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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">Significant neutralizing activities against H2N2 influenza A viruses
in human intravenous immunoglobulin lots manufactured from 1993 to
2010</title>
<author><name sortKey="Kubota Koketsu, Ritsuko" sort="Kubota Koketsu, Ritsuko" uniqKey="Kubota Koketsu R" first="Ritsuko" last="Kubota-Koketsu">Ritsuko Kubota-Koketsu</name>
<affiliation><nlm:aff id="af1-btt-6-245">Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kagawa;</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Yunoki, Mikihiro" sort="Yunoki, Mikihiro" uniqKey="Yunoki M" first="Mikihiro" last="Yunoki">Mikihiro Yunoki</name>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="af3-btt-6-245">Pathogenic Risk Management, Benesis Corporation, Osaka, Japan</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Okuno, Yoshinobu" sort="Okuno, Yoshinobu" uniqKey="Okuno Y" first="Yoshinobu" last="Okuno">Yoshinobu Okuno</name>
<affiliation><nlm:aff id="af1-btt-6-245">Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kagawa;</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Ikuta, Kazuyoshi" sort="Ikuta, Kazuyoshi" uniqKey="Ikuta K" first="Kazuyoshi" last="Ikuta">Kazuyoshi Ikuta</name>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
</author>
</titleStmt>
<publicationStmt><idno type="wicri:source">PMC</idno>
<idno type="pmid">22888217</idno>
<idno type="pmc">3413397</idno>
<idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3413397</idno>
<idno type="RBID">PMC:3413397</idno>
<idno type="doi">10.2147/BTT.S33495</idno>
<date when="2012">2012</date>
<idno type="wicri:Area/Pmc/Corpus">000921</idno>
<idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000921</idno>
</publicationStmt>
<sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">Significant neutralizing activities against H2N2 influenza A viruses
in human intravenous immunoglobulin lots manufactured from 1993 to
2010</title>
<author><name sortKey="Kubota Koketsu, Ritsuko" sort="Kubota Koketsu, Ritsuko" uniqKey="Kubota Koketsu R" first="Ritsuko" last="Kubota-Koketsu">Ritsuko Kubota-Koketsu</name>
<affiliation><nlm:aff id="af1-btt-6-245">Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kagawa;</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Yunoki, Mikihiro" sort="Yunoki, Mikihiro" uniqKey="Yunoki M" first="Mikihiro" last="Yunoki">Mikihiro Yunoki</name>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
<affiliation><nlm:aff id="af3-btt-6-245">Pathogenic Risk Management, Benesis Corporation, Osaka, Japan</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Okuno, Yoshinobu" sort="Okuno, Yoshinobu" uniqKey="Okuno Y" first="Yoshinobu" last="Okuno">Yoshinobu Okuno</name>
<affiliation><nlm:aff id="af1-btt-6-245">Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kagawa;</nlm:aff>
</affiliation>
</author>
<author><name sortKey="Ikuta, Kazuyoshi" sort="Ikuta, Kazuyoshi" uniqKey="Ikuta K" first="Kazuyoshi" last="Ikuta">Kazuyoshi Ikuta</name>
<affiliation><nlm:aff id="af2-btt-6-245">Department of Virology, Research Institute for Microbial Diseases, Osaka University</nlm:aff>
</affiliation>
</author>
</analytic>
<series><title level="j">Biologics : Targets & Therapy</title>
<idno type="ISSN">1177-5475</idno>
<idno type="eISSN">1177-5491</idno>
<imprint><date when="2012">2012</date>
</imprint>
</series>
</biblStruct>
</sourceDesc>
</fileDesc>
<profileDesc><textClass></textClass>
</profileDesc>
</teiHeader>
<front><div type="abstract" xml:lang="en"><p>Influenza A H2N2 virus, also known as the Asian flu, spread worldwide from 1957
to 1967, although there have been no cases reported in humans in the past 40
years. A vaccination program was introduced in Japan in the 1960s. Older
Japanese donors could have been naturally infected with the H2N2 virus or
vaccinated in the early 1960s. Human intravenous immunoglobulin (IVIG) reflects
the epidemiological status of the donating population in a given time period.
Here, the possible viral neutralizing (VN) activities of IVIG against the H2N2
virus were examined. Hemagglutination inhibition (HI) and VN activities of IVIG
lots manufactured from 1993 to 2010 in Japan and the United States were
evaluated against H2N2 viruses. High HI and VN activities against H2N2 viruses
were found in all the IVIG lots investigated. HI titers were 32–64
against the isolate in 1957 and 64–128 against the isolates in 1965. VN
titers were 80–320 against the isolate in 1957 and 1280–5120
against the isolates in 1965. Both the HI and VN titers were higher against the
isolate in 1965 than in 1957. Thus, antibody titers of IVIG against influenza
viruses are well correlated with the history of infection and the vaccine
program in Japan. Therefore, evaluation of antibody titers provides valuable
information about IVIGs, which could be used for immune stimulation when a new
influenza virus emerges in the human population.</p>
</div>
</front>
<back><div1 type="bibliography"><listBibl><biblStruct></biblStruct>
<biblStruct><analytic><author><name sortKey="Hirota, Y" uniqKey="Hirota Y">Y Hirota</name>
</author>
<author><name sortKey="Kaji, M" uniqKey="Kaji M">M Kaji</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Ma, W" uniqKey="Ma W">W Ma</name>
</author>
<author><name sortKey="Vincent, Al" uniqKey="Vincent A">AL Vincent</name>
</author>
<author><name sortKey="Gramer, Mr" uniqKey="Gramer M">MR Gramer</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Yunoki, M" uniqKey="Yunoki M">M Yunoki</name>
</author>
<author><name sortKey="Kubota Koketsu, R" uniqKey="Kubota Koketsu R">R Kubota-Koketsu</name>
</author>
<author><name sortKey="Urayama, T" uniqKey="Urayama T">T Urayama</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Okuno, Y" uniqKey="Okuno Y">Y Okuno</name>
</author>
<author><name sortKey="Tanaka, K" uniqKey="Tanaka K">K Tanaka</name>
</author>
<author><name sortKey="Baba, K" uniqKey="Baba K">K Baba</name>
</author>
<author><name sortKey="Maeda, A" uniqKey="Maeda A">A Maeda</name>
</author>
<author><name sortKey="Kunita, N" uniqKey="Kunita N">N Kunita</name>
</author>
<author><name sortKey="Ueda, S" uniqKey="Ueda S">S Ueda</name>
</author>
</analytic>
</biblStruct>
<biblStruct><analytic><author><name sortKey="Nabel, Gj" uniqKey="Nabel G">GJ Nabel</name>
</author>
<author><name sortKey="Wei, Cj" uniqKey="Wei C">CJ Wei</name>
</author>
<author><name sortKey="Ledgerwood, Je" uniqKey="Ledgerwood J">JE Ledgerwood</name>
</author>
</analytic>
</biblStruct>
</listBibl>
</div1>
</back>
</TEI>
<pmc article-type="research-article"><pmc-dir>properties open_access</pmc-dir>
<front><journal-meta><journal-id journal-id-type="nlm-ta">Biologics</journal-id>
<journal-id journal-id-type="iso-abbrev">Biologics</journal-id>
<journal-id journal-id-type="publisher-id">Biologics: Targets &
Therapy</journal-id>
<journal-title-group><journal-title>Biologics : Targets & Therapy</journal-title>
</journal-title-group>
<issn pub-type="ppub">1177-5475</issn>
<issn pub-type="epub">1177-5491</issn>
<publisher><publisher-name>Dove Medical Press</publisher-name>
</publisher>
</journal-meta>
<article-meta><article-id pub-id-type="pmid">22888217</article-id>
<article-id pub-id-type="pmc">3413397</article-id>
<article-id pub-id-type="doi">10.2147/BTT.S33495</article-id>
<article-id pub-id-type="publisher-id">btt-6-245</article-id>
<article-categories><subj-group subj-group-type="heading"><subject>Short Report</subject>
</subj-group>
</article-categories>
<title-group><article-title>Significant neutralizing activities against H2N2 influenza A viruses
in human intravenous immunoglobulin lots manufactured from 1993 to
2010</article-title>
</title-group>
<contrib-group><contrib contrib-type="author"><name><surname>Kubota-Koketsu</surname>
<given-names>Ritsuko</given-names>
</name>
<xref ref-type="aff" rid="af1-btt-6-245"><sup>1</sup>
</xref>
<xref ref-type="aff" rid="af2-btt-6-245"><sup>2</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Yunoki</surname>
<given-names>Mikihiro</given-names>
</name>
<xref ref-type="aff" rid="af2-btt-6-245"><sup>2</sup>
</xref>
<xref ref-type="aff" rid="af3-btt-6-245"><sup>3</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Okuno</surname>
<given-names>Yoshinobu</given-names>
</name>
<xref ref-type="aff" rid="af1-btt-6-245"><sup>1</sup>
</xref>
</contrib>
<contrib contrib-type="author"><name><surname>Ikuta</surname>
<given-names>Kazuyoshi</given-names>
</name>
<xref ref-type="aff" rid="af2-btt-6-245"><sup>2</sup>
</xref>
<xref ref-type="corresp" rid="c1-btt-6-245"></xref>
</contrib>
</contrib-group>
<aff id="af1-btt-6-245"><label>1</label>
Kanonji Institute, The Research Foundation for Microbial Diseases of Osaka University, Kagawa;</aff>
<aff id="af2-btt-6-245"><label>2</label>
Department of Virology, Research Institute for Microbial Diseases, Osaka University</aff>
<aff id="af3-btt-6-245"><label>3</label>
Pathogenic Risk Management, Benesis Corporation, Osaka, Japan</aff>
<author-notes><corresp id="c1-btt-6-245">Correspondence: Kazuyoshi Ikuta, Department of Virology,
Research Institute for Microbial Diseases, Osaka University, 3-1 Yamadaoka,
Suita, Osaka 565-0871, Japan, Tel +81 6 6879 8307, Fax +81 6
6879 8310, Email <email>ikuta@biken.osaka-u.ac.jp</email>
</corresp>
</author-notes>
<pub-date pub-type="collection"><year>2012</year>
</pub-date>
<pmc-comment>Dove Press titles changed from ppub to collections in 2009.
Fake ppub written to satisfy Coll Date Type=ppub</pmc-comment>
<pub-date pub-type="ppub"><year>2012</year>
</pub-date>
<pub-date pub-type="epub"><day>27</day>
<month>7</month>
<year>2012</year>
</pub-date>
<volume>6</volume>
<fpage>245</fpage>
<lpage>247</lpage>
<history><date date-type="received"><day>26</day>
<month>7</month>
<year>2012</year>
</date>
</history>
<permissions><copyright-statement>© 2012 Kubota-Koketsu et al, publisher and licensee Dove
Medical Press Ltd.</copyright-statement>
<copyright-year>2012</copyright-year>
<license><license-p>This is an Open Access article which permits unrestricted noncommercial use,
provided the original work is properly cited.</license-p>
</license>
</permissions>
<abstract><p>Influenza A H2N2 virus, also known as the Asian flu, spread worldwide from 1957
to 1967, although there have been no cases reported in humans in the past 40
years. A vaccination program was introduced in Japan in the 1960s. Older
Japanese donors could have been naturally infected with the H2N2 virus or
vaccinated in the early 1960s. Human intravenous immunoglobulin (IVIG) reflects
the epidemiological status of the donating population in a given time period.
Here, the possible viral neutralizing (VN) activities of IVIG against the H2N2
virus were examined. Hemagglutination inhibition (HI) and VN activities of IVIG
lots manufactured from 1993 to 2010 in Japan and the United States were
evaluated against H2N2 viruses. High HI and VN activities against H2N2 viruses
were found in all the IVIG lots investigated. HI titers were 32–64
against the isolate in 1957 and 64–128 against the isolates in 1965. VN
titers were 80–320 against the isolate in 1957 and 1280–5120
against the isolates in 1965. Both the HI and VN titers were higher against the
isolate in 1965 than in 1957. Thus, antibody titers of IVIG against influenza
viruses are well correlated with the history of infection and the vaccine
program in Japan. Therefore, evaluation of antibody titers provides valuable
information about IVIGs, which could be used for immune stimulation when a new
influenza virus emerges in the human population.</p>
</abstract>
<kwd-group><kwd>IVIG</kwd>
<kwd>influenza</kwd>
<kwd>H2N2</kwd>
<kwd>neutralization</kwd>
</kwd-group>
</article-meta>
</front>
<body><sec><title>Introduction</title>
<p>The highly pathogenic avian influenza A H5N1 virus has spread among wild birds
worldwide. As of April 2012, there have been 602 cases of human infections, with an
extremely high mortality rate of about 59%.<xref ref-type="bibr" rid="b1-btt-6-245">1</xref>
Therefore, there are public health concerns
regarding the possible global emergence of an H5N1 pandemic virus. However, a
swine-origin pandemic influenza A H1N1 virus suddenly emerged in 2009. This novel
virus spread among human populations within a short period of time because of the
low level of immune responses against the virus, especially among young people.
Thus, a pandemic influenza virus may be transmitted to humans because of limited
immune responses against the virus in humans. The H2N2 virus could be considered one
such virus, because it was prevalent in humans between 1957 and 1967, and a
vaccination program was introduced in Japan in the 1960s.<xref ref-type="bibr" rid="b2-btt-6-245">2</xref>
However, although H2N2 continues to circulate among
birds and pigs, this virus has not infected humans for the last several
decades.<xref ref-type="bibr" rid="b3-btt-6-245">3</xref>
Based on the above
background, it might be worthwhile examining the neutralizing activity of human
intravenous immunoglobulin (IVIG) against the H2N2 virus.</p>
<p>Generally, IVIGs are manufactured in individual lots with serum donations from over
10,000 healthy donors. Therefore, IVIG contains various antibodies against numerous
human pathogens including seasonal influenza viruses. In fact, not only current
(manufactured in 2008) but also previous (manufactured in 1999) IVIGs contain
antibodies with significant neutralizing titers against seasonal and pandemic 2009
influenza viruses.<xref ref-type="bibr" rid="b4-btt-6-245">4</xref>
The antibodies
comprising IVIG therefore reflect the epidemiological status of the donating
population, in a particular time period and geographical area.</p>
<p>In this study, IVIG lots manufactured from 1993 to 2010 were evaluated for
hemagglutination inhibition (HI) and virus neutralizing (VN) activities against the
H2N2 virus.</p>
</sec>
<sec sec-type="materials|methods"><title>Material and methods</title>
<p>Clinical isolates of H2N2 (A/Okuda/1957, A/Izumi/5/1965, A/Kaizuka/2/1965), IVIGs
manufactured from 1993 to 2010 from healthy donors in Japan (currently Kenketsu
Venoglobulin<sup>®</sup>
-IH; Benesis Corporation, Osaka, Japan), and
IVIGs manufactured in 1993 and 1999 from healthy donors in the United States were
used in this study (<xref ref-type="table" rid="t1-btt-6-245">Table 1</xref>
).</p>
<p>The viruses were propagated in Madin–Darby canine kidney (MDCK) cells or in
the allantoic cavity of 11-day-old embryonated chicken eggs. The culture media and
allantoic fluids were stored at −80°C prior to use. Viral
infectivity (FFU/mL) was titrated in MDCK cells using the peroxidase-antiperoxidase
(PAP) staining technique.<xref ref-type="bibr" rid="b5-btt-6-245">5</xref>
The HI
test using 0.7% guinea pig erythrocytes was carried out as described
previously.<xref ref-type="bibr" rid="b5-btt-6-245">5</xref>
HI titers are
expressed as the reciprocal of the highest dilution of the IVIG preparation showing
inhibition. The VN test was also carried out as described previously.<xref ref-type="bibr" rid="b5-btt-6-245">5</xref>
Briefly, each IVIG was serially
diluted two-fold with serum-free minimal essential medium. The IVIG dilutions (30
μL of each) were mixed with 100 FFU (30 μL) of virus. After
incubation for 30 minutes at 37°C, the mixture (30 μL) was applied
to MDCK cells in a 96-well microplate. After incubation for 16 hours, the cells were
fixed with ethanol and stained using PAP technique. The results are expressed as the
reciprocal of the dilution resulting in 50% neutralization
(VN<sub>50</sub>
).</p>
</sec>
<sec sec-type="results"><title>Results</title>
<p>As summarized in <xref ref-type="table" rid="t1-btt-6-245">Table 1</xref>
, the titers
were significant against all three isolates of H2N2: HI titers of 32–64 and
VN<sub>50</sub>
titers of 80–320 against Okuda/1957, HI titers of
64–128 and VN<sub>50</sub>
titers of 1280–5120 against Izumi/5/1965,
and HI titers of 64–128 and VN<sub>50</sub>
titers of 1280–2560
against Kaizuka/2/1965. Both the HI and VN<sub>50</sub>
titers were higher against
the isolate in 1965 than in 1957 (<xref ref-type="table" rid="t1-btt-6-245">Table
1</xref>
). There were no apparent differences in the HI and VN<sub>50</sub>
titers against H2N2 virus isolates among the IVIG products prepared in the United
States and Japan in different years.</p>
</sec>
<sec sec-type="discussion"><title>Discussion</title>
<p>Currently, most of the donor population has not been exposed to the H2N2 virus. IVIG
is manufactured from pooled plasma. In a previous study, HI and VN activities
against seasonal influenza H1N1 and H1N1-pdm2009 were examined in IVIG lots
manufactured from 1999 to 2008. The IVIGs indicated high and stable HI and VN
activity against H1N1. It was taken into account that the donor population may have
been immunized through native infections and/or vaccine programs. Interestingly, the
IVIGs also showed low but significant HI and VN activities against H1N1-pdm2009
despite most of the donor population never having been exposed to swine H1N1 or the
Spanish flu.<xref ref-type="bibr" rid="b4-btt-6-245">4</xref>
</p>
<p>In this study, HI and VN activities against the H2N2 virus, which has not been
included in vaccine programs for the past 45 years, were measured in IVIGs
manufactured from 1993 to 2010 in Japan. Both the HI and VN<sub>50</sub>
titers were
higher against H2N2 isolates in 1965 than in 1957. It has been reported that people
under the age of 50 years have little or no immunity to H2N2 and that people older
than 50 years who have been exposed to the virus show a higher rate of
resistance.<xref ref-type="bibr" rid="b6-btt-6-245">6</xref>
Therefore, older
Japanese donors could have been naturally infected with the H2N2 virus or vaccinated
in the early 1960s. Consequently, antibody titers against the H2N2 virus in the
general population will decrease in the future, which could lead to an H2N2
pandemic. It is not clear why such differences of HI and VN activities of IVIG
preparations were observed between the United States and Japan. One possible
explanation may be derived from the difference in environmental factors and
population for blood donation between the two countries.</p>
</sec>
<sec><title>Conclusion</title>
<p>The results suggest that the antibody titers against influenza viruses in IVIGs
correlate with the history of infection and with vaccination programs. Therefore,
the evaluation of antibody titers provides valuable information about IVIGs, which
could be used for immune stimulation when a new influenza virus emerges in the human
population.</p>
</sec>
</body>
<back><ack><p>This study was partially conducted as a collaborative research project between Osaka
University, The Research Foundation for Microbial Diseases of Osaka University, and
the Benesis Corporation.</p>
</ack>
<fn-group><fn><p><bold>Disclosures</bold>
</p>
<p>Yoshinobu Okuno and Ritsuko Kubota-Koketsu are employed by The Research
Foundation for Microbial Diseases of Osaka University. Mikihiro Yunoki is
employed by the Benesis Corporation. All other authors report no conflicts of
interest in this work.</p>
</fn>
</fn-group>
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<volume>28</volume>
<issue>6</issue>
<fpage>1308</fpage>
<lpage>1313</lpage>
<pub-id pub-id-type="pmid">2380359</pub-id>
</element-citation>
</ref>
<ref id="b6-btt-6-245"><label>6.</label>
<element-citation publication-type="journal"><person-group person-group-type="author"><name><surname>Nabel</surname>
<given-names>GJ</given-names>
</name>
<name><surname>Wei</surname>
<given-names>CJ</given-names>
</name>
<name><surname>Ledgerwood</surname>
<given-names>JE</given-names>
</name>
</person-group>
<article-title>Vaccinate for the next H2N2 pandemic
now</article-title>
<source>Nature</source>
<year>2011</year>
<volume>471</volume>
<issue>7337</issue>
<fpage>157</fpage>
<lpage>158</lpage>
<pub-id pub-id-type="pmid">21390107</pub-id>
</element-citation>
</ref>
</ref-list>
</back>
<floats-group><table-wrap id="t1-btt-6-245" position="float"><label>Table 1</label>
<caption><p>Cross-reactivity of intravenous immunoglobulins to the influenza H2N2
virus</p>
</caption>
<table frame="hsides" rules="groups"><thead><tr><th align="left" valign="top" rowspan="3" colspan="1"><bold>IVIG</bold>
</th>
<th colspan="6" align="left" valign="top" rowspan="1"><bold>H2N2 virus</bold>
<hr></hr>
</th>
</tr>
<tr><th colspan="2" align="left" valign="top" rowspan="1"><bold>Okuda/1957</bold>
<hr></hr>
</th>
<th colspan="2" align="left" valign="top" rowspan="1"><bold>Izumi/5/1965</bold>
<hr></hr>
</th>
<th colspan="2" align="left" valign="top" rowspan="1"><bold>Kaizuka/2/1965</bold>
<hr></hr>
</th>
</tr>
<tr><th align="left" valign="top" rowspan="1" colspan="1"><bold>HI</bold>
</th>
<th align="left" valign="top" rowspan="1" colspan="1"><bold>VN<sub>50</sub>
</bold>
</th>
<th align="left" valign="top" rowspan="1" colspan="1"><bold>HI</bold>
</th>
<th align="left" valign="top" rowspan="1" colspan="1"><bold>VN<sub>50</sub>
</bold>
</th>
<th align="left" valign="top" rowspan="1" colspan="1"><bold>HI</bold>
</th>
<th align="left" valign="top" rowspan="1" colspan="1"><bold>VN<sub>50</sub>
</bold>
</th>
</tr>
</thead>
<tbody><tr><td align="left" valign="top" rowspan="1" colspan="1">1993JP, Lot A</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1993US, Lot B</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">320</td>
<td align="left" valign="top" rowspan="1" colspan="1">128</td>
<td align="left" valign="top" rowspan="1" colspan="1">5120</td>
<td align="left" valign="top" rowspan="1" colspan="1">128</td>
<td align="left" valign="top" rowspan="1" colspan="1">2560</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1999JP-A, Lot C</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1999JP-B, Lot D</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">80</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1999JP-C, Lot E</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1999JP-D, Lot F</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">1999US, Lot G</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">320</td>
<td align="left" valign="top" rowspan="1" colspan="1">128</td>
<td align="left" valign="top" rowspan="1" colspan="1">5120</td>
<td align="left" valign="top" rowspan="1" colspan="1">128</td>
<td align="left" valign="top" rowspan="1" colspan="1">2560</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2000JP, Lot H</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2003JP, Lot I</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">80</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2006JP, Lot J</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2007JP, Lot K</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2008JP, Lot L</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">160</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2009JP, Lot M</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">80</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
<tr><td align="left" valign="top" rowspan="1" colspan="1">2010JP, Lot N</td>
<td align="left" valign="top" rowspan="1" colspan="1">32</td>
<td align="left" valign="top" rowspan="1" colspan="1">80</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
<td align="left" valign="top" rowspan="1" colspan="1">64</td>
<td align="left" valign="top" rowspan="1" colspan="1">1280</td>
</tr>
</tbody>
</table>
<table-wrap-foot><fn id="tfn1-btt-6-245"><p><bold>Abbreviations:</bold>
HI, hemagglutination inhibition titer; IVIG,
intravenous immunoglobulin; JP, Japan; US, United States;
VN<sub>50</sub>
, virus neutralization titer giving 50%
inhibition of infectivity.</p>
</fn>
</table-wrap-foot>
</table-wrap>
</floats-group>
</pmc>
</record>
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