Radwa R Sharaf1, Jonathan Z Li2. 1. Brigham and Women's Hospital, Harvard Medical School, 65 Landsdowne St, Rm 421, Cambridge, Boston, MA, 02139, USA. 2. Brigham and Women's Hospital, Harvard Medical School, 65 Landsdowne St, Rm 421, Cambridge, Boston, MA, 02139, USA. jli@bwh.harvard.edu.
Abstract
PURPOSE OF REVIEW: Despite the success of antiretroviral therapy in suppressing HIV, life-long therapy is required to avoid HIV reactivation from long-lived viral reservoirs. Currently, there is intense interest in searching for therapeutic interventions that can purge the viral reservoir to achieve complete remission in HIV patients off antiretroviral therapy. The evaluation of such interventions relies on our ability to accurately and precisely measure the true size of the viral reservoir. In this review, we assess the most commonly used HIV reservoir assays, as a clear understanding of the strengths and weaknesses of each is vital for the accurate interpretation of results and for the development of improved assays. RECENT FINDINGS: The quantification of intracellular or plasma HIV RNA or DNA levels remains the most commonly used tests for the characterization of the viral reservoir. While cost-effective and high-throughput, these assays are not able to differentiate between replication-competent or defective fractions or quantify the number of infected cells. Viral outgrowth assays provide a lower bound for the fraction of cells that can produce infectious virus, but these assays are laborious, expensive and substantially underestimate the potential reservoir of replication-competent provirus. Newer assays are now available that seek to overcome some of these problems, including full-length proviral sequencing, inducible HIV RNA assays, ultrasensitive p24 assays and murine adoptive transfer techniques. The development and evaluation of strategies for HIV remission rely upon our ability to accurately and precisely quantify the size of the remaining viral reservoir. At this time, all current HIV reservoir assays have drawbacks such that combinations of assays are generally needed to gain a more comprehensive view of the viral reservoir. The development of novel, rapid, high-throughput assays that can sensitively quantify the levels of the replication-competent HIV reservoir is still needed.
PURPOSE OF REVIEW: Despite the success of antiretroviral therapy in suppressing HIV, life-long therapy is required to avoid HIV reactivation from long-lived viral reservoirs. Currently, there is intense interest in searching for therapeutic interventions that can purge the viral reservoir to achieve complete remission in HIV patients off antiretroviral therapy. The evaluation of such interventions relies on our ability to accurately and precisely measure the true size of the viral reservoir. In this review, we assess the most commonly used HIV reservoir assays, as a clear understanding of the strengths and weaknesses of each is vital for the accurate interpretation of results and for the development of improved assays. RECENT FINDINGS: The quantification of intracellular or plasma HIV RNA or DNA levels remains the most commonly used tests for the characterization of the viral reservoir. While cost-effective and high-throughput, these assays are not able to differentiate between replication-competent or defective fractions or quantify the number of infected cells. Viral outgrowth assays provide a lower bound for the fraction of cells that can produce infectious virus, but these assays are laborious, expensive and substantially underestimate the potential reservoir of replication-competent provirus. Newer assays are now available that seek to overcome some of these problems, including full-length proviral sequencing, inducible HIV RNA assays, ultrasensitive p24 assays and murine adoptive transfer techniques. The development and evaluation of strategies for HIV remission rely upon our ability to accurately and precisely quantify the size of the remaining viral reservoir. At this time, all current HIV reservoir assays have drawbacks such that combinations of assays are generally needed to gain a more comprehensive view of the viral reservoir. The development of novel, rapid, high-throughput assays that can sensitively quantify the levels of the replication-competent HIV reservoir is still needed.
Authors: J P Routy; C L Tremblay; J B Angel; B Trottier; D Rouleau; J G Baril; M Harris; S Trottier; J Singer; N Chomont; R P Sékaly; M R Boulassel Journal: HIV Med Date: 2012-01-26 Impact factor: 3.180
Authors: Michael P Busch; Simone A Glynn; David J Wright; Dale Hirschkorn; Megan E Laycock; Joan McAuley; Yongling Tu; Cristina Giachetti; James Gallarda; John Heitman; Steven H Kleinman Journal: Transfusion Date: 2005-12 Impact factor: 3.157
Authors: Adam M Spivak; Adriana Andrade; Evelyn Eisele; Rebecca Hoh; Peter Bacchetti; Namandjé N Bumpus; Fatemeh Emad; Robert Buckheit; Elinore F McCance-Katz; Jun Lai; Margene Kennedy; Geetanjali Chander; Robert F Siliciano; Janet D Siliciano; Steven G Deeks Journal: Clin Infect Dis Date: 2013-12-12 Impact factor: 9.079
Authors: Thomas A Rasmussen; Martin Tolstrup; Christel R Brinkmann; Rikke Olesen; Christian Erikstrup; Ajantha Solomon; Anni Winckelmann; Sarah Palmer; Charles Dinarello; Maria Buzon; Mathias Lichterfeld; Sharon R Lewin; Lars Østergaard; Ole S Søgaard Journal: Lancet HIV Date: 2014-09-15 Impact factor: 12.767
Authors: Carolina Gutiérrez; Sergio Serrano-Villar; Nadia Madrid-Elena; Maria J Pérez-Elías; Maria Elena Martín; Coral Barbas; Javier Ruipérez; Eduardo Muñoz; Maria Angeles Muñoz-Fernández; Trevor Castor; Santiago Moreno Journal: AIDS Date: 2016-06-01 Impact factor: 4.177
Authors: Ole S Søgaard; Mette E Graversen; Steffen Leth; Rikke Olesen; Christel R Brinkmann; Sara K Nissen; Anne Sofie Kjaer; Mariane H Schleimann; Paul W Denton; William J Hey-Cunningham; Kersten K Koelsch; Giuseppe Pantaleo; Kim Krogsgaard; Maja Sommerfelt; Remi Fromentin; Nicolas Chomont; Thomas A Rasmussen; Lars Østergaard; Martin Tolstrup Journal: PLoS Pathog Date: 2015-09-17 Impact factor: 6.823
Authors: Zachary Strongin; Radwa Sharaf; D Jake VanBelzen; Jeffrey M Jacobson; Elizabeth Connick; Paul Volberding; Daniel J Skiest; Rajesh T Gandhi; Daniel R Kuritzkes; Una O'Doherty; Jonathan Z Li Journal: J Virol Date: 2018-05-29 Impact factor: 5.103
Authors: Alexander O Pasternak; Marlous L Grijsen; Ferdinand W Wit; Margreet Bakker; Suzanne Jurriaans; Jan M Prins; Ben Berkhout Journal: JCI Insight Date: 2020-03-26
Authors: Mykola Pinkevych; Christine M Fennessey; Deborah Cromer; Carolyn Reid; Charles M Trubey; Jeffrey D Lifson; Brandon F Keele; Miles P Davenport Journal: Elife Date: 2019-10-25 Impact factor: 8.140
Authors: Maria C Puertas; Ángel Bayón-Gil; Maria C Garcia-Guerrero; Maria Salgado; Víctor Urrea; Sara Morón-López; Ruth Peña; Esther Jiménez-Moyano; Bonaventura Clotet; Julia G Prado; Javier Martinez-Picado Journal: mBio Date: 2021-06-22 Impact factor: 7.867