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<record><TEI><teiHeader><fileDesc><titleStmt><title xml:lang="en">CD63 Is Not Required for Production of Infectious Human Immunodeficiency Virus Type 1 in Human Macrophages<xref ref-type="fn" rid="fn2">▿</xref> </title><author><name sortKey="Ruiz Mateos, Ezequiel" sort="Ruiz Mateos, Ezequiel" uniqKey="Ruiz Mateos E" first="Ezequiel" last="Ruiz-Mateos">Ezequiel Ruiz-Mateos</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Pelchen Matthews, Annegret" sort="Pelchen Matthews, Annegret" uniqKey="Pelchen Matthews A" first="Annegret" last="Pelchen-Matthews">Annegret Pelchen-Matthews</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Deneka, Magdalena" sort="Deneka, Magdalena" uniqKey="Deneka M" first="Magdalena" last="Deneka">Magdalena Deneka</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Marsh, Mark" sort="Marsh, Mark" uniqKey="Marsh M" first="Mark" last="Marsh">Mark Marsh</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author></titleStmt><publicationStmt><idno type="wicri:source">PMC</idno><idno type="pmid">18321974</idno><idno type="pmc">2346747</idno><idno type="url">http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2346747</idno><idno type="RBID">PMC:2346747</idno><idno type="doi">10.1128/JVI.02320-07</idno><date when="2008">2008</date><idno type="wicri:Area/Pmc/Corpus">000178</idno><idno type="wicri:explorRef" wicri:stream="Pmc" wicri:step="Corpus" wicri:corpus="PMC">000178</idno></publicationStmt><sourceDesc><biblStruct><analytic><title xml:lang="en" level="a" type="main">CD63 Is Not Required for Production of Infectious Human Immunodeficiency Virus Type 1 in Human Macrophages<xref ref-type="fn" rid="fn2">▿</xref> </title><author><name sortKey="Ruiz Mateos, Ezequiel" sort="Ruiz Mateos, Ezequiel" uniqKey="Ruiz Mateos E" first="Ezequiel" last="Ruiz-Mateos">Ezequiel Ruiz-Mateos</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Pelchen Matthews, Annegret" sort="Pelchen Matthews, Annegret" uniqKey="Pelchen Matthews A" first="Annegret" last="Pelchen-Matthews">Annegret Pelchen-Matthews</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Deneka, Magdalena" sort="Deneka, Magdalena" uniqKey="Deneka M" first="Magdalena" last="Deneka">Magdalena Deneka</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author><author><name sortKey="Marsh, Mark" sort="Marsh, Mark" uniqKey="Marsh M" first="Mark" last="Marsh">Mark Marsh</name><affiliation><nlm:aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</nlm:aff></affiliation></author></analytic><series><title level="j">Journal of Virology</title><idno type="ISSN">0022-538X</idno><idno type="eISSN">1098-5514</idno><imprint><date when="2008">2008</date></imprint></series></biblStruct></sourceDesc></fileDesc><profileDesc><textClass></textClass></profileDesc></teiHeader><front><div type="abstract" xml:lang="en"><p>During the assembly of human immunodeficiency virus type 1 (HIV-1) particles, the tetraspanin CD63 can be incorporated into the viral membrane. Indeed, cell surface tetraspanin microdomains that include CD63 have been proposed as sites for virus release. In addition, antibodies against CD63 can inhibit HIV infection of macrophages. In this cell type, HIV assembles into intracellularly sequestered plasma membrane domains that contain several other tetraspanins, including CD81, CD9, and CD53. CD63 is recruited to this domain following HIV infection. Together, these observations suggest that CD63 may have some function in the assembly of infectious virus particles and/or the infectivity of assembled virions. Here we have used RNA interference to knock down CD63 expression in monocyte-derived primary macrophages. We show that in the absence of CD63, HIV assembly is quantitatively comparable to that seen in CD63-expressing macrophages and that virus assembly occurs on compartments positive for CD81, CD9, and CD53. Moreover, the infectivity of macrophage-derived virus is unaffected by the loss of CD63. Together, our results indicate that at least in tissue culture, CD63 expression is not required for either the production or the infectivity of HIV-1.</p></div></front></TEI><pmc article-type="research-article"><pmc-comment>The publisher of this article does not allow downloading of the full text in XML form.</pmc-comment>
<front><journal-meta><journal-id journal-id-type="nlm-ta">J Virol</journal-id><journal-id journal-id-type="publisher-id">jvi</journal-id><journal-title>Journal of Virology</journal-title><issn pub-type="ppub">0022-538X</issn><issn pub-type="epub">1098-5514</issn><publisher><publisher-name>American Society for Microbiology (ASM)</publisher-name></publisher></journal-meta><article-meta><article-id pub-id-type="pmid">18321974</article-id><article-id pub-id-type="pmc">2346747</article-id><article-id pub-id-type="publisher-id">2320-07</article-id><article-id pub-id-type="doi">10.1128/JVI.02320-07</article-id><article-categories><subj-group subj-group-type="heading"><subject>Structure and Assembly</subject></subj-group></article-categories><title-group><article-title>CD63 Is Not Required for Production of Infectious Human Immunodeficiency Virus Type 1 in Human Macrophages<xref ref-type="fn" rid="fn2">▿</xref> </article-title></title-group><contrib-group><contrib contrib-type="author"><name><surname>Ruiz-Mateos</surname><given-names>Ezequiel</given-names></name><xref ref-type="aff" rid="aff0"></xref><xref ref-type="fn" rid="fn1">†</xref></contrib><contrib contrib-type="author"><name><surname>Pelchen-Matthews</surname><given-names>Annegret</given-names></name><xref ref-type="aff" rid="aff0"></xref></contrib><contrib contrib-type="author"><name><surname>Deneka</surname><given-names>Magdalena</given-names></name><xref ref-type="aff" rid="aff0"></xref></contrib><contrib contrib-type="author"><name><surname>Marsh</surname><given-names>Mark</given-names></name><xref ref-type="aff" rid="aff0"></xref><xref ref-type="corresp" rid="cor1">*</xref></contrib></contrib-group><aff id="aff0">Cell Biology Unit, MRC Laboratory for Molecular Cell Biology and Department of Cell and Developmental Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom</aff><author-notes><fn id="cor1"><label>*</label><p>Corresponding author. Mailing address: MRC Laboratory for Molecular Cell Biology, University College London, Gower Street, London WC1E 6BT, United Kingdom. Phone: 44 20 7679 7807. Fax: 44 20 7679 7805. E-mail: <email>m.marsh@ucl.ac.uk</email></p></fn><fn id="fn1"><label>†</label><p>Present address: Laboratorio de Inmunovirología. Servicio de Enfermedades Infecciosas, Hospitales Universitarios Virgen del Rocio, Avd. Manuel Siurot s/n, 41013 Sevilla, Spain.</p></fn></author-notes><pub-date pub-type="ppub"><month>5</month><year>2008</year></pub-date><pub-date pub-type="epub"><day>5</day><month>3</month><year>2008</year></pub-date><volume>82</volume><issue>10</issue><fpage>4751</fpage><lpage>4761</lpage><history><date date-type="received"><day>26</day><month>10</month><year>2007</year></date><date date-type="accepted"><day>21</day><month>2</month><year>2008</year></date></history><permissions><copyright-statement>Copyright © 2008, American Society for Microbiology</copyright-statement></permissions><self-uri xlink:title="pdf" xlink:href="zjv01008004751.pdf"></self-uri><abstract><p>During the assembly of human immunodeficiency virus type 1 (HIV-1) particles, the tetraspanin CD63 can be incorporated into the viral membrane. Indeed, cell surface tetraspanin microdomains that include CD63 have been proposed as sites for virus release. In addition, antibodies against CD63 can inhibit HIV infection of macrophages. In this cell type, HIV assembles into intracellularly sequestered plasma membrane domains that contain several other tetraspanins, including CD81, CD9, and CD53. CD63 is recruited to this domain following HIV infection. Together, these observations suggest that CD63 may have some function in the assembly of infectious virus particles and/or the infectivity of assembled virions. Here we have used RNA interference to knock down CD63 expression in monocyte-derived primary macrophages. We show that in the absence of CD63, HIV assembly is quantitatively comparable to that seen in CD63-expressing macrophages and that virus assembly occurs on compartments positive for CD81, CD9, and CD53. Moreover, the infectivity of macrophage-derived virus is unaffected by the loss of CD63. Together, our results indicate that at least in tissue culture, CD63 expression is not required for either the production or the infectivity of HIV-1.</p></abstract></article-meta></front><floats-wrap><fig position="float" id="f1"><label>FIG. 1.</label><caption><p>CD63 knockdown in MDM. Seven days after monocyte isolation (day 0), MDM were nucleofected with scrambled or CD63-specific siRNAs. CD63 knockdown was assessed by immunofluorescence and by Western blot analysis 2, 4, and 6 days after nucleofection. (A) Permeabilized cells were stained with anti-CD63, followed by Alexa Fluor 594 anti-mouse and Hoechst dye for immunofluorescence analysis. Scale bars = 10 μm. (B) For Western blot analysis, 2 × 10<sup>6</sup> MDM were solubilized in lysis buffer. Cell proteins were separated in nonreducing buffer on 10% SDS-polyacrylamide gel electrophoresis gels and immunoblotted for CD63 and clathrin heavy chain (CHC). CD63 runs as a broad smear due to glycosylation (<xref ref-type="bibr" rid="r18">18</xref>). The blots were quantified using Bio-Rad Quantity One software, and band intensities were normalized against clathrin. The loss of CD63 is expressed as a percentage of the CD63 levels in control cells (bottom right in the blots).</p></caption><graphic xlink:href="zjv0100805380001"></graphic></fig><fig position="float" id="f2"><label>FIG. 2.</label><caption><p>Distribution of lysosomal markers in CD63 knockdown MDM. MDM were nucleofected with scrambled or CD63-specific siRNAs and returned to culture for 2 to 12 days. (A) Cells were fixed 6 days after nucleofection, permeabilized, and stained with MAbs against CD63, followed by anti-mouse IgG2b Alexa Fluor 488, and against LAMP-1, followed by anti-mouse IgG1 Alexa Fluor 594. Images show projections of 22 and 23 confocal sections (0.5 μm thick) for the control and CD63 RNAi samples, respectively. Scale bars = 10 μm. Observations similar to those above were made 2 and 4 days after nucleofection. (B) EM immunolabeling of control (panels i and ii) or CD63 knockdown MDM (panels iii and iv) 12 days after nucleofection. Ultrathin cryosections were double labeled with MAbs against LAMP-1/PAG (15 nm) and CD63/PAG (5 nm). Arrows indicate CD63 staining. Scale bars = 200 nm.</p></caption><graphic xlink:href="zjv0100805380002"></graphic></fig><fig position="float" id="f3"><label>FIG. 3.</label><caption><p>CD63 is not required for HIV-1 targeting to the assembly compartment. MDM were nucleofected with scrambled or CD63-specific siRNA as described in the text. Two days later, the cells were infected with HIV-1<sub>BaL</sub>. At 10 dpi the cells were analyzed by immunofluorescence or by EM. (A) For immunofluorescence analysis, cells were fixed, permeabilized, and stained with mouse MAbs against the tetraspanin CD63, CD81, or CD9, followed by Alexa Fluor 594 anti-mouse (red), and costained with rabbit anti-HIV-1 p17 (MA) followed by Alexa Fluor 488 anti-rabbit (green). Single confocal sections are shown. Areas where the markers colocalize appear yellow. Scale bars = 10 μm. (B) For EM analysis, ultrathin cryosections of control (panels i and ii) and CD63 knockdown MDM (panels iii and iv) were double labeled with MAbs against the HIV capsid protein p24 and PAG (15 nm) and with CD63/PAG (5 nm). Black arrows in panels i and iii indicate a virus-filled vacuole. The higher-magnification views in panels ii and iv show mature virus particles as well as immature virions or buds (white arrows). CD63 is incorporated into the HIV particles (arrowheads in panel ii). Scale bars, 1 μm (panels i and iii) and 200 nm (panels ii and iv).</p></caption><graphic xlink:href="zjv0100805380003"></graphic></fig><fig position="float" id="f4"><label>FIG. 4.</label><caption><p>Production of infectious HIV-1 is not affected by CD63 knockdown. MDM were nucleofected with scrambled or CD63-specific siRNA and infected with HIV-1<sub>BaL</sub> as described in the text. (A) CD63 expression levels were assessed by Western blot analysis at 6 and 10 dpi. The clathrin heavy chain (CHC) was analyzed as a loading control. The percentage of knockdown is shown in the relevant lanes. (B to D) Cell-free supernatants from control and CD63 knockdown (KD) HIV-1<sub>BaL</sub>-infected MDM were assayed for p24 levels (B) or infectivity (C), and the number of infectious units per particle were calculated (D). Data from experiments with five different donors were averaged and are represented as relative units after normalizing with respect to control values of p24, infectivity, and infectious units per particle at 6 dpi. For panels B through D, bars represent the average relative units, and error bars represent standard errors of the means. (E) Env and Gag (p24 and pr55) expression levels in control and CD63 knockdown HIV-infected MDM at 6 and 10 dpi were assessed by Western blotting.</p></caption><graphic xlink:href="zjv0100805380004"></graphic></fig><fig position="float" id="f5"><label>FIG. 5.</label><caption><p>Production of infectious HIV-1 is not affected in CD63 knockdown MDM with the same levels of infection. MDM were infected with HIV-1<sub>BaL</sub> 7 days after monocyte isolation (day 0) and nucleofected with scrambled or CD63-specific siRNAs 1 day after infection. Cells were returned to culture in medium containing the entry inhibitor TAK779 to avoid multiple rounds of infection. After 5 days, the medium was changed and virus-containing supernatants were collected 24 h later (i.e., at 6 dpi). (A) CD63 expression levels were assessed by Western blot analysis at 6 dpi. CHC was analyzed as a loading control. KD, CD63 knockdown MDM. (B to D) Cell-free supernatants from CD63 knockdown and control MDM were assayed for p24 levels (B) and infectivity (C), and the number of infectious units per particle were calculated (D). (E) In addition, an <italic>Alu</italic>-LTR-based real-time nested-PCR assay was used to quantify the integrated HIV DNA levels in control and CD63 knockdown MDM. Data corresponding to four experiments with three different donors were averaged and are represented as relative units after normalization with respect to control values at 6 dpi of p24, infectivity, infectious units per particle, or proviral DNA. For panels B through E, bars represent the average relative units, and error bars represent standard errors of the means.</p></caption><graphic xlink:href="zjv0100805380005"></graphic></fig><fig position="float" id="f6"><label>FIG. 6.</label><caption><p>Immunoprecipitation of virus particles from control and CD63 knockdown MDM. Cell-free supernatants from control and CD63 knockdown MDM-derived viruses were incubated at 37°C for 1 h with antibodies against CD63, VSV-G (negative control), and Env (positive control). The virus-antibody mixture was incubated with Pansorbin cells, and viruses were precipitated by centrifugation. The supernatants, containing the unprecipitated viruses, were used to infect TZM-β-gal indicator cells. β-Gal activity was measured 24 h after infection and was expressed as arbitrary units (au). Bars represent averages of values from six different dilutions per sample from a representative experiment. Error bars represent the standard errors of the means. noAb, untreated sample.</p></caption><graphic xlink:href="zjv0100805380006"></graphic></fig><fig position="float" id="f7"><label>FIG. 7.</label><caption><p>Colocalization of the HIV matrix protein with CD63 on HIV-infected MDM. Semithin cryosections from human MDM samples infected with HIV<sub>BaL</sub> for 7 days (A), 14 days (B), or 20 days (C) were stained with a rabbit antiserum against HIV p17 and a mouse MAb to CD63, followed by anti-rabbit Alexa Fluor 594 and anti-mouse Alexa Fluor 488. Sections were examined with an Axioskop fluorescence microscope. The HIV-containing structures (red, left column) costained strongly with CD63 (green, center column), giving a yellow color in the merged images (right column). This distribution was seen for MDM cultures infected for various lengths of time, in single infected cells or syncytia, and for virus-containing compartments of various sizes and morphologies. By contrast, MDM samples infected with a passaged stock of HIV-1<sub>BaL</sub> for 8 days (D) or 13 days (E) did not accumulate CD63 in the virus-containing compartment, which appears red even in the merged images (right column). Scale bars = 10 μm.</p></caption><graphic xlink:href="zjv0100805380007"></graphic></fig><fig position="float" id="f8"><label>FIG. 8.</label><caption><p>Serially passaged HIV-1<sub>BaL</sub> assembles in a CD81- and CD9-containing compartment. Semithin cryosections from MDM infected with the passaged stock of HIV-1<sub>BaL</sub> for 13 days were stained with a rabbit antiserum against HIV p17 and mouse MAbs to CD81 (A) or CD9 (B), followed by anti-rabbit Alexa Fluor 594 and anti-mouse Alexa Fluor 488. The HIV-containing structures (red, left column) costained strongly with CD81 and CD9 (green, center column), giving a yellow color in the merged images (right column). Scale bars = 10 μm.</p></caption><graphic xlink:href="zjv0100805380008"></graphic></fig><table-wrap position="float" id="t1"><label>TABLE 1.</label><caption><p>Analysis of infectious virus production from control and CD63 knockdown MDM at 6 and 10 dpi</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">dpi</th><th colspan="1" rowspan="2" align="center" valign="middle">Experiment no.</th><th colspan="2" rowspan="1" align="center" valign="bottom">p24 level (ng/ml) for:
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Infectivity level (10<sup>5</sup> FFU/ml) for:
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Infectious unit per particle<xref ref-type="table-fn" rid="t1fn2"><italic>b</italic></xref> for:
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD<xref ref-type="table-fn" rid="t1fn1"><italic>a</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD</th><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="char" char="." valign="top">6</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6.21</td><td colspan="1" rowspan="1" align="char" char="." valign="top">7.31</td><td colspan="1" rowspan="1" align="char" char="." valign="top">5.19</td><td colspan="1" rowspan="1" align="char" char="." valign="top">5.54</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0167</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0152</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">2</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.06</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.67</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.32</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.03</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0217</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0165</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">3</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.94</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2.73</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.66</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.89</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0189</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0284</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td><td colspan="1" rowspan="1" align="char" char="." valign="top">13.17</td><td colspan="1" rowspan="1" align="char" char="." valign="top">7.62</td><td colspan="1" rowspan="1" align="char" char="." valign="top">17.40</td><td colspan="1" rowspan="1" align="char" char="." valign="top">8.35</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0264</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0219</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">5</td><td colspan="1" rowspan="1" align="char" char="." valign="top">11.59</td><td colspan="1" rowspan="1" align="char" char="." valign="top">9.60</td><td colspan="1" rowspan="1" align="char" char="." valign="top">7.62</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.96</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0132</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0103</td></tr><tr><td colspan="1" rowspan="1" align="char" char="." valign="top">10</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1</td><td colspan="1" rowspan="1" align="char" char="." valign="top">10.20</td><td colspan="1" rowspan="1" align="char" char="." valign="top">13.40</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.17</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.85</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0062</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0072</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">2</td><td colspan="1" rowspan="1" align="char" char="." valign="top">5.77</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.50</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.86</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.88</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0064</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0083</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">3</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.91</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2.81</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.56</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.16</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0080</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0083</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">4</td><td colspan="1" rowspan="1" align="char" char="." valign="top">10.50</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6.78</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.46</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.88</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0085</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0114</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top"></td><td colspan="1" rowspan="1" align="char" char="." valign="top">5</td><td colspan="1" rowspan="1" align="char" char="." valign="top">13.09</td><td colspan="1" rowspan="1" align="char" char="." valign="top">8.03</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.02</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.03</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0061</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.0100</td></tr></tbody></table><table-wrap-foot><fn id="t1fn1"><label>a</label><p>KD, CD63 knockdown MDM.</p></fn><fn id="t1fn2"><label>b</label><p>The number of infectious units per particle was calculated from the infectivity in FFU/ml and the p24 levels, assuming 5,000 Gag molecules per virion (<xref ref-type="bibr" rid="r4">4</xref>).</p></fn></table-wrap-foot></table-wrap><table-wrap position="float" id="t2"><label>TABLE 2.</label><caption><p>Analysis of infectious virus production from control and CD63 knockdown MDM at 6 dpi</p></caption><table frame="hsides" rules="groups"><thead><tr><th colspan="1" rowspan="2" align="center" valign="middle">Experiment no.</th><th colspan="2" rowspan="1" align="center" valign="bottom">p24 level (ng/ml) for:
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Infectivity level (10<sup>5</sup> FFU/ml) for:
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Infectious unit(s) per particle<xref ref-type="table-fn" rid="t2fn2"><italic>b</italic></xref> (10<sup>−3</sup>)
<hr></hr></th><th colspan="2" rowspan="1" align="center" valign="bottom">Proviral DNA<xref ref-type="table-fn" rid="t2fn3"><italic>c</italic></xref> (au) for:
<hr></hr></th></tr><tr><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD<xref ref-type="table-fn" rid="t2fn1"><italic>a</italic></xref></th><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD</th><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD</th><th colspan="1" rowspan="1" align="center" valign="bottom">Control</th><th colspan="1" rowspan="1" align="center" valign="bottom">KD</th></tr></thead><tbody><tr><td colspan="1" rowspan="1" align="center" valign="top">6</td><td colspan="1" rowspan="1" align="char" char="." valign="top">8.74</td><td colspan="1" rowspan="1" align="char" char="." valign="top">8.13</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.07</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.07</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.15</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.17</td><td colspan="1" rowspan="1" align="char" char="." valign="top">50.3</td><td colspan="1" rowspan="1" align="char" char="." valign="top">41.0</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top">7</td><td colspan="1" rowspan="1" align="char" char="." valign="top">7.58</td><td colspan="1" rowspan="1" align="char" char="." valign="top">7.65</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.22</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.22</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.58</td><td colspan="1" rowspan="1" align="char" char="." valign="top">0.56</td><td colspan="1" rowspan="1" align="char" char="." valign="top">55.9</td><td colspan="1" rowspan="1" align="char" char="." valign="top">45.9</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top">8</td><td colspan="1" rowspan="1" align="char" char="." valign="top">10.55</td><td colspan="1" rowspan="1" align="char" char="." valign="top">9.24</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.97</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.92</td><td colspan="1" rowspan="1" align="char" char="." valign="top">3.74</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.15</td><td colspan="1" rowspan="1" align="char" char="." valign="top">84.0</td><td colspan="1" rowspan="1" align="char" char="." valign="top">83.1</td></tr><tr><td colspan="1" rowspan="1" align="center" valign="top">9</td><td colspan="1" rowspan="1" align="char" char="." valign="top">8.08</td><td colspan="1" rowspan="1" align="char" char="." valign="top">5.97</td><td colspan="1" rowspan="1" align="char" char="." valign="top">1.95</td><td colspan="1" rowspan="1" align="char" char="." valign="top">2.04</td><td colspan="1" rowspan="1" align="char" char="." valign="top">4.84</td><td colspan="1" rowspan="1" align="char" char="." valign="top">6.82</td><td colspan="1" rowspan="1" align="char" char="." valign="top">66.5</td><td colspan="1" rowspan="1" align="char" char="." valign="top">69.3</td></tr></tbody></table><table-wrap-foot><fn id="t2fn1"><label>a</label><p>KD, CD63 knockdown MDM.</p></fn><fn id="t2fn2"><label>b</label><p>The number of infectious units per particle was calculated from the infectivity in FFU/ml and the p24 levels, assuming 5,000 Gag molecules per virion (<xref ref-type="bibr" rid="r4">4</xref>).</p></fn><fn id="t2fn3"><label>c</label><p>Proviral DNA is expressed as arbitrary units (au) based on the ratio of the integrated HIV-1 DNA level to the β-globin gene level.</p></fn></table-wrap-foot></table-wrap></floats-wrap></pmc></record>Pour manipuler ce document sous Unix (Dilib)
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