Abstracts og konklusjoner fra den internasjonale retroviruskonferansen

Posted on 08/06/2011

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Denne hadde jeg helt glemt å publisere, så den kommer nå. Noen notater på norsk, ellers en god oversikt.

Tidligere skrev jeg om en presentasjon/publikasjon fra WPI som viste at XMRV-smittede ME/CFS-pasienter hadde endringer i immunforsvaret. Dette ble presentert på den  15. Internasjonale Konferansen om Human Retrovirus: HTLV og relaterte virus, som avholdes i Belgia nå i disse dager (5.-8.juni).

Det er selvfølgelig flere interessante ting å rapportere fra denne konferansen, som er aktuelt i forhold til XMRV. Her følger en oversikt over de abstracts/oppsummeringer som er lagt ut nå.  Oversikten er en viderutvikling av den som egentlig ble laget av XMRV Global Action, men jeg har redigert den litt og tatt med litt ekstra informasjon.

Fra den 15. Internasjonale Konferansen om Human Retrovirus: HTLV og relaterte virus.

Leuven og Gembloux, Belgia. 5.-8. juni 2011

 

Alle disse er publisert på Retrovirology sine sider.

 

XMRV replicates preferentially in mucosal sites in vivo: Relevance to XMRV transmission?

http://www.retrovirology.com/content/8/S1/A219

Francois Villinger, Jaydip Das Gupta, Nattawat Onlamoon, Ross Molinaro, Suganthi Suppiah, Prachi Sharma, Kenneth Rogers, Christina Gaughan, Eric Klein, Xiaoxing Qiu, Gerald Schochetman, John Hackett Jr og Robert H Silverman

Institusjoner: Emory University (flere avdelinger) (Atlanta, USA), Cleveland Clinic Foundation (Ohio, USA), Mahidol University (Bangkok, Thailand), Abbott Diagnostics (Illinois, USA)

Forskerne ville se på infeksjonsdynamikk og spredning in vivo. Brukte to typer makake-aper i forsøkene.

Resultatene viser at XMRV induserer en kronisk og klinisk stille infeksjon, som likevel er vedvarende og mottakelig for reaksjon in vivo. XMRV ser ut til å gå raskt ut av blodsirkulasjonen, men XMRV protein positive CD4+ T-celler ble funnet i alle lymfoide organer gjennom infeksjonen. Det viste seg at slimhinner generelt og kjønnsorganer hadde høyere frekvens av XMRVgag positive celler, inkludert fordøyelsessystemets slimhinner, lungene og reproduktive organer.  Resultatene av denne forskningen tyder på et potensiale for seksuell overføring av XMRV, og de har vist i forsøk at slik overføring er mulig. (Dette er min korte oversettelse av noen punkter fra sammendraget, les hele her)

 

Development of XMRV producing B Cell lines from lymphomas from patients with Chronic Fatigue Syndrome

http://www.retrovirology.com/content/8/S1/A230

Francis Ruscetti, Vincent C Lombardi, Michael Snyderman, Dan Bertolette, Kathryn S Jones, Judy A Mikovits

Institusjoner: National Cancer Institute, Center for Cancer Research (Maryland, USA), Whittemore Peterson Institute (Nevada, USA), State University of New York, Dept. of Medicine (New York, USA), NCI-Frederick (Maryland, USA)

I en studie av 300 CFS-pasienter, utviklet 13 «lymphoproliferative disorders» (sykdomstilstander hvor det overproduseres lymfocytter, opptrer hos pasienter med immundefekter.) Av disse 13 pasientene ble 11 testet og alle var positive for XMRV. 9 ble testet for «clonal TCR gamma rearrangements», alle var positive. Spontan utvikling av fire B-cellelinjer skjedde under kulturering av celler fra CFS-pasientene. Forskerne konkluderer med at XMRV-infeksjon kan akselerere utviklingen av B-celle maligniteter enten ved indirekte kronisk stimulering av immunsystetem og/eller ved direkte infeksjon av B-celle avstamning. (Slik jeg forstår dette mener de at XMRV-infeksjon gir større risiko for kreft.) Siden virusbeholdningen i blodsystemet er lavt, tyder disse dataene på at B-celler i vev slik som milt og lymfeknuter kan være et in vivo reservoar for XMRV. (Dette er min korte oversettelse av noen punkter fra sammendraget, les hele her)

 

Prevalence of XMRV in blood donors, HTLV and HIV cohorts

http://www.retrovirology.com/content/8/S1/A222

Xiaoxing Qiu, Priscilla Swanson, Ning Tang, Gregor W Leckie, Sushil Devare, Gerald Schochetman, John Hackett Jr

Institusjoner: Abbott Diagnostics, Infectious Diseases og Abbott Molecular Inc. (Illinois, USA)

Forskerne ville se etter antistoffer utløst av XMRV-infeksjon, og studerte blodgivere og retrovirus-infiserte personer for serologisk bevis for en XMRV-infeksjon. De tok blod fra 1000 amerikanske bloddonorer, 100 HIV-1-infiserte kamerunere, 486 HTLV-1-infiserte japanere og 156 ikke-HTLV-infiserte japanske kontroller og screenet dem for antistoffer for XMRV. XMRV seroprevalens varierte fra 0 til 0,6% hos amerikasnke blodgivere, HIV-1-infiserte og HTLV uinfiserte forsøkspersoner. 4,1% av de japanske HTLV-1-infiserte personene var p15E-reaktive. «Inspection of sequence homology between HTLV and XMRV revealed a high level of conservation within the immunodominant region of HTLV gp21 suggesting increased seroreactivity is due to cross-reactive antibodies.» (Dette er min korte oversettelse av noen punkter fra sammendraget, les hele her)


Multi-laboratory evaluations of XMRV nucleic acid detection assays

http://www.retrovirology.com/content/8/S1/A231

Graham Simmons, John M Coffin, Indira K Hewlett, Shyh-Ching Lo, Judy A Mikovits, William M Switzer, Jeffrey M Linnen, Francis Ruscetti, Simone A Glynn, Michael P Busch

Institusjoner: Blood Systems Research Institute (San Francisco, USA),   Department of Laboratory Medicine, University of California,   National Cancer Institute-Frederick (Maryland, USA), Department of Molecular Biology and Microbiology, Tufts University (Boston, USA), Office of Blood Research and Review, FDA (Bethesda, Maryland, USA), Division of Cellular and Gene Therapies and Division of Human Tissues, FDA (Bethesda, Maryland, USA)

(Det vi ofte omtaler på norske blogger som «blodarbeidsgruppen».) «The Blood XMRV Scientific Research Working Group» ble dannet for å fasilitere samarbeidsstudier for betydningen av XMRV for blodgivere (og blodforsyninen). Studiene skal evaluere hvordan man finner XMRV, testmetodene som brukes og hvor sensitive, spesifikke og reproduserbare de er. De skal også teste disse metodene på prøver fra blodbanken og bestemme forekomsten av XMRV i blodgivere. I Fase I testet man og fant ut at alle de seks deltakende laboratoriene har sensitive nukleærsyretester. I fase 2 kjørte man pilotstudier for å sammenligne XMRV-testing ved ulike metoder. To av laboratoriene fant MLV-liknende sekvenser i blodet. Et tredje laboratorium fant ingen virale sekvenser. Fase 2 hadde altså litt blandede resultater. Resultatene fra fase 3 er ventet snart. Fase 4 vil teste et blindet panel av 300 blodgiverprøver. (Dette er min korte oversettelse av noen punkter fra sammendraget, les hele her)

 

 

Immune correlates of XMRV infection

http://www.retrovirology.com/content/8/S1/A221

Vincent Lombardi, Deborah Goetz, Max Pfost, Cassandra Puccinelli, Judy Mikovits

Institusjoner: Whittemore Peterson Institute (Reno, USA), Environmental Sciences, University of Nevada (Reno,USA)

Denne har jeg allerede omtalt ganske grundig her. Vi tar litt klipp og lim:

I oppsummeringen til artikkelen skriver de at CFS-pasienter ofte har en dysfunksjon i det antivirale enzymen RNase L, noe som understreker viktigheten av de uspesifikke immunresponsene i CFS. XMRV-infeksjon kan spille en rolle i patogenesen (sykdomsforståelsen) av CFS, gjennom dysregulering av immunresponsen.

For å teste denne hypotesen brukte de avanserte metoder for å se på cytokiner og chemokiner i plasma hos XMRV-infiserte CFS-pasienter versus uinfiserte kontroller. Seks normale donorer og referanseverdier basert på en frisk populasjon ble brukt som normale baselines. Resultatet var at 16 av de 26 cytokinene/chemokinene målt var signifikant uttrykt annerledes:

11 var oppregulert og 5 ned-regulert, inkludert: IL-8, IL-6, MIP1 alpha, MCP-1, IFN alpha and TNF alpha. XMRV-infiserte CFS-pasienter viste redusert prosent av CD56+ NK og CD19+ B-celler. NK-fenotypen hos XMRV-infiserte pasienter var endret, 80% av pasientene hadde en signifikant redusert CD56+ DIM populasjon.

Konklusjon: XMRV-infeksjon resulterer i dysregulering av immun-responsen, enten direkte ved infeksjon av spesifikke leukocytt-undergrupper eller indirekte gjennom cytokin-modulering.

 

The effects of XMRV gene expression on the mouse prostate

http://www.retrovirology.com/content/8/S1/A223

Daniel Rauch, Sirosh Bokhari, John Harding, Lee Ratner

Institusjon: Division of Molecular Oncology (St Louis, USA)

Forskerne har forsøkt å bestemme om XMRV genuttrykk er tilstrekkelig for å fremme patologi i prostata i en muse-modell. De var i stand til å finne XMRV genuttrykk i prostataen til forsøksmusene.

«Breeding PRO-XMRV mice with PRO-XPR1 mice will allow us to test whether XMRV integration or gene expression can cause more advanced prostate pathology in vivo. With these XMRV mouse models we seek to address the question that remains unanswered to date as to whether XMRV is capable of causing prostate dysplasia or cancer in vivo.»

Les hele sammendraget her.

XMRV: usage of receptors and potential co-receptors

http://www.retrovirology.com/content/8/S1/A224

Mohan K H G Setty, Krishnakumar Devadas, Ragupathy Viswanath, Veeraswamy Ravichandran, Shixing Tang, Owen Wood, Durga S Gaddam, Sherwin Lee, Indira K Hewlett

Institusjon: Center for Biologics Evaluation and Research – Food and Drug Administration (Bethesda, USA)

Forskerne her så på reseptorer og co-reseptorer for å finne ut blant annet mer om smitte, infeksjon og hvordan XMRV er ineffektiv i forskjellige celletyper.  Infeksjon og replikasjon ac XMRV ble sett i flere typer celler. Nivåene varierte i forskjellige celle-linjer.

«XMRV replication was observed in GHOST cells that express CD4, and each of the chemokine receptors ranging from CCR1- CCR8 and Bob suggesting that infectivity in hematopoietic cells could be mediated by use of these receptors. Infection of Lung epithelial cell A549 lacking XPR1 expression clearly indicates usage of other receptors by XMRV for entry into susceptible cells.»

 

Cell line tropism and replication of XMRV

http://www.retrovirology.com/content/8/S1/A225

Krishnakumar Devadas, Mohan K H G Setty, Ragupathy Viswanath, Durga S Gaddam, Owen Wood, Shixing Tang, Jiangqin Zhao, Xue Wang, Veeraswamy Ravichandran, Sherwin Lee, Indira K Hewlett

Institusjoner: Center for Biologics Evaluation and Research, Food and Drug Administration (Bethesda, USA)

Forskerne kunne i denne studien observere replikasjon av XMRV blant annet i celler fra cervix (livmoren) og lungeceller.

«Replication of XMRV could be observed in cervical and lung epithelial cells, T-cell lines Jurkat and H9, B-cell line HL60, U937 cells and in primary PBMC and monocyte-derived macrophages. The levels of XMRV transcripts were lower in primary monocytes compared to T-cell lines suggesting less efficient replication in these cells.»

 

Structure of the xenotropic murine leukaemia virus-related virus matrix protein

http://www.retrovirology.com/content/8/S1/A227

Michal Doležal, Iva Pichová, Tomáš Ruml, Richard Hrabal, Michaela Rumlová

Institusjoner: Institute of Organic Chemistry and Biochemistry, IOCB Research Centre and Gilead Sciences, Academy of Sciences of the Czech Republic, Department of Biochemistry and Microbiology, Institute of Chemical Technology Prague og Laboratory of NMR spectroscopy, Institute of Chemical Technology Prague (alle i Praha, Tsjekkia)

«Although the protein sequence of the XMRV-MA is very similar to that of the murine leukaemia virus matrix protein (MLV-MA), it varies in several amino acid residues. We compared the structures of the XMRV-MA and MLV-MA and found that those changes are localized in a few domains, mostly on the surface of the protein.»

 

Serologic and PCR testing of persons with chronic fatigue syndrome in the United States shows no association with xenotropic or
polytropic murine leukemia virus-related virus

http://www.retrovirology.com/content/pdf/1742-4690-8-s1-a232.pdf

William M Switzer, Hongwei Jia, HaoQiang Zheng, Shaohua Tang, Rebecca A Garcia, Brent C Satterfield

Institusjoner: Division of HIV/AIDS Prevention, CDC (Atlanta, USA), Cooperative Diagnostics, LLC (Greenwood, USA)

«Our findings are consistent with previous negative reports and do not support an association of XMRV or MuLV in the majority of CFS cases across the US.»

 

Detection of MLV-like gag sequences in blood samples from a New York state CFS cohort

http://www.retrovirology.com/content/8/S1/A234

Maureen R Hanson , Li L Lee, Lin Lin, David E Bell, David Ruppert and David S Bell

Institusjoner: Dept. of Molecular Biology and Genetics, Cornell University (Ithaca, USA), Dept. of Medical Anthropology, State University of New York (Buffalo, USA), School of Operations Research and Information Engineering, Cornell University (Ithaca,  USA), Dept. of Pediatrics, State University of New York (Buffalo, USA)

Disse forskerne fant altså tegn på MLV-relaterte virus hos pasientene.

«gag sequences could be amplified from genomic DNA from LNCaP cells of some subjects after 4 or 6 subcultures following incubation with certain subjects’ plasma, indicating the presence of infectious virus in blood. All gag sequences detected in this cohort were more similar to the MLV-like sequences reported by Lo et al. (2010) than to the XMRV sequences reported by Lombardi et al. (2009). Detection of gag sequences in whole blood genomic DNAs that were negative for mouse IAP and mitochondrial DNA provides strong evidence for infection of humans with MLV-like viruses.»

 

Murine leukemia viruses (MuLV) and Xenotropic MuLV-related viruses exhibit inter-tropic complex recombination patterns

 http://www.retrovirology.com/content/8/S1/A235

Mattia CF Prosperi , William M Switzer, Walid Heneine and Marco Salemi

Institusjoner: Department of Pathology, Immunology and Laboratory Medicine, Emerging Pathogens Institute, College of Medicine, University of Florida, (Gainesville, Florida, USA), Laboratory Branch, Division of HIV/AIDS Prevention, National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention, Centers for Disease Control and Prevention (Atlanta, GA, USA)

«Given the evidence of inter-tropic recombination in MuLV, detection and classification of recombination in XMRV using different MuLV tropism prototypes should be interpreted with caution. Despite using a small dataset, a strong phylogenetic signal in the alignments and highly resolved phylogenies inferred both by full-length and sliding-window approaches, different recombination programs reported conflicting results. These results suggest that identification of parental strains of the potential recombinants is difficult and that recombination in the highly genetically related MuLV have been occurring for some time.»

 

In vitro assembly of xenotropic murine leukemia virus-related virus CA-NC protein

http://www.retrovirology.com/content/8/S1/A236

Romana Hadravová, Jitka ¿tokrová, Michal Dole¿al, Iva Pichová, Tomá¿ Ruml and Michaela Rumlová

Institusjoner: Department of Biochemistry, Institute of Organic Chemistry and Biochemistry Czech Academy of Sciences, Department of Biochemistry and Microbiology, Institute of Chemical Technology, (Begge Praha, Tsjekkia)

«We found that purified XMRV full-length CANC, starting with the conserved proline residue at the N-terminus of CA, was not able to assemble into particles. However, a modification of the N-terminus of CANC (modCANC) enabled formation of spherical particles. Moreover, the negative staining of the in vitro assembled particles of XMRV modCANC revealed different organization of protein layers in comparison to CA-NC of M-PMV.»

 

Human infection or lab artifact: will the real XMRV please stand up?

http://www.retrovirology.com/content/8/S1/A241

Robert H Silverman

Institusjoner: Department of Cancer Biology, Lerner Research Institute, Cleveland Clinic, Cleveland, Ohio, USA

«Xenotropic murine leukemia virus-related virus (XMRV) was first identified in 2005 in a study of human prostate cancer patients with genetic variants of the antiviral enzyme, RNase L. Subsequent investigations in North America, Europe and Asia have either observed or failed to detect XMRV in patients [prostate cancer, chronic fatigue syndrome-myalgic encephalomyelitis (CFS-ME), immunosuppressed with respiratory tract infections] or normal, healthy control individuals. Among the confounding factors are the potential for lab contamination with similar or identical viruses or viral sequences originating in mice. In some studies, relatively contamination-resistant methods (e.g. IHC, FISH, and antibody detection) suggest that either XMRV or a similar type of virus is present in some patients. Evidence for and against genuine infections of humans with this intriguing virus (and/or related viruses) will be discussed.»

 

XMRV infection in human diseases

http://www.retrovirology.com/content/8/S1/A238

Otto Erlwein, Mark J Robinson, Steve Kaye, Myra O McClure, Marjorie M Walker, Anup Patel, Wun-Jae Kim, Mongkol Uiprasertkul, Ganesh Gopalakrishnan, Takahiro Kimura and Kikkeri Naresh

Institusjoner: Section of Infectious Diseases, Jefferiss Research Trust Laboratories, Imperial College London, Department of Histopathology, Imperial College London, Department of Urology, Imperial College Healthcare Trust, London, Department of Urology, College of Medicine, Personalised Tumor Engineering Research Centre, Chungbuk National University, Korea, Department of Pathology, Faculty of Medicine, Siriraj Hospital, Mahidol University, Bangkok, Thailand, Vedanayagam Hospital, RS Puram, Coimbatore, India, Department of Urology, Jikei University, School of Medicine 3-25-8, Nishi-Shinbashi, Minato-ku, Tokyo, Japan, Centre for Pathology, Hammersmith Hospital Campus, Imperial College Healthcare NHS Trust, London

«The novel gammaretrovirus xenotropic murine leukemia virus-related virus (XMRV) was identified in human prostate cancer tissue in 2006, confirmed in 2009 and later linked to a second human condition chronic fatigue syndrome, CFS. These investigations, all carried out in the US, have not been reproduced in Europe or in China.

We found no evidence for XMRV infection in CFS. Moreover, we failed to find evidence of XMRV infection in UK prostate cancer patients and in prostate cancer tissue taken from patients in India, Korea, Thailand and Japan, or in cancers other than that of the prostate.

Our UK CFS patients were consistently XMRV-free. We did, however, generate false-positive results from prostate cancer patient tissue, despite the fact that the no-template controls in our PCR were consistently negative and the PCR for murine mitochondrial DNA was often also negative.

Sources of this contamination will be discussed in our presentation.»

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