Function of NK cells from XMRV+ CFS patients Increased in vitro with Ampligen

Kati

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XMRV, Natural Killer Cells and Ampligen: A WPI poster

Characterization of the therapeutic increases in natural killer (NK) activity of xenotropic murine leukemia virus-related virus (XMRV)-positive chronic fatigue syndrome (CFS) patients effected by poly (I): poly (C12U) (Ampligen)


Isabel Barao-Silvestre1, 2, Maria Marshall2, Kathryn S. Hagen1, Max A. Pfost1, David Strayer3, Daniel L. Peterson1, Dorothy Hudig2, Judy A. Mikovits1, 2

1. Whittemore Peterson Institute, Reno, NV, 89557 2. University of Nevada, Reno, 89557 3. Hemispherx Biopharma, Inc., Philadelphia, PA, 19103

ABSTRACT:

Chronic Fatigue Syndrome (CFS) is a debilitating disease of unknown etiology that affects ~17 million people worldwide. Patients suffer persistent viral infections and may develop hematopoietic malignancies. The patients have reduced Natural Killer (NK) cell activities which could contribute to the diseases, and, if restored, potentially reduce symptoms. We characterized the in vitro effects on NK cells of the interferon inducer poly (I): poly (C12U) (Ampligen) which in some CFS patients abates disease symptoms. The 30 CFS patients were infected with the gammaretrovirus xenotropic murine leukemia virus-related virus (XMRV). We cultured their peripheral blood mononuclear cells with the drug for 24 hrs and monitor NK activity to K562 cells by flow cytometry, concurrently measuring degranulation by externalization of CD107a, and expression of Grz B and perforin. Treatment markedly increased CD107a externalization in the NK cell population as indicated by 5-fold increases in CD107a-positive cell frequencies and 3-fold increases in their CD107a MFI, with slight positive shifts in intracellular Grz B and perforin. In contrast, T cells showed little change in CD107a externalization. Our results suggest that degranulation rates may be more affected than the levels of cytotoxic proteins, indicating a novel mechanism by which NK activity was affected by the drug. The increase in degranulation per NK cell indicates a mechanism by which Ampligen treatment can improve NK cell function.
Isabel Barao-Sylvestre will be presenting on May 9 at the Immunology conference in Baltimore.
 

CBS

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Nice find

Hey Kati,

Nice find. This will be interesting and I suspect that it is a prelude to an in vivo study.

Shane
 
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Ok, but by increasing NK cell numbers, are we also making more virus? Or does this drug make new NK cells (from blood marrow)? Wait a minute, I am not even sure NK Cells come from marrow.

Anyway, I remember Mikovits saying that the goal should be improving NK cell function, not make NK cells replicate.

Guys, maybe I am having a CFS cognitive blip. Please help.

tina
 

Lesley

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Tina, I think it's saying ampligen improves NK cell function, not that it increases numbers. Of course, I don't know what half of it means...
 

Dr. Yes

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Don't know if this has been posted yet.. too sick to comment myself, except to note that it is an in vitro study; anyway, here it is (from Co-Cure):

Isabel Barao-Silvestre of WPI to present on May 9
Immunology 2010
Baltimore, Maryland
May 7 - 11, 2010

http://www.wpinstitute.org/news/news_upcoming.html
http://www.wpinstitute.org/news/docs/AAA2010WPIposter.pdf

'Characterization of the therapeutic increases in natural killer (NK)
activity of xenotropic murine leukemia virus-related virus
(XMRV)-positive chronic fatigue syndrome (CFS) patients effected by poly (I): poly (C12U) (Ampligen)'

Isabel Barao-Silvestre1, 2, Maria Marshall2, Kathryn S. Hagen1, Max A. Pfost1, David Strayer3, Daniel L. Peterson1, Dorothy Hudig2, Judy A. Mikovits1, 2
1. Whittemore Peterson Institute, Reno, NV, 89557
2. University of Nevada, Reno, 89557
3. Hemispherx Biopharma, Inc., Philadelphia, PA, 19103


ABSTRACT: Chronic Fatigue Syndrome (CFS) is a debilitating disease of unknown etiology that affects ~17 million people worldwide. Patients suffer persistent viral infections and may develop hematopoietic malignancies. The patients have reduced Natural Killer (NK) cell activities which could contribute to the diseases, and, if restored, potentially reduce symptoms. We characterized the in vitro effects on NK cells of the interferon inducer poly (I): poly (C12U) (Ampligen) which in some CFS patients abates disease symptoms. The 30 CFS patients were infected with the gammaretrovirus xenotropic murine leukemia virus-related virus (XMRV). We cultured their peripheral blood mononuclear cells with the drug for 24 hrs and monitor NK activity to K562 cells by flow cytometry, concurrently measuring degranulation by externalization of CD107a, and expression of Grz B and perforin. Treatment markedly increased CD107a externalization in the NK cell population as indicated by 5-fold increases in CD107a-positive cell frequencies and 3-fold increases in their CD107a MFI, with slight positive shifts in intracellular Grz B and perforin. In contrast, T cells showed little change in CD107a externalization. Our results suggest that degranulation rates may be more affected than the levels of cytotoxic proteins, indicating a novel mechanism by which NK activity was affected by the drug. The increase in degranulation per NK cell indicates a mechanism by which Ampligen treatment can improve NK cell function.
 

Lesley

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Ampligen study participants being tested for XMRV by WPI

Retrospective analyses of patient samples from the completed Phase III trial of Ampligen(R) in potential treatment of Chronic Fatigue Syndrome ("CFS") continued in collaboration with the Whittemore Peterson Institute; these studies are expected to provide a new perspective on the design of a confirmatory Phase III study in this disorder. The samples are being analyzed for the presence of XMRV, a novel retrovirus, reported to be found in approximately two-thirds of CFS patients.
The rest is here: http://www.marketwatch.com/story/hemispherx-biopharma-inc-1st-quarter-2010-financial-results-2010-05-07?reflink=MW_news_stmp
 

Hope123

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I think Ampligen has less side effects then retrovirus drugs.
It depends on who you ask. I've heard that it caused liver problems in some folks and at least as of the last FDA review, Hemispherix had not done a good job of reporting adverse effects in their submission to the FDA.

On the benefits side, I've heard that the benefit was very modest for most participants and it didn't seem clinically signficant except for a few anecdotal reports.

I wish there were more details but my opinion of Hemispherix is very low considering almost all reports I've head from them have been based on the finance side about how their stocks are going up or down rather than the science side of published reports. Details are not easy to get. Private companies are not mandated to publish the results of their studies. This is how we ended up with recent scandals concerning pharma companies not being upfront about side effects of drugs until thousands of people had taken them. When research is sponsored by govermental agencies like NIH, researchers are mandated to publish results, good or bad.

Hopefully the WPI has it written somewhere in their contract with Hemispherix that they are allowed to publish whatever studies they participate in. I am glad they are presenting this data at a conference.

From the abstract, it sounds like they took blood from people who were XMRV+ already? From it, we don't know if NK cell counts or activity was low in these subjects. Increasing NK cell activity is good but is a surrogate end point. That is, the theory goes that increased NK activity should suppress XMRV but does it really? Other than in vivo studies, for in vitro studies, they should try to see if increased NK cell activity suppresses XMRV replication, viral load, or expression of cytokines in the test tube (not that tests for these factors are well-developed yet).