• Korean Journal of Agricultural Science
• /
• v.38 no.1
• /
• pp.71-77
• /
• 2011

Pigs have anatomically and physiologically very similar to human and because of this, pigs are the possible xenotransplantation donors for human organs. PERVs (Porcine Endogenous Retroviruses) are known to be one of the possible obstacles for using porcine organs regardless of the immunological barriers. In order to understand the expression patterns of PERVs in Korean native pigs, we investigated PERV expressions in porcine liver, heart, spleen, and lung samples. After RNA extraction, two types of specific PERV envelope genes (ENV-A and ENV-B) were amplified using specific primers by RT-PCR. The results indicated that the variable PERV expressions were observed in inconsistent patterns among animals and tissues. The PERV expressions were verified with semi-quantitative real-time PCR with three replicates. Even though, these results confirm the previous findings that the PERVs were differentially expressed between animals and tissues. These results also give some valuable information for xenotransplantation when using the Korean native pigs as the organ donor.

• Journal of Microbiology and Biotechnology
• /
• v.24 no.4
• /
• pp.577-583
• /
• 2014

Three major classes of retroviral restriction factors (APOBEC3G, Tetherin, and TRIM5${\alpha}$) have been identified in mammals. Restriction factors are cellular proteins that are able to limit viral replication by targeting specific steps of the viral life cycle. To evaluate which restriction factor is the most effective to inhibit the replication of porcine endogenous retroviruses (PERVs), the antiviral activity of each restriction factor was compared. In pseudotype assay, the antiviral activity of human tetherin against PERV pseudotype was slightly weaker than that of human APOBEC3G (hA3G). A combination of tetherin and hA3G was more potent than each individual restriction factor. We questioned whether a combination of tetherin and hA3G could also inhibit the spreading replication of PERV. In agreement with the pseudotype assay, two restriction factors inhibit infectious PERV replication in a spreading infection. In this study, hA3G could strongly inhibit the replication of PERV, but tetherin modestly restricted it. Based on these results, we concluded that a combination of tetherin and hA3G is the most effective way to restrict PERV. A combination of different restriction factors will encourage the development of a new approach to treat retroviral disease.

• Journal of Microbiology and Biotechnology
• /
• v.18 no.3
• /
• pp.585-590
• /
• 2008

Human tropic Porcine Endogenous Retroviruses (PERVs) are the major concern in zoonosis for xenotransplantation because PERVs cannot be eliminated by specific pathogen-free breeding. Recently, a PERV A/C recombinant with PERV-C bearing PERV-A gp70 showed a higher infectivity (approximately 500-fold) to human cells than PERV-A. Additionally, the chance of recombination between PERVs and HERVs is frequently stated as another risk of xenografting. Overcoming zoonotic barriers in xenotransplantation is more complicated by recombination. To achieve successful xenotransplantation, studies on the recombination in PERVs are important. Here, we cloned and sequenced proviral PERV env sequences from pig gDNAs to analyze natural recombination. The envelope is the most important element in retroviruses as a pivotal determinant of host tropisms. As a result, a total of 164 PERV envelope genes were cloned from pigs (four conventional pigs and two miniature pigs). Distribution analysis and recombination analysis of PERVs were performed. Among them, five A/B recombinant clones were identified. Based on our analysis, we determined the minimum natural recombination frequency among PERVs to be 3%. Although a functional recombinant envelope clone was not found, our data evidently show that the recombination event among PERVs may occur naturally in pigs with a rather high possibility.

• Asian-Australasian Journal of Animal Sciences
• /
• v.20 no.4
• /
• pp.461-465
• /
• 2007

Porcine Endogenous Retroviruses (PERVs) are a major concern in relation to xenotransplantation. Previous research indicated that PERVs are present at about 50 copies in the pig genome and their chromosomal insertion sites are different among pig breeds. We examined nine Korean native pigs and seven Asian Wild Boars for the presence of a PERV-A at SSC 1q2.4 and a PERV-B at SSC 7p1.1-2 previously reported in a Large White pig. The PERV-B at locus 7p1.1-2 displayed insertional variability in Korean native pigs and Asian Wild Boars. Using the primers for the PERV-A at 1q2.4 from Large White pig, we only can amplify an unclassified 798 bp sequence, which showed insertional variability only in Korean native pigs. This study indicates that there are differences within and between Asian and European pigs in PERV insertions and suggests that selection could generate PERV-free lines of pigs more suitable for xenotransplantation.

• Asian-Australasian Journal of Animal Sciences
• /
• v.21 no.11
• /
• pp.1572-1575
• /
• 2008

The xenotransplantation of pig organs and cells can be related with a risk of transmission of infectious diseases to human. Previous findings indicate that the regulatory region of PERV for retroviral transcription, replication and integration into the cellular DNA is located on the 5' Long Terminal Repeat (LTR). The objective of this study is the investigation of methylation and deletion status of the PERV 5' LTR region which can be used for regulating PERV expression. We compared the sequences of genomic DNA and bisulfite-treated genomic DNA from PK-15 cells expressing PERV to observe the methylation status of the 5' LTR. Our results showed that the CpG sites of U3 were methylated and methylation was inconsistent in the R and U5 regions. Also, variable numbers of 18 bp repeats and 21 bp repeats were detected on 5' LTR by sequencing analysis. The consistent U3 methylation might be indicative of host suppression of expression of the retroviruses.

• Korean Journal of Veterinary Research
• /
• v.49 no.3
• /
• pp.215-220
• /
• 2009

The prevalence of potentially xenozoonotic viruses in the reproductive tract of female pigs in Korea was investigated by polymerase chain reaction (PCR). These viruses include porcine endogenous retrovirus (PERV), porcine reproductive and respiratory syndrome virus (PRRSV), swine hepatitis E virus (SHEV), porcine lymphotropic herpesvirus (PLHV), and porcine circovirus type 2 (PCV-2). Histopathological examination and PCR analysis were conducted using the ovaries of 70 slaughtered pigs that were collected from 14 farms in Jeju. Histopathologically, infiltrations of mononuclear inflammatory cells around the thick-walled coiled vessels in the ovarian medulla were observed in 15 cases. Based on the PCR method, PERV, PLHV, PRRSV, SHEV, and PCV-2 were detected in 69 (98.6%), 35 (50%), 5 (7.1%), 4 (5.7%), and 1 sample (1.4%), respectively. These results suggest that PERV and PLHV are the major xenozoonotic viruses in the porcine ovary. This study should aid in the development of a monitoring protocol for potential xenozoonotic agents and in the production of germ-free pigs for xenotransplantation.

• Journal of Animal Science and Technology
• /
• v.47 no.2
• /
• pp.177-186
• /
• 2005

Xenotransplantation may help to overcome the critical shortage of human tissues and organs for human transplantation, Swine represents an ideal source of such organs owing to their anatomical and physiological similarities to human besides their plentiful supply, However, the use of organs across the species barrier may be associated with the risk of transmission of pathogens, specially porcine endogenous retroviruses (PERVs).• Although most of these potential pathogens could be eliminated by pathogen-free breeding, PERVs are not eliminated by this treatment. PERVs are integrated into the genome of all pigs and produced by normal pig cells and infect human cells. They belong to gamma retroviruses and are of three classes viruses: A, B and C. In the present study, PCR based cloning was performed with chromosomal DNA extracted from pigs from domestic pigs in Korea. Amplified PCR fragments of about 1.5 Kb, covering the partial env gene, were cloned into pCR2.l-TOPO vectors and sequenced. A total of 91 env clones were obtained from domestic pigs, Berkshire, Duroc, Landrace and Yorkshire in Korea. Phylogenetic analysis of these genes revealed the presence of only PERV class A and B in the proportion of 58 % and 42 %, respectively. Among these, 28 clones had the correct open reading frame: 18 clones in class A and 10 clones in class B. Since both these PERV classes are polytropic and have the capacity to infect human cells, our data suggest that proviral PERVs have the potential to generate infectious viruses during or after xenotransplantation in human.

• Proceedings of the Korean Society of Life Science Conference
• /
• 2006.09a
• /
• pp.65.1-65.1
• /
• 2006

• Nuclear Engineering and Technology
• /
• v.36 no.4
• /
• pp.357-363
• /
• 2004

A prototype long pulse ion source was developed, and the beam extraction experiments of the ion source were carried out at the Neutral Beam Test Stand (NBTS) of the Korea Superconducting Tokamak Advanced Research (KSTAR). The ion source consists of a magnetic bucket plasma generator, with multi-pole cusp fields, and a set of tetrode accelerators with circular apertures. Design requirements for the ion source were a 120kV/65A deuterium beam and a 300 s pulse length. Arc discharges of the plasma generator were controlled by using the emission-limited mode, in turn controlled by the applied heating voltage of the cathode filaments. Stable and efficient arc plasmas with a maximum arc power of 100 kW were produced using the constant power mode operation of an arc power supply. A maximum ion density of $8.3{\times}10^{11}\;cm^{-3}$ was obtained by using electrostatic probes, and an optimum arc efficiency of 0.46 A/kW was estimated. The accelerating and decelerating voltages were applied repeatedly, using the re-triggering mode operation of the high voltage switches during a beam pulse, when beam disruptions occurred. The decelerating voltage was always applied prior to the accelerating voltage, to suppress effectively the back-streaming electrons produced at the time of an initial beam formation, by the pre-programmed fast-switch control system. A maximum beam power of 0.9 MW (i.e. $70\;kV{\times}12.5\;A$) with hydrogen was measured for a pulse duration of 0.8 s. Optimum beam perveance, deduced from the ratio of the gradient grid current to the total beam current, was $0.7\;{\mu}perv$. Stable beams for a long pulse duration of $5{\sim}10\;s$ were tested at low accelerating voltages.