但是诱导细胞凋亡时间较野毒株长,在感染后24 h仍对细胞凋亡有较明显的诱导作用,但是各病毒诱导的细胞凋亡主要集中在2~12 h。该结果为AMDV的培养、鉴定及致病机理研究提供一定参考。 相似文献
5.
为获得大熊猫犬瘟热病毒株,采集死亡大熊猫的心脏、肺、肝病料,研磨并反复冻融后收集上清液接种Vero细胞,待出现细胞病变(CPE)后,收集病毒液。用逆转录-聚合酶链反应(RT-PCR)鉴定病毒分离株,测定病毒TCID_(50),动物回归试验测定其毒力。结果显示,盲传到第5代时,Vero细胞出现圆缩、聚集、脱落等病变;PCR扩增出287bp片段,与预期相符;测序结果显示,该毒株与已发表的CDV SD(14)11毒株(亚洲-Ⅰ型)的同源性为98%;毒株的TCID_(50)为10~(-5.2)/mL;幼犬感染分离毒株后出现体温升高、鼻头发干、拉稀、眼鼻分泌出水样分泌物等症状。本研究成功分离出大熊猫源犬瘟热病毒株。 相似文献
6.
3种禽类呼肠孤病毒血清学相关性及致细胞病变差异分析 总被引：1，自引：0，他引：1
本研究旨在研究3种不同疾病型禽类呼肠孤病毒间的抗原性关系及病毒的培养特性。作者通过血清中和试验测定了禽呼肠孤病毒(ARV S1133株)、番鸭呼肠孤病毒(MDRV 9710株)、新型鸭呼肠孤病毒(NDRV NP01株)3种禽类呼肠孤病毒的血清学相关性,统计抗原相关性R值;并应用部分禽胚原代细胞及哺乳动物传代细胞对这3种病毒的培养特性进行了初步研究。结果表明,3种病毒株之间的R值很小,抗原相关性较低;三者具有广泛的细胞亲嗜性,能在多种细胞中增殖,并产生细胞病变,但病毒致细胞病变特征有所差异,ARV和NDRV均以巨融合为主,而MDRV则以细胞圆缩坏死为主。上述结果表明导致禽类不同疾病的ARV、MDRV和NDRV三者之间的抗原相关性较低,病毒的细胞培养特性也不同,细胞病变类型的差别提供了一种初步鉴别禽类呼肠孤病毒的方法。 相似文献
7.
为了对疑似牛呼吸道合胞体病毒和巴氏杆菌混合感染的犊牛进行病原鉴定,本研究采用常规病毒经细菌分离鉴定和PCR方法分别进行分离与鉴定。结果表明,该病毒株能在BT细胞上增殖并产生特征性合胞体形态的细胞病变;无血凝性和血吸附特性;能被牛呼吸道合胞体病毒(bovine respiratory syncytial virus,BRSV)标准阳性血清中和;分离的病毒经RT-PCR鉴定为牛呼吸道合胞体病毒;根据菌落形态、细菌染色特性及生化特性,鉴定分离的细菌为巴氏杆菌。提示,该牛场为牛呼吸道合胞体病毒和巴氏杆菌混合感染。 相似文献
8.
9.
本试验将不同浓度的新城疫D90病毒株感染肺癌A549细胞,光学显微镜下,分别在24h、48h、72h观察细胞形态,并通过四甲基偶氮唑盐(MTT)法在不同感染时间,对不同浓度D90作用下的肺癌细胞A549进行细胞生长状况的检测。结果表明,在24h的时间内,不同浓度(1×10-1～1×10-4)的D90毒株作用下的细胞形态学变化具有一个共同的特征:小的气泡出现,且细胞发生圆缩,这是典型的凋亡形态学变化。在48h～72h时间段内,细胞迅速且大量崩解、坏死。在MTT试验中,新城疫病毒D90株对A549肺癌细胞产生了明显的抑制作用,不同浓度的D90毒株对A549肺癌细胞的作用差异较为明显,抑制率随着病毒浓度的增加而增大,即抑制率与病毒的浓度呈正相关。 相似文献
10.
不同动物源新城疫病毒感染不同种属细胞的病变观察 总被引：1，自引：0，他引：1
用10株不同动物源的新城疫病毒F48E8,La Sota、La Sota-GFP、ZJ1、ZJ1-GFP、JS-4-05-Go、Gx-1-05-Ch,JS-1-07-Os、Js-1-07-Du、JS-1-07-Pi等毒株分剐感染鸡胚、鹅胚、鸭胚成纤维细胞、PK-15、Dulac、FK81、MDCK、Vero、Hela多种不同动物来源的传代细胞系,通过对病毒感染细胞后病变的观察、ZJ-GFP及La Sota-GFP两个病毒感染细胞后细胞中绿色荧光蛋白指示的病变,了解不同的毒株对不同细胞感染性的差别.结果表明,不同毒力、不同动物源的新城疫病毒致不同种属细胞病变的作用不同,产生病变的时间有差异,强毒F48E8等毒株感染细胞后48～72 h可以引起几种禽源成纤维细胞全部裂解,而哺乳动物来源的细胞在60～120 h细胞发生病变,且病变也不如禽源成纤维细胞严重,出现了细胞的圆缩.弱毒株在胰酶处理后感染细胞,72 h以后出现明显细胞病变,引起的哺乳动物细胞病变不明显.受试毒株的动物种属来源与细胞的致病性没有明显相关性. 相似文献
11.
Guo A Lu C 《Journal of veterinary medicine. B, Infectious diseases and veterinary public health》2000,47(3):183-190
Apoptosis of Vero cells infected with two canine distemper virus (CDV) vaccine strains was detected using TdT (terminal deoxynucleotidyl transferase)-mediated dUTP nick end-labelling (TUNEL), flow cytometric analysis, agarose gel electrophoresis and electron microscopy (EM). By TUNEL, apoptotic cells were found in CDV-Onderstepoort (CDV-Ond)-infected cells. DNA fragments isolated from infected cells were separated by agarose gel electrophoresis and a 'ladder' pattern appeared. EM observations demonstrated that the cells undergoing cytopathic effect (CPE) possessed morphological characteristics of apoptotic cells. Flow cytometric analysis indicated that CDV could induce apoptosis of Vero cells, but the percentages of the apoptotic cells were correlated with the CPE types. The strain showing the cell-rounding type of CPE produced a much higher percentage of apoptotic cells than CDV-Ond with the syncytium type of CPE (P < 0.01). It was concluded that CDV vaccine strains could induce apoptosis of Vero cells and the apoptosis was virus strain-dependent and cell-dependent. The mechanism remains to be studied. 相似文献
12.
Nishi T Tsukiyama-Kohara K Togashi K Kohriyama N Kai C 《Comparative immunology, microbiology and infectious diseases》2004,27(6):445-455
The Yanaka strain, a field isolate of Canine distemper virus (CDV), caused extensive syncytial cytopathic effects (CPEs) followed by cell death in vitro. Syncytium formation is an important aspect of CDV pathogenicity, but the mechanism of the fusion-induced cell death is still not understood. In this study, the involvement of apoptosis in the CDV-induced CPE was investigated. We also examined apoptosis in cells infected with a persistent strain of CDV, the Yanaka-BP strain derived from the Yanaka strain, because this strain does not cause obvious CPE. DNA laddering together with Terminal transferase dUTP nick endlabeling (TUNEL) assay indicated that the Yanaka strain infection, but not the Yanaka-BP infection induced apoptosis. In addition, flow cytometric analysis similarly indicated that the Yanaka-BP strain induced apoptosis significantly less frequently than the Yanaka strain did. Thus, absence of apoptosis may be implicated in the CPE and establishment of persistent CDV infection. 相似文献
13.
Serageldeen Sultan Nguyen Thi Lan Toshiki Ueda Ryoji Yamaguchi Ken Maeda Kazushige Kai 《Acta veterinaria Scandinavica》2009,51(1):38
Backgrounds
The aim of this study was to confirm the propagation of various canine distemper viruses (CDV) in hamster cell lines of HmLu and BHK, since only a little is known about the possibility of propagation of CDV in rodent cells irrespective of their epidemiological importance.Methods
The growth of CDV in hamster cell lines was monitored by titration using Vero.dogSLAMtag (Vero-DST) cells that had been proven to be susceptible to almost all field isolates of CDV, with the preparations of cell-free and cell-associated virus from the cultures infected with recent Asian isolates of CDV (13 strains) and by observing the development of cytopathic effect (CPE) in infected cultures of hamster cell lines.Results
Eleven of 13 strains grew in HmLu cells, and 12 of 13 strains grew in BHK cells with apparent CPE of cell fusion in the late stage of infection. Two strains and a strain of Asia 1 group could not grow in HmLu cells and BHK cells, respectively.Conclusion
The present study demonstrates at the first time that hamster cell lines can propagate the majority of Asian field isolates of CDV. The usage of two hamster cell lines suggested to be useful to characterize the field isolates biologically. 相似文献14.
15.
Lan NT Yamaguchi R Kai K Uchida K Kato A Tateyama S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(5):491-495
To know growth profiles of canine distemper virus (CDV) on Vero cells stably expressing canine signaling lymphocyte activation molecule (Vero-DogSLAMtag; Vero-DST cells), the propagation of three strains of CDV was tested in Vero-DST cells in comparison with parental Vero cells. Strain MD77 could grow well in both cell lines, but demonstrated no syncytium formation or indistinguishable rounding cytopathic effects (CPE) in Vero cells. Strains Onderstepoort and KDK-1 also grew well in Vero-DST cells with apparent syncytium CPE, while they grew less or no efficiently, respectively, in Vero cells. All three CDV strains demonstrated the peak titers, in Vero-DST cells before reaching to an extensive CPE and drastic decrease of titers at/after full CPE. Immunohistochemistry revealed that viral antigens of all CDV strains were found exclusively in the syncytia in Vero-DST cells, while in Vero cells, viral antigen was identified in their single cells for strain MD77 but none for other strains. Thus, every strain of CDV could grow well in Vero-DST cells and behaved differently against Vero cells. These results would be of practical value for workers of CDV because 1) In Vero-DST cells, by observation of distinct syncytium CPE, the highest titer or the best growth of virus could be identified; 2) In Vero cells, various CDV strains could be readily classified after propagation in Vero-DST cells. 相似文献
16.
对流行病学调查、临床症状检查和ELISA检测为犬瘟热阳性的自然发病犬,取肠内容物为病料,采用同步培养方法接种于犬肾细胞系(MDCK)进行病毒的分离,并对分离株进行了形态学特征、血凝特性、动物感染及RT-PCR鉴定。结果表明:病料接种MDCK细胞产生明显的细胞病变(CPE),电镜负染观察接毒细胞培养物见有典型的犬瘟热病毒粒子。分离株不凝集鸡及人“O”型红细胞,接种犬出现明显的临床症状和病理变化。用RT-PCR技术检测病毒细胞培养液,扩增出的片段长为760 bp,与预期设计的长度相同,由此确证分离株为犬瘟热病毒,命名为CDV-GZ2株。 相似文献
17.
18.
Growth characteristics of canine distemper virus in a new cell line CCT cells originated from canine malignant histiocytosis 总被引：1，自引：0，他引：1
Yamaguchi R Kojimoto A Sakai H Uchida K Sugano S Tateyama S 《The Journal of veterinary medical science / the Japanese Society of Veterinary Science》2005,67(2):203-206
Canine distemper virus (CDV) growth and the morphological characterization were examined in a cell line established from a canine malignant histiocytosis (CCT cell line). The susceptibility of the CCT cells to 3 CDV strains, FXNO, YSA-TC and MD-77 was shown by detection of the antigen in the indirect fluorescent assay. After passaging 4 and 9 times through the CCT cells, only FXNO strain could produce the syncytia where demonstrated the antigens. Titers of 9 passaged viruses through the CCT cells showed slightly higher in the CCT cells than those in Vero cells. Morphological characterization of karyorrhexis and specific DNA ladder by extracted DNA electrophoresis indicated apoptosis in the CDV infected CCT cells. 相似文献
19.
Yufei Guo Chanjuan Shen Anchun Cheng Mingshu Wang Na Zhang Shun Chen Yi Zhou 《Veterinary microbiology》2009,138(3-4):258-265
Anatid herpesvirus 1 (AHV-1) CH virulent strain was first isolated from an infected duck and it was found that this virus strain could induce cytopathic effect (CPE) in duck embryo fibroblast (DEF). Following AHV-1 infection, DEF showed morphological changes such as cell rounding, improved refractivity and detachment from the culture surface. However, its pathological characteristics were not adequately known. Related studies were performed and the results showed that syncytium formation could be observed as the other type of CPE in AHV-1 infection. Hematoxylin-eosin staining and 4’, 6-diamidino-2-phenylindole (DAPI) staining of infected DEF were each used to visualize the shape and distribution of chromatin within nuclei and nuclear fragmentation was observed. Chromatin condensation and margination, as well as formation of apoptotic bodies were observed by transmission electron microscopy (TEM). DNA ladder formation was detected in AHV-1 infected cells and apoptosis of the infected DEF was also detected by flow cytometry analysis of Annexin V-FITC/PI staining method. Therefore, it was suggested that AHV-1 virulent strain can induce syncytium and apoptosis in DEF. Syncytium formation and apoptosis observed in this study may contribute to the elucidation of AHV-1 pathogenesis. 相似文献
20.
Actinomycin D inhibited the cytopathic effects (CPE) of type II strains of porcine enteroviruses in PK-15 cells but had no consistent effect on viral replication. The maximum inhibition of CPE was observed when the antibiotic was present during the early stages of infection. No inhibition of the CPE induced by type I strains was observed. A correlation was found between the activation of lysosomal acid phosphatase and morphological changes in the infected cells and the release of progeny virus. Diffuse enzymatic activity was more pronounced in cells infected with type II strains. The activation of lysosomal acid phosphatase by type II strains was inhibited by actinomycin D, while that of type I strains remained unaffected. 相似文献