Why blood of bird flu survivors is a lifesaver [Discussion]

[Sally Furniss]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

Prick test for allergy to horses is fast, measured in minutes.

[Shannon Bennett]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

Anne, you have relieved my mind considerably. If not then why not then?

[JJackson]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

I have to agree with Mingus that this approach has potential and needs further exploration. Our main defences, as things stand at present, are Tamiflu and …er …. well Tamiflu. Tamiflu has been administered to many of the infected but not always as early in the infection as required on the packet. Despite the mercifully small number of human cases to date there have already been resistant cases and resistance in H3N2 is well documented. In addition while my government stockpiles for 25% of its people regretfully that is not the case for most of the worlds population.

From the articles linked to in this thread it would seem that serotherapy, even when given to the sick, has a significant benefit but the more promising avenue is probably prophylaxis.


From the discussion section of the Meta-Analysis paper (link in post #2)

Our analysis suggests that patients with Spanish influenza pneumonia who received transfusion with influenza-convalescent human blood products may have experienced a clinically important reduction in the risk for death and improvements in clinical signs and symptoms. Adverse effects included chill reactions and possible exacerbations of symptoms in a few seriously ill patients. Our subanalysis indicates that early treatment (after <4 days of pneumonia complications) was superior to late treatment (after 4 days of pneumonia complications). The mortality rate among controls and late-treated patients appeared similar and is consistent with the modern recognition that early definitive therapy for pneumonia and hypoxia is clinically important. Although we calculated a pooled estimate of possible effect, we urge caution in the interpretation of the summary estimate and do not think that it should be generalized to other virulent influenza strains without further study.
Our biological hypothesis for why mortality and morbidity may have been reduced is that the virus was neutralized by anti-influenza antibodies in the blood product. Rapid viral clearance would halt further replication and the stimulus for the cytokine cascade that is responsible for the acute respiratory distress syndrome. Reductions in the mortality rate may have also resulted from fewer secondary cases of bacterial pneumonia, empyema, and septicemia. Several human and animal studies (44–58) have reported protection with use of passively acquired anti-influenza antibodies and provide support for this hypothesis. Successful treatment of a pulmonary influenza virus infection in severe combined immunodeficiency mice with hemagglutinin-specific antibodies with very low virus-neutralizing activity in vitro (51) and in H5N1-infected mice with equine-derived H5N1 F(ab) fragments (52) provides direct evidence that anti-influenza antibodies are therapeutic in a model of severe disease.

Going back to the immune system model.
Scenario 1- An immunologically naive host (i.e. a host with neither the right antibodies or the plasma cells to produce them) is infected then there is a good chance of some virons infecting a host cell (due to a weak challenge from an unprepared host). Once they release lots more virons the hosts immune system should kick in and produce the appropriate response but the virus has already established a head start and is difficult to eradicate.
Scenario 2 – A recovered – or vaccinated - patient is challenged by the virus. This time there are already suitable antibodies present and these bind to their matching antigenic sites and in so doing ‘paint the target’ for an aggressive attack by the hosts other immune cells. The plasma cells are already present and ready to produce more antibodies. In this case the virus fails to establish a bridgehead and is repulsed.
Scenario 3 – This is the case in this paper. The patient is infected as described in scenario 1 when antibodies are injected into the system, these will supplement those already being produced by the plasma cells and will bind to the virus and focus the immune response. The serum will not include live plasma cells this does not immediately increase the rate of production it a one off shot but should by some time.
Scenario 4 – The patient is given serum before being exposed to the virus. What happens here? In theory I would expect these antibodies to bind to the viron and reduce the chance of it binding successfully and subject it to a heightened attack – hopefully stopping or slowing infection.

This poses the question what is the half-life of an antibody? If the serum includes no active plasma cells then the antibodies will not be replaced as they are broken down or excreted. In a patient fighting an infection there may be many disease specific plasma cells, and antibodies, but as they recover most are destroyed and only a small number remain to prime the system against a future infection. Presumably these plasma cells do not live that long so there needs to be a feed back mechanism to maintain a suitable antibody titre. How is this achieved? If antibodies are injected will this stimulate plasma cells to make more (as the antigen would) or will they just tend to zero. If we produce small quantities of vaccine can the vaccinated be used as serum donors? If serum antibodies produce plasma cells can those injected then produce more serum? (As stated in my first post, if my rudimentary immunology is wide of the mark please edit or post corrections)

The research implied that one convalescent patient could provide serum for several sick ones. Would it go further if used as in scenario 4? How much further? We do not want to wait for a devastating first wave to produce lots of donors. In the case of diseases where serum is used (either in human or veterinary medicine) does anyone know if & how volumes can be increased? We only have about 100 recovered H5N1 patients and there are 6.6 billion of us so some kind of amplification is going to be needed.

What was encouraging in the quote from the paper above was the absence of severe side effects despite what I assume to have been pretty crude separation techniques. Mingus’ washing machine based centrifuging system not withstanding, I suspect we should be able to produce a more sophisticated fractionation technique and hence less adverse reactions. Assuming we had a blood supply what do we need to produce acceptable medical grade serum? Is there enough hardware (including in the third world)? What is the storage life of antibody serum?

All thoughts gratefully received.

[wetDirt]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

I have no doubt that this method will be used by the military to keep functional. I think all you need is a good refrigerator to transport the stuff. I think a person's antibody response lasts a considerable time, because the notes about immunity are written down in stem cells in the thymus or bone marrow, which produce the copies circulating in the blood. So temporarily depleting the blood doesn't remove the record of immunity. It grows back. In a month. This is going to take a lot of donors. Maybe it's time to consider clean living a prep.

[JJackson]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

In the post that opened this thread I asked a lot of questions aimed at exploring whether fractionated blood products from convalescent H5N1 patients could be used to enhance the immune response of either of H5N1 patients or prophylactically.

Mingus has been busy reading and posting papers (in the Scientific Library) around the subject and I would like to draw your attention to one of these in particular which provides an extremely promising line of research and therapy using Monoclonal Antibodies (Mabs).Antibody-Based Therapies for Emerging Infectious Diseases http://www.flutrackers.com/forum/showthread.php?t=9864



To recap in 1918 serum was used on H1N1 patients and found to have a measurable beneficial effect however using blood products like this has several major drawbacks. Firstly quantity - With so few H5N1 survivors getting anything like enough serum is unlikely to be possible until a pandemic starts at which point we would be racing against the clock as the antibodies we are after are not produced until late in infection and pandemic waves are scarily fast. Secondly adverse reactions – One of the most attractive aspects of serum therapy is its low-tech nature, basic centrifuging as a means to extract the fraction with the antibodies but in this crude form there are a large numbers of other blood products which may cause adverse reactions. The more sophisticated the preparations the less side effects but the higher the cost and less people helped. Thirdly disease transfer – For a system like this to be practical on a large scale you would need to move to industrial scale production which means mixing blood products from many donors which in turn means careful screening for HIV, vCJD etc. which is already done by Blood transfusion services but is again moving us away from a cheap 3rd world solution. Where does all this leave us? There may still be a role for crude single donor to single patient treatment if a cheap disposable filter was available as envisaged by Mingus

today, syringe filter exist that can collect only serum, it can be done in a ambulance almost costless

here you are trusting the donor does not have any unpleasant blood born disease and may be useful between family members.

How can we scale up?

The short answer is it does not seem to be possible by this method but a Mab cocktail is a distinct possibility.
Each Mab is specific to its matched antigen which makes it extremely effective as long as you have correctly predicted the antigenic site. With flu this is a problem due to the reasonably rapidly changing nature of the antigenic sites on its surface proteins hence the need to produce a cocktail including more than one Mab. H5N1 has seven antigenic sites 5 on the HA protein and the other 2 on NA. If two or three sites can be picked which are common to the strains thought most likely to be pandemic candidates these would form the basis of the cocktail.

Advantages of Mabs.

Production - MAbs are produced by tissue culture techniques. Industrial production of MAbs may utilize immunoglobulin synthesis in yeast, bacteria, or plants. i.e. They are scaleable. (Production on our scale would use Air Lift Fermentation)

Toxicity - Toxicity of serum was due to allergic reactions to animals protein. Human immunoglobulin preparations are well tolerated. Antiidiotypic responses remain a problem for humanized MAb therapy.

Relatively quick to get through regulatory procedures.

Administration is easy and due to large scale batch production product should be consistent.

Disadvantages.

Cost. This seems to be at least in part due to the small scales of production for the diseases which it is currently being used to treat.
Quote from http://www.flutrackers.com/forum/showthread.php?t=9869

The high expense of Ig preparations is related to the fact that these reagents are more fragile than small molecular weight compounds and that they originate from immune donors or cell culture production and hence are costly to obtain, produce, and maintain. In addition, many of the indications for which immunoglobulins are used represent relatively small markets, and the cost efficiency associated with mass production may not apply.
One difficulty that has plagued the development of antibody-based therapies in infectious diseases is that the market size for an antibody reagent is proportional to the prevalence of disease.[3] Since antibody reagents are almost always pathogen specific, the market for antibody-based therapies is often much smaller than that for drugs with broad antimicrobial activity. Small market size combined with high price and the availability of many antimicrobial drugs has not encouraged development of antibody-based therapies for many infectious diseases. However, in considering antibodies for biological defense, the market size equals the potentially vulnerable population. This consideration, combined with the fact that stockpiles would have to be replenished periodically as a result of lot expirations, could make the economic outlook more attractive to industry. Production of sufficient antibody protein for universal protection of the U.S. population against a specific biological agent would involve large-scale production and could result in cheaper unit prices.

For the future there are several other interesting possibilities. If you believe that a large part of the problem with H5N1 and the 1918 pandemic was not so much due to the flu itself as to the immune system’s overreaction then Mabs could be engineered to bind to and inactivate specific cytokines, this potentially allows for a variable dose to dampen the overreaction without interfering with the bodies other immune responses to the disease.

The versatility of antibody-based therapies is illustrated by the ability of digoxin-binding antibodies to reverse digoxin toxicity (36) and recent attempts to treat septic shock by employing MAbs that bind cytokines (37).

The next quote also contains a host of interesting possibilities.

Microbial targets for therapeutic antibody development are not necessarily limited to extracellular pathogens. Although intracellular pathogens are commonly believed to be outside the reach of antibody immunity, several reports have suggested that some MAbs are active against some intracellular microorganisms. Some IgA MAbs can neutralize intracellular viruses (31), and an MAb to Toxoplasma gondii has been reported to interfere with intracellular replication of the parasite (32). It has been proposed that intracellular virus neutralization by IgA occurs by antibodies binding to viral proteins and interfering with viral assembly (31). Additional evidence for intracellular antibody activity comes from the observation that IgG anti-DNA autoantibodies can enter the cytoplasm and nucleus of living cells (33).

The M2 protien would not normally be regarded as having antigenic sites as it is not a surface protein that is likely to come into contact with B cells and so provoke plasma cell generated antibodies but these can be produced synthetically and be introduced into to the cytoplasm they may well act as an ion channel blocker without the same dangers of resistance emergence. This is one potential example there are obviously many other points of attack.

This may also have use in drug resistant diseases like MRSA.

[Mingus]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

OoF:o, such a well writen reflexion text that quote my syringe filter thing just filled me with doubt, that was something someone told me it exist but I did not see it myself...

But I found more info about that...

This is a regular centrifuge tube with
a gel barrier to efficiently separrate serum from blood.

The gel barrier is not recquire to obtain serum but it ensure a better serum clearness.



Here is some filter device to allow serum collection.

• Designed to work with the internal taper in blood collection tubes
• Leak-resistant, one-way valve allows specimens to be stored up to seven days
• One-way valve prevents backflow into primary collection tube

Here is a direct serum collector...

I do not provide the link because it is a lab stuff sale site.

I would not advice anybody to do that desperate action but like JJackson wrote...

There may still be a role for crude single donor to single patient treatment if a cheap disposable filter was available

In such a case, individual trust between donor, patient and medical staff would be crucial.

[Mingus]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

I get the Ok from JJacson to copy this.
He also had a interesting reflexion line to follow...
________________________________________
Quote:
<TABLE cellSpacing=0 cellPadding=6 width="100%" border=0><TBODY><TR><TD class=alt2 style="BORDER-RIGHT: 1px inset; BORDER-TOP: 1px inset; BORDER-LEFT: 1px inset; BORDER-BOTTOM: 1px inset">Yes, in the absence of any other timely treatments, this line of attack might generate more interest.

Have you any knowledge of Air Lift Fermentation or Mab production techniques? The literature keeps saying it is expensive but I am not clear why, most of what I have read has been very small scale (and I can see why that would be costly) but once you have created a bacteria or yeast that produces the Mab you want it seems relatively simple to then produce large quantities. Far more so than setting up large scale vax plants and the inferstructure can be reused to produce any other type of Mab as need arises.

-JJackson
</TD></TR></TBODY></TABLE>

You know, most of the price in biothechnology world are the fact of small production and of R&D and fine tuning cost.

Just as an example there is much more technology in your laserman or in your DVD player than in a PCR thermocycler but the first two one can be found under 100$ while the later cans range from 20000$ to 50000%.

As I can tell, because I work some years ago on a GMO detection research team ( for some months ) the process of transfecting a lab strain bacteria with a choosen protein gene, select the transformed bacteria, culture it like a yogourt and purify it is very simple.
Because they are techniques and knowledge from a very little scientific elite, everything is expensive.

But if we look it without artifice, once the protocol is fixed there is no reason why recombinant protean or Mab production should be more difficult to produce than wine.

The problem is to optimise protocol.
There is also the price of the various kits.
In research, every little tricks or methods have their kits sold by various biothech company at high cost protected by many patents. But it make the research process quicker and less heavy to accomplish.
As they are made for research purpose, there is no "mass scale" kits.

So from this point the problem is to ingeneer the production plant scale "kit".

I think there is more thehnology in many electronic gadget than in most of the Biothech procedures.
Get this out of patent and in massive production, there is few reason why everything biothechly produce should be sold 250$/50ul.

Ming

[JJackson]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

The following post is another part of the conversation with Mingus referred to in his post above.

Your last post put in writing what I already suspected.

Stage 1
Preparation of the organism. This is very much what an academic might do during research in the area, lots of lab time by scientists with specialized knowledge and some expensive kit you would find in a good university or bio-tech co. - Expensive but a one off cost.

Stage 2
Air Lift Fermentation.
Glorified brewery probably sold at exorbitant markup by a bio-tech co. - If this was given to the Russians or Chinese to make I suspect we would get exactly that a slightly modified winery, acceptable but not so high tech. - Cheap per unit production running cost.

Stage 3
Purification, packaging & storage.
Industrial again but needing some rather more specialized equipment. Final product to be paced in single shot disposable syringes (preferably by self administration). - Reasonably cheap

Stage 4
Distribution
Initially to be distributed to the health services of the various governments and then on Pandemic Stage 6 mass immunisation by virtually every HCW in the world capable of giving an injection (or self administration) then the doctors would have to be ready to deal with the adverse reactions.


How to achieve all this.
Stage 1 should be by a new model, commissioned and paid for from the $1.9 billion pledged to the WHO with any resultant patents held by them in thrust for humanity as a whole.
Stage 2 & 3 by open tender and paid for by the governments of the world in the proportion of their UN dues.
Stage 4 responsibility of the individual governments with NGOs, Gates Foundation, public appeals (a la Tsunami) etc. asked to help in the poorest countries.

The truly sad thing is that I would not be surprised to see something very close to this implemented but only after a sever pandemic had been and gone. Until our governments are forced to see the dangers in the status quo there could never be the collective political will to implement something like this. The concepts of patents being held outside of the Corporation and medicine production as a high volume / low margin industry would send the executives in big pharma into shock. The political fallout would not allow the US administration to support it and that pressure would kill it in the UN.

One can but dream.

[gsgs]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

I'm not sure whether I understood all this correctly and I didn't read very carefully - the thread is long - so I'd like to see this summarized in one
question :


can I add some kit to my preps which would enable me
as a layman to transfer a portion of my blood (how much ?)
into the body of a relative or friend, while it is determined
now whether this would be safe enough and the blood is OK
and compatible ?

What would it cost ? Should I transfer the blood prophylactically or only when that relative/friend also becomes ill ? I assume this
almost redoubles his chances to survive.

[JJackson]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

I'm not sure whether I understood all this correctly and I didn't read very carefully - the thread is long - so I'd like to see this summarized in one
question :


can I add some kit to my preps which would enable me
as a layman to transfer a portion of my blood (how much ?)
into the body of a relative or friend, while it is determined
now whether this would be safe enough and the blood is OK
and compatible ?

What would it cost ? Should I transfer the blood prophylactically or only when that relative/friend also becomes ill ? I assume this
almost redoubles his chances to survive.

OK firstly I am not a doctor and am not therefore in a position to give the kind of advice you are asking for. What we have been doing in this thread is exploring the ramifications of research in a number of papers in the hope that someone will jump in and point out why we are wide of the mark.
What we need is expertise on the Immune system, lab & large scale monoclonal antibody production, serum fractionation and an MD.

To try and answer your questions.

The 1918 research showed that serum from recovered patients markedly improved the prognosis of current patients. This is assumed to have been due to the H1N1 antibodies in the serum. What I am less clear on is if this was due to their direct binding to the antigenic sites on the H1N1 (so interfering with binding & endocytosis) or if it also produces an on going immune response.

“can I add some kit to my preps which would enable me as a layman to transfer a portion of my blood (how much ?)”

The thing to add to your kit would be the syringe with integral filter to separate serum from the whole blood (you would have to trust the donor to be free of HIV, Hepatitis etc.), how much is not clear, the article is on the effects from several different trials and does not really address this issue beyond saying “U.S. Food and Drug Administration regulations (62) allow individuals to donate 1000 to 1200 mL of plasma per week. A single H5N1 convalescent donor could provide a weekly volume of plasma sufficient to treat multiple patients with H5N1 influenza”
What should be noted here is they were treating advanced stage patients who had already developed pneumonia. If the injected antibodies are binding to the antigenic sites and inactivating virons then the treatment should be more effective if given just before exposure or as soon as possible after infection. The reasoning being that any given number of antibodies (n) can bind to and disrupt an equivalent number of viral antigenic sites. In theory even a small amount could help reduce the chance of infection if it was in the right place at the right time – or at least decrease net viral load.

would be safe enough and the blood is OK and compatible ?

As I understand it compatibility – or lack there of – is due to blood type (A, B, O etc) and is due to the red blood cells, which are removed by the filter, the remaining serum is not type specific. The serum still contains a wide range of products which could cause adverse reactions but the questions is if you think you have been exposed are you willing to risk these side effects to reduce the risk – or severity - of H5N1 infection. More sophisticated filtration/fractionation techniques will lead to the removal of more of these other serum products and so reduce the risk of adverse reaction (however these are beyond the scope of DIY treatment).

What would it cost ?

Not much but I am not sure where you could buy these items retail. The illustrations Mingus found are from a lab/medical equipment supply catalogue and will not be expensive for a hospital to buy but are not designed for lay use. Needless to say under normal circumstance the kind of thing we are talking about is not exactly encouraged by the medical profession.

The last part of your question is already covered in other parts of this post.

I hope this helps but as I have argued in other posts in this thread this is really a fix in the event that TPTB fail to provide a viable alternative. Mass production of monoclonal antibodies seems a far safer and more accurately targeted solution but is defiantly not a DIY option.

JJ

[JJackson]

Re: Why blood of bird flu survivors is a lifesaver [Discussion]

This is a link to an interesting paper on the merits of Mabs posted in the scientific literature section. It has two main strengths; it shows both the power of Mabs as a tool in elucidating the relationship between the epitope binding and the 3D structure of the HA trimer - and the resultant binding affinity changes of specific point mutations- and the possibility of quickly producing a tailored polyMab cocktail from 'stock' Mabs for any flu - pandemic or seasonal.

http://www.flutrackers.com/forum/showthread.php?t=23457