Hattermann Kim, Roedner Claudia, Schmitt Cornelia, Finsterbusch Tim, Steinfeldt Tobias, Mankertz Annette
Neue Viren/Xenotransplantation, Robert Koch Institut, Nordufer 20, 13353 Berlin, Germany.
Xenotransplantation. 2004 May;11(3):284-94. doi: 10.1111/j.1399-3089.2004.00134.x.
The lack of human donor organs in allotransplantation has led to a proposal for the use of porcine tissues and organs as alternative therapeutic material for humans. Besides immunological problems like graft rejection, one of the major concerns is the transmission of porcine microorganisms as viruses, bacteria and fungi to a human recipient.
Human cell lines have been infected with porcine circovirus type 1 (PCV1) and porcine circovirus type 2 (PCV2) to investigate whether PCV can infect and replicate in human epithelial cells and lymphocytes. Infection of PCV1 was observed with 293, Hela and Chang liver cells, infection with PCV2 only in Rd cells. In addition, religated viral DNA of PCV1 and PCV2 has been used to transfect adherent human cell lines.
PCV1 persisted in most cell lines without causing any visible changes, while PCV2-transfected cells showed a cytopathogenic effect. Presence of PCV DNA was detected in cells and supernatant by PCR, expression of viral proteins by an indirect immune fluorescence assay. A replication assay showed that the replication of PCV DNA was initiated at the origin of replication. When virus-free cells were inoculated with the supernatant of PCV-infected human cells, the infection was not passed.
Although PCV gene expression and replication took place in human cells, the infection is non-productive. Alteration of protein localization suggests that protein targeting may be disturbed in human cells.
同种异体移植中人类供体器官的短缺促使人们提出使用猪的组织和器官作为人类替代治疗材料的建议。除了移植物排斥等免疫问题外,主要担忧之一是猪的微生物如病毒、细菌和真菌向人类受者的传播。
用人细胞系感染1型猪圆环病毒(PCV1)和2型猪圆环病毒(PCV2),以研究PCV是否能在人上皮细胞和淋巴细胞中感染和复制。在293、Hela和Chang肝细胞中观察到PCV1的感染,仅在Rd细胞中观察到PCV2的感染。此外,已使用PCV1和PCV2的环化病毒DNA转染贴壁人细胞系。
PCV1在大多数细胞系中持续存在,未引起任何可见变化,而PCV2转染的细胞表现出细胞病变效应。通过PCR在细胞和上清液中检测到PCV DNA的存在,通过间接免疫荧光试验检测病毒蛋白的表达。复制试验表明,PCV DNA的复制在复制起点开始。当用PCV感染的人细胞的上清液接种无病毒细胞时,感染未传播。
虽然PCV基因在人细胞中表达和复制,但感染是非生产性的。蛋白质定位的改变表明蛋白质靶向在人细胞中可能受到干扰。