Monday, 20 March 2017

What Did the B Cell Video Miss?

Many Years (1994) Ago, 
I made an (immortalised) B cell Line. 
Here are the Pictures (low power on left).  
On cell surface (on the right) there are little blobs. 
What were they?
They were Budding Virus Particles

So Maybe the New B cell Video
didn't miss a trick. 
Here's the B cell containing Little blobs too.
They get infected with Virus in Humans, notably in MS


A Black Swan Idea


Anti-CD20 kills Immature to Memory Cells, 
Alemtuzumab (anti-CD52) really kills the memory B cells
           
Maybe B cell Depletion Kills the Virus

Maybe that is why they work?

Maybe Not a T cell Issue...Food For Thought

57 comments:

  1. Okay, so a basic and probably stupid question from me... Are you saying there's a filthy virus stuck to my B-cells, and it may be this virus which is causing my MS? What is this virus?

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    1. EBV lives in B cells HERVs live in every cell

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    2. So most of your b cells have a filthy virus.Sorry.

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    3. EBV immortalised B cells. Do they create virus factories.

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    4. It may not be the virus itself that causes the problem but the damage to the immortalised cells, changing their behaviour. As an example take a look at how Reed-Sternberg cells in HL affect the immune system.

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  2. Great B-cell "porn" at the right time of the day, too. :-D
    MD, if the cell was full of the virus particles, you blow it up making complement holes and ... the virus particles get out and float around. Or? I am not a virologist or an immunologist, so maybe you need to talk this through with somebody who knows more about these things...

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    1. The figure is an immortalised mouse B cell, EBV is typically latent and so is not going to be shedding but the figure simply makes the point that there can be alternative explanations not in the videos.

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    2. Hey,If it wants to live further it needs to spread, multiply, come to the mucosal surfaces, find it's way out ... and infect new hosts.

      But it gets better, because it would actually fit your B long lived memory cell theory - that is where EBV latently hides before it pops out.

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    3. Reading more in my old notes: EBV DNA has two origins of replication:
      Ori lyt and Ori P. Ori Lyt is used for lytic replication - high copy number.
      Ori P is used for episomal replication in latently infected cells: low copy number.
      But it also says that antibodies and CTLs have problems destroying those infected cells

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  3. So the virus particles might be Epstein Barr Virus. Those are infecting the B cells somehow making them immortal. These images are beautiful, what image is a photo and which is art work? If the images are real how were they taken?

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    1. In the example I show it is not EBV. In most cases EBV is laten and not shedding from the cell surface but I use images to make the point.

      The top two are taken by scanning electron microscopy (you coat the fixed cells with a fine layer of gold using a sputter coater and then us a scanning microscope. The next on down is transmission electromicroscopy which is a section taken at thinner than than a thousandsth of meter, If fact you can see the magnification and size of a bar e.g. 1 micrometer (uM). They are by myself and Peter Munro

      You can colour them in to make beautiful images, check out some of them by a person I worked (Steve G)

      https://render.fineartamerica.com/images/rendered/search/canvas-print/mirror/break/images/artworkimages/medium/1/1-immune-system-cell-sem-steve-gschmeissner-canvas-print.jpg

      Head lice
      https://cosmos-magazine.imgix.net/file/spina/photo/8335/281016_micromonsters_4.jpg?fit=clip&w=835

      One of his images was bought by Damian Hurst...blown up to 2m and sold for a wodge of cash.

      The bottom two images are computer generated from the Roche Video. In their video the blobs are CD20 on the surface of B cells, but I think it makes the alternative point too.

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    2. These images are just soooo nice...

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  4. Okay, so there may be virus shedding somewhere in the B cell lineage that may be contributing to MS. But why is this virus undetectable?

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    1. The problem is that 95% of the population are forever infected with the virus (once it is in you, you have no way of getting rid of it). So most people walking in the street have EBV, but if you take MS population the % is even higher( close to a 100%) - hence forever speculations that it has something to do with pathophysiology of MS.

      And if one could get rid of it by, say, killing all infected B cells - then there are still endless opportunities of being infected a novo from all those 95% of people around us who have not been treated.

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    2. MD, you have convinced me. Love or hate EBV, it is a marvellous machine that is so successful because it knows when to turn on the slow "I'll just stay here in the memery B cell" program and it does not kill the host. That way the host has the potential of carrying and spreading it around the world all its life (literally :-).
      How can something that hasn't even got a nucelus be so clever and speculative? It is not like Ebola which goes for the quick kill.
      Look who's got more hosts "covered"?

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    3. ProfG tells me a story about a neuro saying Hey prof G we have tested one of our alemtuzmab treated patients and they are EBV negative so your idea is rubbish....

      However turns out before alemtuzumab they were positive.

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    4. "then there are still endless opportunities of being infected a novo from all those 95% of people around us who have not been treated."

      This makes me wonder... Say I went on Ocrelizumab or something else which annihilated my EBV-infected B-cells and acquired all benefits of that, but also the risks of an aged immune system, but then got EBV again by kissing my partner. There would be no point in the Ocrelizumab???

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    5. He he he, it is probably more likely that the test is false negative than the MS patients is truly negative. If 95% of the general population are positive this would probably also be true about the rest of the "random people in the street", though. What is the sensitivity and specificity of the tests used, does anybody know?

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    6. Confused, it is a lovely example... it is precisely that the virus is still around us and we are not able to live in a bubble after treatment. Maybe that's why rituximab has to be repeated - in case of RA it takes about 9 months before the patients feel it in the joints, so it is given after 6 months to avoid relapse. As far as I am aware ocrelizumab is also infusions every 6 months?

      I would not worry too much about ocrelizumab aging your system. I would say that the chances of MS aging your body prematurely and reducing function are much higher.

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  5. More and more utter rubbish all useless. Still NO cure says it all.

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    1. I disagree, how do you know it is not a cure? It may have induction potential.

      Please do not use the troll font (Upper case words in lower case sentance) as it increases your chance of being spammed

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    2. Anonymous, if it was all rubbish, we would not be up at night all excited about it, would we? ;-)

      EM pictures put me in a state of awe other people get from climbing a mountain and looking around :-), and viruses are infinitely interesting things with architectural structures that have the capacity of assemblng themselves. Now, how cool is that? Nature making something almost alive out of nothing.

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  6. In 1997 Hauser from UCSF tried to do a NIH trial on Rituximab but
    they told him no because MS was a T cell disease. Couple years later
    he got Genentech to do a trial.

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    1. There you go, bias and this is rife, so now that anti-CD20 clearly works to block relapse, we have the view that it blocks relapses. The arguement is that it stops cells turning into antibody producing cells, but atacicept kills plasma cells and drops antibody levels yet makes MS worse, so it has to be blocking antigen presentation and blocking T cells.

      Whilst I am prepared to accept this view I am likelwise prepared to challenge the view too.

      However if you challenge the view you get stuff in that refereeing treacle because surely at least one of the 4 referees is not going to buy a non-T cell story. Get the idea out do the experiments to disprove the idea, or is it simpler to suppress ideas.

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    2. See both sides then, you (and you convinced me) think that it is the memory B cell harbouring the virus that gets killed. But there is a place for T cells in the story ( Th1 help with fighting viral infections, we do not challenge that) - and everybody is happy, both the T-cell and B-cell religion followers.

      I still struggle explaining what happens with the free virions released from the death of the sacrificed memory B-cell. Don't they have anything new to infect ( because most potentially vulnerable cells have just been punched by the complement?) or isn't there enough of the virus in a latent infection to spread anywhere? or maybe both? It is a long long way to the next lymphocyte when the lymphocyte count is 0 (Lem), but then Mabthera does not get the lymphocytes to 0.
      Have you got a good explanation?

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  7. Excuse me butting in to the interesting exchange between you MD and CD20 cell, but I've a silly question to ask. My professional background is child protection, fostering and adoption, so my world has been human behaviour and psychology. I have an appalling lack of understanding of the world of science and medicine the two of you occupy. Since diagnosis in Nov 2015 I've been accessing this site to attempt to grasp some understanding and insight, and my question is: should I be despondent at you both debating images from 1994 or encouraged that study and debate is moving us forward? I guess, I naively thought that the progress to date with DMTs in the past ten years represented progress in all regards. Some understanding of the terms of reference would be helpful. Thanks.

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    1. Dear Fi
      Don't be despondent, there are more treatmnet options now than there's ever been. It was just steroids when I started my career in MS research. The point of the discussion stems from MD's search to find the specific cell type responsible for MS so this can be dealt with whilst leaving the rest of the immune system intact. Many of the current MS therapeutic agents such as Lemtrada take a shotgun approach to the immune system, good cells are taken out as well as bad. We're looking to make things more specific with less side-effects, such as infections and the development of secondary autoimmunities.
      The ultimate trigger may be Epstein Barr virus infection, meaning vaccination against EBV might stop the development of MS, if only we had a vaccine against EBV. We don't and I do wonder why, though pharma doesn't make much money out of vaccines and if we stopped MS from developing, the money stream for MS therapeutics would dry up, albeit slowly. So not much of an incentive for pharma there.
      Science is always up for discussion and reassessment, it's amazing sometimes that you stumble across some old findings that have been forgotten and see them in a new light which can really move things forward.
      So, don't be despondent be encouraged!

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    2. :-))) I think we are both seeing a bright new dawn here... hopefully it is not the lights of an express train coming at us from far away!

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    3. "We don't and I do wonder why, though pharma doesn't make much money out of vaccines and if we stopped MS from developing, the money stream for MS therapeutics would dry up, albeit slowly. So not much of an incentive for pharma there."

      ...People's lives destroyed by a disease which could have a cure. People are poking around in zebra fish etc. and doing research on half-baked things like biotin when there could be a relatively simple *cure*? Could there not be some more momentum generated to investigate EBV?

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    4. Dear Fi

      Don't be dispondant, the images have been sat in my filing cabinent for the past twenty years doing nothing. In contrast the MS world has not been sat on its bum. In 1994 there were no MS treatments now their a over ten that are licenced, there are more on the way and many more in the pipeline. We have had the first neuroprotection studies and have had the first studies showing an effect in progressive MS and we have had the first positive results from repiar trials. So if you have RRMS you now have the problem of trying to work out which drug to take.

      The images that I made in 1994 was of a mouse cell line that I made and and as soon as I found out it was houching with virus, it was put in the freezer never to be used again. Although we published on the cell line, we never published the pictures and I dont throw stuff away until it is published, as some day we can make it surface.

      However, the story of the B cell video allowed me to dig out these pictures of a cell line (BCTL-1), they were cloned from the original line called Fatboys, because the cells looked abit plump. The point was to show that in the B cell video they had taken the usual dogmatic view that it is all T cells, but the pictures were handy to make us think about viruses and their possible role in MS.

      ProfG's are always looking at ways to try and hit the EBV virus.

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    5. Keep reading the blog and with time, lesss and less will feel like we are speaking a foreign language

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    6. Jesus....1994, the was the year I first started working in the same lab with MD2. He had made a celline line called GP8. Wonder if that was full of virus too.

      In most cases you would never look.

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    7. Thanks for your reply too MD.
      I'm always telling people that I recognise I'm fortunate, for want of a better word, to have RRMS and to have had the range of DMTs available to me (had Alemtuzumab 4 months ago) Their care and concern is then always expressed as 'What about a cure?' or 'What about improvement?' I've often found it useful to reference the science on this site (even if it is a foreign language to me) to highlight the efforts and progress being made. B cells and T cells aside, those images can just make us feel old MD - my son is 21 - 1995!

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    8. Just wanted to add one other thing that enhances the positivity is the mention in your replies to prevention- as a mum - something else that I have focused on.

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  8. Dear Fi,
    I am not a scientist, but an MS patient (and a clinician by training, but that has nothing to do with neurology).

    "Should I be despondent at you both debating..."

    I don't think you should. The progress in all sciences and technology tends to go in hops and jumps rather than a steady line. Steve Jobs could not make an Apple phone or tablet in the 1950, because the parts and the knowledge were not there. Once the parts exist, many companies are making smartphones all over the world within 10 years of the first one being made.

    That said biological systems are considerably more complicated than man-made systems and progress in biology and medicine will always be slower than in other disciplines, if only because of all the regulations/laws/ safety procedures that have to be in place. If MD invented a new gadget he could have it produced in China tomorrow, if he invented new drug it would take 10+ years even if he had all the money needed. I wonder what he answers (... if he does).

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    1. MD2 and CD20cell thank you both very much for your replies. I'm firmly back in the world of being encouraged and having some confidence again that taking a reasonably positive outlook has legitimacy. I'm not being naive and stupid about what this disease does to those of us with it, but I've held on to the important difference a positive (as possible) psychological perspective can make. Working for many years, with children and adults victims of abuse or those unable to have children of their own, really drove this home, especially in respect to the things that can quietly eat away at you - hence my question. I won't be 'haunted' now by the date of some images! Much appreciated.

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    2. Well we did invent a new symptomatic drug we did the first experiment on this drug in November 2002.

      We have treated 60 healthy people in 2014-3015 and will have treated i hope 160 people with MS by some time this year. Hopefully it will work. Then we only have to do a few more trials for MS. It is the beginning of the road for all the other uses we have found.

      PS. We never had the money we need and that is why it is so slow.

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    3. Hopefully you can read the story within the next couple of months.

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    4. Looking forward to it :-)
      As I sit here things suddenly make more sense... because how do interferons work in MS? the mechanism of action for Betaferon according to product information: antiviral and immunomodulatory. We kind of knew it all along, but it is hard to prove when number of exposed to an infectious agent in the population is close to 100%.

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    5. Beta interferons induce protein kinase R to phosphorylates eukaryotic translation initiation factor to block proliferation of viruses, but also blocks replication of cells in the process so it is anti-proliferative, but to hark back to our B cell paper,

      http://www.ebiomedicine.com/article/S2352-3964(17)30045-2/pdf

      It depletes memory B cells (and depletes the virus that they contain), but does not do this very well compared to drugs that work much better If you look at figure 4E few people deplete memory B cells below 20% of CD19 B cells.

      Do you buy it?

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    6. We can absolutely agree that the efficacy of interferons was there, but was never very impressive.
      Now, somebody somewhere thought in the early 90s that problems in MS are, at least in part, viral in origin. (Hence development of beta-interferons as drugs for MS). Now - why and how did that "it's viral" thought get lost in the hiatus of drug-development history?

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    7. In the early 90s, Neuros were clueless having had a sea of failures and were willing to trying all sorts. Alpha interferon was tried and failed (anti-viral idea starts to go on dodgey ground), intrathecal beta interferon gave a response and gamma interferon made things worse. It was an immune problem and became a T cell disease and a monoculture of funding

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    8. I just opened Janeway's ( not bad at 2:52 a.m.:-), and yes "multiple sclerosis is an example of a T-cell mediated chronic neurological disease"(2012 edition). Guess what, RA was a T-cell disease, too :-D. Not any more.

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    9. Ad you comment 5:58
      "Do you buy it?"

      Now I do.

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  9. MD, how do you know that little particles next to B cells in SEM images are viruses, let alone specific viruses (EBV)? Also, how do you know that all these particles were present in the blood rather than appearing (or amplifying) at some stages of sample preparation? It's way to easy to contaminate cell cultures with bacteria, can this also happen with virus contaminants?

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    1. Let me clear the particles are not EBV because EBV cannot infect mouse cells. How do I know they were viruses, They look like a virus.

      The cells were in a test tube and were not from the blood. They had been in culture for at least 3 months and were grown in sterile solutions.

      I am convinced they came from the cell because you can see them inside the cells. Virus need cells to replicate. In different cell types grown in the same solutions we could see differently shaped viruses.

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    2. And then again why do we use interferons in monotherapy for MS if we know they are not very efficient on their own?
      They are not like rituximab that takes your defences down and raises the risk of infections, they could be stacked up like we stack up treatments in RA:... base is methotrexate, if it is not enough you do add on.
      Maybe, if it works as you hope, one drug will be enough to send MS packing.

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    3. Why do we use interferons....at BartsHealth we dont very much these days.
      Stacking $50,000 (interferon) + next $50,000 (next) + next. I think infliximab was the first in RA but its problem was anti-DNA antibodies and SLE, methotrexate gets rid of B cell response.

      The problem however is that all cheap drugs that could be used now have baggage that they failed in MS (The fact that most were tried in SPMS which was not going to respond to drugs in a short trial and teehy were thrown away, I suspect

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    4. You are probably right here, if you don't know what it is about then it is probably about money... Methotrexate costs peanuts, TNF alfa inhibitors are getting cheaper by the minute, too. But interferons also got cheaper?
      I know that there was fear of inducing SLE by using anti-TNFs. In practice it is not a problem we see in the clinic. What plaques us is psoriasis-like reactions in people who never had psoriasis before and psoriasis exacerbating in people who had it before thay started ant-TNF. ( paradoxal, because antiTNFs are also used as treatment for skin Ps and Ps arthritis).

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  10. Crikey, don't look at this blog for a couple of days and things have moved on faster than a lymphocyte in a Roche promotional.
    EBV is the ultimate selfish gene (ok not a gene as such but as in piece of replicating material).
    MD 9.58 sells it to me, but then I've read the paper a fair few times and was already happily convinced (if a little hacked off with Eli Lilly)
    MD and CD20 you were up far too late last night, what would ProfG think? ;-)

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    1. Judy, I do not know what MD is on :-), but I am sailing on a post-steroid high and I have not had a good night's sleep in the last 2 weeks :-). I wake up between 1 and 2 a.m. fresh as a daisy :-).
      And I bet prof G does not sleep much either these days, dancing with aliens ;-) takes a lot of time and effort.

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    2. Judy you may be interested to note that we have a telecom with Lilly so we are moving up the command structure slowly...no promises.

      Still waiting on others to step up to the plate to dish the dirt.

      What I am on?.....The couch watching tele and writing a paper on a role at the moment. It would be easier if the referees would get their act together waiting 2 months for a response is rubbish.

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    3. Good to hear Lilly is on the line again :-).
      Get writing another paper, that way you will always have something in the post ;-)

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    4. MD I've a few old uni friends work for pharma (lovely people) but I've taken this a bit personally. I sincerely hope you get your paws (do mice have paws?) eventually.
      Keep writing the papers! And thanks for sharing them with us, ignore any criticisms I love your posts - they usually make me laugh too :-D

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    5. Yes we have pores, hope to keep you grinning.

      I'll make a post to show you.

      The big questions is What do we have our paws on now. Stay tuned.

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