Is H2H spread of H3N2v occurring in the United States?

[alert]

The CDC has released an MMWR indicating infection in two children with a new reassortant H3N2 made up of 2009 H1N1 and trH3N2:

http://www.flutrackers.com/forum/showthread.php?t=172784

The two cases live in Indiana and Pennsylvania, hundreds of miles apart. The Pennsylvania child did have contact with pigs, and the Indiana child had contact with someone who had contact with pigs.

The question here seems to be: Is this virus spreading efficiently H2H as in April 2009, or is this virus widespread in pigs as H5N1 is in certain places in birds, and simply caused 2 separate animal-to-human transmission? I really can't tell.

Of note is that these two cases do have at least an indirect link to pigs, something we didn't have in April 2009. More importantly, I don't see any report of an increase in ARI or unexplained illness as we had in Mexico in 2009. I think I am still leaning slightly toward two separate incidents, but only slightly now.

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

January 25 this year US CDC had already suggested a possible H2H transmission - limited and unsustained - of swoH3N2: http://www.cdc.gov/flu/swineflu/soiv_cases.htm

Since then, if the virus would be readily transmissible - a widespread outbreak might be originated.

swoH3N2 contains HA and NA from human seasonal strains so - at least in part - but a residual immunity could protect a portion of population from disease.

All the cases of swoH3N2 have recovered.

[alert]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

I'm a little confused on the notation swoH3N2? Is that what we are calling the reassortment between trH3N2 and H1N1? Or is it just shorthand for "swine origin"?

I was pretty sure the trH3N2 from earlier in the year was not transmitting efficiently before this latest MMWR, but I thought this was a new virus as a result of the reassortment.

[Missouriwatcher]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

I recently watched a National Geographic special on hoarding. A woman from Flagler County, FL "collected" and slept with pigs. It wasn't until the U.S. Department of Agriculture stepped in and gassed the 389 pigs did the "animal sanctuary" of 15 years end. The hoarder's last words were something to the effect of "she will begin again". http://channel.nationalgeographic.co.../5331/Overview

Swine-Origin Influenza A (H3N2) Virus Infection in Two Children --- Indiana and Pennsylvania, July--August 2011
"The acquisition of the M gene in these two swine-origin influenza A (H3N2) viruses indicates that they are "reassortants" because they contain genes of the swine-origin influenza A (H3N2) virus circulating in North American pigs since 1998 (2) and the 2009 influenza A (H1N1) virus that might have been transmitted to pigs from humans during the 2009 H1N1 pandemic."

"...the girl was reported to have visited an agricultural fair where she had direct exposure to swine and other animals."

http://www.cdc.gov/mmwr/preview/mmwrhtml/mm60e0902a1.htm?s_cid=mm60e0902a1_w

How can we not expect to transfer viruses back and forth with swine if we have these kinds of people willing to sleep with them?

[sharon sanders]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

Both patients in the report released today are less than 5 years old. Patient A did not have any contact with pigs.

This indicates to me that there is human to human spread.

It appears at this point to be a "mild" infection. Patient A has a history of multiple chronic diseases and he recovered, as well as the other recorded cases.

I can only wonder what is happening elsewhere in the world where surveillance is non-existent or suppressed.

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

Correctly, Lynn Finelli told CIDRAP that swoH3N2 has HA-NA from seasonal human influenza virus circulating in the early '90, so people with more than 20 years of age have likely some level of immune protection.

This is not a secondary element because - first of all - a number of possibile asymptomatic carriers could be around and, secondly, a fully sustained outbreak may not be warranted.

It is a scenario that rensemble 1977 when the Russian flu (A (H1N1) seasonal strain) re-emerged but the outbreaks were limited almost exclusively in children and youngsters.

See the CIDRAP commentary: http://www.cidrap.umn.edu/cidrap/con...0211swine.html

[Vibrant62]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

Looking at the CDC report the following paragraph stands out.

Both of the children recovered, though one was briefly hospitalized, and there is no sign that the viruses spread from the children to others, but any evidence of ongoing transmission would require a rapid response, the CDC said.

The agency also said one of the children had no direct contact with pigs, which suggests he caught it from another person. Both children are under age 5.

The two viruses are similar to eight other swine-origin H3N2 viruses found in humans in the past 2 years, but they are unique in that they contain the matrix (M) gene from the 2009 H1N1 virus, the CDC reported in an early online posting in Morbidity and Mortality Weekly Report (MMWR). The two viruses are similar but not identical.

In other words, it would seem that they believe that we have two different reassortant H3N2s here - so we have two options; a) these were separate reassortant events producing two very similar hybrid viruses, or b) that they both arose from the same reassortant event but the virus is clearly evolving/ adapting to humans producing differences in the two viruses.

Under either option, any further appearance of this virus elsewhere would suggest that we have our next pandemic virus evolving in the wings, but that it is likely that it is still very early in the transmission chain and not yet fully adapted to humans. If the two infections arose from separate events, it indicates a high reassortment potential between pH1N1 and sw H3N2 which I would suggest indicates that it is only a matter of time before a fully fit and transmissible H3N2 arises somewhere in the world given the high level of viral infections of both types in pigs and their proximity to humans.

I don't take too much comfort in the fact that both infections were relatively 'mild' in these children, as this may very possibly be a result of poor human adaptation of the virus at this time.

I hope heightened surveillance is maintained, especially in Asia where there is the added risk factor of potential H5N1 infection in pigs.

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

But swoH3N2 IS NOT A NEW SUBTYPE for most of the human population. Since the donor H3N2 is a seasonal human strain, passed to pigs in the later '90, a large portion of population should have an immune response when challenged with it.

It remains to clarify whether internal genes (poymerase complex, non-structural and matrix segments) would confer to swoH3N2 better transmissibility and/or increased pathogenicity in humans after being mixed with other avian/swine/human strains.

The fact that main antigenic surface proteins are similar to human seasonal pre-2000 strains could partially reassure us, as said by Lyn Finelli in the above cited CIDRAP commentary.

The story could teach us something: H1N1 (2009) had the old spanish flu H1N1 virus as donor, the variant then passed to pigs and that remained almost unchanged for decades, with a parallel evolution with respect to the human H1N1.

Older people have demonstrated to be partially immune to this old ancestor of contemporary seasonal H1N1 and thus the pandemic was clearly much more benign than feared at early stages.

swoH3N2 viruses are also fully susceptible to oseltamivir and zanamivir and I think that a vaccine may be readily prepared even on the basis of the seeds in repositories, for example those employed in pre-2000 H3N2 trivalent vaccine component.

[alert]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

It seems clear to me that this novel combination is circulating in the US, at least in pigs, and has the potential for at least some H2H. That's not good.

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

I think it could be helpful for all of us to read this brief description by R. Webster and other of the internal structure of Influenza A viruses. Full free PDF is available for consultation at the URL indicated below.

Source: http://mmbr.asm.org/cgi/reprint/56/1/152

MICROBIOLOGICAL REVIEWS, Mar. 1992, p. 152-179 Vol. 56, No. 1
0146-0749/92/010152-28$02.00/0
Copyright ? 1992, American Society for Microbiology


Evolution and Ecology of Influenza A Viruses
ROBERT G. WEBSTER,* WILLIAM J. BEAN, OWEN T. GORMAN, THOMAS M. CHAMBERS,t AND YOSHIHIRO KAWAOKA
Department of Virology and Molecular Biology, St. Jude Children's Research Hospital,
332 North Lauderdale, P.O. Box 318, Memphis, Tennessee 38101

(...)


STRUCTURE AND FUNCTION OF THE INFLUENZA VIRUS VIRION

Current knowledge of the molecular biology of influenza
viruses has been recently reviewed in extensive detail by
Lamb (95), and individual references are not given. This
section presents a summary of that knowledge as background
information.


Components of the Virion

Influenza A viruses are members of the Orthomyxoviridae
family. They are differentiated from type B and C influenza
viruses on the basis of the identity of the major internal
protein antigens, the nucleoprotein (NP) and matrix (Ml)
proteins. On initial isolation, influenza A viruses are small
(80 to 120 nm in diameter), pleomorphic particles that later
become generally spherical. These particles consist of a
host-derived lipid bilayer envelope in which the virus-encoded
glycoproteins HA and neuraminidase (NA) and M2
are embedded; an inner shell of matrix protein; and, at the
center, the nucleocapsids of the viral genome (Fig. 1). The
genome of influenza A viruses consists of eight unique
segments of single-stranded RNA, which are of negative
polarity (i.e., complementary to the mRNA sense). The
RNA is loosely encapsidated by multiple NP molecules.

Complexes containing the three viral polymerase proteins
(PB1, PB2, and PA) are situated at the ends of the nucleocapsids.

To be infectious, a single virus particle must contain each
of the eight unique RNA segments. Available evidence
suggests that incorporation of RNAs into virions is at least
partly random. The random incorporation of RNA segments
allows the generation of progeny viruses containing novel
combinations of genes (i.e., genetic reassortment) when cells
are doubly infected with two different parent viruses.

The eight influenza A viral RNA segments encode 10
recognized gene products. These are PB1, PB2, and PA
polymerases, HA, NP, NA, Ml and M2 proteins, and NS1
and NS2 proteins.

PB2 polymerase. PB2 polymerase is encoded by RNA
segment 1, the slowest-migrating RNA species by gel electrophoresis.
It is a member of the protein complex providing
viral RNA-dependent RNA polymerase activity. It is known
to function during initiation of viral mRNA transcription as
the protein which recognizes and binds the 5' capl structures
of host cell mRNAs for use as viral mRNA transcription
primers. Endonucleolytic cleavage of these cap structures
from host mRNAs is also at least in part a function of PB2.

The role of PB2 in the other virus-directed RNA synthetic
processes, i.e., synthesis of full-length template cRNA and
new negative-sense viral RNA (vRNA), is not known since
these processes do not require host cap priming. Newly
synthesized PB2 proteins migrate to the nucleus of infected
cells.

PB1 polymerase. PB1 polymerase is encoded by RNA
segment 2; it functions in the RNA polymerase complex as
the protein responsible for elongation of the primed nascent
viral mRNA and also as elongation protein for template
RNA and vRNA synthesis. PB1 proteins localize in the
nucleus of infected cells.

PA polymerase. PA polymerase is encoded by RNA segment
3. It also localizes in the infected cell nucleus and is a
member of the RNA-dependent RNA polymerase complex
along with PB1 and PB2, but its role in viral RNA synthesis
is unknown. There is evidence for possible roles as a protein
kinase or as a helix-unwinding protein.

Hemagglutinin. The HA protein is an integral membrane
protein and the major surface antigen of the influenza virus
virion. It is responsible for binding of virions to host cell
receptors and for fusion between the virion envelope and the
host cell. HA is encoded by RNA segment 4. It undergoes
three kinds of posttranslational processing: proteolytic
cleavage, glycosylation, and fatty acid acylation. Newly
synthesized HA is cleaved to remove the amino-terminal
hydrophobic sequence of 14 to 18 amino acids, which are the
signal sequence for transport to the cell membrane. Carbohydrate
side chains are added, whose number and position
vary with the virus strain. Palmitic acid is added to cysteine
residues near the HA carboxy terminus. The final processing
step is cleavage of the HA into two subunits, HAl and HA2
(uncleaved HA is called HAO), connected by disulfide linkages.

This cleavage is accomplished by host-produced
trypsinlike proteases and is required for infectivity because
virus-cell fusion is mediated by the free amino terminus of
HA2. The fully processed HA thus consists of HAl of
(typically) about 324 amino acids plus variable carbohydrate,
and HA2 of (typically) about 222 amino acids plus variable
carbohydrate plus 3 palmitate residues.

HA molecules form homotrimers during maturation. The
three-dimensional structure of a complete HA trimer has
been determined. In essence, each HA molecule consists of
a globular head on a stalk. The head is made up exclusively
of HAl and contains the receptor-binding cavity as well as
most of the antigenic sites of the molecule. The stalk consists
of all of HA2 and part of HAl. The carboxy-terminal region
of HA2 contains the hydrophobic transmembrane sequence
and a terminal cytoplasmic anchor sequence where palmitate
is attached.

Owing to error-prone viral RNA polymerase activity,
influenza virus HA is subject to a very high rate of mutation,
estimated at about 2 x 10-3 base substitutions per position
per virus generation, or about one base substitution in the
HA gene per virus generation. Selection for amino acid
substitutions is driven at least in part by immune pressure, as
the HA is the major target of the host immune response.

Although the amino acids making up the receptor-binding
site, as well as cysteine and most proline residues, are highly
conserved, the remainder of the HA molecule is highly
mutable. In nature there are presently 14 recognized subtypes
of HA (called Hi, H2, etc.), which differ by at least
30% in the amino acid sequence of HAl and which are
serologically not cross-reactive (Table 1). Subtypes may
include several variant strains which are partially serologically
cross-reactive.

Nucleoprotein. NP is encoded by RNA segment 5. It is
transported into the infected cell nucleus, where it binds to
and encapsidates viral RNA. In addition to its structural
role, NP is believed to play a role in the switching of viral
RNA polymerase activity from mRNA synthesis to cRNA
and vRNA synthesis. NP is abundantly synthesized in
infected cells and is the second most abundant protein in the
influenza virus virion. It is phosphorylated; the pattern of
phosphorylation is host cell dependent and may be related to
viral host range restriction. NP is also a major target of the
host cytotoxic T-cell immune response.

Neuraminidase. NA, encoded by RNA segment 6, is also
an integral membrane glycoprotein and a second major
surface antigen of the virion. NA cleaves terminal sialic acid
from glycoproteins or glycolipids. Thus, it functions to free
virus particles from host cell receptors, to permit progeny
virions to escape from the cell in which they arose, and so
facilitate virus spread.

NA is glycosylated and possesses an amino-terminal hydrophobic
sequence which functions both as signal for
transport to the cell membrane and as transmembrane domain;
it is not cleaved away. The distribution of NA has not
been conclusively resolved; immunogold-labeling experiments
suggest that the NA tetramers are not evenly distributed
over the virion envelope, as is HA, but aggregate into
patches or caps. The complete three-dimensional structure
of an NA tetramer, bound to antibody, has been determined.

Like HA, NA is highly mutable with variant selection
partly in response to host immune pressure. Nine subtypes
of NA (called Ni, N2, etc.) have been identified in nature;
they are not serologically cross-reactive (Table 1). Different
variants of several subtypes are known.

Ml protein. Influenza virus RNA segment 7 is bicistronic,
encoding both Ml and M2 proteins. Colinear transcription of
segment 7 yields mRNA for the matrix protein. This is the
most abundant protein in the influenza virus virion. Matrix
protein forms a shell surrounding the virion nucleocapsids,
underneath the virion envelope. In the infected cell it is
present in both cytoplasm and nucleus. It has no known
enzymatic activity, although it has been speculated to play
an important role in initiating progeny virus assembly.

M2 protein. The mRNA for M2 is also transcribed from
RNA segment 7. It is derived from the colinear (Ml)
transcript by splicing. M2 is an integral membrane protein,
whose membrane-spanning domain also serves as a signal
for transport to the cell surface. It is present as a tetramer in
large amounts on the infected cell surface, and a small
amount is found in the virion. It is believed to act as a proton
channel to control the pH of the Golgi during HA synthesis
and to allow acidification of the interior of the virion during
virus uncoating.

Nonstructural NS1 and NS2 proteins. RNA segment 8
encodes the two nonstructural proteins NS1 and NS2. NS1
mRNA is colinear with the vRNA, whereas NS2 mRNA is
derived by splicing. These proteins, particularly NS1, are
abundant in the infected cell (NS1 primarily in the nucleus,
NS2 primarily in the cytoplasm) but are not incorporated
into progeny virions. Both proteins play roles in virus
replication, but those roles have not been fully defined. NS2
appears to modulate the synthesis of NS.

(...)

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[sharon sanders]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

This thread is a valuable commentary. Are there any objections to moving it for public viewing? I will wait 12 hours before I move the thread for objections and so everyone has a chance to edit any comments if they wish.

Thanks for the great posts!

[Vibrant62]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

Giuseppe - you have a very valid point; I think that the issue is that we dont know exactly which parts of the genome create the immune response/ virulence. I am also not clear how closely related swine H3N2 are to the human variants and how much genetic divergence there may be; much may depend on whether a viable H3N2/pH1N1 reassortant arises initially in pigs or in humans.

Scanning the data, there have already been numerous incidences of swine and human H3N2 / pH1N1 reassortment, with some evidence to back up the premise that humans may already have a reasonable level of immunity.

FT threads (examples)

1. Reported case in Canada (June) from human dual infection - again the M gene was reassorted (along with the NS) so (pure speculation) perhaps the M gene is the critical element to permit human infections?

New flu virus emerges after child co-infected with H1N1, H3N2; vaccine protective

http://www.flutrackers.com/forum/showthread.php?t=168986&highlight=h3n2+h1n1+reassortment

2. If you remember there were also reports of another H3N2 swine reassortment causing isolated human infections reported back in November of last year.

http://www.flutrackers.com/forum/showthread.php?t=154864&highlight=h3n2+h1n1+reassortment

However, there is also evidence of the potential for reassortment of H3N2 etc in ducks with H5N1 genes transferred; I have no idea how similar this H3N2 is to swine H3N2, although the paper cites similarity but does not indicate to what degree; it could be very different. Whether this type of reassortment is as likely in a mammalian host (pigs) as it is in ducks is unknown.

http://www.flutrackers.com/forum/showthread.php?t=162482&highlight=h3n2+h1n1+reassortment

And there is some evidence that a standard H3N2/ pH1N1 reassortment can give rise to a very mild virus that is attenuated in terms of pathogenicity; however, neraminidase transfer from H3N2 to pH1N1 produces a nastier H1N2 virus.

http://www.flutrackers.com/forum/showthread.php?t=160068&highlight=h3n2+h1n1+reassortment

All in all, the viral soup out there shortens the odds for emergence of a novel virus where there is not widespread human immunity; clearly reassortment is happening easily - and if there IS a high level of pre-existing immunity in the human population any novel variant will 'dead end' because of a shortage of susceptible hosts. i.e any emergent virus that is transmitting will have to have a genetic makeup that is sufficiently genetically divergent to produce an immune escape event for it to become established in a human population.

However as Giuseppe has pointed out there are reasonable grounds to suggest that there may be high levels of partial immunity amongst humans to any such hybrid virus (which should attenuate severity), so unless high virulence H5N1 genes are acquired into the mix or a fit H1N2 virus emerges, it is unlikely to be a major problem IMO.

[Missouriwatcher]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

"The lack of known direct exposure to pigs in one of the two cases described in this report suggests the possibility that limited human-to-human transmission of this influenza virus occurred. Likely transmission of swine-origin influenza A (H3N2) virus from close contact with an infected person has been observed in past investigations of human infections with swine-origin influenza A virus, but has not resulted in sustained human-to-human transmission. Preliminary evidence from the investigation of the Indiana case shows no ongoing transmission. No influenza illness has been identified, but if additional chains of transmission are identified rapid intervention is warranted try to prevent further spread of the virus..."

http://www.cdc.gov/mmwr/preview/mmwrhtml/mm60e0902a1.htm?s_cid=mm60e0902a1_w

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

This thread is a valuable commentary. Are there any objections to moving it for public viewing? I will wait 12 hours before I move the thread for objections and so everyone has a chance to edit any comments if they wish.

Thanks for the great posts!

With respect to my comments, you can surely make them public.

[Giuseppe]

Re: Is H2H spread of a novel swine H3N2 occurring in the United States?

Two just published papers add further details to our discussion:

Since the novel swoH3N2 has acquired M gene from H1N1 (2009):

[Source: US National Library of Medicine, full text: (LINK). Abstract, edited.]

J Virol. 2011 Aug 31. [Epub ahead of print] The M segment of the 2009 new pandemic H1N1 influenza virus is critical for its high transmission efficiency in the guinea pig model.


Chou YY, Albrecht RA, Pica N, Lowen AC, Richt JA, Garc?a-Sastre A, Palese P, Hai R. Source


Department of Microbiology. Abstract


A remarkable feature of the 2009 pandemic H1N1 influenza virus is its efficient transmissibility in humans compared to precursor strains from the triple reassortant swine influenza virus lineage, which cause only sporadic infections in humans. The viral components essential for this phenotype have not been fully elucidated. In this study, we aimed to determine the viral factors critical for aerosol transmission of the 2009 pandemic virus. Single or multiple segment reassortments were made between the pandemic A/California/04/09 (H1N1) [Cal/09] virus and another H1N1 strain A/Puerto Rico/8/34 (H1N1) [PR8]. These viruses were then tested in the guinea pig model to understand which segment of Cal/09 virus conferred transmissibility to the poorly transmissible PR8 virus. We confirmed our findings by generating recombinant A/swine/Texas/1998 (H3N2) [sw/Tx/98] virus, a representative triple reassortant swine virus, containing segments of the Cal/09 virus. The data showed that the M segment of the Cal/09 virus promoted aerosol transmissibility to recombinant viruses with PR8 and sw/Tx/98 virus backgrounds, suggesting that the M segment is a critical factor supporting the transmission of the 2009 pandemic virus. <DL><DT>PMID: <DD>21880744 <DD>[PubMed - as supplied by publisher] </DD></DL>

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... and regarding tissue tropism of H1N1 (2009) with respect its swine counterparts:

[Source: US National Library of Medicine, full text: (LINK). Abstract, edited.]

J Virol. 2011 Aug 31. [Epub ahead of print] Tissue tropism of swine influenza viruses and reassortants in ex vivo cultures of the human respiratory tract and conjunctiva.


Chan RW, Kang SS, Yen HL, Li AC, Tang LL, Yu WC, Yuen KM, Chan IW, Wong DD, Lai WW, Kwong DL, Sihoe AD, Poon LL, Guan Y, Nicholls JM, Peiris JS, Chan MC. Source


Department of Microbiology. Abstract


The 2009 pandemic influenza H1N1 (H1N1pdm) virus was generated by reassortment of swine influenza viruses of different lineages. This was the first influenza pandemic to emerge in over four decades and the first to occur after the realization that influenza pandemics arise from influenza viruses of animals. In order to understand the biological determinants of pandemic emergence, it is relevant to compare the tropism of different lineages of swine influenza viruses and reassortants derived from them with that of 2009 pandemic H1N1 (H1N1pdm) and seasonal influenza H1N1 viruses in ex vivo cultures of the human nasopharynx, bronchus, alveoli and conjunctiva. We hypothesized that virus which can transmit efficiently between humans replicated well in the human upper airways. As previously reported, H1N1pdm and seasonal H1N1 viruses replicated efficiently in the nasopharyngeal, bronchial and alveolar epithelium. In contrast, representative viruses from the classical swine (CS) (H1N1) lineage could not infect human respiratory epithelium; Eurasian avian-like swine (EA) (H1N1) viruses only infected alveolar epithelium and North American triple-reassortant (TRIG) viruses only infected the bronchial epithelium albeit inefficiently. Interestingly, a naturally occurring triple reassortant swine virus, A/SW/HK/915/04 (H1N2) with a matrix gene segment of EA swine derivation (i.e. differing from H1N1pdm only in lacking a neuraminidase (NA) gene of EA derivation) readily infected and replicated in human nasopharyngeal and bronchial epithelia but not in the lung. A recombinant sw915 with the NA from H1N1pdm retained its tropism for the bronchus and acquired additional replication competence for alveolar epithelium. In contrast to H1N1pdm, none of the swine viruses tested nor seasonal H1N1 had tropism in human conjunctiva. Recombinant viruses generated by swapping the surface proteins (HA, NA) of H1N1pdm and seasonal H1N1 virus demonstrated that these two gene segments together are key determinants of conjunctival tropism. Overall, these findings suggest that ex vivo cultures of the human respiratory tract provide a useful biological model for assessing human health risk of swine influenza viruses. <DL><DT>PMID: <DD>21880750 <DD>[PubMed - as supplied by publisher] </DD></DL>

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