Blood Products Advisory Committee Meeting Announcement (BPAC) December 14-15, 2010

[Cort]

"ppl" is just an abbreviation for "people". Been using that one online since long before the Internet!

Thanks!

 

[omerbasket]

Zero? Surely not.

If I understood correctly what Cort wrote in another message of his, it was 2/50 positives in the healthy cohort.

Cort - where are you taking this data from?

 

[Esther12]

Thanks omer. I missed that.

4%... that's consistent.

 

[urbantravels]

Bring on Graham Simmons!

Have I got my time change screwy, or are they running way behind?

 

[CBS]

He does agree with the need to "push your PCR" - this is a rare virus in the blood...and he does question how such a rare virus could be causing disease...of course they've only looked in the blood. In monkeys it appeared - spread through the blood - then basically disappeared. So its maybe a one time infection type of thing. It gets in there - goes to where it will go then is disseminated no more! It's not in the blood anymore - its not going anywhere since that is how viruses move around the body - except in the localized areas where it got in during the initial infection.
...

In light of the Emory macaque studies, Stoye would be foolish to assume that just because at a later stage the copy number on the blood drops that the virus wouldn't be in the lymphatic tissues? bone marrow? reproductive tissues? spinal fluid? respiratory secretions? causing serious pathology.

They are looking in the blood because it's easy, not because it's the only place that a virus might replicate and/or might cause harm. I'm not terribly impressed with his lack of imagination/curiosity. Somebody needs to send him a link to the the Singh patent app.

 

[Cort]

healty control are pos, we have a REAL problem. But no-one has been able to replicate Lombardi study.

XMRV in other contexts - respiratory tract of immuno-competent and im-compromised indivs. Another breast cancer study. Not yet confirmed by other techniques. (we've seen all these studies)

Now sneering about XMRV/autism possible link. Assays look pretty good. I'm surprised if results are 0, if its there in as many patients as has been "claimed."

On to Lo paper. Most appropriate term is "Modified" polytropic MLVs. Wants to make 3 points: (1) No virus isolation or evidence for viral insertion into DNA. (Now talking about the rumor virus crap. Does he do ANYTHING original, except 0/0 studies?)

No replication comp0etent, nonrecombinant endogenous PMVs have been isolated. More detailed characterization of the viruses descri bed by Lo et al is urgently required.

Stoye is pointing out the holes - which is fine. That is how Science proceeds. That's why it works actually. Eventually everything has to get wrapped up one way or the other.

His point about Lo/Alter just emphasizes how much researcher want to see a) an isolated virus and/or evidence that that viruses has gotten into the DNA of a human cell. A problem appears to be that they have never found pMLV's that can do that in humans.

Basically these researchers appear to have two questions - there is the XMRV question and there is the pMLV question. We still need replication of the WPI's results and Hanson/Alter need to show that they have found an actual virus. Neither Stoye nor Coffin regard the Alter study as confirmation of the WPI;s results. Neither does the retrovirologist I recently talked to.....

Still there are differences of opinion.......

Have we heard anything really new yet?

I'm taking it from ValB626 posts on the ME/CFS Forums

 

[urbantravels]

They are looking in the blood because it's easy, not because it's the only place that a virus replicate and/or might cause harm. I'm not terribly impressed with his lack of imagination/curiosity. Somebody needs to send him a link to the the Singh patent app.

Agree with you about Stoye, but I would also point out that the topic of this meeting *is* blood safety, which would explain a certain focus on blood.

 

[Cort]

He is the critic - taking care of and validating XMRV requires answering the critics charges. That's part of the game of science.

He does state that most tests wouldn't detect pMLVs - which is not what I heard. As I remember the CDC stated they thought they would have picked them up. And then gives the requirements for working at the 'limits of detection'

Stoye Now he's talking about the Kozak article on incidence of difference MUlvs in diff parts of the world. Says they're not part of a continuum. If they are diff, then how did each of them get into humans?

Complaining about primers. Says wouldn't detect PMVs. Doesn't believe there is XMRV in any of the samples (Lo, Mikovits, Lombardi).

Review of all the reasons again for discrepant results. Doesn't think all explanations cover all the cases all the time.

"When you have eliminated the impossible, whatever remains, however, improbably, must be the truth" he says. Because of working at the limits of detection.

Amplification of non-viral sequences to give the "right-sized band"
Amplification of enERVs from contaminating mouse NDA (mitoch or IAP controls)
Amplificaton from plasmid DNA (vector controls)Amplification of viral nucleic acide from "outser space" (sporadic cultures will give occasional positives)
Virus infection (signal from culture will rapdily increas with passage)

Having positive controls exacerbates all of the above. Need separate buildings for EACH test..

 

[anciendaze]

Good point about blood safety.

Did anyone else catch that Stoye's answer to the question about positive controls amounted to veto power for naysayers?

 

[Cort]

Not to beat a dead horse but my guess is that this

Lo et al samples do not contain XMRV and so don't corroborate Lombardi

reflects general researcher opinion and Coffin again makes that point.

XMRV levels may be estremely low and hard to detect
Lo et al samples do not contain XMRV and so don't corroborate Lombardi
Reconciliation of diff studies might imply multiple origins of CFS
Remains unclear whether XMRV present any problem to the blood supply

Nancy - good, brilliant questions, as always. Remembering 4 years of improving assays with HIV research led to 4 years' of increasing levels of positives identified....

Annoye: If there were 1/1000, would find it.
Can get cross-reactivity from serology.

New Questioner: Showed studies with same primers. Has anyone tested the same specimens?
Annoye: Will hear of results on the same samples.

Coffin: Our current info is based on 5 diff things/results via diff tests for both PC and CFS. Important not to conflate results together. May not be tips of the same iceberg. Lo et al study doesn't confirm or refute Lombardi study. Looking at 2 different things, each has diff sources of possible problems.

Nancy says, yes, but in HIV we got better with time and we found more and more positives - and Stoye will have nothing to do with that. He appears to have more or less decided....He has gotten off the fence a bit more since the Workshop....

Yes he seems to have decided...

More Coffin discussing artifacts, lack of experimental evidence. Need to keep moving forward, but certainly not at the point yet.

Annoye: 17 env sequences in Danielson, 10 nucleotide diffs in total. Statistical reasons to believe results implausible (he said impossible).

Questioner: How would you define a positive sample? Annoye: Same sample all must receive and all agree are positive.

 

[CBS]

I agree that the focus of this is blood but the Emory macaque studies showed high levels in the blood that then dropped. In addition to the existence of a retrovirus, the BWG is trying to establish correlation with, and causation of, disease states - two of their stated conditions for screening the blood supply.

And what does this mean?

"When you have eliminated the impossible, whatever remains, however, improbably, must be the truth" he says. Because of working at the limits of detection.

If you've eliminated the impossible (referring to what?), all you've done is eliminated the impossible. "Eliminating the impossible" tells you nothing about the possible alternative hypotheses (pretty much the bedrock of science). Believing that CFS could not have an infectious origin (the impossible?) tells you more about the "scientist" than it does the science.

 

[Cort]

Monkey study - I wonder if any of this is new??? This could signal where else XMRV is in humans. In lots of organs... The retro I talked to did note that they essentially flooded the monkeys with virus and that it might not spread as readily in nature; still if it can get going it can go far....

Interesting that it's in the GI tract of females. It could lead to chronic immune activation..hmmmm......and marked activation of the T and NK cells.....they are responding......they note that immune activation does fall over time...No evidence of fatigue in the monkeys, though.

If looking for FISH tech, find cells that are still pos. Not cleared from prostate, no protein production. Difficult to localize.

Besides prostate, acute inf: pancreas, lung, testis (acute and chronic).
Liver, kidney - FISH, signal there.

Female animals - lots of virus in GI. In cervix isolate cells and vaginal wall.
Cervix, 2 diff types of cells affected.
Lung epithelial may be infected in additional to alveolar macrophages.

Reproductive organs positive in both males and females. Hints of transmission.

chronic, persistent, can be reactivated. don't see much in blood past acute infection unless is reactivated.

Transient activation of B, T and NK cells - studying now. Could have chronic immune activation which could lead to oncogenesis by itself.

Does XMRV cross the mucosa to induce infection? - studying now.

Went back to infected macaques, infected into urethra. Antibody titers in new rhesus infections could infect mucosa, went through roof. Clearly a delay before could find antibodies (days).

Dr. Nelson: Did you study any CNS issues? If prob w/fatigue, could be an issue? Did anyone modify activity level of the animals? Behavior?
Villinger: Yes, can find occasional FISH pos cell in CNS, no proteins. Not a good medium for virus to replicate.
During the day, were as active as other group members.
Dr. Nelson: Could you monitor how much animals sleep? Villinger - no evidence of behav change. Late-stage HIV monkeys, become sluggish.

Hanson: Do you have any sequence to compare to what you found in animals? What you used to infect them vs what was in the tissues? Vill - haven't done that yet.

Nancy - degree of immune activation?

Vill - jumps from 2% to about 10-20% of CD4 and CD8 and marked activation of NK cells, while virus in blood. If could induce virus to replicate on chronic basis, want to see how activation plays out over time. Falls after acute.

 

[Esther12]

It sounded like Stoye knew the BWG results and felt they backed him up. But the others must know the results too.

 

[CBS]

Why do I feel like we went to visit the summer cabin in August and found the building demolished, all the surrounding trees in massive heap at the bottom of the avalanche shoot (never a good place to build your cabin!) and we're trying to find the snow?

Just because the snow is two miles down the canyon in the lake doesn't mean that the snow wasn't the culprit (don't forget to allot some share of the blame to gravity)!

 

[Otis]

Stoye is pointing out the holes - which is fine. That is how Science proceeds. That's why it works actually. Eventually everything has to get wrapped up one way or the other.

Disagree. One can't present "Summary of Current Research on MLV-related Human Retroviruses and Disease Association" and play devils's advocate at the same time. This is an effort to "balance out the story" is quite the opposite.

And to repeat my previous request: Bring a complete theory for contamination or leave the friggin accusation and innuendo out of it.

His point about Lo/Alter just emphasizes how much researcher want to see a) an isolated virus and/or evidence that that viruses has gotten into the DNA of a human cell. A problem appears to be that they have never found pMLV's that can do that in humans.

Elaborate please. Who is the "they" who have "never found pMLV's that can do that in humans."

Yes it needs sequenced. The notion of seeing insertion of DNA of a human cell came from a PNAS reviewer. Who's calling for that today?

 

[Cort]

Here's where they are now:

.

Animal Studies: Potential Transfusion Transmission of MLV-related Human Retroviruses, Francois Villinger, Emory University (20)
E. Update of Blood XMRV Working Group Activities, Graham Simmons, Ph.D., BSRI (15)
F. Prospective and Retrospective U.S. Donor Surveillance Studies, Michael Busch, M.D., Ph.D., Blood Systems Research Institute (15)
G. Assay Development Efforts on MLV-related Human Retroviruses, Rachel Bagni, Ph.D., National Cancer Institute (20)

BWG is next!!!!!!!!

 

[Cort]

CDC actually finds XMRV - but only on days 2 and 4 (and from what samples?) and WPI is in accordance with CDC. Interesting consistency on those days. NCI can't find it.

WB (?) does not work- plasma does......(?)

Simmons - BWG activities overview. Mission, group composition, evaluate blood safety risks. Review of the 4 stages.

These are the slides I posted earlier, before session began.

We've seen Phase I results. WB is good. Plasma a little worse. But not much diff in diff labs.

NAT detection assays were highly sensitive. Caveats I'll post later.

CDC results all negative for WB. 2/4 positive on plasma, but only on day 2 and day 4 samples. Appeared to be XMRV-related. (I really hope they post these. I couldn't read the copies, can't see the projected slides).

WPI - WB also all tested negative. Similar results from plasma. Better detection at days 2 and 4, too

NCI/DRP results, all were negative at all time points

So, 2/3 labs found it in clinical samples. WB better. No explanation for why Day 2, 4 better, except cells may be dying and releasing particles.
More caveats.

Phase IIb - more structured to overcome limitations of Phase IIa. Interesting that one patients started ARV therapy about the time the first study started.

NCI/DRP all negative, all time points yet.

 

[Sasha]

No evidence of fatigue in the monkeys, though.

But the monkeys only had XMRV for a few months before they were killed, if memory serves - I thought the working model for CFS was that you get XMRV and then viruses that you catch later cause the fatigue.

 

[Cort]

CDC found all plasma and PBMC samples negative at both time points. Added some assays.

Gen-Probe (TIGRIS System) All plasma and PBMC samples at both time points negative.

(Not reporting WB results here becuz 1 lab isn't done with testing)

WPI results - No virus in plasma. Found it in 2 of PBMCs. Problem with one sample.

Hard to decipher - they are all looking at the same samples presumably. Now WPI finds it in 2 (of 4 PBMC's) but not plasma while CDC cannot find it

 

[Esther12]

Well that's cleared things up. Now we know.