The Experts below are selected from a list of 7593 Experts worldwide ranked by ideXlab platform

Axel Haverich - One of the best experts on this subject based on the ideXlab platform.

  • Acellularized porcine heart valve scaffolds for heart valve tissue engineering and the risk of cross-species transmission of porcine Endogenous Retrovirus
    The Journal of Thoracic and Cardiovascular Surgery, 2003
    Co-Authors: Rainer G. Leyh, Tobias Herden, Axel Haverich, Michaela Wilhelmi, Thorsten Walles, Klaus Kallenbach, P Rebe, A Oberbeck, Heike Mertsching
    Abstract:

    Abstract Objective Acellularized porcine heart valve scaffolds have been successfully used for heart valve tissue engineering, creating living functioning heart valve tissue. However, there is concern about the possibility of porcine Endogenous Retrovirus transmission. In this study we investigated whether acellularized porcine heart valve scaffold causes cross-species transmission of porcine Endogenous Retrovirus in a sheep model. Methods Acellularized porcine pulmonary valve conduits (n = 3) and in vitro autologous repopulated porcine pulmonary valve conduits (n = 5) were implanted into sheep in the pulmonary valve position. Surgery was carried out with cardiopulmonary bypass support. The animals were killed 6 months after the operation. Blood samples were collected regularly up to 6 months after the operation and tested for porcine Endogenous Retrovirus by means of polymerase chain reaction and reverse transcriptase-polymerase chain reaction. In addition, explanted tissue-engineered heart valves were tested for porcine Endogenous Retrovirus after 6 month in vivo. Results Porcine Endogenous Retrovirus DNA was detectable in acellularized porcine heart valve tissue. However, 6 months after implantation of in vitro and in vivo repopulated acellularized porcine heart valve scaffolds, no porcine Endogenous Retrovirus sequences were detectable in heart valve tissue and peripheral blood. Conclusion Acellularized porcine matrix scaffolds used for creation of tissue-engineered heart valves do not transmit porcine Endogenous Retrovirus.

  • Heart valves from pigs and the porcine Endogenous Retrovirus: experimental and clinical data to assess the probability of porcine Endogenous Retrovirus infection in human subjects.
    The Journal of Thoracic and Cardiovascular Surgery, 2001
    Co-Authors: Ajay K Moza, Tobias Herden, Augustinus Bader, Heike Mertsching, Axel Haverich
    Abstract:

    Abstract Objective: Replacement of heart valves in human subjects has become a routine procedure in cardiac operations. We sought to investigate whether commercially available glutaraldehyde-fixed porcine heart valve prostheses cause porcine Endogenous Retrovirus infection in human subjects because recent studies revealed that human cells can be infected with porcine Endogenous Retrovirus. Methods: Blood samples of 18 patients who underwent aortic or mitral valve replacement with porcine heart valves were collected 6 months to 3 years after operation and tested for porcine Endogenous Retrovirus by means of polymerase chain reaction and reverse transcriptase–polymerase chain reaction. In addition, we tried to trace porcine Endogenous Retrovirus in 3 commercially available, glutaraldehyde-fixed, porcine heart valves. Results: Porcine Endogenous Retrovirus can be easily detected in native porcine heart valves and degrades completely within 1 week of fixation in glutaraldehyde. In all 3 commercially available porcine heart valves, no traces of porcine Endogenous Retrovirus were found. All blood samples showed negative test results for the porcine Endogenous Retrovirus genome. Conclusion: Our results indicate that glutaraldehyde fixation of porcine heart valves reliably prevents cross-species transmission of porcine Endogenous Retrovirus. (J Thorac Cardiovasc Surg 2001;121:697-701)

  • expression of pig Endogenous Retrovirus by primary porcine endothelial cells and infection of human cells
    The Lancet, 1998
    Co-Authors: Ulrich Martin, Axel Haverich, Verena Kiessig, Jurgen H Blusch, Klaus Von Der Helm, Tanja Herden, Gustav Steinhoff
    Abstract:

    Summary Background The risk of interspecies transmission of Retroviruses during xenotransplantation is suggested by reports of pig Endogenous Retrovirus (PERV) released from porcine cell lines productively infecting human cell lines in vitro and of infectious PERV being released from pig peripheral blood mononuclear cells after mitogenic stimulation. Endothelial cells are the main interface between a xenograft and the recipient's leucocytes and tissues. Methods We have analysed pig primary aortic endothelial cells (PAEC) together with other transplantation-relevant porcine cells and tissues for expression of PERV mRNA. Release of virus particles by PAEC was monitored by reverse transcriptase (RT) activity in the medium of cultured PAEC. Infectivity for human cells was tested by co-cultivation of irradiated PAEC with the human embryonal kidney cell line HEK293 and looking for virus release from the human cells. Findings PAECs, hepatocytes, lung, and skin from a variety of pig strains and breeds expressed PERV mRNA. PAEC released infectious particles. Co-cultivation of PAEC and HEK293 led to productive infection of the human cells and expression of PERV types A and B. Interpretation Release of infectious virus from PAEC occurred without mitogenic stimulation, suggesting a serious risk of Retrovirus transfer after xeno-transplantation.

  • expression of pig Endogenous Retrovirus by primary porcine endothelial cells and infection of human cells
    The Lancet, 1998
    Co-Authors: Ulrich Martin, Axel Haverich, Verena Kiessig, Jurgen H Blusch, Tanja Herden, Klaus Von Der Helm, Gustav Steinhoff
    Abstract:

    Summary Background The risk of interspecies transmission of Retroviruses during xenotransplantation is suggested by reports of pig Endogenous Retrovirus (PERV) released from porcine cell lines productively infecting human cell lines in vitro and of infectious PERV being released from pig peripheral blood mononuclear cells after mitogenic stimulation. Endothelial cells are the main interface between a xenograft and the recipient's leucocytes and tissues. Methods We have analysed pig primary aortic endothelial cells (PAEC) together with other transplantation-relevant porcine cells and tissues for expression of PERV mRNA. Release of virus particles by PAEC was monitored by reverse transcriptase (RT) activity in the medium of cultured PAEC. Infectivity for human cells was tested by co-cultivation of irradiated PAEC with the human embryonal kidney cell line HEK293 and looking for virus release from the human cells. Findings PAECs, hepatocytes, lung, and skin from a variety of pig strains and breeds expressed PERV mRNA. PAEC released infectious particles. Co-cultivation of PAEC and HEK293 led to productive infection of the human cells and expression of PERV types A and B. Interpretation Release of infectious virus from PAEC occurred without mitogenic stimulation, suggesting a serious risk of Retrovirus transfer after xeno-transplantation.

  • porcine Endogenous Retrovirus perv was not transmitted from transplanted porcine endothelial cells to baboons in vivo
    Transplant International, 1998
    Co-Authors: Ulrich Martin, Verena Kiessig, Gustav Steinhoff, M G Chikobava, Marcel Anssar, Torsten Morschheuser, B A Lapin, Axel Haverich
    Abstract:

    The discussion about the clinical risk of zoonoses in xenotransplantation has recently culminated in the demand for a moratorium on clinical organ transplantation using pig donors. The basis for this discussion was a recent report showing a possible trans-species transmission of pig Endogenous Retrovirus (PERV) by in vitro transfer to human cell lines. At present, it remains unclear if this could also happen in vivo or in the setting of xenotransplantation. Potential in vivo transfer of PERV after xenotransplantation was investigated in an experimental pig-to-baboon cell transplantation model. Baboons were immunosuppressed with high-dose cyclophosphamide (total 45–150 mg/kg) and transplanted with primary porcine aortic endothelial cells (PAEC). Tissue samples (skin, lymph nodes, lung) and peripheral blood leukocytes of 15 baboons, taken about 12–24 months after transplantation of PAEC, were then analyzed by PCR and showed no PERV infection. PERV expression in PAEC was also analyzed: PERV mRNA and reverse transcriptase in the culture supernatant could be detected. In spite of the release of retroviral particles from cultured PAEC, transplantation of these cells into baboon recipients did not result in virus transmission, not even under heavy immunosuppression.

Jay A Fishman - One of the best experts on this subject based on the ideXlab platform.

  • genomic presence of recombinant porcine Endogenous Retrovirus in transmitting miniature swine
    Virology Journal, 2006
    Co-Authors: Stanley I Martin, Robert A. Wilkinson, Jay A Fishman
    Abstract:

    The replication of porcine Endogenous Retrovirus (PERV) in human cell lines suggests a potential infectious risk in xenotransplantation. PERV isolated from human cells following cocultivation with porcine peripheral blood mononuclear cells is a recombinant of PERV-A and PERV-C. We describe two different recombinant PERV-AC sequences in the cellular DNA of some transmitting miniature swine. This is the first evidence of PERV-AC recombinant virus in porcine genomic DNA that may have resulted from autoinfection following exogenous viral recombination. Infectious risk in xenotransplantation will be defined by the activity of PERV loci in vivo.

  • identification of a full length cdna for an Endogenous Retrovirus of miniature swine
    Journal of Virology, 1998
    Co-Authors: Donna E Akiyoshi, Maria Denaro, Julia L Greenstein, Papia T Banerjee, Jay A Fishman
    Abstract:

    Endogenous Retroviruses of swine are a concern in the use of pig-derived tissues for xenotransplantation into humans. The nucleotide sequence of porcine Endogenous Retrovirus taken from lymphocytes of miniature swine (PERV-MSL) has been characterized. PERV-MSL is a type C Retrovirus of 8,132 bp with the greatest nucleic acid sequence identity to gibbon ape leukemia virus and murine leukemia virus. Constitutive production of PERV-MSL RNA has been detected in normal leukocytes and in multiple organs of swine. The copy numbers of full-length PERV sequences per genome (approximately 8 to 15) vary among swine strains. The open reading frames for gag, pol, and env in PERV-MSL have over 99% amino acid sequence identity to those of Tsukuba-1 Retrovirus and are highly homologous to those of Endogenous Retrovirus of cell line PK15 (PK15-ERV). Most of the differences in the predicted amino acid sequences of PK15-ERV and PERV-MSL are in the SU (cell attachment) region of env. The existence of these PERV clones will enable studies of infection by Endogenous Retroviruses in xenotransplantation. The use of nonhuman species as sources of organs for human transplantation, i.e., xenotransplantation, is considered a potential solution to the shortage of human organs and tissues

  • identification of a full length cdna for an Endogenous Retrovirus of miniature swine
    Journal of Virology, 1998
    Co-Authors: Donna E Akiyoshi, Maria Denaro, Julia L Greenstein, Papia T Banerjee, Haihong Zhu, Jay A Fishman
    Abstract:

    Endogenous Retroviruses of swine are a concern in the use of pig-derived tissues for xenotransplantation into humans. The nucleotide sequence of porcine Endogenous Retrovirus taken from lymphocytes of miniature swine (PERV-MSL) has been characterized. PERV-MSL is a type C Retrovirus of 8,132 bp with the greatest nucleic acid sequence identity to gibbon ape leukemia virus and murine leukemia virus. Constitutive production of PERV-MSL RNA has been detected in normal leukocytes and in multiple organs of swine. The copy numbers of full-length PERV sequences per genome (approximately 8 to 15) vary among swine strains. The open reading frames for gag, pol, and env in PERV-MSL have over 99% amino acid sequence identity to those of Tsukuba-1 Retrovirus and are highly homologous to those of Endogenous Retrovirus of cell line PK15 (PK15-ERV). Most of the differences in the predicted amino acid sequences of PK15-ERV and PERV-MSL are in the SU (cell attachment) region of env. The existence of these PERV clones will enable studies of infection by Endogenous Retroviruses in xenotransplantation.

  • identification of a full length cdna for an Endogenous Retrovirus of miniature swine
    Journal of Virology, 1998
    Co-Authors: Donna E Akiyoshi, Maria Denaro, Julia L Greenstein, Papia T Banerjee, Jay A Fishman
    Abstract:

    Endogenous Retroviruses of swine are a concern in the use of pig-derived tissues for xenotransplantation into humans. The nucleotide sequence of porcine Endogenous Retrovirus taken from lymphocytes of miniature swine (PERV-MSL) has been characterized. PERV-MSL is a type C Retrovirus of 8,132 bp with the greatest nucleic acid sequence identity to gibbon ape leukemia virus and murine leukemia virus. Constitutive production of PERV-MSL RNA has been detected in normal leukocytes and in multiple organs of swine. The copy numbers of full-length PERV sequences per genome (approximately 8 to 15) vary among swine strains. The open reading frames for gag, pol, and env in PERV-MSL have over 99% amino acid sequence identity to those of Tsukuba-1 Retrovirus and are highly homologous to those of Endogenous Retrovirus of cell line PK15 (PK15-ERV). Most of the differences in the predicted amino acid sequences of PK15-ERV and PERV-MSL are in the SU (cell attachment) region of env. The existence of these PERV clones will enable studies of infection by Endogenous Retroviruses in xenotransplantation. The use of nonhuman species as sources of organs for human transplantation, i.e., xenotransplantation, is considered a potential solution to the shortage of human organs and tissues

Jens Mayer - One of the best experts on this subject based on the ideXlab platform.

  • Nomenclature for Endogenous Retrovirus (ERV) loci
    Retrovirology, 2018
    Co-Authors: Robert J. Gifford, Jens Mayer, Jonas Blomberg, John M. Coffin, Hung Fan, Thierry Heidmann, Jonathan Stoye, Michael Tristem, Welkin E. Johnson
    Abstract:

    Retroviral integration into germline DNA can result in the formation of a vertically inherited proviral sequence called an Endogenous Retrovirus (ERV). Over the course of their evolution, vertebrate genomes have accumulated many thousands of ERV loci. These sequences provide useful retrospective information about ancient Retroviruses, and have also played an important role in shaping the evolution of vertebrate genomes. There is an immediate need for a unified system of nomenclature for ERV loci, not only to assist genome annotation, but also to facilitate research on ERVs and their impact on genome biology and evolution. In this review, we examine how ERV nomenclatures have developed, and consider the possibilities for the implementation of a systematic approach for naming ERV loci. We propose that such a nomenclature should not only provide unique identifiers for individual loci, but also denote orthologous relationships between ERVs in different species. In addition, we propose that—where possible—mnemonic links to previous, well-established names for ERV loci and groups should be retained. We show how this approach can be applied and integrated into existing taxonomic and nomenclature schemes for Retroviruses, ERVs and transposable elements.

  • (Ursus maritimus) Endogenous Retrovirus Identified from Next Generation Sequence Data
    2015
    Co-Authors: Polar Bear, Jens Mayer, Kyriakos Tsangaras, David E. Alquezar-planas, Alex D. Greenwood, Johnson Mak, Peter Walker, Marcus Thomas Gilbert
    Abstract:

    Abstract: Transcriptome analysis of polar bear (Ursus maritimus) tissues identified sequences with similarity to Porcine Endogenous Retroviruses (PERV). Based on these sequences, four proviral copies and 15 solo long terminal repeats (LTRs) of a newly described Endogenous Retrovirus were characterized from the polar bear draft genome sequence. Closely related sequences were identified by PCR analysis of brown bear (Ursus arctos) and black bear (Ursus americanus) but were absent in non-Ursinae bear species. The virus was therefore designated UrsusERV. Two distinct groups of LTRs were observed including a recombinant ERV that contained one LTR belonging to each group indicating that genomic invasions by at least two UrsusERV variants have recently occurred. Age estimates based on proviral LTR divergence and conservation of integration sites among ursids suggest the viral group is only a few million years old. The youngest provirus was polar bear specific, had intact open reading frames (ORFs) and could potentially encode functional proteins. Phylogenetic analyses of UrsusERV consensus protein sequences suggest that it is part of a pig, gibbon and koala Retrovirus clade. The young age estimates and lineage specificity of the virus suggests UrsusERV is a recent cross species transmission from an unknown reservoir and places th

  • transcriptional profiling of human Endogenous Retrovirus group herv k hml 2 loci in melanoma
    Genome Biology and Evolution, 2013
    Co-Authors: Katja Schmitt, Eckart Meese, Jorg Reichrath, Alexander Roesch, Jens Mayer
    Abstract:

    Recent studies suggested a role for the human Endogenous Retrovirus (HERV) group HERV-K(HML-2) in melanoma because of upregulated transcription and expression of HERV-K(HML-2)-encoded proteins. Very little is known about which HML-2 loci are transcribed in melanoma. We assigned >1,400 HML-2 cDNA sequences generated from various melanoma and related samples to genomic HML-2 loci, identifying a total of 23 loci as transcribed. Transcription profiles of loci differed significantly between samples. One locus was found transcribed only in melanoma-derived samples but not in melanocytes and might represent a marker for melanoma. Several of the transcribed loci harbor ORFs for retroviral Gag and/or Env proteins. Env-encoding loci were transcribed only in melanoma. Specific investigation of rec and np9 transcripts indicated transcription of protein encoding loci in melanoma and melanocytes hinting at the relevance of Rec and Np9 in melanoma. UVB irradiation changed transcription profiles of loci and overall transcript levels decreased in melanoma and melanocytes. We further identified transcribed HML-2 loci formed by reverse transcription of spliced HML-2 transcripts by L1 machinery or in a retroviral fashion, with loci potentially encoding HML-2-like proteins. We reveal complex, sample-specific transcription of HML-2 loci in melanoma and related samples. Identified HML-2 loci and proteins encoded by those loci are particularly relevant for further studying the role of HML-2 in melanoma. Transcription of HERVs appears as a complex mechanism requiring specific studies to elucidate which HERV loci are transcribed and how transcribed HERVs may be involved in disease.

  • an n terminally truncated envelope protein encoded by a human Endogenous Retrovirus w locus on chromosome xq22 3
    Retrovirology, 2010
    Co-Authors: Christina Roebke, Jens Mayer, Marlies Sauter, Nikolaus Muellerlantzsch, Silke Wahl, Georg Laufer, Christine Stadelmann, Klemens Ruprecht
    Abstract:

    Background We previously showed that the envelope (env) sequence of a human Endogenous Retrovirus (HERV)-W locus on chromosome Xq22.3 is transcribed in human peripheral blood mononuclear cells. The env open reading frame (ORF) of this locus is interrupted by a premature stop at codon 39, but otherwise harbors a long ORF for an N-terminally truncated 475 amino acid Env protein, starting at an in-frame ATG at codon 68. We set out to characterize the protein encoded by that ORF.

  • genomic organization of the human Endogenous Retrovirus herv k hml 2 hom ervk6 on chromosome 7
    Genomics, 2001
    Co-Authors: Katrin Reus, Jens Mayer, Marlies Sauter, Daniela Scherer, Nikolaus Mullerlantzsch, Eckart Meese
    Abstract:

    Recently, we reported an almost intact human Endogenous Retrovirus (HERV-K(HML-2.HOM); HGMW-approved symbol ERVK6) located on human chromosome 7, with open reading frames for all retroviral genes and a mutation only within the reverse transcriptase. We further characterized the genomic organization of this Endogenous Retrovirus by subcloning and sequencing of the proviral insert contained within a chromosome 7-specific cosmid clone and found HERV-K(HML-2.HOM) to be organized as a tandem repeat. Examination of various human DNA samples for this specific proviral repeat suggests a relatively ubiquitous distribution of the HERV-K(HML-2.HOM) tandem structure. However, we identified two human samples having only a single provirus at this locus. In addition, we investigated the presence of HERV-K(HML-2.HOM) alleles having an intact YXDD motif within the reverse transcriptase domain by sequencing the corresponding polymerase gene from various human DNA samples. We identified a HERV-K(HML-2.HOM) polymerase with an intact YXDD motif in two samples, thus potentially coding for an active reverse transcriptase. Our results show for the first time an Endogenous Retrovirus tandem repeat in human populations and suggest the existence of alleles harboring an intact human Endogenous Retrovirus including an intact polymerase gene.

Joachim Denner - One of the best experts on this subject based on the ideXlab platform.

  • expression of the human Endogenous Retrovirus k transmembrane envelope rec and np9 proteins in melanomas and melanoma cell lines
    Melanoma Research, 2006
    Co-Authors: Kristina Buscher, Silvia Hahn, Maja A Hofmann, Muhsin Ozel, Wolfram Sterry, Johannes Lower, Roswitha Lower, Reinhard Kurth, Uwe Trefzer, Joachim Denner
    Abstract:

    The human Endogenous Retrovirus-K encodes two potential tumor proteins, Rec and Np9. Rec is related to the Rev protein of HIV-1 and has been shown to be associated with tumor development in nude mice. Having shown the expression of human Endogenous Retrovirus-K in human melanomas and melanoma cell l

  • expression of human Endogenous Retrovirus k in melanomas and melanoma cell lines
    Cancer Research, 2005
    Co-Authors: Kristina Buscher, Maja A Hofmann, Wolfram Sterry, Reinhard Kurth, Uwe Trefzer, Joachim Denner
    Abstract:

    The human Endogenous Retrovirus K family (HERV-K) comprises 30 to 50 closely related proviruses, most of which are defective. In contrast to all other human Endogenous Retroviruses, some HERV-K proviruses have maintained open reading frames for all viral proteins. In addition to the structural proteins Gag and Env and the reverse transcriptase, two regulatory proteins (Rec and Np9) have been described. Malignant melanoma has the highest mortality among skin cancers and is particularly aggressive. To study the expression of HERV-K, a set of seven primers was developed that allows discrimination between full-length and spliced mRNA and mRNA from deleted and undeleted proviruses. Expression of full-length mRNA from deleted and undeleted proviruses was detected in all human cells investigated. Expression of spliced env and rec was detected in a teratocarcinoma cell line, in 45% of the metastatic melanoma biopsies, and in 44% of the melanoma cell lines. In normal neonatal melanocytes, spliced rec was detected but not spliced env. Viral proteins were shown to be expressed in primary melanomas, metastases, and melanoma cell lines by immunohistochemistry, immunofluorescence, and Western blot analyses using specific antisera. For the first time, antibodies against HERV-K were found in melanoma patients. Melanomas are, in addition to teratocarcinomas and human breast cancer, the third tumor type with enhanced expression of HERV-K.

Yasuhiro Takeuchi - One of the best experts on this subject based on the ideXlab platform.

  • Suboptimal Porcine Endogenous Retrovirus Infection in Non-Human Primate Cells: Implication for Preclinical
    2013
    Co-Authors: Giada Mattiuzzo, Yasuhiro Takeuchi
    Abstract:

    Background: Porcine Endogenous Retrovirus (PERV) poses a potential risk of zoonotic infection in xenotransplantation. Preclinical transplantation trials using non-human primates (NHP) as recipients of porcine xenografts present the opportunity to assess the zoonosis risk in vivo. However, PERV poorly infects NHP cells for unclear reasons and therefore NHP may represent a suboptimal animal model to assess the risk of PERV zoonoses. We investigated the mechanism responsible for the low efficiency of PERV-A infection in NHP cells. Principal Findings: Two steps, cell entry and exit, were inefficient for the replication of high-titer, human-tropic A/C recombinant PERV. A restriction factor, tetherin, is likely to be responsible for the block to matured virion release, supported by the correlation between the levels of inhibition and tetherin expression. In rhesus macaque, cynomolgus macaque and baboon the main receptor for PERV entry, PERV-A receptor 1 (PAR-1), was found to be genetically deficient: PAR-1 genes in these species encode serine at amino acid 109 in place of the leucine in human PAR-1. This genetic defect inevitably impacts in vivo sensitivity to PERV infection of these species. In contrast, African green monkey (AGM) PAR-1 is functional, but PERV infection is still poor. Although the mechanism is unclear, tunicamycin treatment, which removes N-glycosylated sugar chains, increases PERV infection, suggesting a possible role for the glycosylation of the receptors. Conclusions: Since cynomolgus macaque and baboon, species often used in pig-to-NHP xenotransplantation experiments

  • Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species-4
    2011
    Co-Authors: Giada Mattiuzzo, Magda Matouskova, Yasuhiro Takeuchi
    Abstract:

    Copyright information:Taken from "Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species"http://www.retrovirology.com/content/4/1/93Retrovirology 2007;4():93-93.Published online 14 Dec 2007PMCID:PMC2241639. PCR products on genomic DNA were aligned using Clustal W software [48]. and have the same identical a.a. sequence in the ECL2 (boxed) of other murine species. RatPAR from different rat cell lines have identical a.a. sequences

  • Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species-3
    2011
    Co-Authors: Giada Mattiuzzo, Magda Matouskova, Yasuhiro Takeuchi
    Abstract:

    Copyright information:Taken from "Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species"http://www.retrovirology.com/content/4/1/93Retrovirology 2007;4():93-93.Published online 14 Dec 2007PMCID:PMC2241639.ic receptors huPAR-2 with L109P mutation and muPAR with P109L mutation (B) were incubated with 1 ml of medium (grey filled) or with the supernatant of 293T containing N-terminal c-myc tagged soluble PERV-A 14/220 SU ENV (bold line). The cells were then immunostained with an anti- human c-myc antibody and a PE conjugated anti-mouse IgG secondary antibody. The histograms show a representative result of at least three independent experiments

  • Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species-1
    2011
    Co-Authors: Giada Mattiuzzo, Magda Matouskova, Yasuhiro Takeuchi
    Abstract:

    Copyright information:Taken from "Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species"http://www.retrovirology.com/content/4/1/93Retrovirology 2007;4():93-93.Published online 14 Dec 2007PMCID:PMC2241639.s huPAR-1 (grey bar) and the mutant H1M g were introduced into QT6 cells by MLV-based retroviral vectors. 50–70% of the QT6 cell population showed PAR expression as confirmed by anti-HA staining. These cultures were infected with EGFP(PERV-A). Cells were harvested 48 hours later and PERV-A infection was measured by flow cytometry as percentage of EGFP-positive cells. Arrows indicate infection below detectable levels. Results are expressed as average ± SEM from three independent experiments

  • Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species-2
    2011
    Co-Authors: Giada Mattiuzzo, Magda Matouskova, Yasuhiro Takeuchi
    Abstract:

    Copyright information:Taken from "Differential resistance to cell entry by porcine Endogenous Retrovirus subgroup A in rodent species"http://www.retrovirology.com/content/4/1/93Retrovirology 2007;4():93-93.Published online 14 Dec 2007PMCID:PMC2241639. B-D is depicted. . HuPAR-2 bearing an N- or C-terminal HA-tag was transiently transfected into 293T cells. After 48 hours cells were treated with saponin (intracellular staining) or without (surface staining). Following immunostaining using an anti-HA antibody and a FITC-conjugated secondary antibody, the samples were visualised either by confocal microscopy (B) or processed by flow cytometry (C). Immunostaining of the cells with anti-human CD71 was used as cell surface protein control. The cells nuclei were counter stained with propidium iodide. . Cell lysates from 293T transiently transfected with an empty pcDNA3 (-), HuPAR-2 (wild type), HA-tagged HuPAR-2 wild type (C-HA wild type) or glycosylation mutant (C-HA N178A) were either treated (+) or untreated (-) with an enzyme removing N-linked oligosaccharide chains (PNGase F) and analysed by western blotting