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<meta name="description" content="West Nile virus (WNV), a mosquito-borne flavivirus, has recently emerged in North America, and the elderly are particularly susceptible to severe neurological disease and death from infection with this virus. We have investigated the innate immune response of primary human macrophages to WNV in vitr …"><meta name="keywords" content="pmid:18508883, PMC2493309, doi:10.1128/JVI.00618-08, Research Support, N.I.H., Extramural, Kok-Fai Kong, Karine Delroux, Ruth R Montgomery, Adult, Age Factors, Aged, Aged, 80 and over, Cell Adhesion Molecules / metabolism, Cell Line, Cells, Cultured, Cytokines / biosynthesis, Female, Humans, Immunity, Innate*, Lectins, C-Type / metabolism, Macrophages / immunology*, Male, Middle Aged, North America, Protein Binding, Receptors, Cell Surface / metabolism, STAT1 Transcription Factor / metabolism, Toll-Like Receptor 3 / biosynthesis, Toll-Like Receptor 3 / immunology*, Viral Envelope Proteins / metabolism, West Nile Fever / immunology*, West Nile virus / immunology*, PubMed Abstract, NIH, NLM, NCBI, National Institutes of Health, National Center for Biotechnology Information, National Library of Medicine, MEDLINE"><meta name="robots" content="index,nofollow,noarchive"><meta property="og:title" content="Dysregulation of TLR3 impairs the innate immune response to West Nile virus in the elderly - PubMed"><meta property="og:url" content="https://pubmed.ncbi.nlm.nih.gov/18508883/"><meta property="og:description" content="West Nile virus (WNV), a mosquito-borne flavivirus, has recently emerged in North America, and the elderly are particularly susceptible to severe neurological disease and death from infection with this virus. We have investigated the innate immune response of primary human macrophages to WNV in vitr …"><meta property="og:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:image:secure_url" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg"><meta property="og:type" content="website"><meta property="og:site_name" content="PubMed"><meta name="twitter:domain" content="pubmed.ncbi.nlm.nih.gov"><meta name="twitter:card" content="summary_large_image"><meta name="twitter:title" content="Dysregulation of TLR3 impairs the innate immune response to West Nile virus in the elderly - PubMed"><meta name="twitter:url" content="https://pubmed.ncbi.nlm.nih.gov/18508883/"><meta name="twitter:description" content="West Nile virus (WNV), a mosquito-borne flavivirus, has recently emerged in North America, and the elderly are particularly susceptible to severe neurological disease and death from infection with this virus. We have investigated the innate immune response of primary human macrophages to WNV in vitr …"><meta name="twitter:image" content="https://cdn.ncbi.nlm.nih.gov/pubmed/persistent/pubmed-meta-image-v2.jpg">
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<label for="email-citation-format" class="action-panel-label">
Format:
</label>
<select id="email-citation-format" name="citation-format" class="action-panel-selector email-citation-format">
<option selected="selected" value="summary">Summary</option>
<option value="summary-text">Summary (text)</option>
<option value="abstract">Abstract</option>
<option value="abstract-text">Abstract (text)</option>
</select>
</div>
<div class="include-supplemental-container">
<input type="checkbox" aria-label="Include MeSH and other data" name="include-supplemental" id="email-include-supplemental" class="email-include-supplemental">
<label for="email-include-supplemental" class="email-include-supplemental-label">MeSH and other data</label>
</div>
<div class="form-field recaptcha ">
<div class="g-recaptcha" id="id-recaptcha" data-sitekey="6LfsWHMdAAAAAClKbtOpjQ2pMjgsGxvv7NdZW9uI"></div>
</div>
<div id="captcha-error-message" class="usa-input-error-message captcha-validation-message" role="alert"></div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="send">
Send email
</button>
<button class="action-panel-cancel"
aria-label="Close 'Email citations' panel"
ref="linksrc=close_email_panel"
aria-controls="email-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="email"
data-ga-label="cancel">
Cancel
</button>
</div>
<input type="hidden" name="email-search-details" value="" />
<input type="hidden" name="email-search-details-hash" value="0e42663a6c3bd85498fcb88798998fed7bfdc45d457db35281e41afe13cc0524" />
</form>
</div>
</div>
<div id="collections-action-panel"
class="collections-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-collections-open-panel-enabled="false"
data-collections-open-panel-url-hash="#open-collections-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to Collections
</h3>
<form id="collections-action-panel-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-myncbi-max-collection-name-length="100"
data-add-to-collection-max-amount="1000"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/">
<input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif">
<div class="choice-group" role="radiogroup">
<ul class="radio-group-items">
<li>
<input type="radio"
id="collections-action-panel-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_new">
<label for="collections-action-panel-new">Create a new collection</label>
</li>
<li>
<input type="radio"
id="collections-action-panel-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="collections_radio_existing">
<label for="collections-action-panel-existing">Add to an existing collection</label>
</li>
</ul>
</div>
<div class="controls-wrapper">
<div class="action-panel-control-wrap new-collections-controls">
<label for="collections-action-panel-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label>
<input
type="text"
name="add-to-new-collection"
id="collections-action-panel-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="save_share"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection">
<div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
</div>
</div>
<div class="action-panel-control-wrap existing-collections-controls">
<label for="collections-action-panel-add-to-existing" class="action-panel-label">
Choose a collection:
</label>
<select id="collections-action-panel-add-to-existing"
class="action-panel-selector collections-action-add-to-existing"
data-ga-category="save_share"
data-ga-action="select_collection"
data-ga-label="($('#collections-action-add-to-existing').val() === 'Favorites') ? 'Favorites' : 'non_favorites_collection'">
</select>
<div class="collections-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your collection due to an error<br>
<a href="#">Please try again</a>
</div>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="add">
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to Collections' panel"
ref="linksrc=close_collections_panel"
aria-controls="collections-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="collections"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="bibliography-action-panel"
class="bibliography-action-panel action-panel in-progress-dots-panel"
aria-hidden="true"
data-bibliography-open-panel-enabled="false"
data-bibliography-open-panel-url-hash="#open-bibliography-panel">
<div class="inner-wrap">
<h3 class="action-panel-heading">
Add to My Bibliography
</h3>
<form id="bibliography-action-panel-form"
class="bibliography-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-add-to-bibliography-max-amount="100"
data-add-to-bibliography-batch-size="10"
data-bibliography-delegates-url="/list-bibliography-delegates/"
data-add-to-bibliography-url="/add-to-bibliography/"
data-get-article-ids-by-search-url="/get-article-ids-by-search/"
data-mybib-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/mybibliography/">
<input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif">
<div class="action-panel-control-wrap bibliographies-controls">
<div class="choice-group">
<ul class="bibliographies-action-add radio-group-items">
<li>
<input name="bibliography" id="my-bibliography" class="my-bibliography" type="radio" checked/>
<label for="my-bibliography">My Bibliography</label>
</li>
</ul>
</div>
</div>
<div class="bibliographies-retry-load-on-error usa-input-error-message selection-validation-message">
Unable to load your delegates due to an error<br>
<a href="#">Please try again</a>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Adding..."
data-pinger-ignore>
Add
</button>
<button class="action-panel-cancel"
aria-label="Close 'Add to bibliography' panel"
ref="linksrc=close_bibliography_panel"
aria-controls="bibliography-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="mybib"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="saved-search-action-panel" class="saved-search-action-panel action-panel " aria-hidden="true"
data-saved-search-open-panel-enabled="false"
data-saved-search-open-panel-url-hash="#open-saved-search-panel">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your saved search
</h2>
<form id="saved-search-action-panel-form"
class="saved-search-action-panel-form action-panel-content action-form"
data-create-saved-search-url="/create-saved-search/"
data-try-search-terms-url="/try-search-term/"
data-saved-search-root-url="https://www.ncbi.nlm.nih.gov/myncbi/searches/">
<input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif">
<div class="action-panel-control-wrap">
<label for="saved-search-name" class="action-panel-label saved-search-name-label required-field-asterisk">
Name of saved search:
</label>
<input maxlength="200"
type="text"
name="saved-search-name"
id="saved-search-name"
class="saved-search-name"
value=""
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-term" class="action-panel-label required-field-asterisk">
Search terms:
</label>
<textarea name="saved-search-term" id="saved-search-term" class="saved-search-term" required></textarea>
</div>
<div class="test-search-term-wrap">
<a href="#" class="try-search-term">Test search terms</a>
</div>
<div class="choice-group action-panel-extra-margin-top">
<span class="action-panel-label" id="fieldset-label">
Would you like email updates of new search results?
</span>
<fieldset id="saved-search-alert" aria-describedby="fieldset-label">
<legend class="usa-sr-only">Saved Search Alert Radio Buttons</legend>
<ul class="radio-group-items">
<li>
<input type="radio" id="saved-search-alert-yes" class="saved-search-alert-yes" name="saved-search-alert" value="yes" checked>
<label for="saved-search-alert-yes" class="action-panel-label">Yes</label>
</li>
<li>
<input aria-label="No radio input" type="radio" id="saved-search-alert-no" class="saved-search-alert-no" name="saved-search-alert" value="no">
<label for="saved-search-alert-no" class="action-panel-label">No</label>
</li>
</ul>
</fieldset>
</div>
<div class="alert-schedule-wrap">
<div class="action-panel-control-wrap">
<label class="action-panel-label">
Email:
</label>
<span aria-label="Email address" id="saved-search-email" class="action-panel-label"><span class="action-panel-label-bold"></span> (<a class="myncbi-account-settings" href="https://www.ncbi.nlm.nih.gov/account/settings/">change</a>)</span>
</div>
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="saved-search-frequency" class="action-panel-label">
Frequency:
</label>
<select id="saved-search-frequency" class="no-border-panel-selector saved-search-frequency">
<option value="monthly">Monthly</option>
<option value="weekly">Weekly</option>
<option value="daily">Daily</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-monthly-additional">
<label for="saved-search-monthly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-monthly-on-day" class="no-border-panel-selector">
<option value="Sunday">The first Sunday</option>
<option value="Monday">The first Monday</option>
<option value="Tuesday">The first Tuesday</option>
<option value="Wednesday">The first Wednesday</option>
<option value="Thursday">The first Thursday</option>
<option value="Friday">The first Friday</option>
<option value="Saturday">The first Saturday</option>
<option value="day">The first day</option>
<option value="weekday">The first weekday</option>
</select>
</div>
<div class="action-panel-control-wrap saved-search-weekly-additional">
<label for="saved-search-weekly-on-day" class="action-panel-label">
Which day?
</label>
<select id="saved-search-weekly-on-day" class="no-border-panel-selector saved-search-weekly-on-day">
<option value="Sunday">Sunday</option>
<option value="Monday">Monday</option>
<option value="Tuesday">Tuesday</option>
<option value="Wednesday">Wednesday</option>
<option value="Thursday">Thursday</option>
<option value="Friday">Friday</option>
<option value="Saturday">Saturday</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-report" class="action-panel-label">
Report format:
</label>
<select id="saved-search-report" class="no-border-panel-selector saved-search-report">
<option value="DocSum">Summary</option>
<option value="DocSumText">Summary (text)</option>
<option value="Abstract">Abstract</option>
<option value="AbstractText">Abstract (text)</option>
<option value="MEDLINE">PubMed</option>
</select>
</div>
<div class="action-panel-control-wrap">
<label for="saved-search-amount" class="action-panel-label">
Send at most:
</label>
<select id="saved-search-amount" class="no-border-panel-selector saved-search-amount">
<option value="1">1 item</option>
<option value="5" selected>5 items</option>
<option value="10">10 items</option>
<option value="20">20 items</option>
<option value="50">50 items</option>
<option value="100">100 items</option>
<option value="200">200 items</option>
</select>
</div>
<div>
<input type="checkbox" id="saved-search-send-if-no-result" class="saved-search-send-if-no-result" name="saved-search-send-if-no-result">
<label for="saved-search-send-if-no-result" class="action-panel-label smaller-checkbox">
Send even when there aren't any new results
</label>
</div>
<div class="action-panel-control-wrap option-text-in-email-wrap">
<label for="saved-search-email-text" class="action-panel-label">
Optional text in email:
</label>
<textarea name="saved-search-email-text"
id="saved-search-email-text"
class="saved-search-email-text"></textarea>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Saving..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Save
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your saved search' panel"
ref="linksrc=close_saved_search_panel"
aria-controls="saved-search-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="citation-manager-action-panel" class="citation-manager-action-panel action-panel" aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Create a file for external citation management software
</h2>
<form id="citation-manager-action-panel-form"
class="action-panel-content action-form"
action="/results-export-ids/"
data-by-search-action="/results-export-search-data/"
data-by-ids-action="/results-export-ids/"
method="post"
data-by-search-method="post"
data-by-ids-method="post">
<input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif">
<input name="results-format" type="hidden" value="pubmed"/>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Sending..."
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="save">
Create file
</button>
<button class="action-panel-cancel"
aria-label="Close 'Send citations to citation manager' panel"
ref="linksrc=close_citation_manager_panel"
aria-controls="citation-manager-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="citation_manager"
data-ga-label="cancel">
Cancel
</button>
</div>
</form>
</div>
</div>
<div id="rss-action-panel" class="rss-action-panel action-panel " aria-hidden="true">
<div class="inner-wrap">
<h2 class="action-panel-heading">
Your RSS Feed
</h2>
<form id="rss-action-panel-form"
class="rss-action-panel-form action-panel-content action-form"
data-create-rss-feed-url="/create-rss-feed-url/"
data-search-form-term-value="">
<input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif">
<div class="action-panel-control-wrap">
<label for="rss-name" class="action-panel-label required-field-asterisk">
Name of RSS Feed:
</label>
<input maxlength="200"
placeholder="Name"
type="text"
name="rss-name"
id="rss-name"
class="rss-name"
value=''
required
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/">
</div>
<div class="rss-limit-wrap">
<div class="action-panel-control-wrap action-panel-extra-margin-top">
<label for="rss-limit" class="action-panel-label">
Number of items displayed:
</label>
<select id="rss-limit" class="no-border-panel-selector rss-limit">
<option value="5">5</option>
<option value="10">10</option>
<option value="15" selected="selected">15</option>
<option value="20">20</option>
<option value="50">50</option>
<option value="100">100</option>
</select>
</div>
</div>
<div class="action-panel-actions">
<button class="action-panel-submit"
type="submit"
data-loading-label="Creating..."
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="save">
Create RSS
</button>
<button class="action-panel-cancel"
aria-label="Close 'Your RSS' panel"
ref="linksrc=close_rss_panel"
aria-controls="rss-action-panel"
aria-expanded="false"
data-ga-category="save_share"
data-ga-action="alert"
data-ga-label="cancel">
Cancel
</button>
</div>
<div class="action-panel-control-wrap rss-link-copy-wrap">
<label for="rss-link" class="usa-sr-only">RSS Link</label>
<input placeholder="Your RSS Feed Link" type="text" name="rss-link" id="rss-link" class="rss-link" title="RSS Link">
<button
type="button"
disabled
class="rss-link-copy-button disabled"
data-ga-category="save_share"
data-ga-action="rss"
data-ga-label="copy">
Copy
</button>
</div>
</form>
</div>
</div>
</div>
</div>
<div class="article-page" id="article-page" data-article-pmid="18508883">
<aside class="page-sidebar">
<div class="inner-wrap">
<div class="full-text-links">
<div class="full-view">
<h3 class="title">
Full text links
</h3>
<div class="full-text-links-list">
<a class="link-item
dialog-focus"
href="https://journals.asm.org/doi/10.1128/JVI.00618-08?url_ver=Z39.88-2003&amp;rfr_id=ori:rid:crossref.org&amp;rfr_dat=cr_pub 0pubmed"
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ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=False&amp;PrId=4487&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=18508883&amp;db=pubmed&amp;nlmid=0113724"
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pmc
"
href="https://pmc.ncbi.nlm.nih.gov/articles/pmid/18508883/"
target="_blank"
rel="noopener"
ref="linksrc=fulltextorjournal_fulltext&amp;is_pmc=True&amp;PrId=3494&amp;itool=Abstract-def&amp;log$=linkouticon&amp;uid=18508883&amp;db=pubmed&amp;nlmid=0113724"
title="Free full text at PubMed Central"
data-ga-category="full_text"
data-ga-action="PMC"
data-ga-label="18508883"
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Free PMC article
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</div>
</div>
<div class="short-view">
<a href="#" class="full-text-links-button full-text-links-dialog-trigger">
Full text links
</a>
</div>
</div>
<div class="actions-buttons sidebar"><h3 class="title">Actions</h3><div class="inner-wrap"><button class="citation-button citation-dialog-trigger"
aria-label="Open dialog with citation text in different styles"
data-ga-category="save_share"
data-ga-action="cite"
data-ga-label="open"
data-all-citations-url="/18508883/citations/"
data-citation-style="nlm"
data-pubmed-format-link="/18508883/export/"><span class="button-label">Cite</span></button><link type="text/css" href="ncbi-overlay-block/src/overlay-block.css"><div class="collections-button-container" data-article-id="18508883" data-article-db="pubmed"><button class="collections-button collections-dialog-trigger"
aria-label="Save article in MyNCBI collections."
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_button"
data-collections-open-dialog-enabled="false"
data-collections-open-dialog-url="https://account.ncbi.nlm.nih.gov/?back_url=https%3A%2F%2Fpubmed.ncbi.nlm.nih.gov%2F18508883%2F%23open-collections-dialog"
data-in-collections="false"><span class="button-label">Collections</span></button><div class="overlay" role="dialog"><div id="collections-action-dialog"
class="dialog collections-dialog"
aria-hidden="true"><div class="title">Add to Collections</div><div class="collections-action-panel action-panel"><form id="collections-action-dialog-form"
class="collections-action-panel-form action-panel-content action-form action-panel-smaller-selectors"
data-existing-collections-url="/list-existing-collections/"
data-add-to-existing-collection-url="/add-to-existing-collection/"
data-create-and-add-to-new-collection-url="/create-and-add-to-new-collection/"
data-myncbi-max-collection-name-length="100"
data-collections-root-url="https://www.ncbi.nlm.nih.gov/myncbi/collections/"><input type="hidden" name="csrfmiddlewaretoken" value="ndlx9ux5Q7wCIVLUAvkUwJuNJgRh0oKVE05CMk0rqIj6CmRKdOXW0R0rhkztTZif"><div class="choice-group" role="radiogroup"><ul class="radio-group-items"><li><input type="radio"
id="collections-action-dialog-new"
class="collections-new"
name="collections"
value="new"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_new"><label for="collections-action-dialog-new">Create a new collection</label></li><li><input type="radio"
id="collections-action-dialog-existing"
class="collections-existing"
name="collections"
value="existing"
checked="true"
data-ga-category="collections_button"
data-ga-action="click"
data-ga-label="collections_radio_existing"><label for="collections-action-dialog-existing">Add to an existing collection</label></li></ul></div><div class="controls-wrapper"><div class="action-panel-control-wrap new-collections-controls"><label for="collections-action-dialog-add-to-new" class="action-panel-label required-field-asterisk">
Name your collection:
</label><input
type="text"
name="add-to-new-collection"
id="collections-action-dialog-add-to-new"
class="collections-action-add-to-new"
pattern="[^&quot;&amp;=&lt;&gt;\/]*" title="The following characters are not allowed in the Name field: &quot;&amp;=&lt;&gt;/"
maxlength="100"
data-ga-category="collections_button"
data-ga-action="create_collection"
data-ga-label="non_favorties_collection"><div class="collections-new-name-too-long usa-input-error-message selection-validation-message">
Name must be less than 100 characters
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Abstract
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<p>
West Nile virus (WNV), a mosquito-borne flavivirus, has recently emerged in North America, and the elderly are particularly susceptible to severe neurological disease and death from infection with this virus. We have investigated the innate immune response of primary human macrophages to WNV in vitro and have found significant differences between the responsiveness of macrophages derived from younger donors and that from older donors. Binding of the glycosylated WNV envelope protein to the C-type lectin dendritic cell-specific intercellular adhesion molecule 3 (ICAM3) grabbing nonintegrin (DC-SIGN) leads to a reduction in the expression of Toll-like receptor 3 (TLR3) in macrophages from young donors via the signal transducer and activator of transcription 1 (STAT1)-mediated pathway. This signaling is impaired in the elderly, and the elevated levels of TLR3 result in an elevation of cytokine levels. This alteration of the innate immune response with aging may contribute to the permeability of the blood-brain barrier and suggests a possible mechanism for the increased severity of WNV infection in older individuals.
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data-image-width="400"
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data-pmc-id="PMC2493309"
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<img class="figure-thumb" itemprop="thumbnail"
id="article-image-0"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/bba6cdc02dd8/zjv0150808450001.gif"
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<strong class="figure-label">
<p> FIG. 1. </p>
</strong>
<div class="figure-caption-contents"><p> TLR3 levels are reduced by… </p></div>
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</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 1. </p>
</strong>
<div class="figure-caption-contents"><p> TLR3 levels are reduced by WNV infection in macrophages from young but not… </p></div>
</div>
<figcaption id="figure-caption-0" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 1.
</strong>
<div class="figure-caption-contents">TLR3 levels are reduced by WNV infection in macrophages from young but not elderly donors. (A) Primary macrophages from healthy young and older individuals were infected with WNV (MOI, 0.0, 0.1, 1.0 and 10.0) and incubated for 1 h (young, <i>n</i> = 7; elderly, <i>n</i> = 8; data shown are the means ± the standard errors of the means; **, ANOVA, <i>P</i> &lt; 0.05). (B) Primary macrophages from healthy young and older individuals were infected with WNV (MOI, 1) for 0, 1, and 3 h. Total proteins were harvested, and the protein level of TLR3 was detected via immunoblotting. β-Actin was used as a control to ensure equal loading. Data are representative of three subjects in each cohort.</div>
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itemprop="contentUrl"
aria-describedby="figure-caption-1"
role="button"
data-image-width="450"
data-image-height="575"
data-image-alt="FIG. 2."
data-pmc-id="PMC2493309"
data-figure-id="f2">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-1"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/2665e348c80d/zjv0150808450002.gif"
alt="FIG. 2." />
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<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="450">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="575">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
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<strong class="figure-label">
<p> FIG. 2. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between the glycosylated WNV-E… </p></div>
</div>
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<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 2. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between the glycosylated WNV-E and DC-SIGN suppresses TLR3 in macrophages from young… </p></div>
</div>
<figcaption id="figure-caption-1" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 2.
</strong>
<div class="figure-caption-contents">Interaction between the glycosylated WNV-E and DC-SIGN suppresses TLR3 in macrophages from young but not from elderly donors. (A) Primary macrophages derived from young adults (<i>n</i> = 9; data are means ± standard errors of the means) were infected with native WNV strain CT-2741 and glycosylation-deficient WNV strain NY99-E154 for 1 h (**, ANOVA, <i>P</i> of &lt;0.05 for uninfected vs. infection with CT2741 and for infection with CT2741 vs. E154). (B) Primary macrophages from young and older adults were treated with WNV-E (30 to 120 ng/10<sup>6</sup> cells) for 1 h and assessed by qPCR (young, <i>n</i> = 6; elderly, <i>n</i> = 8; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for macrophages of the young vs. the elderly. (C) Primary macrophages from young and older adults were treated with WNV-E protein (30 ng/10<sup>6</sup> cells) for 0, 1, and 3 h. Protein data are representative of three subjects in each cohort.</div>
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</figure>
<figure class="figure-item "
itemscope itemtype="http://schema.org/ImageObject"
itemprop="associatedMedia"
data-slide-index="2"
data-label-slug="fig-3">
<a class="figure-link"
href="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/c02b08c8aedc/zjv0150808450003.jpg"
itemprop="contentUrl"
aria-describedby="figure-caption-2"
role="button"
data-image-width="450"
data-image-height="733"
data-image-alt="FIG. 3."
data-pmc-id="PMC2493309"
data-figure-id="f3">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-2"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/70d6f9080dcf/zjv0150808450003.gif"
alt="FIG. 3." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="450">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="733">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG. 3. </p>
</strong>
<div class="figure-caption-contents"><p> Macrophages with reduced levels of… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 3. </p>
</strong>
<div class="figure-caption-contents"><p> Macrophages with reduced levels of DC-SIGN fail to suppress the expression of TLR3.… </p></div>
</div>
<figcaption id="figure-caption-2" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 3.
</strong>
<div class="figure-caption-contents">Macrophages with reduced levels of DC-SIGN fail to suppress the expression of TLR3. Primary macrophages from young individuals were transfected with siRNA targeting DC-SIGN or L-SIGN or without siRNA (Mock), using nucleofection technology. (A) The efficiency of RNA interference was assessed via qPCR (<i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for mock-treated vs. DC-SIGN-treated cells). (B) The efficiency of siRNA was assessed by immunoblotting using anti-DC-SIGN monoclonal antibody. β-Actin was used as the loading control. Data are representative of at least three subjects. (C) The level of TLR3 expression was assessed by qPCR in macrophages transfected with siRNA as shown and treated with the WNV-E protein (30 ng/10<sup>6</sup> cells) for 1 h (<i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for mock-treated vs. DC-SIGN-treated cells).</div>
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data-slide-index="3"
data-label-slug="fig-4">
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itemprop="contentUrl"
aria-describedby="figure-caption-3"
role="button"
data-image-width="560"
data-image-height="376"
data-image-alt="FIG. 4."
data-pmc-id="PMC2493309"
data-figure-id="f4">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-3"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/598e211b73ec/zjv0150808450004.gif"
alt="FIG. 4." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="560">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="376">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG. 4. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between DC-SIGN and WNV-E… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 4. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between DC-SIGN and WNV-E attenuates STAT1 activity through Lyn in a Jak-independent… </p></div>
</div>
<figcaption id="figure-caption-3" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 4.
</strong>
<div class="figure-caption-contents">Interaction between DC-SIGN and WNV-E attenuates STAT1 activity through Lyn in a Jak-independent pathway. (A) Primary macrophages derived from young and older individuals were treated with the WNV-E protein at a concentration of 30 ng/10<sup>6</sup> cells. Total proteins were harvested at 0, 5, 10, 20, and 30 min. Cell lysates were immunoprecipitated (IP) with anti-DC-SIGN monoclonal antibody, and the products were immunoblotted with antibodies against phospho-Lyn (Tyr507), phospho-Src (Tyr416), and total Lyn. DC-SIGN was used as a control to ensure equal loading. Data are representative of three subjects from each cohort. (B) Lysates of infected macrophages were immunoblotted with monoclonal antibodies against phospho-STAT1 (Tyr701), phospho-STAT1 (Ser727), total STAT1, phosphor-Jak1 (Tyr1022/1023), total Jak1, phospho-Tyk2 (Tyr1054/1055) and Tyk2. Data are representative of three subjects from each cohort. (C) Primary macrophages derived from young and older individuals were treated with the WNV strain NY99-E154 at an MOI of 1. Total proteins were harvested at 0, 5, 10, 20, and 30 min. Cell lysates were immunoblotted with monoclonal antibodies as for panel B. Data are representative of three subjects in each cohort.</div>
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data-slide-index="4"
data-label-slug="fig-5">
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itemprop="contentUrl"
aria-describedby="figure-caption-4"
role="button"
data-image-width="500"
data-image-height="1636"
data-image-alt="FIG. 5."
data-pmc-id="PMC2493309"
data-figure-id="f5">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-4"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/a0c007810b13/zjv0150808450005.gif"
alt="FIG. 5." />
</a>
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<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG. 5. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between DC-SIGN and WNV… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 5. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction between DC-SIGN and WNV reduces the induction of antiviral response in an… </p></div>
</div>
<figcaption id="figure-caption-4" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 5.
</strong>
<div class="figure-caption-contents">Interaction between DC-SIGN and WNV reduces the induction of antiviral response in an age-dependent manner. (A) Primary macrophages derived from young and older individuals were incubated with the WNV-E protein at a concentration of 30 ng/10<sup>6</sup> cells for 1 h. Chromatin immunoprecipitation of STAT1 was performed as described in Materials and Methods. The immunoprecipitated products are referred to as output, and samples collected prior to immunoprecipitation are noted as input. Differential binding in the absence and presence of WNV-E was determined by PCR using primers specific to the promoters of TLR3, IRF1, and ICAM1 (Table 1). Data are representative of three subjects from each cohort. (B) The expression of the STAT1-regulated RNaseL, CIITA, and OAS1 genes was measured by qPCR using RNA obtained from primary macrophages of healthy young and older individuals infected with WNV (MOI, 0, 0.1, 1, and 10) for 1 h. Stimulation in the presence of 1 ng/ml of IFN-γ was used as a positive control (young, <i>n</i> = 7; elderly, <i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for the young vs. the elderly at the MOI indicated).</div>
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data-label-slug="fig-6">
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itemprop="contentUrl"
aria-describedby="figure-caption-5"
role="button"
data-image-width="560"
data-image-height="412"
data-image-alt="FIG. 6."
data-pmc-id="PMC2493309"
data-figure-id="f6">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-5"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/f87c322d41a2/zjv0150808450006.gif"
alt="FIG. 6." />
</a>
<meta itemprop="width" itemtype="http://schema.org/ImageObject" content="560">
<meta itemprop="height" itemtype="http://schema.org/ImageObject" content="412">
<div class="figure-caption figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="caption" aria-hidden="true">
<div class="caption-wrap">
<strong class="figure-label">
<p> FIG. 6. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction of DC-SIGN and WNV-E… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 6. </p>
</strong>
<div class="figure-caption-contents"><p> Interaction of DC-SIGN and WNV-E prevents STAT1-regulated cytokine release in the young but… </p></div>
</div>
<figcaption id="figure-caption-5" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 6.
</strong>
<div class="figure-caption-contents">Interaction of DC-SIGN and WNV-E prevents STAT1-regulated cytokine release in the young but not in the elderly. (A) Primary macrophages from healthy young and older individuals were infected with WNV (MOI, 1) for 0, 1, and 3 h. Total proteins were harvested and immunoblotted with monoclonal antibodies against phospho-STAT1 (Tyr701), phospho-STAT1 (Ser727), total STAT1, phospho-Jak1 (Tyr1022/1023), total Jak1, phospho-Tyk2 (Tyr1054/1055), and total Tyk2. Data are representative of three subjects from each cohort. (B) Primary macrophages from young and older individuals were incubated in the absence and presence of WNV-E (30 ng/10<sup>6</sup> cells) for 3 h prior to fixation. Fixed cells were stained with specific antibody against STAT1 (red) and the nuclear stain TO-PRO3 (blue). White arrowheads indicate the nuclear translocation of STAT1 when cells were treated with the WNV-E protein. Cells were imaged by confocal microscopy, and images were recorded at a magnification of ×63. Data are representative of at least three subjects from each cohort. (C) Primary macrophages from young and older individuals were infected with WNV (MOI, 1) for 3 h. The mRNA levels of IL-8, TNF-α, IL-6, IFN-β1, and WNV-E and protein levels of IL-8, TNF-α, and IL-6 cytokines were measured by qPCR, using total RNA, and ELISA, using culture supernatants (young, <i>n</i> = 6; elderly, <i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for the young vs. the elderly for the cytokine indicated).</div>
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data-label-slug="fig-7">
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itemprop="contentUrl"
aria-describedby="figure-caption-6"
role="button"
data-image-width="664"
data-image-height="388"
data-image-alt="FIG. 7."
data-pmc-id="PMC2493309"
data-figure-id="f7">
<img class="figure-thumb" itemprop="thumbnail"
id="article-image-6"
src="https://cdn.ncbi.nlm.nih.gov/pmc/blobs/86d0/2493309/70fc46e442df/zjv0150808450007.gif"
alt="FIG. 7." />
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<strong class="figure-label">
<p> FIG. 7. </p>
</strong>
<div class="figure-caption-contents"><p> Infection with WNV produces significantly… </p></div>
</div>
</div>
<div class="figure-caption-medium figure-caption-text" aria-hidden="true">
<strong class="figure-label">
<p> FIG. 7. </p>
</strong>
<div class="figure-caption-contents"><p> Infection with WNV produces significantly higher levels of cytokines from macrophages of the… </p></div>
</div>
<figcaption id="figure-caption-6" class="figure-caption-full figure-caption-text" itemtype="http://schema.org/ImageObject" itemprop="description">
<strong class="figure-label">
FIG. 7.
</strong>
<div class="figure-caption-contents">Infection with WNV produces significantly higher levels of cytokines from macrophages of the elderly than from those of the young. Primary macrophages from young and older individuals were infected with WNV (MOI, 1) over the course of 3 days, and cytokines were measured by qPCR using total RNA (A) and by ELISA using culture supernatants (B) (young, <i>n</i> = 7; elderly, <i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for macrophages of the young vs. the elderly for the time points indicated). (C) The infection rate of primary macrophages represented by the WNV-E gene level and TLR3 level were measured using qPCR. Poly(I:C) was used as a positive control (young, <i>n</i> = 7, elderly, <i>n</i> = 7; data are means ± standard errors of the means; **, ANOVA, <i>P</i> of &lt;0.05 for macrophages of the young vs. the elderly at the time points indicated).</div>
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