Expression of pig endogenous retrovirus by primary porcine endothelial cells and infection of human cells

Martin, Ulrich; Kiessig, Verena; Blusch, Jürgen; Haverich, Axel; Helm, K. von der; Herden, Tanja; Steinhoff, Gustav

doi:10.1016/s0140-6736(98)07144-x

Cited by 297 publications

(185 citation statements)

References 11 publications

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“…The ability of PERV produced by pig cell lines or primary cell cultures to infect human cells in vitro has been documented [8,9,13,14,15,16], and serial passages of PERV on a human cell line have produced retroviruses with a higher tropism for human cells [17]. Our previous work found that pancreas expressed the lowest PERV mRNA levels in SPF pig tissues intended for grafting [5] and that long-term follow-up failed to detect in vitro transmission of PERV from SPF pig islets to a panel of human cells (including very permissive 293 tumour cells), even after repeated and optimised co-incubations [8].…”

Section: Discussionmentioning

confidence: 99%

“…However, there is also a risk of transmitting porcine endogenous retroviruses (PERV) [9,10,11,12]. The ability of PERV [10] produced by pig cells to infect human cells in vitro has been documented [8,9,13,14,15,16,17]. PERV mRNA and full-length endogenous retroviral genome were expressed in all pigs tested [9,15,18,19], including SPF pigs [5,20].…”

Section: Pcr-derived Monitoring Of Perv Infection and Microchimerism mentioning

confidence: 99%

“…The ability of PERV [10] produced by pig cells to infect human cells in vitro has been documented [8,9,13,14,15,16,17]. PERV mRNA and full-length endogenous retroviral genome were expressed in all pigs tested [9,15,18,19], including SPF pigs [5,20]. Thus, the use of SPF pigs does not seem to be able to prevent PERV transmission to xenograft recipients.…”

Section: Pcr-derived Monitoring Of Perv Infection and Microchimerism mentioning

confidence: 99%

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Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

et al. 2002

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Aims/hypothesis. Pig islets could transmit porcine endogenous retroviruses (PERV) to diabetic patients. Our previous work showed that pig islets expressed low levels of PERV mRNA and were not likely to transmit PERV to human cells in vitro. The real risk of infection during pig tissue xenografts can only be evaluated by in vivo experiments. Methods. Nude mice bearing tumours containing human 293 cells were grafted with specific pathogenfree pig islets or PERV-producing pig PK15 cells to determine whether pig cells could transmit PERV to mouse and human cells in vivo. Infection was monitored by PCR, long PCR, RT-PCR and long RT-PCR. As detection of PERV sequences could be due to the presence of residual pig cells, we looked for pig mitochondrial (mt) DNA. Quantitative PCR for PERV and pig mt DNA was done to compare the PERV-topig mt (P-to-M) ratio of each sample with the reference ratio for grafted pig cells. Results. Among 78 mouse tissues from PK15-grafted mice, 54 and 72 were positive for gag and pig mt DNA, respectively. Human tumours developed in these mice were positive for PERV (78%) and pig mt (89%). The P-to-M ratios for mouse tissues and PERV-positive human tumours from PK15-grafted mice were higher than the ratio in PK15 cells. Among 41 tissues from pig islet cell-grafted mice, 7 were positive for PERV (3 lymph nodes, 1 kidney, 2 salivary glands, 1 ovary), and 14 were positive for pig mt DNA. Three of these samples (1 lymph node, 1 kidney and 1 salivary gland) were positive for gag DNA, but negative for pig mt DNA. One human tumour in these mice was positive for PERV DNA. P-to-M reference ratio in grafted islet cells was 0.05±0.03. The three PERVpositive lymph nodes contained 78 gag/3 mt copies (P-to-M: 26), 101 gag/3 mt copies (P-to-M: 34), and 4 gag/0 mt copies. The two PERV-positive salivary glands contained 14 gag/1 mt copies, and 28 gag/0 mt copies. The ovary and the kidney contained 46 gag/ 3 mt and 69 gag/0 mt copies, respectively. The PERV-positive human tumour contained 47 gag/3 mt copies. Conclusions/interpretation. Microchimerism and PERV transmission were frequently observed in both mouse and human tissues during grafting of pig PK15 cells into nude mice bearing human tumours, and sometimes during pig islet xenograft in this model. This strengthens the notion that there is a risk of transmitting PERV during pig islet xenograft. [Diabetologia (2002) 45:914-923] Keywords Xenograft, pig endogenous retrovirus, pig islet cell, PK15 cells, specific pathogen free pig, human cells, mouse. vided by E. Gouin (Ploufragan and Nantes, France) from Large-White SPF pigs (80-120 kg), aged 20 weeks, as previously described [1,2]. Pancreases were removed in betadine solution and inflated with 100 ml ice-cold sterile modified University of Wisconsin (mUW) solution and transported in mUW. Pancreases were then inflated again with 0.5 ml/g of mUW solution containing 2 mg/ml Liberase (Roche Diagnostics, Meylan, France) and placed in a digestion chamber filled with mUW solution kept at 36°C. Crude isle...

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Section: Discussionmentioning

confidence: 99%

Section: Pcr-derived Monitoring Of Perv Infection and Microchimerism mentioning

confidence: 99%

Section: Pcr-derived Monitoring Of Perv Infection and Microchimerism mentioning

confidence: 99%

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Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

et al. 2002

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“…9 Along with PK15 cells, other cultured porcine cells have been shown to produce PERV based on the presence of PERV RNA 10 or porcine RT 11 in their supernatant. These other porcine cells include hepatocytes, 10 lung, 10 skin, 10 endothelium, 10 and peripheralblood mononuclear cells. 11 Under in vitro co-culture conditions, both porcine endothelial cells 10 and porcine peripheral-blood mononuclear cells 11 have been shown to produce PERV capable of infecting human HEK293 cells.…”

confidence: 99%

Influence of human fulminant hepatic failure sera on endogenous retroviral expression in pig hepatocytes

et al. 2000

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A porcine endogenous retrovirus (PERV) has been shown to infect human embryonic kidney 293 (HEK293) cells in vitro. The PERV proviral sequence exists in the genome of all porcine cells, including hepatocytes used in a bioartificial liver (BAL). We examined the possibility of PERV infection in HEK293 cells during exposure to supernatant from cultured pig hepatocytes. Pig hepatocytes were cultured in media supplemented with serum from patients in fulminant hepatic failure (FHF) to simulate conditions of an extracorporeal BAL. Pig hepatocytes were cultured in serum-free media for 24 hours and then exposed to fresh medium containing serum from a patient with FHF (22 patients tested). Twenty-four hours later, supernatant was collected and analyzed by polymerase chain reaction (PCR), with and without reverse transcriptase. Primers targeting the pol gene of PERV were used for PCR. Products of amplification were detected by an enzyme-linked immunosorbent assay-based technique using an internal capture probe also targeting the pol gene. Levels of PERV sequences were estimated by serial dilution. All positive samples were tested for infectivity in HEK293 cells. Porcine kidney 15 cell supernatant and fresh culture media were studied as positive and negative controls, respectively. Pig hepatocytes were also studied in the absence of FHF sera and in the presence of mitogenic stimulation with phytohemagglutinin (PHA) and phorbol 12-myristate-13-acetate (PMA). PERV DNA and PERV RNA were detected in all supernatants of cultured pig hepatocytes. The level of PERV RNA in the supernatant of pig hepatocytes was not altered by exposure to human FHF serum or stimulation with PHA and PMA. In addition, PERV RNA was undetectable in the supernatant of HEK293 cells for up to 50 days after exposure to pig hepatocyte supernatant (with or without FHF sera). These findings show that production of PERV by cultured pig hepatocytes was unaffected by exposure to growth factors and cytokines present in human FHF sera.

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“…Host range analysis by the vector transduction assay has showed that PERV-A and PERV-B viruses have wider host ranges, including several human cell lines than PERV-C viruses, which infect only two pig cell lines and one human cell line. Two of the three identifi ed receptor classes of PERV, distinguished by their envelope sequence and tropism, have been shown to be capable of replicating in human cells in vitro [3][4][5][6]. Clinical trials to date with live pig xenograft tissues include perfusion with pig livers or porcine hepatocytes as a bridging strategy for hepatic failure, use of pancreatic islet cells as a treatment for chronic diabetics and implantation of fetal neuronal tissue as a therapy for Parkinson's disease [7].…”

Section: Introductionmentioning

confidence: 99%

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

Prabha

Verghese

2009

Indian J Microbiol

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Pigs offer an unlimited source of xenografts for humans. The use of transplants from animal origin offers a potential solution to the limited supply of human organs and tissues. However, like many other mammalian species, pigs harbor porcine endogenous retrovirus (PERV), which are encoded in their genomic DNA and are assumed to have been integrated into the porcine germline. The ability of PERV to infect human cells in vitro has heightened safety concerns regarding the transmission of PERV to pig xenograft recipients. Porcine tissues were analyzed using validated assays specifi c for PERV: polymerase chain reaction (PCR) (for PERV DNA) and reverse transcriptase (RT)-PCR (for PERV RNA). PERV-specifi c gag sequences were found in the porcine heart tissue samples using DNA-PCR and RT-PCR. PCR is a rapid and specifi c test for the detection of PERV from xenografts. These fi ndings have demonstrated that the presence of both DNA and RNA forms of PERV in porcine tissues needs to be carefully considered when the infectious disease potential of xenotransplantation is being assessed.

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Expression of pig endogenous retrovirus by primary porcine endothelial cells and infection of human cells

Cited by 297 publications

References 11 publications

Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

Microchimerism and transmission of porcine endogenous retrovirus from a pig cell line or specific pathogen-free pig islets to mouse tissues and human cells during xenografts in nude mice

Influence of human fulminant hepatic failure sera on endogenous retroviral expression in pig hepatocytes

Polymerase chain reaction in detection of porcine endogenous retrovirus (PERV) from porcine tissues

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