JUDY MIKOVITS' NEW STUDY

Xenotropic Murine Leukemia Virus-related Virus-associated Chronic Fatigue Syndrome Reveals a Distinct Inflammatory Signature 

Published IN VIVO

VINCENT C. LOMBARDI, KATHRYN S. HAGEN, KENNETH W. HUNTER, JOHN W. DIAMOND, JULIE SMITH-GAGEN, WEI YANG and JUDY A. MIKOVITS

Abstract. Background: The recent identification of xenotropic

murine leukemia virus-related virus (XMRV) in the blood of

patients with chronic fatigue syndrome (CFS) establishes that a

retrovirus may play a role in the pathology in this disease.

Knowledge of the immune response might lead to a better

understanding of the role XMRV plays in this syndrome. Our

objective was to investigate the cytokine and chemokine

response in XMRV-associated CFS. Materials and Methods:

Using Luminex multi-analyte profiling technology, we

measured cytokine and chemokine values in the plasma of

XMRV-infected CFS patients and compared these data to those

of healthy controls. Analysis was performed using the Gene

Expression Pattern Analysis Suite and the Random Forest tree

classification algorithm. Results: This study identifies a

signature of 10 cytokines and chemokines which correctly

identifies XMRV/CFS patients with 93% specificity and 96%

sensitivity. Conclusion: These data show, for the first time, an

immunological pattern associated with XMRV/CFS.

HAVE YOU NO SENSE OF DECENCY?

Here is my testimony at the Chronic Fatigue Syndrome Advisory Committee meeting on Wednesday at Health and Human Services in Washington, D.C.  Below the written testimony is the video clip.

My name is Mindy Kitei.  I’m a science reporter who’s covered ME/CFS for twenty years.  Last June, I began my blog, CFS Central, in honor of my friend Nancy Kaiser.

I met Nancy in 1994, while working on an investigative piece for Philadelphia magazine called “The AIDS Drug No One Can Have” about the experimental HIV and ME drug, Ampligen.

Nancy had a severe case of ME. She had multiple seizures every day. When she tried to sit or stand, her blood pressure plummeted; she often crawled instead. She tried many experimental treatments to get well.

Nancy died on June 15, 2008.  I naively thought she’d never succumb to the illness, as if by sheer will she’d keep herself alive.

Three other ME patients whom I interviewed in 1994 have also died of the disease.

Despite its gravity, despite ample evidence that ME is an infectious disease, the government treats it like a joke. The CDC and parts of the NIH have been playing a shell game:  studying patients with simple fatigue or chronic fatigue or depression—but labeling them CFS patients. 

Even when the CDC conducted its XMRV study, it studied the wrong cohort and refused to do an actual replication of the Science study.  It’s just a different kind of shell game from the bogus psychological CFS studies that are the agency’s trademark.

To the CDC and NIH scientists who’ve been doing this ludicrous research for three decades and sweeping a worldwide human catastrophe of 17 million people under the carpet, I say to you:  Have you no sense of decency at long last?

ME patients are suffering from a serious infection— most likely a retrovirus—but are told by charlatans to exercise and have a positive attitude. 

Researchers in government and at universities, as well as the CFIDS Association, admonish desperate patients that taking anti-retroviral drugs is medically indefensible.  When the healthy reprove the sick that they’re impatient and reckless and foolish and need to wait for treatment, I say there is no treatment, and where are the drug trials?  Thirty years and not one approved drug and none in the offing.

ME patients should have the same freedom to try medications that AIDS patients had in the early days.  The AIDS patients became their own advocates because there was no one advocating for them.  The same holds true for ME patients now. Patients are gravely ill, and they have the right to treatment.  To say that they don’t—that’s what is medically indefensible.

The U.S. government conducted the Tuskegee Syphilis Experiment from 1932 to 1972.  The study tracked the progression of untreated syphilis among poor African American men but didn’t tell them they had syphilis. The men got sicker and many died.

In 1997, President Clinton apologized to the remaining Tuskegee men.  Clinton said:  “What was done cannot be undone. But we can end the silence. We can look at you in the eye and finally say on behalf of the American people, what the United States government did was shameful, and I am sorry.”

The United States government has watched ME patients suffer and die for 30 years, and has done nothing, and that is shameful.

In less than a year, more than 125 thousand patients from 108 countries and territories have found my blog, CFS Central.  Patients write to me asking for help every day.  Toward that end, I request a meeting with Kathleen Sebelius, Howard Koh, Francis Collins, Tony Fauci and Thomas Frieden to discuss how to turn this situation around, by funding good studies and finding effective medications.

About funding ME, Dr. Dennis Mangan said during this meeting:  “We’ll use one dollar and try to make two.”   I’m sure Dr. Mangan means well, but it isn’t enough.  As AIDS activist Larry Kramer said years ago about HIV patients:  “We are not crumbs.” After thirty years of neglect, ME needs research parity with HIV. We also need a czar who will oversee ME and report directly to President Obama. 

Finally, we need to enact the ME/CFS Care Act.  Much like the Ryan White Care Act for HIV patients, the ME/CFS Care Act will provide health coverage to needy patients.

In closing, I ask you, Dr. Wanda Jones, to ensure that this meeting occurs.  Dr. Jones, will you help me? 

BLOGGER LOST ALL THE COMMENTS ON THE POST.  SHOULD ANY OF YOU WANT TO COMMENT AGAIN, I WILL OF COURSE POST THEM AGAIN.

BLOOD VERSUS TISSUE

4generations commented on yesterday’s post on Dr. Ila Singh's XMRV-negative ME/CFS study: 

Singh's primate study found that XMRV left the blood of the infected primates after a few weeks (6, I believe). Isn't that finding plus the finding of XMRV in prostate cancer patient tissue good enough evidence to justify a study looking for XMRV/MLV in the tissues of patients with ME/CFS?

4generations, the primate study at Emory University wasn’t Singh’s. But your point is well taken, as XMRV quickly left the blood and settled in the tissues in the macaques. In addition, in the CDC’s new XMRV study on prostate cancer this week, the three patients who tested PCR positive to XMRV in tissue had no virus in plasma by PCR or Western blot.  

If that can occur in prostate cancer, perhaps that can occur in ME/CFS as well.  Moreover, as some readers have pointed out, Dr. Kenny de Meirleir in Belgium is taking tissue samples in the gut of ME/CFS patients and finding them positive for XMRV.

DR. ILA SINGH:

She believes XMRV isn't in ME/CFS patients but that there's evidence for the retrovirus in prostate cancer

CFS Central emailed University of Utah's Dr. Ila Singh about her new XMRV study, which found no evidence of XMRV in ME/CFS patients. In 2009, Singh found evidence of the retrovirus in prostate-cancer tissue.

CFS Central:  Will you revisit your prostate-cancer XMRV findings in light of this XMRV negative CFS study?  From what you’ve learned in this latest CFS study, do you now believe that the XMRV that you found in prostate cancer is a human infection or just contamination?

Dr. Ila Singh: Prostate cancer and chronic fatigue syndrome are completely different illnesses.  I recognize that recent studies have cast doubts on the prostate cancer association as well, but there is still considerable data supporting the link to prostate cancer that cannot be easily explained by contamination. We will present some of this work at the Cold Spring Harbor meeting later this month.  But clearly more work needs to be done before that question can be settled. 

CFS Central:  Do you believe scientists should be looking for XMRV in CFS patients’ tissues instead of blood?
 

Singh: The original study by Mikovits' group reported finding XMRV only from blood.  They did not examine other tissues.  So blood is where the focus should be. Now if one found it in blood, then of course you'd be interested in finding out where else the virus is.  And then it would be interesting to look at tissues.  But looking at tissues is not trivial and not something to be attempted without good evidence of the virus being present in the body first. 

CFS Central: Did you test the new assays/methods used in the new study against any XMRV positive samples from your prostate-cancer study?

Singh: Our prostate cancer study was entirely on prostate tissues. These were archived in tissue banks in a de-identified manner, so there was no way to go back to those patients and obtain blood samples.  So, we could not test some of the new tests we developed on our material from prostate cancer patients. 

CFS Central:  In your new study it says that “2 positive controls were also included” (line 349 of your study). Were the two positive controls clones or clinical samples from prostate-cancer studies or from Mikovits’s positive CFS patients? 

Singh:  The samples from Mikovits' patients were all tested in a completely blinded fashion.  We did not know which of them were positive, so could not use them as positive controls.  But more accurately, there are no real patient 'positive controls' for XMRV.  In order to use patient samples as controls, you'd have to first be absolutely certain that these patients have XMRV.  How could you do that right now?  So we used what you call a 'clone' for our PCR studies.  But remember, this clone was isolated from a patient (a prostate cancer patient).  And this is over 99% identical to the isolates from CFS patients described in Lombardi et al.  For the viral culture studies, we used very small amounts of titrated virus that was grown in the lab as positive controls. And all of these positive controls were always positive. 

CFS Central:  You grew XMRV in the prostate-cancer cell line LNCaP and the breast-cancer cell line MCF-7 in your study “Raltegravir is a potent inhibitor of XMRV, a virus implicated in prostate cancer and chronic fatigue syndrome.”  It’s unclear to me why the new study didn’t culture XMRV from those two positives. 

Singh:  Yes, we did.  And these grew just fine.  Apologies for not being clearer in the paper.  None of the patient samples tested positive, but the positive controls were always positive. 

CFS Central:  Some patients on the forums find this sentence from the new study problematic:  “We are forced to conclude that prescribing antiretroviral agents to CFS patients is insufficiently justified and potentially dangerous.”  The Science and the Alter/Lo studies have reported XMRV and related MLVs in CFS patients.  CFS patients are very ill, many for decades, and there are no approved treatments. They believe the decision to try antiretroviral drugs should be between a patient and his or her doctor. Why did you feel the need to put that statement in the study and press release? 

Singh:  The patient and their doctor did not make the decision to try antiretroviral drugs in a vacuum.  It was based on reports of finding XMRV in CFS patients.  We are now convinced that there is no XMRV in CFS patients--so the reason for starting those drugs does not exist.  And there is no good evidence for continuing to use drugs that could lead to serious side effects of liver or bone-marrow failure.

****

I wrote to Singh to ask her to clarify two of her answers:
 

CFS Central:  In your prostate-cancer study, “XMRV is present in malignant prostatic epithelium and is associated with prostate cancer, especially high-grade tumors,” you found 4 percent of healthy controls with evidence of XMRV.  If you’re finding a background rate in controls in your prostate-cancer studies, why do you think you didn’t find a background rate in CFS patients and controls?  

Singh:  Not entirely sure, but there were different assays (e.g. immunohistochemistry) and different sample types (blood vs prostate tissue).

CFS Central: To clarify one of my previous questions, beginning on line 347 of your new study, it states: “We inoculated LNCaP cells with 100 [microliters] of plasma from 31 patients and 34 healthy volunteers, and passaged the cells weekly for 6 weeks. 13 negative controls and 2 positive controls were also included. Only one culture was handled at a time to prevent any cross-contamination. After weeks 2, 4 and 6, cultures were lysed and analyzed by Western blots (Fig. 4) and by qPCR for XMRV. No XMRV protein or DNA was detected in any of the cultures. [emphasis added]  

My question is this: Is the bolded sentence correct, or did the 2 positive controls grow in these cultures and/or did the positive samples continue to test positive in culture?  If the positive controls didn’t grow or if the positives controls didn’t test positive in culture, why didn’t they?  You grew XMRV in the prostate-cancer cell line LNCaP and the breast-cancer cell line MCF-7 in your study “Raltegravir is a potent inhibitor of XMRV, a virus implicated in prostate cancer and chronic fatigue syndrome.”

Singh:  That was our poor wording in the paper (the words in bold).  All positive controls grew XMRV--as one would expect.  None of the samples from the healthy controls or CFS patients grew any virus in culture.  And of course the negative controls did not grow any XMRV.