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International Meeting On Controversial Bird Flu Research Draws Near

The World Health Organization has just one week left to prepare for a highly anticipated meeting on controversial bird flu research. One official says that 22 invitations have gone out and the WHO is still waiting to hear back from some of the invitees.

Recent experiments involving the H5N1 bird flu virus have caused a furor in the science community, and the WHO was urged to convene an international discussion.

The scientists, journal editors and others who attend are expected to review the facts and the most pressing issues related to this specific work, rather than have a broader discussion about the possibility of international oversight of potentially worrisome biological research.

Critics of the experiments say scientists took a potentially deadly bird flu virus and tweaked it in ways that could make it contagious between people. They worry that the altered virus might escape the lab and cause a global pandemic, and that openly publishing details of the work in a scientific journal could provide terrorists with a recipe for a bioweapon.

Other scientists say the possible risks have been exaggerated and that the research is important for public health.

On Jan. 20, top influenza researchers announced that they were putting a voluntary 60-day moratorium on doing any further experiments with these viruses or creating any new ones. Publication of manuscripts describing the work is on hold as well.

Twenty-two people have been invited to an initial meeting at WHO headquarters in Geneva, which will be held Feb. 16 and 17, says Keiji Fukuda, assistant director-general for Health Security and Environment at the WHO. But on Wednesday, he said, they were still not fully sure of all of the people who will be coming.

The public isn't invited. "We won't be able to have it open to the public because of the nature of the information to be gone over," says Fukuda, noting that attendees will be discussing unpublished details of the experiments.

He described it as a "fact-finding, context-setting" meeting aimed at identifying the most pressing issues related to this research. "What we've tried to do is invite people who are directly involved with either conducting the research or who have, for example, a role in publishing the research," Fukuda says.

Bruce Alberts, editor-in-chief of the journal Science, which wants to publish one of the bird flu manuscripts in some form, says the deputy editor from the journal will attend the event. He said on Wednesday morning he had received an email with a list of some attendees. "They're from all around the world," Alberts said.

Other attendees will be people who have formally reviewed the research, such as Paul Keim, acting chair of the National Science Advisory Board for Biosecurity. Late last year, in an unprecedented move, the NSABB recommended that key details of the work should not be publicly revealed when researchers publish on their findings.

On the day of the meeting, Fukuda says, the WHO will post the names of the attendees. And soon after the meeting, the organization will post a report on what was discussed and any consensus that was reached.

The most urgent, practical issues that could be discussed at the WHO meeting include things like how the research could be published without revealing sensitive information, while still allowing the full findings to be available to public health researchers around the world. Discussions may also cover what additional research should go forward on the lab-created viruses — and under what conditions.

Some experts say the viruses should be moved to a lab with the highest possible security and that any future experiments should be tightly controlled. Others argue that's unnecessary and that allowing work to go forward will reveal information about flu viruses and how they evolve that's important for protecting the public's health.

http://www.npr.org/blogs/health/2012/02 ... draws-near

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To publish? What to publish? Those will be the questions at WHO bird flu meeting
By: Helen Branswell, The Canadian Press
Posted: 02/9/2012 1:38 PM

Vials of H5N1 flu vaccine by Beijing-based drug maker Sinovac Biotech Ltd. are seen during production at Sinovac facilities in Beijing, Nov.24, 2011. To publish and what to publish? Those will be the questions on the table when the World Health Organization convenes a special meeting next week about controversial bird flu studies. THE CANADIAN PRESS/AP-Andy Wong

To publish and what to publish? Those will be the questions on the table when the World Health Organization convenes a special meeting next week about controversial bird flu studies.

The meeting will be small, with only 22 invitees from outside the WHO, senior communications staff of the Geneva-based global health agency said.

Potentially among them will be representatives of the Vietnamese and Indonesian laboratories that provided the original viruses on which the research was done.

People looking for solutions to broader issues such as whether and where similar research should be conducted in future may need to dial down their expectations for the meeting.

"In this first meeting, we've got to sort out the deadlock, I think, that we're in with the scientific community. I don't think anyone anticipates that to happen very quickly," says Christy Feig, the WHO's director of communications.

The session, to be held Feb. 16 and 17 at the WHO's headquarters, will focus on the most basic problems that have arisen as a result of the controversy. In essence: Should the two papers at the heart of the dispute be published? In full or in abbreviated form? If they must be abbreviated, which sections must be held back?

That disclosure suggests the tentative agreement on the part of the research teams and the journals involved to publish only redacted versions is not yet set in stone.

In fact, the belief in Geneva is that the distance between the meeting participants is wide. As a result, expectations for how much can be achieved next week are the opposite.

"The focus is very narrow," Feig said from Geneva. "There's only so much ground you can cover in two days."

The controversy relates to two studies, done by Dutch and American researchers, which reportedly show how H5N1 flu viruses could be made to spread like seasonal flu viruses among ferrets. Such viruses could not be tested in humans and ferrets are considered the best — though not foolproof — animal model for predicting how flu viruses will act in people.

A U.S. panel of biosecurity experts is urging that the studies not be published in full, arguing the details of how the work was done would be dangerous in the wrong hands.

Invitations to the meeting are directed to individuals in some cases. In others, they have been issued to an institution that is playing a role in the controversy.

Representatives of the Vietnamese and Indonesian labs that provided the source viruses have not yet indicated whether they'll attend, Feig said.

Vietnam provided viruses to Yoshihiro Kawaoka of the University of Wisconsin-Madison while Ron Fouchier, of Erasmus Medical Center in Rotterdam, the Netherlands, received his viruses from Indonesia.

Their inclusion is a nod on the WHO's part to the Pandemic Influenza Preparedness Framework, a multi-country agreement forged after years of painful negotiations. The framework, ratified last spring, is meant to ensure countries that share viruses with foreign scientists in turn will share in the benefits that flow from that research.

The talks that led to the framework were triggered by a 2007 move by Indonesia to stop sharing H5N1 viruses with the broader world unless it was guaranteed access to pandemic vaccine and other benefits.

While the transfer of viruses from Vietnam and Indonesia to the American and Dutch labs occurred well before the PIP agreement was put in place, the WHO wants to observe the spirit of that agreement going forward, said spokesperson Gregory Hartl.

The director of the U.S. scientific agency that funded the research — Dr. Anthony Fauci of the National Institute of Allergy and Infectious Diseases — will be there. So too will be a representative of the National Science Advisory Board on Biosecurity, the expert panel charged by the U.S. government with reviewing potential dual-use research, legitimate science that could be put to illegitimate use.

Others attending the meeting will be the researchers themselves, people who reviewed the research proposals to see if they triggered dual-use concerns (both when the work was in the proposal stage and when the studies were being readied for publication) and representatives of the journals Science and Nature, which plan to publish the studies.

At least several of the directors of laboratories in the network of WHO collaborating labs for influenza have been invited and are expected to attend.

A list of the participants and their declarations of interest will be posted on the WHO website the day the meeting begins. Hartl said the list can only be posted after the meeting participants formally go through and accept one another's declarations of interests.

One of the conditions the researchers and journals made before tentatively agreeing to redact the studies was that the full details of how the work was done would be shared with other scientists and public health officials on a need-to-know basis.

The system for doing that — vetting applications to see the work, figuring out how to share the material — has not yet been devised. And that work probably won't happen at this meeting, said Feig, adding that people with the expertise needed to set up that type of system won't be in the room.

The WHO has repeatedly said that it envisages helping to organize a second meeting with broader goals and wider international involvement after this initial gathering is completed. Hartl said that is still the plan, but planning won't begin until this meeting occurs.

http://www.winnipegfreepress.com/world/ ... 34654.html

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The above comments confirm suspicions on the H5N1 targets used in the transmission studies. Previously Fouchier has mentioned his collaboration with Indonesia and Kawaoka has published extensively on clade 1 receptor binding domain changes.
Human clade 1 and clade 2.1 sequences have been been known since 2004/2005 respectively and both countries had many examples of clusters, signaling human to human transmission.

WHO almost delared an H5N1 pandemic in 2005 at a special meeting in Jakarta, and the meeting noted receptor binding domain changes such as S227N, which is still circulating in Cambodia, along with PB2 E627K. Both were in the most recent public sequence from a fatal case in 2011.

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HA S227N and PB2 E627K In Fatal H5N1 Cambodia Case
Recombinomics Commentary 11:00
January 23, 2012

A man in southwest China who contracted the bird flu virus died on Sunday, health authorities said, the second human death from the virulent disease in the country in just under a month.

The news comes after neighbouring Vietnam, Cambodia and Indonesia also reported deaths from avian influenza
The above comments note the recent increase in fatal H5N1 cases and clusters in southeastern Asia. Although sequences have not yet been released for any of the above cases, recently released sequences from 2011 Cambodia cases include A/Cambodia/V0606311/2011, which is the most recent 2011 sequence from Cambodia, where all 8 reported cases for 2011 were fatal. Clade 1 continues to circulate in southeast Asia, including the above sequence, which has HA S227N and PB2 E627K. Both are these changes are likely to be among the 5 changes in 2 genes linked to aerosol transmission of H5N1 in a ferret model, which has been censored by Nature and Sciences per recommendations by the US NSABB, even though 4 of the 5 changes have also been found in a single published sequence, raising serious concerns about the understanding of influenza evolution by the NSABB board or the understanding of the mission of Nature and Science by editorial boards of the journals.

Both S227N and PB2 E627K are well known changes associated with increased transmission of H5N1......

http://www.recombinomics.com/News/01231 ... bodia.html

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Positive selection at the receptor-binding site of haemagglutinin H5 in viral sequences derived from human tissues
Alita Kongchanagul,1,2 Ornpreya Suptawiwat,1 Pumaree Kanrai,1
Mongkol Uiprasertkul,3 Pilaipan Puthavathana1 and Prasert Auewarakul1
Correspondence
Prasert Auewarakul
sipaw@mahidol.ac.th
1Department of Microbiology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok
10700, Thailand
2Department of Immunology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok
10700, Thailand
3Department of Pathology, Faculty of Medicine Siriraj Hospital, Mahidol University, Bangkok 10700,
Thailand

RNA sample No. synonymous
substitutions
No. non-synonymous
substitutions
List of non-synonymous substitutions (frequency)
Patient A
nasopharyngeal
aspirate
17 138 S128R (1), L133V (54), S136G (1), A138V (53), A138E (1), Q142R (1), K157R
(3), Y161H (7), N172D (1), N186D (3), E190D (1), L194H (1), N224S (3),
N224D (2), S227N (2), S227G (1), N240S (1), N250S (2)
Patient B lung 98 94 S128G (1), A131V (1), A131T (3), Y141C (1), K144R (1), V152A (1), W153R
(1), I155V (1), N158D (1), Y161H (1), T163A (11), I164V (1), N169S (1),
V178A (2), D187G (1), D187N (2), E190G (1), K193R (18), K193S (3), K193E
(1), L194P (13), N197D (1), P198L (1), T200A (1), I202V (1), N210S (1),
I217V (1), K222E (2), G228E (2), M230T (10), F233S (1), F223L (1), N244D
(1), F245S (1), I252V (3), E255G (1)
Patient B intestine 35 62 E134V (2), S136G (2), R149K (2), S167G (1), S167C (1), Y168* (1), N172S (1),
H183R (1), H184R (1), N186D (2), R193K (11), L194P (2), Y195* (1), Q196R
(2), P198S (1), T199A (1), V204A (1), T206A (2), K222E (7), N224H (1),
G225R (1), S227N (2), M230T (1), F233S (1), P239S (1), N244S (1), I252V
(8), I268V (2), K270E (2)
Patient C lung 20 145 S128N (1), H129Q (1), L133V (6), S136G (4), A138V (55), S146P (1), W153*
(1), I155T (1), N158D (1), Y161H (3), Y161R (1), Y168C (2), N169H (1),
I182V (1), N186D (1), T199I (2), T206A (1), T208A (1), T219I (1), E222K
(56), R229G (1), S247N (2), F251S (2)
Patient C intestine 31 189 S127A (1), H129Q (3), A131D (1), L133V (19), L133F (1), S136G (1), A138V
(20), S146P (1), W153* (1), N158S (3), Y161C (1), Y161R (1), L177S (1),
W180R (2), I182T (1), N186D (37), K193R (1), Y201* (1), N210D (1), E222K
(51), S227N (36), I252V (4), K262R (1)

http://vir.sgmjournals.org/content/89/8/1805.full.pdf

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E627K Increases H5N1 Replication at Lower Temperatures
Recombinomics Commentary
October 5, 2007

Birds usually have a body temperature of 106 degrees F, and humans are 98.6 degrees F usually. The human nose and throat, where flu viruses usually enter, is usually around 91.4 degrees F.

"So usually the bird flu doesn't grow well in the nose or throat of humans," Kawaoka said. This particular mutation allows H5N1 to live well in the cooler temperatures of the human upper respiratory tract.

H5N1 caused its first mass die-off among wild waterfowl in 2005 at Qinghai Lake in central China, where hundreds of thousands of migratory birds congregate.

That strain of the virus was carried across Asia to Africa and Europe by migrating birds. Its descendants carry the mutation, Kawaoka said.

"So the viruses circulating in Europe and Africa, they all have this mutation. So they are the ones that are closer to human-like flu," Kawaoka said.

Luckily, they do not carry other mutations, he said.
The above comments on the paper describing PB2 E627K replication in experimental mice and cells highlight the effect of the change on the ability of the Qinghai strain to replicate at lower temperatures.....
http://www.recombinomics.com/News/10050 ... _Temp.html

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Nature 444, 378-382 (16 November 2006) | doi:10.1038/nature05264; Received 20 August 2006; Accepted 21 September 2006


Haemagglutinin mutations responsible for the binding of H5N1 influenza A viruses to human-type receptors

Shinya Yamada1,3, Yasuo Suzuki3,4, Takashi Suzuki3,5, Mai Q. Le6, Chairul A. Nidom7, Yuko Sakai-Tagawa1,3, Yukiko Muramoto1,3, Mutsumi Ito1,3, Maki Kiso1,3, Taisuke Horimoto1,3, Kyoko Shinya8, Toshihiko Sawada9, Makoto Kiso9, Taiichi Usui10, Takeomi Murata10, Yipu Lin11, Alan Hay11, Lesley F. Haire11, David J. Stevens11, Rupert J. Russell11,13, Steven J. Gamblin11, John J. Skehel11 & Yoshihiro Kawaoka1,2,3,12

1. Division of Virology, Department of Microbiology and Immunology, and,
2. International Research Centre for Infectious Diseases, Institute of Medical Science, University of Tokyo, Tokyo 108-8639, Japan
3. Core Research for Evolutional Science and Technology, Japan Science and Technology Agency, Kawaguchi, Saitama 332-0012, Japan
4. College of Life and Health Sciences, Chubu University, Kasugai, Aichi 487-8501, Japan
5. Department of Biochemistry, University of Shizuoka, School of Pharmaceutical Sciences and COE Program in the 21st Century, Yada, Shizuoka 422-8526, Japan
6. National Institute of Hygiene and Epidemiology, Hanoi, Vietnam
7. Avian Influenza Laboratory, Tropical Disease Centre, Airlangga University, Surabaya, Indonesia
8. The Avian Zoonosis Research Centre, Tottori University, Tottori 680-8553, Japan
9. Department of Applied Bioorganic Chemistry, The United Graduate School of Agricultural Science, Gifu University, Yanagido, Gifu 501-1193, Japan
10. Department of Applied Biological Chemistry, Shizuoka University, Shizuoka 422-8529, Japan
11. MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK
12. Department of Pathobiological Sciences, University of Wisconsin-Madison, Madison, Wisconsin 53706, USA
13. Present address: Centre for Biomolecular Sciences, University of St Andrews, St Andrews KY16 9ST, UK.

Correspondence to: Yoshihiro Kawaoka1,2,3,12 Correspondence and requests for materials should be addressed to Y.K. (Email: kawaoka@ims.u-tokyo.ac.jp). Coordinates for the H5 structure have been deposited in the Protein Data Bank under accession code 2IBX.


Top of page
H5N1 influenza A viruses have spread to numerous countries in Asia, Europe and Africa, infecting not only large numbers of poultry, but also an increasing number of humans, often with lethal effects1, 2. Human and avian influenza A viruses differ in their recognition of host cell receptors: the former preferentially recognize receptors with saccharides terminating in sialic acid-2,6-galactose (SA2,6Gal), whereas the latter prefer those ending in SA2,3Gal (refs 3–6). A conversion from SA2,3Gal to SA2,6Gal recognition is thought to be one of the changes that must occur before avian influenza viruses can replicate efficiently in humans and acquire the potential to cause a pandemic. By identifying mutations in the receptor-binding haemagglutinin (HA) molecule that would enable avian H5N1 viruses to recognize human-type host cell receptors, it may be possible to predict (and thus to increase preparedness for) the emergence of pandemic viruses. Here we show that some H5N1 viruses isolated from humans can bind to both human and avian receptors, in contrast to those isolated from chickens and ducks, which recognize the avian receptors exclusively. Mutations at positions 182 and 192 independently convert the HAs of H5N1 viruses known to recognize the avian receptor to ones that recognize the human receptor. Analysis of the crystal structure of the HA from an H5N1 virus used in our genetic experiments shows that the locations of these amino acids in the HA molecule are compatible with an effect on receptor binding. The amino acid changes that we identify might serve as molecular markers for assessing the pandemic potential of H5N1 field isolates.

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Introduction of the Gln192Arg mutation, but not the Ser223Asn mutation, into the HA of VN1194 appreciably enhanced the capacity of the HA to recognize SA2,6Gal, and introduction of both mutations increased the binding capacity further. This finding implicates Gln192Arg as a possible determinant of the shift to recognition of the human receptor by VN/3028IIcl3 (Fig. 2a). The Thai/KAN virus also showed two amino acid changes in HA1 as compared with VN1194: Gly139Arg and Asn182Lys. Introduction of either mutation into the VN1194 HA enhanced SA2,6Gal binding (Fig. 2b), and an additional increase in binding capacity was observed when both mutations were substituted simultaneously. Thus, both Gly139Arg and Asn182Lys seem to contribute to recognition of the human-type receptor.

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H3 numbering is N186K and Q196R

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N186K In H5N1 in Azerbaijan Patients

Recombinomics Commentary
December 2, 2006

Recently released human H5N1 sequences from Azerbaijan patients shed additional light on HA changes affecting the affinity of binding to 2,6 gal receptors, which are found in the human upper respiratory tract. Interest in such changes has been high, since an increased affinity is likely to lead to more efficient human-to-human transmission. The acquisition of one such change, S227N, was predicted because donor sequences were present in H9N2 in birds in the Middle East and Qinghai H5N1 migrated into the region in the fall of 2005.

S227N was present in the index case for the outbreak in Turkey as well as a second case in Turkey and a case in Egypt. A recent paper in Nature indicated S227N could synergize with another HA change, Q196R which was present patients from Iraq and or Azerbaijan. Moreover, another change that increased affinity for human receptor, N186K, was as present in these patients. However, these changes were not present in the public human sequences from these two countries and the description in the Nature paper was ambiguous. It was unclear if patients had one or both changes, and it was unclear which changes were in which patients. The only change in these positions in the public sequences was N186S in the index case for Iraq. Thus, only WHO consultants who could access the private database could confirm the statements in the Nature paper.

The release of five human H5N1 from Iraq clarified the distribution in Iraq. All six sequences had N186S and two had Q196R. The effect of N186S in receptor binding was not described in the Nature paper. However, this change is only in the human sequences from Iraq. It was not present in a goose or cat sequence from Iraq.

In addition to the five human sequences from Iraq, four additional human sequences from Azerbaijan were released. Those isolates are listed below. Two of the newly released sequences, A/Azerbaijan/001-161/2006 and A/Azerbaijan/011-162/2006 have N186K.......
http://www.recombinomics.com/News/12020 ... aijan.html

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