Synchronous infection of SIV and HIV in vitro for virology, immunology and vaccine-related studies

被引:28
作者
Sacha, Jonah B. [1 ]
Watkins, David I. [1 ]
机构
[1] Univ Wisconsin, Dept Pathol & Lab Med, Madison, WI 53706 USA
关键词
CD8(+) T-CELLS; VIRAL PROTEIN; VIRUS; RECOGNIZE; MAGNETOFECTION; REPLICATION; LYMPHOCYTES; CULTURES; ESCAPE;
D O I
10.1038/nprot.2009.227
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
the development of an HIV vaccine will require a more precise understanding of the immunological and virological underpinnings of HIV infection. Magnetofection, the process of magnetizing HIV by coupling it to ferrous nanoparticles and coordinating infection using a magnetic field, synchronizes the viral replication cycle at attachment while recapitulating the events of natural infection. although spinoculation also concentrates virus onto target cells to increase infection, it does not synchronize infection. the synchronization of HIV infection in vitro facilitates the study of events in the viral replication cycle and the antiviral immune response on timelines previously impossible. Furthermore, by infecting a high percentage of cells in a short time frame, magnetofection increases the throughput of in vitro assays. once a virus stock is generated, magnetofection of target cells is rapid, requiring only 1-2 h. Here we present a detailed protocol for this assay and review its applications for studying the immune response to HIV.
引用
收藏
页码:239 / 246
页数:8
相关论文
共 18 条
[1]   Efficient transfection of DNA or shRNA vectors into neurons using magnetofection [J].
Buerli, Thomas ;
Pellegrino, Christophe ;
Baer, Kristin ;
Lardi-Studler, Barbara ;
Chudotvorova, Ilona ;
Fritschy, Jean-Marc ;
Medina, Igor ;
Fuhrer, Christian .
NATURE PROTOCOLS, 2007, 2 (12) :3090-3101
[2]   Synchronized infection of cell cultures by magnetically controlled virus [J].
Haim, H ;
Steiner, I ;
Panet, A .
JOURNAL OF VIROLOGY, 2005, 79 (01) :622-625
[3]   Time frames for neutralization during the human immunodeficiency virus type 1 entry phase, as monitored in synchronously infected cell cultures [J].
Haim, Hillel ;
Steiner, Israel ;
Panet, Amos .
JOURNAL OF VIROLOGY, 2007, 81 (07) :3525-3534
[4]   Soluble CD4 and CD4-Mimetic Compounds Inhibit HIV-1 Infection by Induction of a Short-Lived Activated State [J].
Haim, Hillel ;
Si, Zhihai ;
Madani, Navid ;
Wang, Liping ;
Courter, Joel R. ;
Princiotto, Amy ;
Kassa, Aemro ;
DeGrace, Marciella ;
McGee-Estrada, Kathleen ;
Mefford, Megan ;
Gabuzda, Dana ;
Smith, Amos B., III ;
Sodroski, Joseph .
PLOS PATHOGENS, 2009, 5 (04)
[5]   The antiviral efficacy of simian immunodeficiency virus-specific CD8+ T cells is unrelated to epitope specificity and is abrogated by viral escape [J].
Loffredo, John T. ;
Burwitz, Benjamin J. ;
Rakasz, Eva G. ;
Spencer, Sean P. ;
Stephany, Jason J. ;
Vela, Juan Pablo Giraldo ;
Martin, Sarah R. ;
Reed, Jason ;
Piaskowski, Shari M. ;
Furlott, Jessica ;
Weisgrau, Kim L. ;
Rodrigues, Denise S. ;
Soma, Taeko ;
Napoe, Gnankang ;
Friedrich, Thomas C. ;
Wilson, Nancy A. ;
Kallas, Esper G. ;
Watkins, David I. .
JOURNAL OF VIROLOGY, 2007, 81 (06) :2624-2634
[6]   AIDS virus-specific CD8+ T lymphocytes against an immunodominant cryptic epitope select for viral escape [J].
Maness, Nicholas J. ;
Valentine, Laura E. ;
May, Gemma E. ;
Reed, Jason ;
Piaskowski, Shari M. ;
Soma, Taeko ;
Furlott, Jessica ;
Rakasz, Eva G. ;
Friedrich, Thomas C. ;
Price, David A. ;
Gostick, Emma ;
Hughes, Austin L. ;
Sidney, John ;
Sette, Alessandro ;
Wilson, Nancy A. ;
Watkins, David I. .
JOURNAL OF EXPERIMENTAL MEDICINE, 2007, 204 (11) :2505-2512
[7]   Lytic Granule Loading of CD8+ T Cells Is Required for HIV-Infected Cell Elimination Associated with Immune Control [J].
Migueles, Stephen A. ;
Osborne, Christine M. ;
Royce, Cassandra ;
Compton, Alex A. ;
Joshi, Rohan P. ;
Weeks, Kristin A. ;
Rood, Julia E. ;
Berkley, Amy M. ;
Sacha, Jonah B. ;
Cogliano-Shutta, Nancy A. ;
Lloyd, Margaret ;
Roby, Gregg ;
Kwan, Richard ;
McLaughlin, Mary ;
Stallings, Sara ;
Rehm, Catherine ;
O'Shea, Marie A. ;
Mican, JoAnn ;
Packard, Beverly Z. ;
Komoriya, Akira ;
Palmer, Sarah ;
Wiegand, Ann P. ;
Maldarelli, Frank ;
Coffin, John M. ;
Mellors, John W. ;
Hallahan, Claire W. ;
Follman, Dean A. ;
Connors, Mark .
IMMUNITY, 2008, 29 (06) :1009-1021
[8]   The Mamu B*17-restricted SIV NefIW9 to TW9 mutation abrogates correct epitope processing and presentation without loss of replicative fitness [J].
Minang, Jacob T. ;
Trivett, Matthew T. ;
Coren, Lori V. ;
Barsov, Eugene V. ;
Piatak, Michael, Jr. ;
Chertov, Oleg ;
Chertova, Elena ;
Ott, David E. ;
Ohlen, Claes .
VIROLOGY, 2008, 375 (01) :307-314
[9]   Efficient inhibition of SIV replication in rhesus CD4+ T-cell clones by autologous immortalized SIV-specific CD8+ T-cell clones [J].
Minang, Jacob T. ;
Barsov, Eugene V. ;
Yuan, Fang ;
Trivett, Matthew T. ;
Piatak, Michael, Jr. ;
Lifson, Jeffirey D. ;
Ott, David E. ;
Ohlen, Claes .
VIROLOGY, 2008, 372 (02) :430-441
[10]   Nef-mediated MHC class I down-regulation unmasks clonal differences in virus suppression by SIV-specific CD8+ T cells independent of IFN-γ and CD107a responses [J].
Minang, Jacob T. ;
Trivett, Matthew T. ;
Coren, Lori V. ;
Barsov, Eugene V. ;
Piatak, Michael, Jr. ;
Ott, David E. ;
Ohlen, Claes .
VIROLOGY, 2009, 391 (01) :130-139