Author Topic: Lombardi et al 2011  (Read 24126 times)

since

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Re: Lombardi et al 2011
« Reply #225 on: May 14, 2011, 10:20:52 PM »
The following was presented at the Centennial Retrovirus Meeting in Prague, Czech Republic last spring:

Characterization of innate immune responses in Xenotropic Murine Leukemia Retrovirus-Related Virus (XMRV) infected individuals

J. Mikovits 1 , V. Lombardi 1 , K. Hagen 1 , D. Goetz 1 , M. Marshall 2 , I. Barao-Silvestre 1

Chronic fatigue syndrome (CFS) is a debilitating disease manifested by inflammatory sequelae including innate immune activation, low natural killer cell numbers and function. We recently demonstrated the first direct isolation of an infectious gammaretrovirus, XMRV, from the blood of CFS patients. We have developed quantitative assays to detect XMRV replication and methods for infection in culture. Moreover, we found evidence of XMRV infection in >95% of the 141 CFS patients tested to date. These data implicate a role for XMRV infection in the pathogenesis of CFS. We studied the interaction of XMRV with the immune system, first by identifying cellular targets of XMRV ex vivo by flow cytometry (FCM). We hypothesized that NK cells could be a target of infection and that infection of other PBMC cell types could lead to dysfunction of NK cells either directly or indirectly through the upregulation of inflammatory cytokines and chemokines, which are also a hallmark of this disease. We isolated leukocytes from XMRV infected individuals analyzed phenotypic differences from uninfected controls by FCM. Using purified primary B, T monocyte macrophage and dendritic cells and cell line models we infected with XMRV and profiled cytokine and chemokine expression by Luminex multiplex assays. We analyzed XMRV infection by quantitative RT-PCR and Intracellular FCM. Infection of PBMC in vitro results in a cytokine signature similar to that signature seen in the plasma of CFS patients. These data advance our understanding of the pathogenesis of CFS and may ultimately lead to new therapeutic interventions.

"Infection of PBMC in vitro results in a cytokine signature similar to that signature seen in the plasma of CFS patients."
Quality over quantity.

subtr4ct

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Re: Lombardi et al 2011
« Reply #226 on: May 14, 2011, 10:21:41 PM »
;)  Whenever someone says you people this clip flashes in my mind

Oh great.  Thanks, Keith.  Now I have to run to the store for some Booty Sweat and Bust-a-nuts.   ;D
Disclaimer: I am not a medical doctor.  This post is not medical advice.  Consult your physician before taking any action.

Patricia

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Re: Lombardi et al 2011
« Reply #227 on: May 14, 2011, 10:24:18 PM »
Quote
Infection of PBMC in vitro results in a cytokine signature similar to that signature seen in the plasma of CFS patients.

Thank you, Since.  Is there a way to determine whether that was the same cytokine signature as the one in the current paper?
There may be times when we are powerless to prevent injustice. But let there never be a time when we fail to protest. --Revenge

since

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Re: Lombardi et al 2011
« Reply #228 on: May 14, 2011, 10:33:10 PM »
Thank you, Since.  Is there a way to determine whether that was the same cytokine signature as the one in the current paper?

I would bet that it is, but we won't know until they publish that data.
Quality over quantity.

Tango

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Re: Lombardi et al 2011
« Reply #229 on: May 14, 2011, 10:38:29 PM »
That's interesting.  Makes a lot of sense.

Thank you Since
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Dr. Yes

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Re: Lombardi et al 2011
« Reply #230 on: May 14, 2011, 10:50:10 PM »
Hi omer, I'm not sure the following is clear because I'm reeeally sleepy, but for what it's worth:

It's true that because this study only used an XMRV positive cohort of CFS patients it cannot draw definitive conclusions about the specificy of this cytokine profile to XMRV infection.  [However, just because it cannot do so definitively does not mean that it does not provide evidence in support of that conclusion; the fact that the particular bias of the profile is consistent with retroviral infection is evidence that this profile is specific for XMRV positive CFS.]   

The value of this study, aside from revealing an underlying cytokine pattern, is as a crucial first step to developing a diagnostic tool for XMRV+ CFS (and perhaps, in the future, other forms of X-associated disease), which requires very accurately defining the terms used.  There are serious scientific problems with trying to generalize any conclusions to traditionally defined CFS, as it is not a rigorously defined term.  So the WPI has here established a foothold for XMRV by characterizing "XMRV+ CFS" patients, in this case by immune parameters. 

For that reason the scope of this study was narrower; it confined itself to identifying a cytokine profile that is predictive of at least XMRV+ CFS.  This approach is understable given the difficulty of being certain that someone is X negative; given those circumstances, this was a rational first step in developing a potentially diagnostic profile, which requires initial characterization with a tightly defined cohort.

Does this study 'prove' that the cytokine profile it has found is caused by XMRV?  No, but that was not its intention and at this stage it would not be expected to.  Even if an XMRV negative Fukuda CFS cohort had been included as a control, and had different results, we wouldn't be able to conclusively say that XMRV is the determining factor because Fukuda CFS is such a heterogeneous label that the difference could be explained by other variables.  If instead a CCC, acute onset CFS cohort was to be used as a control, there would be the problem of finding true XMRV negatives in that cohort.  How would you find, say, 100 such patients who are XMRV negative if by a combination of WPI assays only 5% or so of such patients are XMRV negative.. and we don't know for certain if even those are true negatives?  As a result, you couldn't be certain that there aren't XMRV infected individuals in such a cohort, and therefore the value of its overall immune profile as a control would be questionable.  So to initially develop a baseline from which to assess the sensitivity and specificity of the profile as a diagnostic tool, this study had to exclude such cohorts.

What this study does do, like the Lombardi 2009 study, is document an association between XMRV and a particular characteristic (in this case a particular cytokine/chemokine profile in CFS patients).  It makes no comment about the nature of this association, such as if XMRV causes the profile.  It is a necessary first step to clarifying that issue, however.  Such clarification will not be easy, though, as I mentioned above, because of the difficulty in isolating a cohort of well-defined CFS cases who are truly XMRV negative.   

In this case, it is a cytokine/chemokine profile, and the association is so strong that the profile generated can be used to predict XMRV/CFS status with high accuracy as opposed to healthy controls.  We do not know yet what happens when it is used on CFS populations defined by one criteria or another.

Had they been able to find an XMRV negative cohort of CFS patients and test them as well, they might have been able to make a more definitive association between XMRV and this cytokine profile, or make a strong case for the abandonment of vague Fukuda or Reeves cohorts, as well as build evidence for a causal role for XMRV in X+ patients.  That was all outside the scope of this study...However, I guarantee you that such studies are in the works.  This is just a first step, and you can see some of its value within the context of other WPI research; for example, one way to look at it is as an extension of the Science paper: a further investigation of the CFS patients found positive for XMRV as compared with healthy controls revealing a distinct cytokine profile.  The WPI clearly has more data which they will publish soon enough that will investigate the potential causal role of XMRV in generating this cytokine profile.
« Last Edit: May 15, 2011, 08:01:42 AM by Dr. Yes »
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Patricia

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Re: Lombardi et al 2011
« Reply #231 on: May 14, 2011, 10:55:03 PM »
Thank you, Dr. Yes.  This is an excellent, very clear explanation.
There may be times when we are powerless to prevent injustice. But let there never be a time when we fail to protest. --Revenge

Tango

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Re: Lombardi et al 2011
« Reply #232 on: May 14, 2011, 11:01:01 PM »
If there were another disease with high levels of XMRV positives, it would be interesting to see the profile they have in comparison.  More future work and why we now need billions.
"I suspect there have been a number of conspiracies that never were described or leaked out. But I suspect none of the magnitude and sweep of Watergate." Woodward

"I would favor any name that does not impose (or give the appearance of imposing) taxonomic preconceptions on the nomenclature." Coffin

Jaz

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Re: Lombardi et al 2011
« Reply #233 on: May 14, 2011, 11:56:51 PM »
Yep, we need billions of dollars for research into ME.  Look what AIDS gets:

Quote
From the 1990s until 2009, funding for the AIDS epidemic increased substantially. In 2008, an estimated $15.6 billion was spent on HIV/AIDS1 compared to $300 million in 1996.2 From 2002-2008, in particular, funding increased six-fold. Since 2009, however, total global funding for HIV/AIDS has remained flat. This means that the funding gap (the difference in the amount of money needed and the amount actually allocated) was $7.7 billion in 2009, compared to $6.5 billion in 2008.3
http://www.avert.org/aids-funding.htm

Now look at how much the CFSAC asks for.  How much did they end up asking for this time?  Did they ever come up with an amount?

And what do we normally get for ME?  Just a few million, right?   And yet, ME is estimated to be costing the U.S. Government 23 billion dollars in lost revenues annually.
« Last Edit: May 15, 2011, 12:05:23 AM by starryeyes »
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dipic

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Re: Lombardi et al 2011
« Reply #234 on: May 15, 2011, 12:12:14 AM »
Thank you, Dr. Yes.  This is an excellent, very clear explanation.
Seconded. :)
"We're not going to go, you know, doing more and more tests to find out, well, what was the virus because, frankly, even if we found it, there's nothing we're going to do about it. We are in the business of 'rehabilitation'." -Simon Wessely (BMJ Podcast, Mar 5, 2010 - http://goo.gl/XNlMM)

Ginger.Tea

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Re: Lombardi et al 2011
« Reply #235 on: May 15, 2011, 12:22:42 AM »
Thirded.

Thanks Dr Yes!  :)

B8b_E

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Re: Lombardi et al 2011
« Reply #236 on: May 15, 2011, 12:29:30 AM »
The only explanation for the Cytokine profle in the CFS XMRV+ is a shared infection. creating an immune response.

It's unlikely to be shared EBV, HHV-6, CMV,  Parvovirus, Lyme because they wouldn't produce identical Cytokine profiles as different viruses and bacteria/infections work (affect the body) in differing ways and thus pathology and not all the patients would have the infections, like in AIDS. Thus the Cytokine/Chemokine and other markers wouldn't be universal. But they were, almost, universal. HIV is required to blow CD4 away in AIDS, and it is evident that XMRV is required to ramp up Inflammatory Cytokines/Chemokines and other markers in ME.

That is very telling.

To have a shared profile in 93% of people with a political label  of 'CFS' who all have a retrovirus XMRV, it is clear the answer is  shared HGRV's. There is no realistic logical explanation  other than this.

The infection has to be shared to create the same immune response, and we know the infection is shared as everyone had HGRV infection.

Klimas talked about anti TNF-Drugs, Mikovits mentioned non steroidal anti-inflammatorys. Lets see some in action  in clinical trials and see what they do to XMRV+ CFS.
« Last Edit: May 15, 2011, 01:09:20 AM by X+BäbY »

B8b_E

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Re: Lombardi et al 2011
« Reply #237 on: May 15, 2011, 01:11:11 AM »
Reality check for the people who still don't believe the WPI have blown the lid off Pandora's Box. Get it? :P


The Lombardi et al 2011 paper of XMRV Retrovirus also had two abnormal markers seen in AIDS, another Retrovirus.

Namely,  Rantes and Eotaxin

Quote
Role of Th2 Cytokines, RANTES and Eotaxin in AIDS-associated Eosinophilic Folliculitis
Amerio et al
Acta Derm Venereol 2001; 81: 92–95

Folliculitis is a symptom of ME. I have Eosinophilic Folliculitis  for 20 years since a kid with no explanation.
I am XMRV+

Quote
[Eotaxin receptor CCR3 is required for HIV-1 entry into target cells such as microglia and eotaxin inhibits the infection of HIV-1
He J, Chen Y, Farzan M, et al. CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia.
Nature. 1997;385(6617):645-9

Eotaxin and MIP-1 beta inhibits HIV-1 infection of Microglia

CCR3 and CCR5 are co-receptors for HIV-1 infection of microglia.
He J, Chen Y, Farzan M, Choe H, Ohagen A, Gartner S, Busciglio J, Yang X, Hofmann W, Newman W, Mackay CR, Sodroski J, Gabuzda D.
Nature. 1997 Feb 13;385(6617):645-9.

Microgial activation is a feature of ME and directly linked to MLV's.
Microglia in my brain is kicking out MIP-1 at over 1200% abnormal levels in me.
I am XMRV+

Quote
Eoxtaxin is associated to CD4 loss in AIDS

Plasma cytokine levels during acute HIV-1 infection predict HIV disease progression.
Roberts L, Passmore JA, Williamson C, Little F, Bebell LM, Mlisana K, Burgers WA, van Loggerenberg F, Walzl G, Djoba Siawaya JF, Karim QA, Karim SS.
AIDS. 2010 Mar 27;24(6):819-31.

CD4 loss causes gross immunodeficiency and death in HIV AIDS.

For sure, this is not a feature of ME!

But another immunodeficiency cell surface marker is..........

CD3 T-cell loss loss is also reported in ME literature.
I have CD3 loss myself and am XMRV+
Low CD3 is associated with immunodeficiency.
Retroviruses cause immunodeficiency.

Quote
MCP-1 and Eotaxin is associated to HIV transmission

MCP-1-MCP-3-Eotaxin gene cluster influences HIV-1 transmission.
Modi WS, Goedert JJ, Strathdee S, Buchbinder S, Detels R, Donfield S, O'Brien SJ, Winkler C.
AIDS. 2003 Nov 7;17(16):2357-65.


MCP-1 and Eoxtain is elevated in XMRV+ CFS

Quote
''RANTES can slow the progression to Aids in HIV-positive individuals''

Dr Anthony Fauci
US National Institute of Allergy and Infectious Diseases
Published: Journal of Aids

Rantes is elevated in XMRV+ CFS.




If I can find this out in 20 mins, what can the WPI and other scientists do in 2 more years..............
« Last Edit: May 15, 2011, 01:16:50 AM by X+BäbY »

cath

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Re: Lombardi et al 2011
« Reply #238 on: May 15, 2011, 04:36:06 AM »
IL 13 is downregulated.

Quote
http://pfam.janelia.org/family/PF03487#Interleukin-13

IL 13 stimulates growth and differentiation of B cells (IgE), inhibits TH1-cells and the production of macrophage inflammatory cytokines (e.g. IL-1, IL-6), ↓ IL-8, IL-10, IL-12

Not enough IL 13 will give less growth and differentiation of B cells, less inhibition of Th1 cells and more production of IL 1. IL 6. IL 8, Il 10, IL 12.

WPI found that IL 6, IL 8 and IL 12 are upregulated.



Angela Kennedy

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Re: Lombardi et al 2011
« Reply #239 on: May 15, 2011, 06:55:48 AM »
Seconded. :)

Thirded.  :)

Edit: I mean fourthed!  :-[