Ex vivo analysis of SIV‐infected cells by flow cytometry
Deciphering the complex interactions between human and simian immunodeficiency viruses (HIV/SIV) and their host cells is crucial to the development of improved therapies and vaccines. Investigating these relationships has been complicated by the inability to directly analyze infected cells among fre...
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| Veröffentlicht in: | Cytometry. Part A 2010-11, Vol.77A (11), p.1059-1066 |
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| container_title | Cytometry. Part A |
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| creator | Reynolds, Matthew R Piaskowski, Shari M Weisgrau, Kimberly L Weiler, Andrea M Friedrich, Thomas C Rakasz, Eva G |
| description | Deciphering the complex interactions between human and simian immunodeficiency viruses (HIV/SIV) and their host cells is crucial to the development of improved therapies and vaccines. Investigating these relationships has been complicated by the inability to directly analyze infected cells among freshly isolated peripheral blood lymphocytes. Here, we describe a method to detect cells productively infected with SIVmac239 ex vivo from the blood or lymph nodes by flow cytometry. Using this method, we show a close correlation between the frequency of productively infected cells in both sample type and the plasma viral load. We define that the minimum threshold for detecting productively infected cells in lymph nodes by flow cytometry requires a plasma virus concentration of ∼2.5 × 10⁴ vRNA copy Equivalents (Eq)/ml. Conversely, an approximately 2 logs higher plasma viral load is needed to detect productively infected cells in the peripheral blood. This novel protocol provides a direct analytical tool to assess interactions between SIV and host cells, which is of key importance to investigators in AIDS research. © 2010 International Society for Advancement of Cytometry. |
| doi_str_mv | 10.1002/cyto.a.20960 |
| format | Article |
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Investigating these relationships has been complicated by the inability to directly analyze infected cells among freshly isolated peripheral blood lymphocytes. Here, we describe a method to detect cells productively infected with SIVmac239 ex vivo from the blood or lymph nodes by flow cytometry. Using this method, we show a close correlation between the frequency of productively infected cells in both sample type and the plasma viral load. We define that the minimum threshold for detecting productively infected cells in lymph nodes by flow cytometry requires a plasma virus concentration of ∼2.5 × 10⁴ vRNA copy Equivalents (Eq)/ml. Conversely, an approximately 2 logs higher plasma viral load is needed to detect productively infected cells in the peripheral blood. 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Part A</title><addtitle>Cytometry A</addtitle><description>Deciphering the complex interactions between human and simian immunodeficiency viruses (HIV/SIV) and their host cells is crucial to the development of improved therapies and vaccines. Investigating these relationships has been complicated by the inability to directly analyze infected cells among freshly isolated peripheral blood lymphocytes. Here, we describe a method to detect cells productively infected with SIVmac239 ex vivo from the blood or lymph nodes by flow cytometry. Using this method, we show a close correlation between the frequency of productively infected cells in both sample type and the plasma viral load. We define that the minimum threshold for detecting productively infected cells in lymph nodes by flow cytometry requires a plasma virus concentration of ∼2.5 × 10⁴ vRNA copy Equivalents (Eq)/ml. Conversely, an approximately 2 logs higher plasma viral load is needed to detect productively infected cells in the peripheral blood. 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Piaskowski, Shari M ; Weisgrau, Kimberly L ; Weiler, Andrea M ; Friedrich, Thomas C ; Rakasz, Eva G</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c4840-f62f295187741460302b2b16ccfbea080d4be2644da3e4df8ed0a8982aed141c3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2010</creationdate><topic>Acquired immune deficiency syndrome</topic><topic>Animals</topic><topic>Cell Separation</topic><topic>ex vivo detection</topic><topic>Flow cytometry</topic><topic>Flow Cytometry - methods</topic><topic>Host-Pathogen Interactions - physiology</topic><topic>Human immunodeficiency virus</topic><topic>Immunodeficiency</topic><topic>Limit of Detection</topic><topic>Lymph nodes</topic><topic>Lymph Nodes - pathology</topic><topic>Lymph Nodes - virology</topic><topic>Lymphocytes</topic><topic>Lymphocytes - pathology</topic><topic>Lymphocytes - virology</topic><topic>Macaca mulatta</topic><topic>Peripheral blood</topic><topic>RNA, Viral - analysis</topic><topic>Simian Acquired Immunodeficiency Syndrome - blood</topic><topic>Simian Acquired Immunodeficiency Syndrome - pathology</topic><topic>Simian immunodeficiency virus</topic><topic>Simian Immunodeficiency Virus - genetics</topic><topic>Simian Immunodeficiency Virus - isolation & purification</topic><topic>SIV infected cells</topic><topic>Vaccines</topic><topic>Viral Load</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Reynolds, Matthew R</creatorcontrib><creatorcontrib>Piaskowski, Shari M</creatorcontrib><creatorcontrib>Weisgrau, Kimberly L</creatorcontrib><creatorcontrib>Weiler, Andrea M</creatorcontrib><creatorcontrib>Friedrich, Thomas C</creatorcontrib><creatorcontrib>Rakasz, Eva G</creatorcontrib><collection>AGRIS</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Virology and AIDS Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>AIDS and Cancer Research Abstracts</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><collection>PubMed Central (Full Participant titles)</collection><jtitle>Cytometry. Part A</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Reynolds, Matthew R</au><au>Piaskowski, Shari M</au><au>Weisgrau, Kimberly L</au><au>Weiler, Andrea M</au><au>Friedrich, Thomas C</au><au>Rakasz, Eva G</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>Ex vivo analysis of SIV‐infected cells by flow cytometry</atitle><jtitle>Cytometry. Part A</jtitle><addtitle>Cytometry A</addtitle><date>2010-11</date><risdate>2010</risdate><volume>77A</volume><issue>11</issue><spage>1059</spage><epage>1066</epage><pages>1059-1066</pages><issn>1552-4922</issn><issn>1552-4930</issn><eissn>1552-4930</eissn><abstract>Deciphering the complex interactions between human and simian immunodeficiency viruses (HIV/SIV) and their host cells is crucial to the development of improved therapies and vaccines. Investigating these relationships has been complicated by the inability to directly analyze infected cells among freshly isolated peripheral blood lymphocytes. Here, we describe a method to detect cells productively infected with SIVmac239 ex vivo from the blood or lymph nodes by flow cytometry. Using this method, we show a close correlation between the frequency of productively infected cells in both sample type and the plasma viral load. We define that the minimum threshold for detecting productively infected cells in lymph nodes by flow cytometry requires a plasma virus concentration of ∼2.5 × 10⁴ vRNA copy Equivalents (Eq)/ml. Conversely, an approximately 2 logs higher plasma viral load is needed to detect productively infected cells in the peripheral blood. 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| subjects | Acquired immune deficiency syndrome Animals Cell Separation ex vivo detection Flow cytometry Flow Cytometry - methods Host-Pathogen Interactions - physiology Human immunodeficiency virus Immunodeficiency Limit of Detection Lymph nodes Lymph Nodes - pathology Lymph Nodes - virology Lymphocytes Lymphocytes - pathology Lymphocytes - virology Macaca mulatta Peripheral blood RNA, Viral - analysis Simian Acquired Immunodeficiency Syndrome - blood Simian Acquired Immunodeficiency Syndrome - pathology Simian immunodeficiency virus Simian Immunodeficiency Virus - genetics Simian Immunodeficiency Virus - isolation & purification SIV infected cells Vaccines Viral Load |
| title | Ex vivo analysis of SIV‐infected cells by flow cytometry |
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