Abstracts from the 1st XMRV Conference

[aruschima]

Georgieeeeea, as usual you sniffed out the interesting stuff. There is so much info here, where is the link to the info on EBV / XMRV pairing up ?

 

[Rrrr]

joey posted this:
From Abstract P_04 by Cleveland Clinic Foundation:

The chemokine IL-8 is one of the most highly induced genes in response to XMRV infection of prostate cancer cell line DU145

______

in response to joey's first point (above) i will repost this info below. as you will see, the IL-8 is sky high in CFS patients vs health controls.
_____

Treating cytokine up/down-regulation


WPI & NCI Paper

"Type 1 IFN Pathyway Response to Viral Infection in Chronic Fatigue Syndrome"

Mikovits,J. Hagen,K. Peterson,D. Stephens,R. Lombardi,V.
Whittemore Peterson Institute, Reno, NV, USA. Laboratory of Experimental Immunology, National Cancer Institute-Frederick, Frederick, MD, USA.

118 CFS patients
138 Controls

Cytokines/Chemokines:

CFS:

IL-8: 1045
MIP-1a: 763
MIP-1b: 1985
IL-6: 336
TNF-a: 148
IL-1b: 500
IP-10: 98
IFN-a: 35
IL-13: 28
IL-7 160

Controls:

IL-8: 13
MIP-1a: 91
MIP-1b: 164
IL-6: 29
TNF-a: 13
IL-1b: 56
IP-10: 32
IFN-a: 60
IL-13: 86
IL-7: 60

Guide to Chemokines/Cytokines:

IL-8: RNase L & CMV activated
MIP-1a: Elevated in neurodegenerative disease
MIP-1b: Elevated in neurodegenerative disease
IL-6: Stimulates chronic inflammation
TNF-a: Stimulates chronic inflammation
IL-1b: Stimulates chronic inflammation
IP-10: Interferon response protein
IFN-a:Stimulates macrophages and NK cells to elicit an anti viral response
IL-13: Inhibits inflammatory cytokine production
IL-7: Stimulates proliferation of B & T lymphocytes & NK cells

Summary and conclusions:

Cytokine and Chemokine profiling in combination with machine logic algorythms reveals an inflammatory signature consistent with an over-expression of herpes virus and is useful for diagnosis for CFS

 

[leela]

Rrrrr,

prrrrrrrrrrr

How do we get media attention on all this? I presume Mindy Kitei is summarizing all this gold for publication? Where do we go from here?
Science Daily, which I read for kicks (?!) has NO section on CFS, and only negative articles about the doubtful or nonexistent link between CFS and XMRV.
I guess it's mostly up to the patient community to shift the media slant now? I'm not in the know on how to do that.

 

[mojoey]

I emailed Amy Dockser Marcus from WSJ the info this morning.

 

[leela]

Great! Did you forward the whole lot of abstracts? What's the correct protocol for contacting journalists? Do we have names for the health/science reporters at the other biggies? Like I said, I'm a moron in this department. I'm imagining most of them await press releases from the CDC or NIH to write articles on subjects like these?

 

[Otis]

Here's a cleaned up version of the abstracts in MS Word format

This version is much easier to read. It's not formatted much but it's much more readable and will make cutting and pasting an abstract easier.

Here's the link to download:

http://www.sendspace.com/file/0wsvpr.

Scroll to the bottom of the page and click the download button. You may get a popup - just close it if you do. That's the price for a "free" service. I ran a virus scan and it's a clean document. PM me if you have any troubles.

I suggest starting an individual thread for each abstract you want to discuss, and use the abstract "name" (e.g. 'O_01') and the title (e.g "Basic virology Screening mouse genomes for XMRV-like elements") in the thread title. I think we'll get more participation if we break out the discussions that way.

 

[leela]

Great work and great suggestion, Otis.

 

[Rrrr]

Rrrrr,

prrrrrrrrrrr

How do we get media attention on all this? I presume Mindy Kitei is summarizing all this gold for publication? Where do we go from here?
Science Daily, which I read for kicks (?!) has NO section on CFS, and only negative articles about the doubtful or nonexistent link between CFS and XMRV.
I guess it's mostly up to the patient community to shift the media slant now? I'm not in the know on how to do that.

great question, leela! i think we need to support the effort that are out there already, like the buying of a HUGE ad in the washington post (see advocacy section for that: http://www.forums.aboutmecfs.org/sh...nched-This-WILL-Get-the-Attention-of-MILLIONS!)

and i think individuals need to hold their own public protests and call their local media to come attend it. sorta like this one woman protest this person did: http://www.youtube.com/watch?v=ME764z_4vEY

and if you can't do an outdoor protest, then a good video of you in yr bed, sick, sent to HHS secretary sebelius or NIH director collins are great alternatives. like this video this person did: http://www.youtube.com/watch?v=8t1Xqp1LDxM

those are my thoughts.

if we don't act to change things, they won't get changed. it is in our hands.

 

[natasa778]

"Since gammaretroviruses including MLV have been shown to be unable to infect terminally-differentiated, non-dividing cells including macrophages and dendritic cells, we are currently investigating whether the infection observed represents the infection of a small number of dividing cells in these cultures, or whether XMRV is not subject to the same restriction as MLV in these cells."

XMRV presence in those cells could be down to coinfections/cofactors. Have a look at these:

Lancet. 1994 Jan 29;343(8892):255-8.
Co-infection and synergy of human immunodeficiency virus-1 and herpes simplex virus-1.
Heng MC, Heng SY, Allen SG. Department of Medicine, UCLA San Fernando Valley Internal Medicine Program, Veterans Affairs Medical Center, Sepulveda 91343.
The human immunodeficiency virus (HIV-1) uses the CD4 molecule, expressed by T helper cells and activated macrophages, as a receptor for entry into host cells. In tissues co-infected with herpes simplex type 1 (HSV-1), HIV-1 virions were observed to infect keratinocytes, which, because they lack the CD4 molecule, are normally incapable of being infected by HIV-1. Although a number of other viruses have been reported to enhance HIV-1 viral transcription in vitro, this is the first in-vivo report to our knowledge of reciprocal enhancement of viral replication associated with co-infection of keratinocytes and macrophages by HIV-1 and HSV-1 in patients with AIDS and non-genital herpes simplex lesions. The virions in the co-infected cells were larger, morphologically atypical, and appear to be hybrids; most contain the HIV-1 envelope necessary for infectivity. The increased viral load and the proximity of the virions to the cutaneous surface may lead to increased risk of transcutaneous transmission of both viruses. These findings point to the need for incorporation of suppressive treatment for herpes simplex in the treatment of AIDS. PMID: 7905094


AIDS Res Hum Retroviruses. 1990 May;6(5):641-7.
Human immunodeficiency virus type 1 (HIV-1) and herpes simplex virus type 2 (HSV-2) can coinfect and simultaneously replicate in the same human CD4+ cell: effect of coinfection on infectious HSV-2 and HIV-1 replication.
Kucera LS, Leake E, Iyer N, Raben D, Myrvik QN. Department of Microbiology and Immunology, Bowman Gray School of Medicine, Wake Forest University, Winston-Salem, North Carolina 27103.
Experiments were designed to determine whether HIV-1 and herpes simplex virus type 2 (HSV-2) coinfection leads to simultaneous replication of both viruses in the same human CD4+ cell (MT-4 cell line) and the possible effects of coinfection on infectious virus production. Results from transmission electron microscopy analysis revealed replication of typical HSV-2 nucleocapsids in the nucleus and budding of HIV-1 particles through the plasma membrane and through intracytoplasmic vacuoles containing enveloped HSV-2 particles in the same coinfected cell. Coinfection of HIV-1 persistently infected H9IIIB or promonocytic U1 cells with HSV-2 did not alter total production of infectious HSV-2 or the percentage of HSV-2 infectious centers compared with control H9 and U937 cells infected with HSV-2 alone. However, in coinfected promonocytic U1 cells HSV-2 induced infectious HIV-1 production measured by syncytial plaque assay. In summary, both HIV-1 and HSV-2 can coinfect and simultaneously replicate in the same human CD4+ cell. Interactions between HIV-1 and HSV-2 appear to be unidirectional, resulting in accelerated replication of HIV-1 as reported by Albrecht et al. (J Virol 1989;63:1861-1868), but not HSV-2 as shown by us. PMID: 1972888

 

[garcia]

A printable PDF of the abstracts

Hi all,
I managed to find a way around the pdf security settings on the original abstract pdf. My version looks exactly like the original, except it can be printed. Also you can copy and paste text from it. Thought it might be useful to someone out there.

http://www.divshare.com/download/12629255-deb

Please ignore any pop-up windows (the price of using free file hosting), and excuse the 15 second wait-time.

Enjoy!

garcia.

 

[Cort]

My take on XMRV from the Buzz Page
http://aboutmecfs.org/Rsrch/XMRVBuzz.aspx

XMRV International Workshop Abstracts

German Researcher Draws Blanks Again - Norbert Bannert is not having an easy time with XMRV. First he tried to find it in prostate cancer and failed and now he's tried to find it in ME/CFS and failed. Unlike other researchers he activated the PBMC's he sampled and then cultured them in order to boost the viral presence but he used a different process than the WPI and Dr. Hansen did. Then he tried to transfer the virus to the LnCap cells the WPI does use in at least some samples but to no avail.He did show, though, that XMRV is infectious; that is he was able to produce the virus from a cell line and showed that it could infect cells. His particular population - a small group of 39 CFS patients - was negative, as were his healthy controls and a group of MS patients. if

Swedish Study Comes Up Empty - Dr. Blomberg and his 'Swedish XMRV Study Group' - used quantitative PCR - the same type of PCR Dr. Singh favors, to look for XMRV. Dr. Blomberg used the same LnCap cells the WPI recommends to culture the virus for five days for the same period of time the WPI recommends. Blomberg found no XMRV in 50 people with ME/CFS, 400 prostate cancer samples or 200 controls using what he called his 'novel' PCR methods. He did get a few weakly positive (or 'uncertain') samples from ME/CFS patients. We can see from both these studies and the Mikovits and Hansen studies that as time goes on studies are getting more complex and are starting to hue more towards using the WPI's techniques. Dr. Singh is currently engaged in an XMRV study in Salt Lake City which is using LnCap cells as well.

XMRV Blood Working Group is Halfway There - Key Problem Identified? - Phase II of the project, which compared different labs ability to find XMRV in WPI identified positive patients is complete! One thing they looked was whether the 2-4 day delay in processing that is common in many labs affects a labs ability to find the virus. Dr. Mikovits' stated that blood preparation techniques are important and it brings to mind Dr. Singh's painstaking attention to this factor in her study. (What an astonishing finding that would be if it was something as simple as blood storage all along....) In Phase III they will determine the best assay using blinded tests and in phase IV they will look at 300 samples from the Western United States. They will be done by the end of the year.

Finding XMRV's Tissue Reservoir - Dr. Ruscetti stated that the results from the Science paper suggest that XMRV is not very active in T and B cells in the peripheral blood - the main players in the adaptive immune response. His study looked further into the immune system and found that it can infect two major players in the innate immune response - monocytes/macrophages and antigen presenting dendritic cells. Interestingly, while XMRV appears to be able to infect and replicate in these cells, MLV's cannot - the reason being that they cannot infect 'non-dividing cells'. Ruscetti is currently investigating whether XMRV is infecting a small number of macrophages/dendritic cells that are dividing.

XMRV and the immune System - MLV's have a history of producing cancer and neurological disorders in rodents and other mammals but what about those immunologically disturbed ME/CFS patients? Does XMRV do anything to disturb the immune system? This study, which looked at how the gene expression in prostate cells changes when they become infected with the virus, found a good deal of abnormal immune and other gene expression. The authors stated their findings suggest the virus could have a 'profound effect' on fundamental cellular physiology and inflammation.

XMRV and EBV Team Up?- a Spanish team found that XMRV in a significant percentage of EBV transformed (infected?) Cell Lines from a small group of ME/CFS patients and healthy controls suggested that EBV infected B cells could be a tissue reservoir for the virus.

XMRV Present in Leukemia and Mantle Cell Lymphoma Cells - Dr. Mikovits and Dr. Ruscetti and others found XMRV in leukemia and Mantle Cell Lymphoma cells from two patients. The abstract suggested that the development of these cancers 'coincides' with three factors; the growth of the gamma delta T-cells, XMRV infection and a 'distinct inflammatory profile'. They appear to suggest that these factors could place patients at risk for the development of these cancers. (Interestingly, neither EBV, nor HHV-6 or CMV appeared to coincide with cancer development). They also noted that antivirals can help in HIV derived cancers and that AZT and Raltegravir stopped and significantly reduced previously rising lymphocyte counts in one XMRV infected ME/CFS patient with cancer. This suggests antiretrovirals may be helpful in this set of ME/CFS cancer patients.

Dr. Cheny (ahem Cheney's) Study - Dr. Cheney's study probably did little to answer any of the more pressing questions about XMRV because the tests were not done at an independent lab but it should prove quite valuable when the WPI's XMRV test is validated. Some of his stats were alarming. Dr. Cheney has said he probably sees a sicker population and judging from this sample he probably is; the average duration was almost 20 years! He found XMRV in almost 75% of them. Just as the Phoenix Rising Forums his patients had a very high incidence of CFS or CFS-like illnesses in their family (45%); 48% had cancer in their families or themselves and 28% had autoimmune diseases in the family. (One wonders what the norms are.) The most startling fact was 50% XMRV positivity in non-CFS family members. If Dr. Cheney is right this virus is spreading in families.

XMRV Positivity Rates at VIP Dx - In the 7 months between November and May VIP Dx Labs received 712 samples (about 100/month) from 46 of the 50 states in the US and about 50 from Europe and the UK. Of those only 35% tested positive for XMRV. The discrepancy between Dr. Cheney's 75% and VIP Dx's 35% is surely one reason why Dr. Mikovits stated the type of patient being tested is a major factor in the positive rates for a test. Of course, Dr. Cheney with his 20 year ME/CFS patients was undoubtedly testing a severe subset. We'll have to wait for more comprehensive testing of ME/CFS populations to determine just what type of ME/CFS patient tests postive...It will be fascinating!!!

Children with XMRV - the Ruscetti/Mikovits juggernaut rolled with their (fourth?) abstract, this time indicating that XMRV was present in no less that 82% of 17 autistic children (!) The finding that 16 of the 17 families with autism spectrum disorder, CFS, FM, Lupus, etc. had at least one member infected with XMRV suggested that it may run in these diseases, although, of course, much more work needs to be done.

 

[Cort]

Second part

XMRV International Workshop Abstracts Part I - the abstracts are not the video's - they should be out soon - but they do give essentials about the talks and they are INTERESTING.

Those Monkeys - Five Rhesus macaques were infected with XMRV; two of them were sacrificed quickly and the others several months later. Creating only low levels of antibodies, the immune system did not react strongly to the virus possibly because it was able to quickly remove the virus from the bloodstream. This apparently suggested to the investigators that the virus showed low infectivity and virulence, but low and behold when they opened the animals up, they were surprised to find that the virus had infected many organs and they were able to detect evidence of ongoing replication. Interestingly, the virus hung out in the lymphoid organs - which give it a clear pathway to the bloodstream. (Dr. Mikovits has suggested that the lymphoid organs could be a tissue reservoir for the virus). They found that XMRV was also replicating in all the sexual glands - which, of course, suggests that sexual transmission is a distinct possibility.

XMRV is Different in the Prostate (and the Prostate) - we've heard that different XMRV variants may exist in the blood and the prostate but this study suggested that the XMRV may be different within the prostate itself. This study found different forms of XMRV which, the authors speculated, could enhance XMRV's ability to cause prostate cancer (if indeed it does that).

XMRV Is Not Present in Prostate Cancer in the US - While several positive prostate cancer XMRV studies have occurred this fairly large John Hopkins study, which used immunohistochemistry, found zero evidence of XMRV in almost 200 samples...But...

XMRV IS Present in Prostate Cancer in the US - a Baylor study found XMRV was present in 22% of 144 prostate cancer patients - so much for consistency at this stage of the game. Still a positive report should trump a negative one, all things considered. Two more US studies will alternately not find and find XMRV in prostate cancer patients.

Hanson XMRV CFS Study Strongly Positive - This is the first study to use LnCap Cells (aka WPI) to culture at least some of the blood samples. Reports from the conference suggested that Dr. Hanson found polytropic MLV's but no XMRV but her abstract stated her study was strongly XMRV positive with XMRV gag sequences showing up in just over half the CFS patients. The abstract ends "Our results corroborate those of Lombardi et al (Science paper)"! Dr. Hanson is the first recipient of an NIH grant to study XMRV in CFS patients and her extensive study to examine the immune factors and the effects of exercise on XMRV is projected to take at least a year.

Yes, There is XMRV in the UK (and Quite a Bit of It!) - This UK study with Dr. Mikovits in the lead and Dr. Ruscetti the senior author, states that 'more sensitive' methods to detect XMRV have been developed since the Science paper was published last October. The blood was collected in England and shipped to the National Cancer Institute in the US. They came right out and said it's not about the methods - that the most important factor in the negative results in the type of patient studied (eg, "It's the patients stupid") a theory which seems to have gained some strength lately. The paper states "Since the most important variable in detecting XMRV in CFS is patient selection" - an interesting statement, if there ever was one.

They used five methods, the most prominent of which was culturing in the LnCap cells that Dr. Hanson used but they also used antibody tests and various kinds of PCR tests on different types of cells. The WPI has been working hard on the diagnostic end as they cited their new antibody tests and new primers that can detect both XMRV and polytropic MLV env sequences. They also showed, as did the Science paper, that cell free media from patients contained the virus (it was present in the blood outside of the cells) and that the virus could infect other cells once they were placed in it; no wonder Dr. Ruscetti reportedly claimed his work on the diagnostic end was over and he was moving onto the next stsge of research. This was a very strong study - with multiple overlapping tests - just as occurred in the original Science study.

A New Method For Detecting XMRV - God knows what this technical abstract means but apparently Dr. Ruscetti has has developed 'DERSE' cells that can quickly and sensitively detect the presence of XMRV - another diagnostic improvement in a Workshop that was chock full of them.

'High Throughput' Antibody Tests Available - want to test large quantities of blood for antibodies to XMRV? This abstract, if I read it right, states the authors think they can now do that.

Blood Working Group Has Been 'Working' - Not very fast but they are working. They are now very clear that they have a good test to detect XMRV in spiked vs unspiked whole blood, PBMC's and plasma. Next step is looking for XMRV in people!

 

[jimm]

why is no one picking up on the fact that the infected wild mice had panleukopenia????? this is a lowered immune system i.e low white cells accross the board....leaving them open to more infections on top...like HIV people...I am a patient who HAS panleukopenia and have always thought it was part of what ever is wrong with me...I am suprised more people havent picked up on this it seems really significant

 

[FunkOdyssey]

why is no one picking up on the fact that the infected wild mice had panleukopenia????? this is a lowered immune system i.e low white cells accross the board....leaving them open to more infections on top...like HIV people...I am a patient who HAS panleukopenia and have always thought it was part of what ever is wrong with me...I am suprised more people havent picked up on this it seems really significant

Yeah, I've had a low white cell count forever too. Bone marrow results were normal, doctor shrugged, congratulated me on not having cancer, and sent me home. I think its related to or caused by the high rates of neutrophil apoptosis that are found in CFS patients. Specifically my neutrophils are always low and seem to be dragging down the overall WBC count.

 

[jimm]

Yeah, I've had a low white cell count forever too. Bone marrow results were normal, doctor shrugged, congratulated me on not having cancer, and sent me home. I think its related to or caused by the high rates of neutrophil apoptosis that are found in CFS patients. Specifically my neutrophils are always low and seem to be dragging down the overall WBC count.

exactly the same here....i didnt get bone marrow test tho but haematologist just wanst interested, apart from ironically giving me an HIV test and all the hepatitis tests, once they were negative he just sent me on my way....which i found ironic....like it could have been hiv or hepatitis or NOTHING...seems a bit of an extreme range....but my neutrophils and lymphocytes are always low so I think it must be signigficant, hence it jumped out at me when i saw this about the wild mice,...sounds so plausable

 

[alex3619]

Hi George,

I completely agree with this. Let me give you a quotable bite: Science by Sound Bites!

Yep, yep and yep. The whole Q and A was about the politics not about the science. The Q and A will be watched by the media and can be dissected and writen up in nasty little sound bites. The Abstracts will most likely not be read by anyone in the media, too much work. (grins) plus what the heck is the fun of reporting on "science". (big grins)

Bye
Alex