"In Vitro Infidelium" on the WPI-RED-VIPdx-Connection and Conflict of Interest

"In Vitro Infidelium" on the WPI-RED-VIPdx-Connection and Conflict of Interest.

This looks like a rats-nest to me.

Longitudinal investigation of natural killer cells and cytokines in chronic fatigue syndrome/myalgic encephalomyelitis

Longitudinal investigation of natural killer cells and cytokines in chronic fatigue syndrome/myalgic encephalomyelitis
Ekua W Brenu, Mieke L van Driel, Donald R Staines, Kevin J Ashton, Sharni L Hardcastle, James Keane, Lotti Tajouri, Daniel Peterson, Sandra B Ramos and Sonya M Marshall-Gradisnik

Journal of Translational Medicine 2012, 10:88 doi:10.1186/1479-5876-10-88
Published: 9 May 2012

Abstract

Background
Chronic Fatigue Syndrome/Myalgic Encephalomyelitis (CFS/ME) is an etiologically unexplained disorder characterised by irregularities in various aspects of the immunological function.

Presently, it is unknown whether these immunological changes remain consistent over time.

This study investigates Natural Killer (NK) cell cytotoxic activity, NK cell subsets (CD56brightCD16- and CD56dimCD16+) and cytokines, over the course of a12 month period in patients with CFS/ME.


Methods
The participants in the study comprised 65 (47.2+/-11.5 years) CFS/ME participants and 21 (45.2 +/-9.3 years) non-fatigued controls.

Flow cytometry protocols were used to assess NK subsets and NK cytotoxic activity at various time points that included baseline (T1), 6 (T2) and 12 months (T3).

Cytokine secretions were measured following mitogenic stimulation of peripheral blood mononuclear cells.


Results
NK cytotoxic activity was significantly decreased in the CFS/ME patients at T1, T2 and T3 compared to the non-fatigued group.

Additionally, in comparison to the non-fatigued controls, the CFS/ME group had significantly lower numbers of CD56brightCD16- NK cells at both T1 and T2.

Interestingly, following mitogenic stimulation, cytokine secretion revealed significant increases in IL-10, IFN-gamma and TNF-alpha at T1 in the CFS/ME group.

A significant decrease was observed at T2 in the CFS/ME group for IL-10 and IL-17A while at T3, IL-2 was increased in the CFS/ME group in comparison to the non- fatigued controls.

Overall cytotoxic activity was significantly decreased at T3 compared to T1 and T2.

CD56brightCD16- NK cells were much lower at T2 compared to the T1 and T3.

IL-10 and IL-17A secretion was elevated at T2 in comparison to the T1 and T3.


Conclusion
These results confirm decreases in immune function in CFS/ME patients, suggesting an increased susceptibility to viral and other infections.

Furthermore NK cytotoxic activity may be a suitable biomarker for diagnosing CFS/ME as it was consistently decreased during the course of the 12 months study.

Now that at least looks like a proper longitudinal study with the participation of Dan Peterseon – unlike the cytokine BS from Mikovits/WPI. How close this study is to the reality of ME/CFS and how useful it will prove for research, diagnosis and treatment of ME/CFS is another question…

Ampligen ("Rintatolimod") Phase III study published

Finally!

[Update: This was not a new study, just a re-analysis http://phoenixrising.me/archives/8882]

A Double-Blind, Placebo-Controlled, Randomized, Clinical Trial of the TLR-3 Agonist Rintatolimod in Severe Cases of Chronic Fatigue Syndrome

David R. Strayer 1*, William A. Carter 1, Bruce C. Stouch 2, Staci R. Stevens 3, Lucinda Bateman 4, Paul J. Cimoch 5, Charles W. Lapp 6, Daniel L. Peterson 7, the Chronic Fatigue Syndrome AMP-516 Study Group¶, William M. Mitchell 8*

1 Hemispherx Biopharma, Inc., Philadelphia, Pennsylvania
2 BCS Consulting, Philadelphia, Pennsylvania
3 University of the Pacific, Stockton, California
4 Fatigue Consultation Clinic, Salt Lake City, Utah
5 Center for Special Immunology, Fountain Valley
6 Hunter-Hopkins Center, Charlotte, North Carolina
7 Sierra Internal Medicine Associates, Incline Village, Nevada
8 Vanderbilt University School of Medicine, Nashville, Tennessee

Abstract

Background
Chronic fatigue syndrome/myalgic encephalomyelitis (CFS/ME) is a severely debilitating disease of unknown pathogenesis consisting of a variety of symptoms including severe fatigue.

The objective of the study was to examine the efficacy and safety of a TLR-3 agonist, rintatolimod (Poly I: C12U), in patients with debilitating CFS/ME.

Methods and Findings
A Phase III prospective, double-blind, randomized, placebo-controlled trial comparing twice weekly IV rintatolimod versus placebo was conducted in 234 subjects with long-standing, debilitating CFS/ME at 12 sites.

The primary endpoint was the intra-patient change from baseline at Week 40 in exercise tolerance (ET).

Secondary endpoints included concomitant drug usage, the Karnofsky Performance Score (KPS), Activities of Daily Living (ADL), and Vitality Score (SF 36).

Subjects receiving rintatolimod for 40 weeks improved intra-patient placebo-adjusted ET 21.3% (p = 0.047) from baseline in an intention-to-treat analysis.

Correction for subjects with reduced dosing compliance increased placebo-adjusted ET improvement to 28% (p = 0.022).

The improvement observed represents approximately twice the minimum considered medically significant by regulatory agencies.

The rintatolimod cohort vs. placebo also reduced dependence on drugs commonly used by patients in an attempt to alleviate the symptoms of CFS/ME (p = 0.048).

Placebo subjects crossed-over to receive rintatolimod demonstrated an intra-patient improvement in ET performance at 24 weeks of 39% (p = 0.04).

Rintatolimod at 400 mg twice weekly was generally well-tolerated.

Conclusions/Significance
Rintatolimod produced objective improvement in ET and a reduction in CFS/ME related concomitant medication usage as well as other secondary outcomes.

Revisiting oldies but goodies - Another one

Revisiting oldies but goodies.

http://news.sciencemag.org/scienceinsider/2011/10/xmrv-researcher-fired.html
…

Mikovits's collaborator, Francis Ruscetti of the National Cancer Institute (NCI) in Frederick, Maryland, who ran all of the Western blots, confirms that the Ottawa slide uses the same image that appears in Lombardi et al.

So wait, did I get his right, Ruscetti ran all of the Western Blots in Lombardi et al. 2009?

So what did the WPI do, except write one sentence about the 68 PCR positive patient samples?

Ruscetti and Mikovits, in a joint e-mail to Science for this article, said many patients and their doctor, Daniel Peterson (who since has had a falling out with WPI), knew the original coded numbers, so the researchers changed them for the Science publication to "protect the patient privacy." Ruscetti says it was a mistake for Mikovits to have used the original patient codes in Ottawa. "We were under so much pressure, we missed it," says Ruscetti.

Yeah, but at least one of the so called "original coded numbers", the number 1282 that appear on the Mikovits' Slide of Shame (which I shall rename Ruscetti/Mikovits Slide of Shame) appeared as well in the study itself – in the Addendum! Either Ruscetti/Mikovits don't recall what they did – or the words "bear false witness" come to my mind.

(And please note how someone is trying to throw Daniel Peterson under the bus.)

As far as the use of 5-azacytidine, Ruscetti and Mikovits stressed in their e-mail that "there was no attempt in the original paper to hide anything." They say for the purposes of Lombardi et al., the use of 5-azacytidine was not germane: They were simply trying to demonstrate that CFS patients had viral proteins not seen in controls. By the time of the Ottawa meeting, they say they realized that this experiment did not in fact show XMRV but proteins from a broader family of gammaretroviruses.

In fact they had to realize that they forgot to run a control for the usage of 5-AZA! Oh, the rush of writing something for Science!

Trying to help and getting back-stabbed

With help from Peterson, UCSF virologist Jay Levy examined 43 patients who tested XMRV-positive at WPI. Peterson says he wanted to help his former colleagues at the institute. “I went to Jay Levy to prove them right,” he says. All samples tested negative. Again, Mikovits said the study was flawed.

Even after he left the WPI, Dan Peterson believed in XMRV and Mikovits' work, invested his time and resources in order to try to help. And for what what? To be backstabbed by Judy Mikovits…

99% similarity to VP62 “seems very unlikely and may indicate contamination, despite the evidence presented to the contrary”

The manuscript didn't convince Science. After reviews by three referees and members of the Board of Reviewing Editors (provided for this article by Mikovits), the editors rejected the paper. “Although the referees were intrigued by your findings, they had a number of serious reservations,” read a 4 June letter, which included excerpts from reviewers.

The rejection letter noted that Science would re-review the paper if the authors could both retain the “novelty of its main message” and “address the referees' concerns with new data rather than with counter-arguments.” … An otherwise enthusiastic referee wrote that the “one major caveat I have is that the issue of potential contamination has not been completely dealt with.” A second referee found it odd that the genetic sequence of XMRV derived from CFS patients and the virus earlier discovered in prostate cancer were 99% similar. This “seems very unlikely and may indicate contamination, despite the evidence presented to the contrary,” the referee warned. One also wondered why they omitted Peterson as a co-author.

On working with Dr. Mikovits (2)

Oddly, Peterson, who had supplied the patient samples, was not one of the authors. His name was left off—and he was kept out of the loop on the study's results—because of worries that he might prematurely tell his patients, Mikovits says.

A prediction for 2012: Mikovits will try to throw Lipkin under the bus

She is just that kind of fun person.

Just look at who she has thrown under the bus: A post-doc at the WPI went under the bus for Judy, so did Robert Silverman, Vincent Lombardi and Annette Whittemore. Basically she threw the whole WPI under the bus. And that is just the people she directly worked with (and I am most certainly forgetting some). Not finding XMRV in your study? Under the bus you go! Suggesting that Dr. Mikovits checks for contamination? Under the bus you go!

And I do want to note that Konnie Knox was fired while Dr. Judy Mikovits was working on her XMRV VP62 plasmid thingy. And that Dr. Daniel Peterson left the WPI after Dr. Judy Mikovits has done the major part of her "research".

Update:

Judy Mikovits also asked me to go to the Applied Research Facility ("ARF" Building) and get all files, papers and documents on Dr. Dan Peterson. Dr. Mikovits stated that she believed Peterson had committed Medicare Fraud.

http://s3.documentcloud.org/documents/268454/exh-2-reply-iso-motion-for-preliminary-injunction.pdf (via)

"The change in XMRV copy number was detected with qRT-PCR"

Methods for addressing the NK cell dysregulation

• PBMCs from XMRV infected patients with low NK cell function were activated with the mitogen PHA and treated with Ampligen
• The effects on NK cell (CD56+) phenotype were determined by flow cytomentry
• Signaling changes due to the treatment were detected via cytokine analysis
• The change in XMRV copy number was detected with qRT-PCR.

…
Preliminary Results Cont.

XMRV copy number is modulated by Ampligen
When treated with Ampligen, qRT-PCR indicates a decrease in some patients and an increase in others.

What the fruck?  First of all, when did the WPI use qRT-PCR? Why did they kept rather quiet about it? And secondly, this seems to have been done in the context of treatment? Were are the results of these studies?

What research has the WPI done since October 2009?

The only thing that comes to my mind is the 2011 Vincent Lombardi cytokine study.

The cancer study thing didn't seem to have gone anywhere.

The 33 PCR negative patients from the Science paper? Done by 10/2009.

The claim to have found XMRV in autism (6/15)? Done by 10/2009.

The claim to have found XMRV in atypical MS (3/3)? Done by 10/2009.

The claim to have found XMRV in fibromyalgia (12/20)? Done by 10/2009.

Usage of qRT-PCR in the context of treatment? Done by 10/2009.

All presented by Daniel Peterson on the CFSAC October 2009 meeting.

The Blood Working Group (BWG)? Done at VIPDx.

Any other research regarding "XMRV" at the WPI? Anything? Am I missing anything?

There is one list of studies that seems to have gone nowhere:

Current Research Program (June 2010)

Role of Chronic Inflammatory and Immune Stimulation by Active Herpesvirus Infection in the Development of Immune Dysfunction in CFS
Studies of immune abnormalities in this CFS cohort will involve phenotypic analysis of NK, DC, and T cell populations. States of activation and differentiation will be studied to determine whether the cells are immune activators or suppressors due to response to infection. The goal is to understand the imbalance in the immune system that leads to unregulated virus expression.
Collaborators: Francis Ruscetti, PhD, NIC, Dennis Taub PhD, NIA (INIP Research Award)

Novel Viruses/Co-infections in Subgroups of CFS
A Virus DNA Microarray (Virochip) was used to screen a cohort of CFS with Immunological defects and increased incidence of Mantle cell lymphoma (MCL).
Collaborators: Francis Ruscetti, PhD, Cancer Inflammation Program, NCI

Role of the Interferon/RNaseL Antiviral Pathway in CFS
The goal of this study is to characterize defects in the Interferon/RNaseL antiviral pathway which result in viral persistence.
Collaborators: Robert Silverman, PhD, Cleveland Clinic

Role of Inflammatory Cytokines and Chemokines in CFS
The purpose of this study is to identify and develop the serum biomarker patterns of cytokine and chemokines, which correlate with clinical disease and can be used to monitor intervention.
Collaborators: Dennis D. Taub, PhD, NIA

Host Susceptibility in CFS
The goal of this study is to elucidate genetic factors of susceptibility and the dysregulation of the host defense system in order to develop biomarkers for diagnostic development and thus predict response to immune modulating therapy and vaccines.
Collaborators: Jonathan R. Kerr, MD, PhD, St. Georges University, London UK,
Mary Carrington, PhD, NCI, Mike Dean, PhD, NCI

Enumeration and Function of Natural Killer (NK) Cells in CFS
The purpose of this project is to develop improved NK diagnostic tests for both the number and function of NK cells.
Collaborators Doug Redelman, PhD, UNR, Dept. of Microbiology & Immunology

Epidemiologic Evaluation of Lymphoma and Cancer Incidence in Nevada CFS Cohort
The goal of this study is to determine if a possible cancer cluster exists in the cohort of Nevada CFS patients.
Collaborators: Paul Levine, MD, George Washington University

Study of Clonal T Cell Receptor-gamma Rearrangements in Nevada CFS Cohort
The purpose of this study is to determine the significance of clonal T cell receptor-gamma rearrangements in the pathogenesis of CFS.
Collaborators: Dorothy Hudig, PhD, UNR, Dept. of Microbiology & Immunology

Development of a Whole Genome Transcriptome Assay for HHV6
The goal is to develop a whole genome transcriptome assay for HHV6 and a sub array for HHV6A specific detection to be used as a diagnostic tool or research tool to understand the viral life cycle in disease.
Collaborators: Rachel Bagni, PhD, Applied Technology Program, NCI

HHV6 Latency in CFS
The goal of this study is to understand the role of HHV6A latency in bone marrow hematopoietic stem cells in the pathogenesis of CFS.
Collaborators: Stephen St. Joer, PhD
Marianna Bego, PhD, UNR, Dept. of Microbiology & Immunology

http://forums.phoenixrising.me/showthread.php?5387-XMRV-Current-Research-Program

And there is this (from 2009?):

Clinical/Translational Research Program

Study Title and Description		Collaborators
Development of Clinical Application for Cytokine Profiling of ME/CFS Patients Patients with particular cytokine and chemokine profiles will be evaluated to identify specific clinical traits of characteristics with the goal to correlate these traits with specific environmental or infectious triggers.
Investigation of Active Human Cytomegalovirus (HCMV) Infections in ME/CFS Patients The goal of these studies is to determine whether active HCMV infections can be detected in the blood of ME/CFS patients, and if so, to determine its prevalence and clinical manifestations.		Konnie Knox, PhD Don Carrigan, PhD, Wisconsin Viral Research Group
Confirm and Extend Observations of Persistent Enterovirus Infection in Subpopulation of ME/CFS Patients The goal of this study is to correlate EV positivity with clinical phenotype, cytokine profile, "leaky gut," co-infections.		Konnie Knox, PhD Don Carrigan, PhD, Wisconsin Viral Group
Define Predictors of Positive Clinical Response to Treatment of ME/CFS Patients with Antivirals, e.g., Valcyte and Visitide The goal of this study is to determine the differences in clinical phenotype between antiviral Responders and Non Responders. Compare R and NR Cytokines, viral infections, RNaseL activity, blood counts, NK cell activity, etc.		Konnie Knox, PhD Don Carrigan, PhD, Wisconsin Viral Research Group

http://www.wpinstitute.org/research/research_clintrans.html

I guess throwing people under the bus is more important than science.

[Update]
I found Dr. Mikovits stash of unpublished studies!

Cancer in Lombardi et al. 2009?

At least 4 patients from the Science study had cancer, according to Peterson’s CFSAC presentation (WPI-1118, WPI-1125, WPI-1150, WPI-1199). How many other patients had cancer? WPI states: “Blood samples from the WPI repository were chosen at random and there were no patients chosen with lymphoma or mention of lymphoma in this study. Another preliminary study was done at a later date that had nothing to do with the XMRV Science publication.”

I don't know about that CFSAC presentation, should check. These cancer claims are just nuts