東華大學圖書館 |

Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies.

紀錄類型:	書目-電子資源 : Monograph/item
正題名/作者:	Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies./
作者:	Wang, Zheng.
出版者:	Ann Arbor : ProQuest Dissertations & Theses, : 2018,
面頁冊數:	76 p.
附註:	Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
Contained By:	Dissertations Abstracts International80-10B.
標題:	Virology. -
電子資源:	http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13890040
ISBN:	9781392067567

Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies.
Wang, Zheng.

Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies. - Ann Arbor : ProQuest Dissertations & Theses, 2018 - 76 p.

Source: Dissertations Abstracts International, Volume: 80-10, Section: B.

Thesis (Ph.D.)--The Johns Hopkins University, 2018.

This item must not be sold to any third party vendors.

Despite effective antiretroviral therapy (ART), HIV-1 persists in all infected individuals as proviral DNA integrated within long-lived resting memory CD4+ T cells. The population of infected CD4+ T cells carrying replication-competent proviruses is the major barrier to HIV-1 cure. Several lines of evidence have demonstrated that cellular proliferation of infected cells contributes to HIV-1 persistence. This proliferative process is complicated by the fact that most infected cells carry defective proviruses and cells harboring replication-competent HIV die quickly upon viral reactivation. To elucidate mechanisms that drive proliferation of HIV-1-infected CD4+ T cells, we followed proliferation of cells carrying replication-competent HIV-1 induced by antigen stimulation or cytokine treatment and demonstrated that latently infected cells carrying replication-competent HIV can proliferate in response to both stimuli. To study the dynamics of cells carrying replication-competent HIV-1, we sampled infectious virus from p24+ wells of the quantitative viral outgrowth assay at multiple time points spanning 2-3 years. Sequencing of replication-competent HIV-1 at multiple time points revealed that expanded cellular clones containing replication-competent HIV-1 is common. While some clones persist for 2-3 years, other clones wax and wane overtime. A similar pattern is observed with virus clones in the residual viremia. This observation with residual viremia supports our hypothesis that viruses in plasma are produced by activation of latently infected cells carrying replication-competent HIV-1 rather than ongoing cycles of virus replication. In addition, this supports our hypothesis that antigens drive proliferation of cells carrying replication-competent HIV-1 and activate some of the cells, leading to virus production. The observed patterns with proviruses in the latent reservoir and viruses in the residual viremia do not support a continuous proliferative process related to HIV-1 integration into cancer-associated genes. These studies are also being extended to cells carrying defective proviruses. A previous study has demonstrated that defective HIV-1 proviruses can be transcribed, translated or even recognized by HIV-1 specific CD8+ T cells. We hypothesized that defective proviruses may have a proliferative advantage that allows the cells carrying defective proviruses to proliferate more than cells carrying intact proviruses. Given that intact proviruses only account for 2% of total proviruses in patients on long-term suppressive ART, infected cells carrying defective proviruses greatly outnumber the cells harboring intact proviruses in vivo. We hypothesized that in vitro, cells carrying defective proviruses would proliferate upon T cell activation while cells harboring intact proviruses capable of viral gene expression, would die upon T cell activation due to viral cytopathic effects. Therefore, cells carrying either defective proviruses and or intact proviruses would show different proliferation dynamics upon TCR activation in vitro. To determine whether cells infected with intact or defective proviruses would proliferate to the same extent, we subjected single HIV-1 infected cells to 4 rounds of anti-CD3/CD28 stimulation in a microculture system and used a novel droplet digital PCR assay (intact proviral DNA assay; IPDA) to quantitate the number of intact and defective proviral DNA sequences in infected cells. We demonstratethat cells carrying defective proviruses were capable of enormous expansion with in vitro anti-CD3/CD28 stimulation, while cells harboring intact proviruses were rarely detected and showed little proliferative potential. Integration site analysis of clones expanded in vitro demonstrated that HIV-1 provirus integration into cancer-associated gene is not required for proliferation of HIV-1-infected cells. Additionally, we sequenced the cell clones that proliferated the most in vitro and found that proviruses in these clones were highly defective. These microculture experiments revealed a profound proliferation defect for cells carrying intact proviruses upon anti-CD3/CD28 stimulation. To explore whether cells carrying intact and defective proviruses show similar dynamics in vivo, we examined longitudinal samples collected 2-8 years apart using the IPDA. We found that the half-life of infected was ~44 months, consistent with a previous measurements of the latent reservoir as measured with a quantitative viral outgrowth assay (QVOA), while cells carrying defective proviruses showed greater variability among patients. Collectively, my thesis has measured the dynamics of CD4+ T cells carrying different types of proviruses and has provided insight into mechanisms that may contribute to proliferation of HIV-infected cells in vivo.

ISBN: 9781392067567Subjects--Topical Terms:

642304
Virology.
Subjects--Index Terms:

Clonal expansion

Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies.
LDR:06047nmm a2200361 4500 001 2267607
005 20200724103012.5
008 220629s2018 ||||||||||||||||| ||eng d
020 $a 9781392067567
035 $a (MiAaPQ)AAI13890040
035 $a (MiAaPQ)0098vireo:3998Wang
035 $a AAI13890040
040 $a MiAaPQ $c MiAaPQ
100 1 $a Wang, Zheng. $3 660235
245 1 0 $a Dynamics of HIV-Infected CD4+ T Cells: Implications for Cure Strategies.
260 1 $a Ann Arbor : $b ProQuest Dissertations & Theses, $c 2018
300 $a 76 p.
500 $a Source: Dissertations Abstracts International, Volume: 80-10, Section: B.
500 $a Publisher info.: Dissertation/Thesis.
500 $a Advisor: Siliciano, Robert F.
502 $a Thesis (Ph.D.)--The Johns Hopkins University, 2018.
506 $a This item must not be sold to any third party vendors.
506 $a This item must not be added to any third party search indexes.
520 $a Despite effective antiretroviral therapy (ART), HIV-1 persists in all infected individuals as proviral DNA integrated within long-lived resting memory CD4+ T cells. The population of infected CD4+ T cells carrying replication-competent proviruses is the major barrier to HIV-1 cure. Several lines of evidence have demonstrated that cellular proliferation of infected cells contributes to HIV-1 persistence. This proliferative process is complicated by the fact that most infected cells carry defective proviruses and cells harboring replication-competent HIV die quickly upon viral reactivation. To elucidate mechanisms that drive proliferation of HIV-1-infected CD4+ T cells, we followed proliferation of cells carrying replication-competent HIV-1 induced by antigen stimulation or cytokine treatment and demonstrated that latently infected cells carrying replication-competent HIV can proliferate in response to both stimuli. To study the dynamics of cells carrying replication-competent HIV-1, we sampled infectious virus from p24+ wells of the quantitative viral outgrowth assay at multiple time points spanning 2-3 years. Sequencing of replication-competent HIV-1 at multiple time points revealed that expanded cellular clones containing replication-competent HIV-1 is common. While some clones persist for 2-3 years, other clones wax and wane overtime. A similar pattern is observed with virus clones in the residual viremia. This observation with residual viremia supports our hypothesis that viruses in plasma are produced by activation of latently infected cells carrying replication-competent HIV-1 rather than ongoing cycles of virus replication. In addition, this supports our hypothesis that antigens drive proliferation of cells carrying replication-competent HIV-1 and activate some of the cells, leading to virus production. The observed patterns with proviruses in the latent reservoir and viruses in the residual viremia do not support a continuous proliferative process related to HIV-1 integration into cancer-associated genes. These studies are also being extended to cells carrying defective proviruses. A previous study has demonstrated that defective HIV-1 proviruses can be transcribed, translated or even recognized by HIV-1 specific CD8+ T cells. We hypothesized that defective proviruses may have a proliferative advantage that allows the cells carrying defective proviruses to proliferate more than cells carrying intact proviruses. Given that intact proviruses only account for 2% of total proviruses in patients on long-term suppressive ART, infected cells carrying defective proviruses greatly outnumber the cells harboring intact proviruses in vivo. We hypothesized that in vitro, cells carrying defective proviruses would proliferate upon T cell activation while cells harboring intact proviruses capable of viral gene expression, would die upon T cell activation due to viral cytopathic effects. Therefore, cells carrying either defective proviruses and or intact proviruses would show different proliferation dynamics upon TCR activation in vitro. To determine whether cells infected with intact or defective proviruses would proliferate to the same extent, we subjected single HIV-1 infected cells to 4 rounds of anti-CD3/CD28 stimulation in a microculture system and used a novel droplet digital PCR assay (intact proviral DNA assay; IPDA) to quantitate the number of intact and defective proviral DNA sequences in infected cells. We demonstratethat cells carrying defective proviruses were capable of enormous expansion with in vitro anti-CD3/CD28 stimulation, while cells harboring intact proviruses were rarely detected and showed little proliferative potential. Integration site analysis of clones expanded in vitro demonstrated that HIV-1 provirus integration into cancer-associated gene is not required for proliferation of HIV-1-infected cells. Additionally, we sequenced the cell clones that proliferated the most in vitro and found that proviruses in these clones were highly defective. These microculture experiments revealed a profound proliferation defect for cells carrying intact proviruses upon anti-CD3/CD28 stimulation. To explore whether cells carrying intact and defective proviruses show similar dynamics in vivo, we examined longitudinal samples collected 2-8 years apart using the IPDA. We found that the half-life of infected was ~44 months, consistent with a previous measurements of the latent reservoir as measured with a quantitative viral outgrowth assay (QVOA), while cells carrying defective proviruses showed greater variability among patients. Collectively, my thesis has measured the dynamics of CD4+ T cells carrying different types of proviruses and has provided insight into mechanisms that may contribute to proliferation of HIV-infected cells in vivo.
590 $a School code: 0098.
650 4 $a Virology. $3 642304
653 $a Clonal expansion
653 $a HIV-1
653 $a Virology
690 $a 0720
710 2 $a The Johns Hopkins University. $b Pharmacology and Molecular Sciences. $3 3544864
773 0 $t Dissertations Abstracts International $g 80-10B.
790 $a 0098
791 $a Ph.D.
792 $a 2018
793 $a English
856 4 0 $u http://pqdd.sinica.edu.tw/twdaoapp/servlet/advanced?query=13890040