No Definite Evidence for Human Endogenous Retroviral HERV-W and HERV-H RNAS in Plasma of Latvian Patients Suffering from Multiple Sclerosis and Other Neurological Diseases / Nav Apstiprinošu Pierādījumu Par Endogēno Retrovīrusu HERV-W Un HERV-H Rns Sastopamību Latvijas Pacientu Ar Multiplo Sklerozi Un Citām NeiroloĢiskām Slimībām Asins Plazmā

Open access


Multiple sclerosis (MS) is a neurological disease of unknown aetiology. Several research groups reported an increased level of human endogenous retroviruses HERV-W and HERV-H RNAs in cerebrospinal fluid, plasma and supernatants of cell cultures from MS individuals. To quantify the abundance of extracellular virion-associated HERV, RNAs in blood, plasma samples from Latvian MS patients, patients with other neurological diseases (OND), as well as blood donors (BD), were retrospectively studied by using both our previously published and newly developed quantitative Real-time reverse transcription PCR assays (QPCRs) targeting different polymerase (pol) gene regions of HERV-W and HERV-H. Unspecific signals due to incomplete removal of DNA were monitored by running the assays with and without reverse transcription (RT±) in parallel. According to our score, a few MS, OND and healthy controls gave borderline signals simultaneously with both newly developed HERV-H and HERV-W QPCRs, but the rest were negative. All borderline positive samples also had small amounts of non-retroviral cellular mRNA with possible origin from cell-free circulating RNA fragments, apoptotic bodies or exosomes, which can mimic the previously described virus-like particles. The results do not confirm the previous reports on prevalence of HERV-H or -W virion-associated RNA in plasma of MS patients.

If the inline PDF is not rendering correctly, you can download the PDF file here.

  • Altschul S. F. Madden T. L. Schaffer A. A. Zhang J. Zhang Z. Miller W. Lipman D. J. (1997). Gapped BLAST and PSI-BLAST: A new generation of protein database search programs. Nucleic Acids Res. 25 (17) 3389-3402. Available at: (accessed 27.05.2016).

  • Alvarez-Lafuente R. Garcia-Montojo M. De Las Heras V. Dominguez- Mozo M. I. Bartolome M. Benito-Martin M. S. Arroyo R. (2008). Herpesviruses and human endogenous retroviral sequences in the cerebrospinal fluid of multiple sclerosis patients. Mult. Scler. 14 (5) 595-601.

  • Andersson A. C. Yun Z. Sperber G. O. Larsson E. Blomberg J. (2005). ERV3 and related sequences in humans: Structure and RNA expression. J. Virol. 79 (14) 9270-9284.

  • Antony J. M. Izad M. Bar-Or A. Warren K. G. Vodjgani M. Mallet F. Power C. (2006). Quantitative analysis of human endogenous retrovirus- W env in neuroinflammatory diseases. AIDS Res. Hum. Retroviruses 22 (12) 1253-1259.

  • Antony J. M. Zhu Y. Izad M. Warren K. G. Vodjgani M. Mallet F. Power C. (2007). Comparative expression of human endogenous retrovirus- W genes in multiple sclerosis. AIDS Res. Hum. Retroviruses 23 (10) 1251-1256.

  • Arias M. Fan H. (2014). The saga of XMRV: A virus that infects human cells but is not a human virus. Emerg. Microbes Infect. 3 (4) e.

  • Arru G. Leoni S. Pugliatti M. Mei A. Serra C. Delogu L. G. Manetti R. Dolei A. Sotgiu S. Mameli G. (2014). Natalizumab inhibits the expression of human endogenous retroviruses of the W family in multiple sclerosis patients: A longitudinal cohort study. Mult. Scler. 20 (2) 174-182.

  • Arru G. Mameli G. Astone V. Serra C. Huang Y. M. Link H. Fainardi E. Castellazzi M. Granieri E. Fernandez M. Villoslada P. Fois M. L. Sanna A. Rosati G. Dolei A. Sotgiu S. (2007). Multiple Sclerosis and HERV-W/MSRV: A Multicentric Study. Int. J. Biomed. Sci. 3 (4) 292-297.

  • Asbury A. K. Cornblath D. R. (1990). Assessment of current diagnostic criteria for Guillain-Barre syndrome. Ann. Neurol. 27 Suppl. S21-24. Available at: (accessed 27.05.2016).

  • Balaj L. Lessard R. Dai L. Cho Y. J. Pomeroy S. L. Breakefield X. O. Skog J. (2011). Tumour microvesicles contain retrotransposon elements and amplified oncogene sequences. Nat. Commun. 2 180.

  • Balestrieri E. Pica F. Matteucci C. Zenobi R. Sorrentino R. Argaw- Denboba A. Cipriani C. Bucci I. Sinibaldi-Vallebona P. (2015). Transcriptional activity of human endogenous retroviruses in human peripheral blood mononuclear cells. Biomed. Res. Int. 2015 164529.

  • Blomberg J. Ushameckis D. Jern P. (2005). Evolutionary aspects of human endogenous retroviral sequences (HERVs) and disease. In: Sverdlov E. D. (Ed.). Retroviruses and Primate Genome Evolution. Landes Bioscience Austin TX USA pp. 204-238.

  • Bronson D. L. Fraley E. E. Fogh J. Kalter S. S. (1979). Induction of retrovirus particles in human testicular tumor (Tera-1) cell cultures: An electron microscopic study. J. Natl. Cancer Inst. 63 (2) 337-339. Available at: (accessed 27.05.2016).

  • Bustin S. A. Benes V. Garson J. A. Hellemans J. Huggett J. Kubista M. . . . Wittwer C. T. (2009). The MIQE guidelines: Minimum information for publication of quantitative real-time PCR experiments. Clin. Chem. 55 (4) 611-622.

  • Chomczynski P. Mackey K. (1995). Substitution of chloroform by bromo-chloropropane in the single-step method of RNA isolation. Anal. Biochem. 225 (1) 163-164.

  • Christensen T. (2005). Association of human endogenous retroviruses with multiple sclerosis and possible interactions with herpes viruses. Rev. Med. Virol. 15 (3) 179-211.

  • Christensen T. Dissing Sorensen P. Riemann H. Hansen H. J. Munch M. Haahr S. Moller-Larsen A. (2000). Molecular characterization of HERV-H variants associated with multiple sclerosis. Acta Neurol. Scand. 101 (4) 229-238.

  • Christensen T. Jensen A. W. Munch M. Haahr S. Sorensen P. D. Riemann H. Hansen H. J. Moller-Larsen A. (1997). Characterization of retroviruses from patients with multiple sclerosis. Acta Neurol. Scand. Suppl. 169 49-58.

  • Christensen T. Pedersen L. Sorensen P. D. Moller-Larsen A. (2002). A transmissible human endogenous retrovirus. AIDS Res. Hum. Retroviruses 18 (12) 861-866.

  • Christensen T. Tonjes R. R. zur Megede J. Boller K. Moller-Larsen A. (1999). Reverse transcriptase activity and particle production in B lymphoblastoid cell lines established from lymphocytes of patients with multiple sclerosis. AIDS Res. Hum. Retroviruses 15 (3) 285-291.

  • Compston A. Coles A. (2008). Multiple sclerosis. Lancet 372 (9648) 1502-1517.

  • Contreras-Galindo R. Kaplan M. H. Leissner P. Verjat T. Ferlenghi I. Bagnoli F. Dosik M.. H. Hayes D. F. Gitlin S. D. Markovitz D. M. (2008). Human endogenous retrovirus K (HML-2) elements in the plasma of people with lymphoma and breast cancer. J. Virol. 82 (19) 9329-9336.

  • Cooper A. Poinar H. N. (2000). Ancient DNA: Do it right or not at all. Science 289 (5482) 1139.

  • de Villiers J. N. Treurnicht F. K. Warnich L. Carr J. van Rensburg S. J. Kotze M. J. (2006). Analysis of viral and genetic factors in South African patients with multiple sclerosis. Metab. Brain Dis. 21 (2-3) 163-169.

  • Diem O. Schaffner M. Seifarth W. Leib-Mosch C. (2012). Influence of antipsychotic drugs on human endogenous retrovirus (HERV) transcription in brain cells. PLoS One 7 (1) e30054.

  • Dolei A. Serra C. Mameli G. Pugliatti M. Sechi G. Cirotto M. C. Rosati G. Sotgiu S. (2002). Multiple sclerosis-associated retrovirus (MSRV) in Sardinian MS patients. Neurology 58 (3) 471-473.

  • El-Hefnawy T. Raja S. Kelly L. Bigbee W. L. Kirkwood J. M. Luketich J. D. Godfrey T. E. (2004). Characterization of amplifiable circulating RNA in plasma and its potential as a tool for cancer diagnostics. Clin. Chem. 50 (3) 564-573.

  • Elfaitouri A. Shao X. Mattsson Ulfstedt J. Muradrasoli S. Bolin Wiener A. Golbob S. Ohrmalm C. Matousek M. Zachrisson O. Gottfries C. G. Blomberg J. (2011). Murine gammaretrovirus group G3 was not found in Swedish patients with myalgic encephalomyelitis/chronic fatigue syndrome and fibromyalgia. PLoS One 6 (10) e24602.

  • Flockerzi A. Maydt J. Frank O. Ruggieri A. Maldener E. Seifarth W. Medstrand P. Lengauer T. Meyerhans A. Leib-Mösch C. Meese E. Mayer J. (2007). Expression pattern analysis of transcribed HERV sequences is complicated by ex vivo recombination. Retrovirology 4 39.

  • Forsman A. Yun Z. Hu L. Uzhameckis D. Jern P. Blomberg J. (2005). Development of broadly targeted human endogenous gammaretroviral pol-based real time PCRs Quantitation of RNA expression in human tissues. J. Virol. Methods 129 (1) 16-30.

  • Forsman A. Uzameckis D. Ronnblom L. Baecklund E. Aleskog A. Bindra A. Pipkorn R. Lejniece S. Kozireva S. Murovska M. Blomberg J. (2003). Single-tube nested quantitative PCR: A rational and sensitive technique for detection of retroviral DNA. Application to RERV-H/HRV-5 and confirmation of its rabbit origin. J. Virol. Methods 111 (1) 1-11.

  • Freeman G. H. Halton J. H. (1951). Note on an exact treatment of contingency goodness of fit and other problems of significance. Biometrika 38 (1-2) 141-149. Available at: (accessed 27.05.2016).

  • Froussard P. (1995). Is LM7 a new human retrovirus or a mycoplasma virion? Mult. Scler. 1 (2) 88-94.

  • Garcia-Montojo M. de la Hera B. Varade J. de la Encarnacion A. Camacho I. Dominguez-Mozo M. Árias-Leal A. García-Martínez A. Casanova I. Izquierdo G. Lucas M. Fedetz M. Alcina A. Arroyo R. Matesanz F. Urcelay E. Alvarez-Lafuente R. (2014). HERV-W polymorphism in chromosome X is associated with multiple sclerosis risk and with differential expression of MSRV. Retrovirology 11 2.

  • Garcia-Montojo M. Dominguez-Mozo M. Arias-Leal A. Garcia-Martinez A. De las Heras V. Casanova I. Faucard R. Gehin N. Madeira A. Arroyo R. Curtin F. Alvarez-Lafuente R. Perron H. (2013). The DNA copy number of human endogenous retrovirus-W (MSRV-type) is increased in multiple sclerosis patients and is influenced by gender and disease severity. PLoS One 8 (1) e53623.

  • Garson J. A. Tuke P. W. Giraud P. Paranhos-Baccala G. Perron H. (1998). Detection of virion-associated MSRV-RNA in serum of patients with multiple sclerosis. Lancet 351 (9095) 33.

  • Gefrides L. A. Powell M. C. Donley M. A. Kahn R. (2010). UV irradiation and autoclave treatment for elimination of contaminating DNA from laboratory consumables. Forensic Sci. Int. Genet. 4 (2) 89-94.

  • Gifford R. Tristem M. (2003). The evolution distribution and diversity of endogenous retroviruses. Virus Genes 26 (3) 291-315.

  • Greijer A. E. Dekkers C. A. Middeldorp J. M. (2000). Human cytomegalovirus virions differentially incorporate viral and host cell RNA during the assembly process. J. Virol. 74 (19) 9078-9082. Available at: (accessed 27.05.2016).

  • Griffiths D. J. Voisset C. Venables P. J. Weiss R. A. (2002). Novel endogenous retrovirus in rabbits previously reported as human retrovirus 5. J. Virol. 76 (14) 7094-7102.

  • Haahr S. Sommerlund M. Christensen T. Jensen A. W. Hansen H. J. Moller-Larsen A. (1994). A putative new retrovirus associated with multiple sclerosis and the possible involvement of Epstein-Barr virus in this disease. Ann. NY Acad. Sci. 724 148-156.

  • Hasselmann D. O. Rappl G. Tilgen W. Reinhold U. (2001). Extracellular tyrosinase mRNA within apoptotic bodies is protected from degradation in human serum. Clin. Chem. 47 (8) 1488-1489. Available at: (accessed 27.05.2016).

  • Jern P. Sperber G. O. Ahlsen G. Blomberg J. (2005). Sequence variability gene structure and expression of full-length human endogenous retrovirus H. J. Virol. 79 (10) 6325-6337.

  • Jern P. Sperber G. O. Blomberg J. (2004). Definition and variation of human endogenous retrovirus H. Virology 327 (1) 93-110.

  • Kamm R. C. Smith A. G. (1972). Nucleic acid concentrations in normal human plasma. Clin. Chem. 18 (6) 519-522. Available at: (accessed 27.05.2016).

  • Kim H. S. (2001). Sequence and phylogeny of HERV-W pol fragments. AIDS Res. Hum. Retroviruses 17 (17) 1665-1671.

  • Knapp M. Clarke A. C. Horsburgh K. A. Matisoo-Smith E. A. (2012). Setting the stage - building and working in an ancient DNA laboratory. Ann. Anat. 194 (1) 3-6.

  • Komurian-Pradel F. Paranhos-Baccala G. Bedin F. Ounanian-Paraz A. Sodoyer M. Ott C. Rajoharison A. Garcia E. Mallet F. Mandrand B. Perron H. (1999). Molecular cloning and characterization of MSRV-related sequences associated with retrovirus-like particles. Virology 260 (1) 1-9.

  • Kurtzke J. F. (2000). Epidemiology of multiple sclerosis. Does this really point towards an etiology? Neurol. Sci 21 383-403.

  • Lander E. S. Linton L. M. Birren B. Nusbaum C. Zody M. C. Baldwin J. ... International Human Genome Sequencing Consortium (2001). Initial sequencing and analysis of the human genome. Nature 409 (6822) 860-921.

  • Laufer G. Mayer J. Mueller B. F. Mueller-Lantzsch N. Ruprecht K. (2009). Analysis of transcribed human endogenous retrovirus W env loci clarifies the origin of multiple sclerosis-associated retrovirus env sequences. Retrovirology 6 37.

  • Leonard J. A. Shanks O. Hofreiter M. Kreuz E. Hodges L. Ream W. Wayne R. K. Fleischer R. C. (2007). Animal DNA in PCR reagents plagues ancient DNA research. J. Archaeol. Sci. 34 (9) 1361-1366.

  • Liggett T. Melnikov A. Tilwalli S. Yi Q. Chen H. Replogle C. Feng X. Reder A. Stefoski D. Balabanov R. Levenson V. (2010). Methylation patterns of cell-free plasmaDNAin relapsing-remitting multiple sclerosis. J. Neurol. Sci. 290 (1-2) 16-21.

  • Lindeskog M. Medstrand P. Cunningham A. A. Blomberg J. (1998). Coamplification and dispersion of adjacent human endogenous retroviral HERV-H and HERV-E elements; presence of spliced hybrid transcripts in normal leukocytes. Virology 244 (1) 219-229.

  • Liu C. Chen Y. Li S. Yu H. Zeng J. Wang X. Zhu F. (2013). Activation of elements in HERV-W family by caffeine and aspirin. Virus Genes 47 (2) 219-227.

  • Lo Y. M. Chan K. C. (2006). Setting up a polymerase chain reaction laboratory. Methods Mol. Biol. 336 11-18.

  • Longo M. C. Berninger M. S. Hartley J. L. (1990). Use of uracil DNA glycosylase to control carry-over contamination in polymerase chain reactions. Gene 93 (1) 125-128.

  • Munch M. Moller-Larsen A. Christensen T. Morling N. Hansen H. J. Haahr S. (1995). B-lymphoblastoid cell lines from multiple sclerosis patients and a healthy control producing a putative new human retrovirus and Epstein-Barr virus. Mult. Scler. 1 (2) 78-81.

  • Muralidharan-Chari V. Clancy J. Plou C. Romao M. Chavrier P. Raposo G. D'Souza-Schorey C. (2009). ARF6-regulated shedding of tumor cell-derived plasma membrane microvesicles. Curr. Biol. 19 (22) 1875-1885.

  • Nellaker C. Li F. Uhrzander F. Tyrcha J. Karlsson H. (2009). Expression profiling of repetitive elements by melting temperature analysis: Variation in HERV-W gag expression across human individuals and tissues. BMC Genomics 10 532.

  • Nowak J. Januszkiewicz D. Pernak M. Liwen I. Zawada M. Rembowska J. Nowicka K. Lewandowski K. Hertmanowska H.Wender M. (2003). Multiple sclerosis-associated virus-related pol sequences found both in multiple sclerosis and healthy donors are more frequently expressed in multiple sclerosis patients. J. Neurovirol. 9 (1) 112-117.

  • Ou C. Y. Moore J. L. Schochetman G. (1991). Use of UV irradiation to reduce false positivity in polymerase chain reaction. Biotechniques 10 (4) 442 444 446.

  • Perron H. Garson J. A. Bedin F. Beseme F. Paranhos-Baccala G. Komurian-Pradel F. Mallet F. Tuke P. W. Voisset C. Blond J. L. Lalande B. Seigneurin J. M. Mandrand B. (1997). Molecular identification of a novel retrovirus repeatedly isolated from patients with multiple sclerosis. The Collaborative Research Group on Multiple Sclerosis. Proc. Natl. Acad. Sci. USA 94 (14) 7583-7588.

  • Perron H. Geny C. Laurent A. Mouriquand C. Pellat J. Perret J. Seigneurin J. M. (1989). Leptomeningeal cell line from multiple sclerosis with reverse transcriptase activity and viral particles. Res. Virol. 140 (6) 551-561.

  • Perron H. Lalande B. Gratacap B. Laurent A. Genoulaz O. Geny C. Mallaret M. Schuller E. Stoebner P. Seigneurin J. M. (1991). Isolation of retrovirus from patients with multiple sclerosis. Lancet 337 (8745) 862-863.

  • Perron H. Perin J. P. Rieger F. Alliel P. M. (2000). Particle-associated retroviral RNA and tandem RGH/HERV-W copies on human chromosome 7q: Possible components of a 'chain-reaction' triggered by infectious agents in multiple sclerosis? J. Neurovirol. 6 (Suppl 2) S67-75.

  • Polman C. H. Reingold S. C. Edan G. Filippi M. Hartung H. P. Kappos L. Lublin F. D. Metz L. M. McFarland H. F. O'Connor P. W. Sandberg-Wollheim M. Thompson A. J. Weinshenker B. G. Wolinsky J. S. (2005). Diagnostic criteria for multiple sclerosis: 2005 revisions to the “McDonald Criteria”. Ann. Neurol. 58 (6) 840-846.

  • Pruvost M. Grange T. Geigl E. M. (2005). Minimizing DNA contamination by using UNG-coupled quantitative real-time PCR on degraded DNA samples: Application to ancient DNA studies. Biotechniques 38 (4) 569-575.

  • Pugliatti M. Rosati G. Carton H. Riise T. Drulovic J. Vecsei L. Milanov I. (2006). The epidemiology of multiple sclerosis in Europe. Eur. J. Neurol. 13 (7) 700-722.

  • Schmitt K. Richter C. Backes C. Meese E. Ruprecht K. Mayer J. (2013). Comprehensive analysis of human endogenous retrovirus group HERV-W locus transcription in multiple sclerosis brain lesions by high-throughput amplicon sequencing. J. Virol. 87 (24) 13837-13852.

  • Sciortino M. T. Suzuki M. Taddeo B. Roizman B. (2001). RNAs extracted from herpes simplex virus 1 virions: Apparent selectivity of viral but not cellular RNAs packaged in virions. J. Virol. 75 (17) 8105-8116. Available at: (accessed 27.05.2016).

  • Seifarth W. Skladny H. Krieg-Schneider F. Reichert A. Hehlmann R. Leib-Mosch C. (1995). Retrovirus-like particles released from the human breast cancer cell line T47-D display type B- and C-related endogenous retroviral sequences. J Virol 69 (10) 6408-6416. Available at: (accessed 27.05.2016).

  • Serra C. Sotgiu S. Mameli G. Pugliatti M. Rosati G. Dolei A. (2001). Multiple sclerosis and multiple sclerosis-associated retrovirus in Sardinia. Neurol. Sci. 22 (2) 171-173.

  • Shah K. V. (2007). SV40 and human cancer: A review of recent data. Int. J. Cancer 120 (2) 215-223.

  • Siegel S. R. Mackenzie J. Chaplin G. Jablonski N. G. Griffiths L. (2012). Circulating microRNAs involved in multiple sclerosis. Mol. Biol. Rep. 39 (5) 6219-6225.

  • Soper D. S. (2015). Fisher's Exact Test Calculator for a 3X3 Contingency Table [Software]. Available at: (accessed 27.05.2016).

  • Sperber G. Lovgren A. Eriksson N. E. Benachenhou F. Blomberg J. (2009). RetroTector online a rational tool for analysis of retroviral elements in small and medium size vertebrate genomic sequences. BMC Bioinformatics 10 (Suppl 6) S4.

  • Sperber G. O. Airola T. Jern P. Blomberg J. (2007). Automated recognition of retroviral sequences in genomic data-RetroTector. Nucleic Acids Res. 35 (15) 4964-4976.

  • Tamariz J. Voynarovska K. Prinz M. Caragine T. (2006). The application of ultraviolet irradiation to exogenous sources of DNA in plasticware and water for the amplification of low copy number DNA. J. Forensic Sci. 51 (4) 790-794.

  • Thery C. Amigorena S. Raposo G. Clayton A. (2006). Isolation and characterization of exosomes from cell culture supernatants and biological fluids. Curr. Protoc. Cell Biol. Chapt. 3 Unit 3 22.

  • Thompson J. D. Higgins D. G. Gibson T. J. (1994). CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting position-specific gap penalties and weight matrix choice. Nucleic Acids Res. 22 (22) 4673-4680. Available at: (accessed 27.05.2016).

  • Tristem M. (2000). Identification and characterization of novel human endogenous retrovirus families by phylogenetic screening of the human genome mapping project database. J. Virol. 74 (8) 3715-3730.

  • Tuke P. W. Perron H. Bedin F. Beseme F. Garson J. A. (1997). Development of a pan-retrovirus detection system for multiple sclerosis studies. Acta Neurol. Scand. Suppl. 169 16-21. Available at: (accessed 27.05.2016).

  • Unger M. Wettergren A. Clausen J. (1985). Characterization of DNA and RNA in circulating immunocomplexes in multiple sclerosis amyotrophic lateral sclerosis and normal controls. Acta Neurol. Scand. 72 (4) 392-396. Available at: (accessed 27.05.2016).

  • Vandesompele J. De Preter K. Pattyn F. Poppe B. Van Roy N. De Paepe A. Speleman F. (2002). Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 3 (7) RESEARCH0034. Available at: (accessed 27.05.2016).

  • Wieczorek A. J. Rhyner C. Block L. H. (1985). Isolation and characterization of an RNA-proteolipid complex associated with the malignant state in humans. Proc. Natl. Acad. Sci. USA 82 (10) 3455-3459. Available at: (accessed 27.05.2016).

  • Wieczorek A. J. Sitaramam V. Machleidt W. Rhyner K. Perruchoud A. P. Block L. H. (1987). Diagnostic and prognostic value of RNA-proteolipid in sera of patients with malignant disorders following therapy: First clinical evaluation of a novel tumor marker. Cancer Res. 47 (23) 6407-6412. Available at: (accessed 27.05.2016).

Journal information
Impact Factor

CiteScore 2018: 0.3

SCImago Journal Rank (SJR) 2018: 0.137
Source Normalized Impact per Paper (SNIP) 2018: 0.192

Cited By
All Time Past Year Past 30 Days
Abstract Views 0 0 0
Full Text Views 379 209 5
PDF Downloads 155 94 3