1. NEWSFOCUS
Downloaded from www.sciencemag.org on September 22, 2011
A report in Science 2 years ago that linked a mouse retrovirus, XMRV, to chronic fatigue
syndrome astonished scientists and patients alike. But the theory soon began to take hits,
and now, to all but a few researchers, it has completely unraveled
Done. Case closed. Finito, lights off, The End. must have occurred. The leader of the team that conducted the study,
For the past 2 years, a controversy has roiled around the purported Judy Mikovits of the Whittemore Peterson Institute for Neuro-Immune
link between a mouse retrovirus, XMRV and chronic fatigue syndrome
, Disease (WPI) in Reno, Nevada, resolutely maintained that her lab had
(CFS), a baffling, debilitating disease with no known origin. Many no evidence of contamination and that it could repeatedly find the virus
researchers who have followed this saga closely thought that a definitive with its techniques. Millions of dollars have gone into clarifying the
study, published online this week by Science (http://scim.ag/xmrv-cfs) question, which has had far-reaching consequences for people with CFS
and conducted by nine labs—including the main proponents of the the- and, if the virus lurked in the blood supply, the public at large.
CREDIT: M. HICKS/SCIENCE
sis—would finally bring a halt to the impassioned debate. The study just published found that none of the nine labs could repro-
Think again. ducibly detect XMRV or relatives of the virus in blood samples distrib-
The uproar began with an October 2009 paper in Science that found uted under a blinded code. Pounding another nail into the coffin, Science
XMRV in the blood of two-thirds of the CFS patients examined. A is also running a partial retraction (http://scim.ag/R-H-S) of the origi-
steady assault on the report soon began, with more than a dozen labs nal paper, as a contributing lab found that it in fact had a contamination.
failing to replicate it to date and several asserting that contamination In an unexpected twist to this operatic saga, Mikovits co-authored
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2. NEWSFOCUS
the Science Express paper and has no quarrel with the results. Her col- ciation of America, a patient group in Charlotte, North Carolina.
laborator, Francis Ruscetti, a retrovirologist at the U.S. National Cancer Many retrovirologists wish the entire controversy, which has
Institute (NCI) in Frederick, Maryland, who is a co-author of both the ensnarled dozens of labs and cost millions of dollars, would simply
original Science report and the new one, concurs. “Where there is dis- disappear. “All of it’s a waste of money and it’s wrong,” says Robert
agreement is in the interpretation,” Ruscetti says. Gallo, head of the Institute of Human Virology in Baltimore, Mary-
By their lights, the new study—conducted by the Blood XMRV land. “It’s like a bad dream.”
Scientific Research Working Group sponsored by the U.S. National
Heart, Lung and Blood Institute—does not rule out the possibility
that mouse retroviruses infect people with CFS. “The conclusion of
the Blood Working Group was that
X MRV owes its discovery to a little-known enzyme called
RNase L that helps the body battle viruses—and has ties to both
prostate cancer and CFS.
we don’t have a reproducible In the early 1990s, a few CFS
assay to detect XMRVs in the researchers reported that their
blood—not that they weren’t patients had higher levels of RNase
in the patients at all,” Mikov- “ I don’t care if L than healthy people had, sug-
its says. Ruscetti adds that the nobody else in gesting that this natural viricide
working group analyzed their signaled an undiagnosed infec-
the world wants
Downloaded from www.sciencemag.org on September 22, 2011
original patients but used samples tion causing the disease. RNase L
taken a few years later. to work on it. levels, they contended, provided a
If this seems like wordsmith- long-sought “biomarker” for CFS,
ing and splitting hairs, welcome to
Fine, leave us which has caused much confusion
the confusing, maddening world alone! ” because clinicians use varying cri-
of XMRV Mikovits and Ruscetti,
. —JUDY MIKOVITS, teria to make a diagnosis. It also
who have become increasingly WHITTEMORE PETERSON could potentially help researchers
isolated from the broader scien- INSTITUTE gauge the effectiveness of treat-
tific community, now say their ments, as a drop in the enzyme
original paper erred by focusing would mark a decrease in under-
on a single XMRV isolate that lying, undetected infections. This
turned out to be a contaminant. idea became central to tests using
They say that isolate is but one Ampligen, an immune modulator
of many XMRVs, which belong made by Hemispherx Biopharma
to a still larger family of gamma- in Philadelphia, Pennsylvania, to
retroviruses. They also contend treat CFS.
that the virus may lurk in tissues, One of the clinicians testing
only traveling to the blood occa- Ampligen was Daniel Peterson
sionally. “We still stand by our of Incline Village, a town on the
data that we isolated gamma- northern shore of Lake Tahoe,
retroviruses from patients with the popular tourist destination on
CFS and also from healthy con- Nevada’s border with California.
trols,” says Mikovits, who has taken a more public role than Ruscetti in Peterson had investigated a mysterious outbreak of CFS that began in
battling critics and reaching out to supporters. Incline Village in 1984 and had become one of the world’s best-known
Mikovits has become something of a savior in the community of CFS clinicians. His work drew the attention of Harvey Whittemore, a
people with CFS (also known as myalgic encephalomyelitis, or ME), prominent Reno attorney who lobbied on behalf of the gaming industry
who for decades have endured charges that the disease is psycho- and later became a major real estate developer, and his wife, Annette, a
somatic. The 2009 Science paper shouted out that CFS may well teacher of children with cognitive disorders. The Whittemores’ daugh-
have a clear biological cause, and, in turn, raised hopes of effec- ter, Andrea, had developed CFS at age 12, and at Peterson’s suggestion
tive treatments and even a cure. The new she began receiving Ampligen.
findings give her “great pause,” yet she sus-
pects they’re but a speed bump. “I haven’t Online
sciencemag.org
With the Whittemores’ financial support, Kenny De Meirleir, a
Belgian physician who had an RNase testing lab in his own country,
changed my thinking at all,” she says. And helped them open RedLabs USA in Reno to monitor the enzyme in
she worries that the Blood Working Group Podcast interview U.S. patients taking Ampligen. Vincent Lombardi, who would soon
with co-author
conclusions will confuse people with CFS, Jon Cohen. play a prominent role in the XMRV saga, co-founded the new lab and
some of whom got wind of the results early served as its director of operations.
in the blogosphere and contacted her in a panic. “I had 15 suicidal Lombardi was a late bloomer in science. In 1990, while he worked
patients call me last week,” she says. as a securities trader in Lake Tahoe, he decided to pursue an under-
In scientific circles, Mikovits has developed a less flattering reputa- graduate degree at a local school. As part of a biostatistics class, he
CREDIT: J. COHEN/SCIENCE
tion. Critics have accused her and her backers of stubbornly wedding worked with Peterson, analyzing immune parameters of CFS patients.
themselves to a thesis and moving the goalposts with each study that After his graduation in 1995, he ran investment companies and also
challenges their conclusions. Even disease advocates who welcome the worked as a marketing director for a pawnbroker business. He went
attention XMRV has brought to CFS believe the time has come to put on to pursue a Ph.D., first studying RNase L but switching to peptides
this line of research to rest. “It’s hard to say that this has not received a in the tobacco hornworm, which he completed at the University of
fair appraisal,” says Kimberly McCleary, president of the CFIDS Asso- Nevada, Reno, 2 years after starting RedLabs USA.
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In 2002, meanwhile, a team of prostate
cancer researchers had discovered that a muta-
tion in the RNase L gene frequently occurs in
families in which men are prone to the early- “ I began comparing
onset form of that disease. A few years later,
one of those investigators, Robert Silverman
Judy Mikovits to Joan
of the Cleveland Clinic in Ohio, began probing of Arc. The scientists
for a viral link: If RNase L fights viruses, he will burn her at the
reasoned, then perhaps having a crippled form
of the enzyme opened the door for a cancer- stake, but her faithful
causing infection. following will have
Silverman teamed up with Joseph DeRisi
and Don Ganem, two veteran virus hunters at her canonized. ”
the University of California, San Francisco —JOHN COFFIN,
(UCSF), who had developed a microarray TUFTS UNIVERSITY
called ViroChip that had proved its mettle
Downloaded from www.sciencemag.org on September 22, 2011
in identifying new viruses. The assay con-
firmed Silverman’s hunch: In the prostate
cancer tumors, ViroChip found a novel retro-
virus, apparently a cousin of a known mouse
virus and therefore given the ungainly name
of xenotropic murine leukemia virus–related
virus (XMRV).
Silverman first reported the existence of
XMRV and its prostate cancer link in April
2005 at an HIV meeting in a mountain resort
in Banff, Canada. “My talk was well received, people were clearly while the Whittemores’ alma mater, the University of Nevada, Reno,
interested, but it did not receive that much attention,” Silverman wrote offered about 1400 square meters of lab space within a new, gleam-
to Science in an e-mail delivered through a public affairs manager— ing $78 million Center for Molecular Medicine. It would take 4 years
the only form of communication the Cleveland Clinic would allow. for that building to open, so the nascent Whittemore Peterson Insti-
(XMRV has become such a radioactive topic that several institutions tute (WPI) set up shop in borrowed space on campus in 2006. Annette,
restricted what their researchers could discuss, and both Ganem and WPI’s president, hired Mikovits to run the lab.
DeRisi declined to talk to Science about their XMRV work.) Mikovits had come into the CFS world by a circuitous route. She had
In March 2006, Silverman, DeRisi, Ganem, and colleagues pub- spent more than 20 years at NCI, first as a technician studying HTLV-I
lished their report in PLoS Pathogens, a respected but decidedly low- and HIV with Ruscetti, who served as her Ph.D. adviser. In 2001, she
key choice for the description of a new retrovirus that appeared to married and moved to southern California, becoming chief scientific
infect humans—the only three others are HIV and HTLV-I and -II— officer at EpiGenX Biosciences in Santa Barbara, which aimed to use a
and had ties to a serious disease. Although they wrote that there was a new epigenetic approach to develop drugs and diagnostics.
“strong association” between XMRV and the RNase L mutation, the In 2006, Mikovits became a consultant to a CFS-related nonprofit
researchers still had misgivings about whether the virus contributed to foundation that Annette Whittemore co-founded, which explored
prostate cancer. the link between the disease and another virus, human herpesvirus 6
In a commentary in the 30 January 2007 issue of the Proceedings of (HHV-6), that had been discovered in Gallo’s lab. At a meeting in Bar-
the National Academy of Sciences (PNAS), mouse retrovirologist Hung celona, Spain, that year, Mikovits spoke to Whittemore for the first time
Fan of UC Irvine called it an “exciting” discovery. But others were and heard Peterson give a talk. Peterson focused on a non-Hodgkin’s
skeptical. Gallo, whose lab played a central role in the discovery of lymphoma in some of his CFS patients, and Mikovits smelled a virus.
all three known human retroviruses, contacted PNAS’s editor to com- She offered to work with him, and Whittemore helped set up a collabo-
plain. The commentary was “substantially over the top,” says Gallo, ration. Later that year, Mikovits joined WPI.
who saw no compelling evidence that this supposedly new human She soon enlisted Ruscetti, who had worked in Gallo’s lab when it
retrovirus caused disease and doubted that a mouse retrovirus could discovered HTLV-I, to screen blood samples from Peterson’s patients
even infect humans. “Once claims of etiology were made, I just gasped for viruses. Intrigued by the RNase L link to XMRV Mikovits and
,
for breath,” Gallo says. “My own experience argued to me that it’s best Lombardi—who by then had joined WPI as well—met Silverman in
to stay away from this one.” October 2007 at a prostate cancer conference in Lake Tahoe, where
they discussed the possible role of XMRV in CFS. Silverman was
A
CREDIT: KELVIN MA/TUFTS UNIVERSITY
round the time Silverman first publicly described XMRV Annette happy to collaborate and sent WPI a clone of the virus, known as VP62.
,
Whittemore took on one of the most ambitious pursuits of her The institute could use it as a reference to start hunting for the virus in
career: using her family’s wealth and powerful contacts to build a full- CFS patient blood samples that Peterson had stored.
fledged research and clinical institute devoted to CFS. Peterson would A little over a year later, on 18 November 2008, Mikovits had the
head a medical research team to elucidate the causes of the disease and first evidence of XMRV in CFS patients. Working with Ruscetti and
provide state-of-the-art care. Nevada Governor Kenny Guinn and the Silverman, the group amassed evidence that the virus occurred in 67%
current U.S. Senate Majority Leader, Harry Reid (D–NV)—both fam- of 101 CFS patients and 3.7% of 218 healthy controls. If the latter
ily friends of the Whittemores—helped arrange government funding, number was representative of the general population, up to 10 million
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4. NEWSFOCUS
healthy people in the United States alone might Seattle, who had studied XMRV left the meet-
,
be infected, and the virus might be spreading ing convinced that the link to CFS was real.
through blood donations and organ transplan- The group had not only detected the virus using
tation—a silent epidemic of frightening pro- PCR but also grown it from patients’ cells and
portions. Mikovits, Ruscetti, Silverman, and found antibodies to it. “It sounded really good
their co-authors submitted a paper to Science to me because they had all these different lines
in May 2009. The paper, of which Lombardi of evidence,” Miller says. The fact that an NCI
was first author, did not claim that XMRV veteran such as Ruscetti endorsed it helped
caused CFS, noting that the disease might sim- convince him. “Frank said, ‘I grew it with my
ply make patients more vulnerable to infection. own hands,’ ” Miller recalls. “At the time, it
Causality “is probable but not definitive at this sounded really exciting.”
time,” Lombardi et al. stated. Coffin, who chaired the meeting, thought
But what they were asserting was stunning Mikovits and her colleagues had persuasive
enough: WPI, NCI, and the Cleveland Clinic evidence, especially because XMRV formed
had all found evidence of XMRV in the same its own branch on a genetic family tree of
CFS patients. Some of these people had par- mouse retroviruses, strengthening the case that
Downloaded from www.sciencemag.org on September 22, 2011
ticipated in a 2007 NCI drug study, too, and it was not a contaminant.
their blood samples, stored separately, also The next day, as Mikovits sat at Washing-
tested positive, “ruling out the possibility of ton, D.C.’s Dulles International Airport wait-
lab contamination as a source,” the authors ing for her flight back home, she received a
wrote. When the team calculated the so-called phone call from Science, indicating for the first
p-value—which generally needs to be under time serious interest in publishing the paper.
0.05 for a finding to be considered signifi- Science accepted the Lombardi paper on
cant—they arrived at the astonishingly 31 August and published it online on 8 October
low number of 8.1 × 10–35. 2009, accompanied by a supportive commen-
Oddly, Peterson, who had supplied “Here was this tary that Coffin co-authored. “There are several
the patient samples, was not one of the lines of evidence that transmission happened
authors. His name was left off—and he was
mysterious disease, … in the outside world and was not a laboratory
kept out of the loop on the study’s results— and along comes a new contaminant,” the commentary declared. The
because of worries that he might prematurely publication catapulted Mikovits into the spot-
tell his patients, Mikovits says.
virus. … It seemed to … light, where she has remained ever since.
have the makings of
T he manuscript didn’t convince Science.
After reviews by three referees and
members of the Board of Reviewing Editors
a medical
breakthrough.”
“H ere we go again.” That’s what
crossed Kimberly McCleary’s
mind when she read the first headlines
(provided for this article by Mikovits), the —ROBERT SILVERMAN, about the paper that week in her office
editors rejected the paper. “Although the ref- CLEVELAND CLINIC in Charlotte. In her 20-plus years at
erees were intrigued by your findings, they CFIDS, McCleary had seen many infec-
had a number of serious reservations,” read a 4 June letter, which tious agents fingered as a potential cause of
included excerpts from reviewers. CFS. HHV-6. Epstein-Barr virus. Mycoplasma. Adenovirus. Cyto-
The rejection letter noted that Science would re-review the paper megalovirus. HTLV-I and -II. And on and on. Each time, hopes were
if the authors could both retain the “novelty of its main message” and dashed as scientists closely evaluated the suspects.
“address the referees’ concerns with new data rather than with counter- The string of disappointments had made McCleary cautious,
arguments.” But the criticisms were substantial. “Chronic Fatigue Syn- and CFIDS urged patients not to get carried away. “We tried to tem-
drome is full of false alarms,” wrote one advisory board member, “and per things early on, and we were criticized heavily for raining on the
the detection of XMRV could be false positive PCR.” An otherwise parade,” she says. Still, many patients went wild with joy. “ME/CFS
enthusiastic referee wrote that the “one major caveat I have is that the patients have never seen anything like this,” wrote Cort Johnson in an
issue of potential contamination has not been completely dealt with.” A item, “Game Changer,” posted on his popular blog Phoenix Rising.
second referee found it odd that the genetic sequence of XMRV derived Lombardi et al. had made XMRV a superstar. “Here was this myste-
from CFS patients and the virus earlier discovered in prostate cancer rious disease (CFS-ME) with no known cause, all of these patients suf-
were 99% similar. This “seems very unlikely and may indicate con- fering, and along comes a new virus that associates with the disease,”
tamination, despite the evidence presented to the contrary,” the referee Silverman says. “It seemed to some at the time to have the makings of
warned. One also wondered why they omitted Peterson as a co-author. a medical breakthrough.”
Mikovits and her co-workers addressed many of the critiques, and But many scientists were skeptical. Simon Wessely, a psychiatric
CREDIT: LISA DEJONG/LANDOV
on 14 July, they resubmitted the paper, this time with Peterson’s name researcher at King’s College London who has long studied CFS, says
on it. The next week, a committee—which included John Coffin, a the virology went over his head, but the fact that fully two-thirds of CFS
prominent retrovirologist at Tufts Sackler School of Graduate Biomed- patients harbored the virus was an alarm bell. CFS, whose definition
ical Sciences in Boston, and Silverman—organized a workshop at NCI has been the subject of years of debate, is far too heterogeneous a phe-
with Mikovits and Ruscetti to help the institute better understand the nomenon for that, he says.
unanswered scientific questions and the potential public health ramifi- With a few U.K. colleagues, Wessely started penning a letter to
cations. Dusty Miller, a retrovirologist at the University of Washington, Science to address what they saw as methodological flaws in the paper.
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One problem was the unclear patient selection, they wrote. If all patients hope,” Lombardi said. “We’re not the doctors ordering these tests for
came from Nevada and healthy controls were from elsewhere, then per- our patients.” Whittemore stressed that all profits would go back into
haps XMRV wasn’t a CFS-related virus, Wessely says, but something the institute. “This wasn’t set up to make money, and it never has,” she
that happened to be more prevalent in the gambling state. said. Whittemore also dismissed critics—a group that included Peter-
Wessely also contacted Jonathan Weber, a retrovirologist at Imperial son—who cringed at CFS patients taking antiretrovirals. “How many
College London, to set up a study of British CFS patients. “Our reac- years does this patient population have to be impacted and their lives
tion was, ‘It’s probably wrong, but if it’s true, it’s a pretty big advance. destroyed?” she asked.
So it’s worth testing,’ ” he says. Andrea Whittemore was one of the CFS patients who tested posi-
The first published critique appeared a few weeks later—and it tive for XMRV .
would prove to be prophetic. On 18 November, Patrick Moore, a cancer
virologist at the University of Pittsburgh in Pennsylvania, and his post-
doc Masahiro Shuda published a blistering commentary on Faculty of T he flood of negative data started in January 2010, when
Wessely, Weber, and their colleagues reported in PLoS ONE that
1000, a Web site that evaluates reports in what it calls postpublication they couldn’t find any trace of XMRV in 186 British CFS patients. By the
peer review. end of February, two more negative reports
“Unfortunately, in my field, there’s a were published, one of them by Jonathan
tendency with any new virus to hope that it Stoye, Coffin’s co-author on the favorable
Downloaded from www.sciencemag.org on September 22, 2011
is causing a disease,” says Moore, who co- Science commentary that accompanied the
discovered HHV-8 and helped prove that it Lombardi paper.
causes Kaposi’s sarcoma. “One has to be so Back in Reno, Mikovits and Lombardi
cautious about that. A lot of stories sound began feeling besieged. “After the first neg-
good, but they’re built on a house of cards.” ative study, it was a dog pile,” Lombardi
As Moore and Shuda noted, Mikov- says. “Let’s be honest: A number of peo-
its’s group used a technique called “nested ple in the mainstream medical community
PCR” to identify XMRV infections, which, heard chronic fatigue, and they rolled their
they wrote, “is inherently prone to inter- eyes and laughed.”
mittent false positivity that has occurred The problem, as they saw it, was that
in our lab and many others.” What’s more, nobody was following their recipe exactly.
the researchers had not randomized and Many labs failed to find XMRV using PCR
blinded patient and control samples, a because the virus exists in scant amounts,
standard way to protect against bias and to they said—so much so that a patient can
detect errors. Together, Moore and Shuda test positive on one bleed and negative
wrote, this was “a recipe for uncontrolled on the next. Mikovits said they overcame
PCR contamination.” this problem by first mixing patient blood
The next month, a study co-authored samples with an uninfected cell line that
by Silverman and published online by is especially permissive to the virus and
Virology showed that AZT, an anti-
HIV drug, worked against XMRV in
“ We tried to temper coculturing for 8 weeks. Other researchers,
including XMRV discoverer Silverman,
test tubes. Again, CFS patients were things early on, and we questioned the need for this step.
overjoyed. Not only had a likely culprit of were criticized heavily for Seven months after Lombardi et al.,
their suffering been found, drugs already
on the market might treat it. raining on the parade. ” Science published Wessely’s critique and
two others, which discussed patient
Of course, no one had proved that the —KIMBERLY MCCLEARY, selection problems and the growing
virus actually caused CFS. But by the THE CFIDS ASSOCIATION OF AMERICA list of negative studies, including three
start of 2010, three commercial labs, one that failed to find XMRV in prostate
of which was WPI’s VIP Dx (formerly RedLabs USA), offered an cancer. None of the Technical Comments,
XMRV test. however, mentioned the possibility of contamination.
Peterson, sitting in one of his practice’s waiting rooms in Incline Despite a vigorous defense by Mikovits and Ruscetti in the same
Village in June, explained that his patients split into two groups. One issue of Science, the three comments further eroded confidence in the
had ridden the CFS roller coaster so many times that these patients findings. But then, 2 weeks later, a research group unconnected to WPI
wanted validation and replication before buying the XMRV story. would rekindle the fading hope that the link between XMRV and CFS
“There’s another group that, for whatever reason, has made this ill- was real. It would also spark a peculiar new controversy.
ness into a religion and becomes polarized into who believes and who
CREDIT: COURTESY OF KIMBERLY MCCLEARY
doesn’t believe,” he said. Increasing numbers of believers began taking
XMRV tests, which cost $500 or more, and, if they were positive, ask- T hat study originated with Shyh-Ching Lo, an infectious-disease
researcher at the U.S. Food and Drug Administration (FDA) who
ing their physicians to prescribe anti-HIV drugs, says Peterson, who by took a keen interest in CFS. In the early 1990s, Lo had tested a theory
then had severed ties with WPI over a contractual issue. that a microbe called Mycoplasma was involved in CFS. For that study,
When a Science reporter visited WPI the same month, Annette he had received blood samples from Anthony Komaroff, a physician
Whittemore and Lombardi both strongly defended the decision to sell who had treated hundreds of CFS patients at Brigham and Women’s
an XMRV test before evidence existed of a causal link. (The group has Hospital in Boston. Like so many others, the lead had proved spurious.
filed related patent applications, too.) “Every physician who requests After XMRV surfaced as a new candidate, Lo suggested to
patient testing is aware of contamination and the potential of false Komaroff they test the same patient samples, stored at –80°C and
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6. NEWSFOCUS
aggressive, ACT UP–style pro-
“Once claims of tests at blood banks to demand
publication: “I believe we need
etiology were to act quickly, before the FDA/
made, I just NIH paper is killed.” Some scien-
gasped for breath. tists thought the federally ordered
delay impinged on scientific
My own freedom. Why not publish the
experience competing papers and let other
researchers scrutinize them? “It
argued to me was very strange business,” Cof-
that it’s best to fin says. Alter says nothing sinis-
stay away from ter was afoot peril. Retrovirology
were never in
and that the papers
this one. ” published the CDC study on
—ROBERT GALLO, 1 July 2010, and the Lo-
Downloaded from www.sciencemag.org on September 22, 2011
INSTITUTE OF Alter study ran in PNAS on
HUMAN VIROLOGY 23 August.
Once they saw it, many
researchers concluded that
untouched for almost 2 decades. Komaroff was enthusiastic. Lo also the Lo-Alter data didn’t confirm the Lombardi paper at all. They had
contacted Harvey Alter, an infectious-disease specialist who works on found that 86.5% of samples from CFS patients harbored DNA from
transfusion medicine at the U.S. National Institutes of Health’s Clinical mouse retroviruses as did 6.8% of healthy controls, but it was not
Center in Bethesda, Maryland. Alter—who won the prestigious Lasker XMRV; the sequences were more closely related to a different, well-
Award for his role in the discovery of hepatitis C—provided Lo with known group, the murine leukemia viruses (MLVs). It was as if a new
blood samples from healthy people to serve as controls. suspect suddenly had been nabbed. Miller says the findings actually
It was Alter who accidentally dropped the bombshell news. At a argued against Lombardi et al. “If XMRV is everywhere and these
blood bank meeting in the Croatian capital Zagreb in late May 2010, guys are doing the same experiment, why didn’t they find XMRV?”
he presented an overview of current blood safety issues, and his Power- he asks. Alter now says he regrets asserting that the paper confirmed
Point presentation had one slide on what he called the “Agent du jour,” the XMRV results and that it was “naïve” to show the slide in Zagreb.
XMRV Although Alter didn’t present any evidence, the slide was blunt:
. “That is something I shouldn’t have said because we really hadn’t
“The data in the Lombardi et al. Science manuscript are extremely found it,” he says. “I somehow got to be the spokesperson for this. I
strong and likely true, despite the controversy,” one bullet point said. had no idea what it would mean.” Lo agrees that the team didn’t find
“We (FDA & NIH) have independently confirmed the Lombardi group the virus that Mikovits and Lombardi had reported.
findings,” read another. Mikovits to this day contends that the Lo-Alter paper confirms
After the editor of a Dutch newsletter discovered the slide on Lombardi et al. and insists that from the beginning, she viewed XMRV
the meeting’s Web site and sent out a press release, the CFS world as one of many gammaretroviruses, which includes the MLVs, involved
exploded, again. “Is this it, then, the big one???? Holy ****!!!!!” one with CFS. In the Lombardi study, some patients tested negative in PCR
blogger wrote on the Phoenix Rising forum. tests for XMRV and yet produced MLV-related proteins, she claims,
Some researchers had doubts. True, Alter had a stellar reputa- but they decided to count them as negatives. She has another serious
tion, but he wasn’t a retrovirologist. And Lo in the 1990s had pushed regret about the paper. “I’d not put XMRV in the title,” Mikovits says.
the widely discredited theory that a Mycoplasma infection played an “We never considered that it would be a single sequence.”
important role in HIV causing AIDS.
Yet news of the impending confirmation had a big impact on a spe-
cial XMRV panel at the AABB, an international association focused on
transfusion medicine. The panel, which had heard about the Lo-Alter
A s more negative data poured in, Mikovits and Lombardi became
ever more ardent. Improved techniques now found the virus in
almost every CFS patient, they said.
study before the news broke, recommended on 18 June that AABB Patient groups began to see Mikovits as a martyr—a Galileo-like
members discourage CFS patients from donating blood. figure fighting an all-powerful scientific establishment to expose the
Patients and scientists alike were eager to see the data. Lo and Alter truth. But many of her initial supporters had joined the growing camp
had written a paper accepted by PNAS, but they ran headfirst into an of skeptics. “I began comparing Judy Mikovits to Joan of Arc,” Coffin
institutional roadblock: Retrovirologists at the U.S. Centers for Disease says. “The scientists will burn her at the stake, but her faithful follow-
Control and Prevention (CDC) had their own paper in preparation that ing will have her canonized.”
showed no evidence of XMRV antibodies, proteins, or DNA in well- Mikovits has her own theory about when Coffin changed his mind.
characterized CFS patients. Faced with contradictory results from two She and Lombardi had found evidence, not included in the Science
CREDIT: GAIL BURTON/AP IMAGES
teams of researchers under its purview, the U.S. Department of Health paper, that XMRV was also linked to autism. On 11 November 2009,
and Human Services (HHS) ordered the groups to delay publishing Lombardi presented those data at a meeting at the Cleveland Clinic.
until each could review the other’s data. “You don’t talk about autism in the U.S.—it’s too politically charged,”
Patients cried foul. Many had long distrusted the federal govern- Mikovits claims Coffin told him. She believes Coffin turned against her
ment, and especially CDC, for not taking CFS seriously and suppress- that very day. Coffin confirms he was upset that Lombardi presented
ing research results. Now, they said, CDC was trying to bury another such preliminary data on such a fraught topic but says, “I did not ‘turn
theory it didn’t like. The author of a blog, CFS Central, called for against’ Judy at that or any other point.”
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7. NEWSFOCUS
Mikovits further lost credibility with her contribution to an early Retroviruses and Opportunistic Infections in Boston. Working with
study organized by the Blood XMRV Scientific Research Working Oya Cingöz in Coffin’s lab, Pathak had analyzed the 22Rv1 cell line
Group, formed by HHS 1 month after the publication of Lombardi all the way back to its origins, a prostate cancer patient at Case West-
et al. It included safety experts such as Susan Stramer of the Ameri- ern Reserve University in Cleveland, Ohio. To create the cell line,
can Red Cross, as well as Mikovits, Alter, Coffin, and other players researchers had used a common technique called passaging in which
in the XMRV saga. The group had devised a four-stage program that human tumor cells are grown repeatedly in mice and then harvested.
included sending blinded samples to various labs to see whether they Early versions of the cell line, still stored in university freezers, had
could detect XMRV At a meeting on 14 December 2010, the work- no XMRV But two different strains of mice used in the later experi-
. .
ing group discussed the results of the second stage of its study, which ments each harbored DNA that matched half of the virus. This sug-
again did not support Mikovits’s findings. Her lab had found XMRV gested XMRV was created in the 1990s when two viruses combined
in a sample from a healthy person who all labs agreed beforehand was in a lab culture, and the widely used cell line spread the virus in many
negative for the virus. labs, infecting other cell lines, too. The finding furthered doubts that
Mikovits had an explanation. The false positive was caused by a XMRV had ever infected a human being—let alone had a role in either
postdoc who mistakenly used the same needle twice to lyse cells and CFS or prostate cancer.
shear DNA, contaminating the sample. She dismisses the error as triv- The accidental-origin evidence hasn’t convinced Mikovits. “Yes,
ial, the result of working late at night on a weekend because of repeated that can be an origin of an XMRV she says. “But it could have arisen
,”
Downloaded from www.sciencemag.org on September 22, 2011
power failures in a new building, coupled with intense pressure from multiple times. It’s not one sequence.” Pathak contended the chances
the working group to get results quickly. “People make mistakes, and that the exact same virus would arise twice were the same as dropping
we reported it as a mistake,” she says. a quarter from a helicopter flying over the Grand Canyon and having it
But many researchers threw up their hands: How could WPI botch land on a quarter on the ground.
a study essential to the survival of its theory? The University of Wash- “It’s all contamination,” Coffin concluded, which outraged Miko-
ington’s Miller was astonished that Mikovits vits. “How can John Coffin shut down research like that?” Mikov-
didn’t do the critical tests herself and that its shouted during one interview, her blue-gray
they were rushed. eyes shooting fire. “He’s not God!” She specu-
Ruscetti is not bothered that most of “ The virus is real. … lates that perhaps the U.S. government, afraid
his peers think he’s wrong. “I’ve been there
before,” he says. But he thinks Mikovits has
I have isolated it from of the huge consequences oftoadiscredit her
XMRV outbreak, was trying
widespread
been treated unfairly. “I’ve been in science for patients. I know it’s work. “We can’t afford another public health
35 years, and she’s as honest a scientist as I’ve
ever met,” he says.
there. Believe me. ” crisis,” she said.
Eventually, two studies that looked for
More setbacks followed. A week after the —JUDY MIKOVITS, XMRV in fresh blood samples taken from
WHITTEMORE PETERSON INSTITUTE
working group meeting, Retrovirology pub- patients in the Lombardi paper failed to
lished four devastating papers online that find it. Ila Singh, a virologist at the Univer-
together made a compelling case that a contaminant marred both the sity of Utah in Salt Lake City, consulted with
Lombardi et al. and Lo-Alter studies. Two reports co-authored by Mikovits and Lombardi to copy their protocols precisely, and among
Coffin looked for XMRV and MLV in both CFS and prostate cancer 100 CFS patients in her study, 14 had tested XMRV-positive by WPI.
patients; every positive sample they found also harbored mouse DNA, Singh and co-workers found 5% of both patients and controls to be
suggesting that a reagent had become contaminated. Another report XMRV-positive—a finding they traced back to a contaminated enzyme
put a finer point on it: Researchers discovered MLV—at least 97% sim- used in a PCR reaction.
ilar to the Lo-Alter viruses—in a commercially available enzyme used With help from Peterson, UCSF virologist Jay Levy examined 43
in a PCR kit. Some speculated that perhaps the patient samples were patients who tested XMRV-positive at WPI. Peterson says he wanted to
handled more than the controls. help his former colleagues at the institute. “I went to Jay Levy to prove
The fourth study, led by Greg Towers at University College Lon- them right,” he says. All samples tested negative. Again, Mikovits said
don, was even more damning. It was a follow-up to an earlier report, the study was flawed.
published in the April 2009 issue of the Journal of Virology by “They didn’t do one thing we did,” Mikovits says.
Miller’s group, that described the discovery of XMRV in a cell line “We did it exactly the way they did it,” Levy says.
called 22Rv1, which was derived from a human prostate cancer patient. On 31 May, Science published the Levy report and the Coffin-
At the time, Miller thought the finding supported the theory that the Pathak origins study online, along with a so-called Editorial Expres-
virus infects humans, as he assumed the original prostate tumor used sion of Concern by Science Editor-in-Chief Bruce Alberts. “The study
to make the cell line harbored XMRV Now Towers’s team had com- by Lombardi et al. attracted considerable attention, and its publication
.
pared the genetic sequence of XMRV from different 22Rv1 lines with in Science has had a far-reaching impact on the community of CFS
reported sequences from patients. patients and beyond,” he wrote, before stating that “the validity of the
The cell line viruses proved ancestral to those in patients. There study … is now seriously in question.” “What Science giveth, Science
was also more genetic diversity in viruses from different 22Rv1 lines taketh away,” Johnson blogged on Phoenix Rising.
than different patients, precisely the opposite of what should happen if Alberts and Science Executive Editor Monica Bradford had first
the virus infected humans: Immune system pressure typically forces a suggested that Mikovits and her co-authors retract the paper volun-
virus to diversify as a means of escaping attack. Towers concluded that tarily. “Science feels it would be in the best interest of the scientific
XMRV never infected humans and that the cell line virus had somehow community,” they wrote in a 26 May letter. Mikovits was livid and
contaminated the patient samples. questioned Alberts’s motives. “Who wrote that letter? I don’t think it
In March 2011, Coffin and NCI virologist Vinay Pathak delivered was Science,” she says. The co-authors thought the retraction request
what many thought was the final blow to XMRV at the Conference on was premature, too. “What if we walk away from this based on con-
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8. NEWSFOCUS
tamination and it’s not contamination?”
Lombardi asked. “You’ve got to give us time
to figure this out.”
Alberts stresses that they floated the retrac- “People will
tion idea because Science already planned to
publish the Expression of Concern. “It wasn’t rather go over
a public call for retraction,” he notes, empha- the Niagara in a
sizing that the recipients shared it with the
media. He also does not think it would have
barrel than ever
been premature, although he says it’s often a getting involved in
tough call whether to retract a paper. “Ulti-
mately, it requires expert judgment and a lot of
CFS again.”
sensitivity to the issues,” he says. “We had lost —SIMON WESSELY,
KING’S COLLEGE
confidence in the results.”
LONDON
T he growing rift between XMRV believers
Downloaded from www.sciencemag.org on September 22, 2011
and doubters became painfully obvious at
a June retrovirology meeting in Leuven, Bel-
gium, where the two camps literally kept their
distance. In the scientific sessions, questions
were mostly polite and informative. During
coffee breaks and a poster session with Bel-
gian beer and cheese, Mikovits, her old friend
Ruscetti, and De Meirleir, who helped open
RedLabs USA, stuck together and barely talked to their scientific oppo- But he emphasizes that many scientific unknowns remain about XMRV .
nents. The scientific debate was grinding to a halt. He points to a study in the Journal of Virology in May that intentionally
“I don’t care if nobody else in the world wants to work on it!” infected macaques with XMRV It shows that XMRV moved out of the
.
Mikovits exclaimed at one point, rolling her eyes. “Fine, leave us blood of the monkeys but stayed in tissue reservoirs and that antibod-
alone!” When Mikovits’s anger subsided, she appeared earnest and ies disappeared. “We know nothing about the viral life cycle,” he says.
even confused by all the criticism. “The virus is real. … I have isolated Blood Working Group member Michael Busch, head of the Blood
it from patients. I know it’s there,” she said. “Believe me.” Systems Research Institute in San Francisco, California, says Ruscetti,
It wasn’t just that scientists were growing tired of the debate, Mikovits, Lo, and Alter deserve kudos for participating fully in the
Wessely says. Some were put off by the “appalling, unforgivable study. “I commend them on their scientific integrity and commitment
attacks” by some patient advocates on those who criticized WPI’s to the scientific process,” Busch says. “This has been a difficult and
findings. Wessely says he has received death threats in recent years. disappointing process for them and for CFS patients, but hopefully we
“People will rather go over the Niagara in a barrel than ever getting have all learned lessons that will guide future research and lead to dis-
involved in CFS again,” he says. covery of the cause and cure of this disease.”
Silverman notified Science and his collaborators on Lombardi et al. The results of yet another multilab study are due early next year.
of his discovery that the data he supplied for the paper were wrong. Led by Ian Lipkin of Columbia University, the $2.3 million project
Resequencing samples he tested for the study revealed that somewhere plans to test 150 samples from CFS patients and create a repository
along the line, VP62 had contaminated them. Without casting blame or of their blood so that future putative causes of the disease can more
explaining how the contamination occurred, the partial retraction says easily be tested. Lipkin says the study will continue despite the Blood
two figures and a table that reported viral sequences, including one that Working Group’s negative results. “Our study designs are different, our
showed XMRV in the family tree, were “spurious.” power is different, our subjects are different, and our assays are differ-
In late August, the Blood Working Group completed its roughly ent,” Lipkin says. “Whether our results will differ remains to be seen.”
$500,000 study, which conclusively determined that no one need worry Mikovits and Ruscetti are soldiering on. “As long as there are
about XMRV or MLVs in the blood supply. The nine labs—which scientific reasons to continue, I think it’s incumbent upon me to do
included WPI, Ruscetti, and Lo at FDA—each had received blinded it,” Ruscetti says. “I owe it to both the scientific community and the
samples from 15 negative controls and 15 others who had tested posi- patient community.”
tive for a gammaretrovirus in Lombardi et al. or Lo-Alter. Different Mikovits says it is irresponsible to dismiss the link between XMRVs
teams cultured the virus, looked for antibodies to it, and used PCR to and CFS at this point. “Anyone who says this is a lab contaminant has
fish for DNA. All labs could use whatever assays they chose. drawn the wrong conclusion and has done a disservice to the public,”
Only WPI and Ruscetti found intermittent evidence of viruses or she says. Okay, maybe not as many CFS patients have XMRVs as they
antibodies in patients, but they also reported similar numbers of pos- initially reported, but she’s still convinced that a gammaretrovirus is in
CREDIT: KING’S COLLEGE LONDON
itive responses in the negative controls. What’s more, there was no at least 20% or 30% of them. “I know of hundreds if not thousands of
agreement between the two labs on which patient samples tested posi- people with evidence of this infection, from what we’ve done over the
tive. “What this says, at the very minimum, is that we can’t find it reli- last 3 years,” she says. “I don’t know what it means. And I’m gong to
ably in the blood of patients we found it in before,” Ruscetti says. keep looking for in vivo reservoirs like the ones seen in the macaques,
Ruscetti understands why many of his peers think Lombardi et al. and I’m going to try to figure out mechanisms of pathogenesis, epi-
and Lo-Alter have now run their scientific course. “It is quite legitimate genetics, or other things. I’m not going to stop studying it.”
for those people to say maybe these two papers were wrong,” he says. –JON COHEN AND MARTIN ENSERINK
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