Expression of Chimeric Receptor CD4<i>ζ</i>by Natural Killer Cells Derived from Human Pluripotent Stem Cells Improves In Vitro Activity but Does Not Enhance Suppression of HIV Infection In Vivo

Ni, Zhenya; Knorr, David A.; Bendzick, Laura; Allred, Jeremy; Kaufman, Dan S.

doi:10.1002/stem.1611

Cited by 82 publications

(83 citation statements)

References 77 publications

Supporting

Mentioning

Contrasting

Unclassified

Order By: Relevance

“…We found that the newly designed CAR-T cells, here referred to as VC-CAR-T cells, were able to induce T cell-mediated cytolysis after coculture with gp120-expressing cells and wild-type HIV-1-infected CD4 ϩ T cells. We also found that VC-CAR-T cells display superior potency compared to the CD4-CAR described previously (26,27,(31)(32)(33)(34)(35)(36). Importantly, we have also confirmed that they can effectively kill the reactivated HIV-1-infected CD4 ϩ T lymphocytes isolated from HIV-1-infected patients.…”

supporting

confidence: 84%

“…Therefore, we chose to use VRC01-28BBZ-3-expressing T cells, here referred to as VC-CAR-T cells, for subsequent experiments. Previous studies have reported that CD4-based CAR-T cells, referred to here as CD4-CAR-T cells, display direct cytotoxic activity against HIV-1 Env-expressing target cells (26,27,(31)(32)(33)(34)(35)(36). To compare the efficiency of CD4-CAR and VC-CAR, we generated the former by replacing the VC-CAR scFv with the human CD4 extracellular domain and transduced this construct into CD8 ϩ T lymphocytes (Fig.…”

Section: Selection Of the Most Efficient Car Moiety To Identify The mentioning

confidence: 99%

“…Previous reports have described the generation of HIV-1-specific CAR-T cells by connection of an extracellular antigenbinding domain to intracellular T cell activation domains (e.g., CD3 and CD28). The former can be a single-chain variable fragment (scFv) (26)(27)(28)(29)(30) or a natural molecule, such as CD4 (26,27,(31)(32)(33)(34)(35)(36), capable of directly inducing the death of cells expressing the viral envelope glycoprotein (Env). However, it remains to be determined whether these CAR-T cells can be used to kill the reactivated HIV-1 latently infected cells from the infected individuals receiving suppressive cART.…”

mentioning

confidence: 99%

See 2 more Smart Citations

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Liu

Zou

et al. 2016

J Virol

View full text Add to dashboard Cite

Despite the advent of combined antiretroviral therapy (cART), the persistence of viral reservoirs remains a major barrier to curing human immunodeficiency virus type 1 (HIV-1) infection. Recently, the shock and kill strategy, by which such reservoirs are eradicated following reactivation of latent HIV-1 by latency-reversing agents (LRAs), has been extensively practiced. It is important to reestablish virus-specific and reliable immune surveillance to eradicate the reactivated virus-harboring cells. In this report, we attempted to reach this goal by using newly developed chimeric antigen receptor (CAR)-T cell technology. To generate anti-HIV-1 CAR-T cells, we connected the single-chain variable fragment of the broadly neutralizing HIV-1-specific antibody VRC01 to a third-generation CAR moiety as the extracellular and intracellular domains and subsequently transduced this into primary CD8 ؉ T lymphocytes. We demonstrated that the resulting VC-CAR-T cells induced T cell-mediated cytolysis of cells expressing HIV-1 Env proteins and significantly inhibited HIV-1 rebound after removal of antiviral inhibitors in a viral infectivity model in cell culture that mimics the termination of the cART in the clinic. Importantly, the VC-CAR-T cells also effectively induced the cytolysis of LRA-reactivated HIV-1-infected CD4؉ T lymphocytes isolated from infected individuals receiving suppressive cART. Our data demonstrate that the special features of genetically engineered CAR-T cells make them a particularly suitable candidate for therapeutic application in efforts to reach a functional HIV cure. IMPORTANCEThe presence of latently infected cells remains a key obstacle to the development of a functional HIV-1 cure. Reactivation of dormant viruses is possible with latency-reversing agents, but the effectiveness of these compounds and the subsequent immune response require optimization if the eradication of HIV-1-infected cells is to be achieved. Here, we describe the use of a chimeric antigen receptor, comprised of T cell activation domains and a broadly neutralizing antibody, VRC01, targeting HIV-1 to treat the infected cells. T cells expressing this construct exerted specific cytotoxic activity against wild-type HIV-1-infected cells, resulting in a dramatic reduction in viral rebound in vitro, and showed persistent effectiveness against reactivated latently infected T lymphocytes from HIV-1 patients receiving combined antiretroviral therapy. The methods used in this study constitute an improvement over existing CD4-based CAR-T technology and offer a promising approach to HIV-1 immunotherapy. HIV-1 replication can be efficiently suppressed with combined antiretroviral therapy (cART). However, treatment must be maintained throughout the lifetime of the patient, as this virus can persist in a stable latent reservoir, constituting a major barrier to the establishment of an HIV-1 cure. To date, many strategies have been proposed for the eradication of HIV-1 reservoirs (1-3). Recent efforts have focused on the reactivation of ...

show abstract

supporting

confidence: 84%

Section: Selection Of the Most Efficient Car Moiety To Identify The mentioning

confidence: 99%

mentioning

confidence: 99%

See 1 more Smart Citation

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Liu

Zou

et al. 2016

J Virol

View full text Add to dashboard Cite

show abstract

“…Early clinical successes with CARs against leukemia and lymphoma (28,29) have garnered particular recognition (30), and the strategy has been proposed for use against viruses, including HIV (reviewed in references 31, 32, 33, and 34). Indeed, previous reports demonstrated favorable in vitro results with CARs targeted by single-chain variable-region antibody (Ab) constructs directed against the gp120 or gp41 subunits of HIV-1 Env (35)(36)(37)(38) as well as with CARs employing extracellular CD4 domains (35,36,(39)(40)(41)(42). Clinical studies were conducted more than a decade ago with CD4 CARs (43)(44)(45); see also https://clinicaltrials.gov/ct2/show/NCT01013415), and subsequent analyses of peripheral blood samples verified long-term persistence as well as retention of CAR expression and the proliferative potential of adoptively transferred CAR-transduced T cells (CAR-T) (46).…”

mentioning

confidence: 99%

Novel CD4-Based Bispecific Chimeric Antigen Receptor Designed for Enhanced Anti-HIV Potency and Absence of HIV Entry Receptor Activity

Liu

Patel

Ghanem

et al. 2015

J Virol

103

133

View full text Add to dashboard Cite

Adoptive transfer of CD8 T cells genetically engineered to express "chimeric antigen receptors" (CARs) represents a potential approach toward an HIV infection "functional cure" whereby durable virologic suppression is sustained after discontinuation of antiretroviral therapy. We describe a novel bispecific CAR in which a CD4 segment is linked to a single-chain variable fragment of the 17b human monoclonal antibody recognizing a highly conserved CD4-induced epitope on gp120 involved in coreceptor binding. We compared a standard CD4 CAR with CD4-17b CARs where the polypeptide linker between the CD4 and 17b moieties is sufficiently long (CD4-35-17b CAR) versus too short (CD4-10-17b) to permit simultaneous binding of the two moieties to a single gp120 subunit. When transduced into a peripheral blood mononuclear cell (PBMC) or T cells thereof, all three CD4-based CARs displayed specific functional activities against HIV-1 Env-expressing target cells, including stimulation of gamma interferon (IFN-␥) release, specific target cell killing, and suppression of HIV-1 pseudovirus production. In assays of spreading infection of PBMCs with genetically diverse HIV-1 primary isolates, the CD4-10-17b CAR displayed enhanced potency compared to the CD4 CAR whereas the CD4-35-17b CAR displayed diminished potency. Importantly, both CD4-17b CARs were devoid of a major undesired activity observed with the CD4 CAR, namely, rendering the transduced CD8 ؉ T cells susceptible to HIV-1 infection. Likely mechanisms for the superior potency of the CD4-10-17b CAR over the CD4-35-17b CAR include the greater potential of the former to engage in the serial antigen binding required for efficient T cell activation and the ability of two CD4-10-17b molecules to simultaneously bind a single gp120 subunit. IMPORTANCEHIV research has been energized by prospects for a cure for HIV infection or, at least, for a "functional cure" whereby antiretroviral therapy can be discontinued without virus rebound. This report describes a novel CD4-based "chimeric antigen receptor" (CAR) which, when genetically engineered into T cells, gives them the capability to selectively respond to and kill HIV-infected cells. This CAR displays enhanced features compared to previously described CD4-based CARs, namely, increased potency and avoidance of the undesired rendering of the genetically modified CD8 T cells susceptible to HIV infection. When adoptively transferred back to the individual, the genetically modified T cells will hopefully provide durable killing of infected cells and sustained virus suppression without continued antiretroviral therapy, i.e., a functional cure. C ombination antiretroviral therapy (cART) (1) today offers the promise of near-normal life expectancy for HIV-infected individuals (2), most of whom would previously have succumbed to the lethal consequences of immune system demise. Nevertheless, even under conditions of plasma viral load suppression below the limits of detection, CD4 T-cell recovery is often incomplete. The pathogenic sequel...

show abstract

“…Recently, Ni et al took a novel strategy to utilize the anti-HIV specificity of TCRξ by engineering it into hESCs/iPSCs. After differentiation to NK cells, these modified counterparts were effective in inhibiting HIV-1 replication in NSG mice 53 .…”

Section: Revolution In Gene Modification:-mentioning

confidence: 99%

Cell-based gene therapy against HIV

Dey

Pillai

2015

Gene Ther

View full text Add to dashboard Cite

The ability to integrate inside the host genome lays a strong foundation for HIV to play hide and seek with the host's immune surveillance mechanisms. Present anti-viral therapies, although successful in suppressing the virus to a certain level, fail to wipe it out completely. However, recent approaches in modifying stem cells and enabling them to give rise to potent/resistant T-cells against HIV holds immense hope for eradication of the virus from the host. In this review, we will briefly discuss previous landmark studies on engineering stem cells or T-cells that have been explored for therapeutic efficacy against HIV. We will also analyze potential benefits and pitfalls of some studies done recently and will share our opinion on emerging trends.

show abstract

Expression of Chimeric Receptor CD4ζby Natural Killer Cells Derived from Human Pluripotent Stem Cells Improves In Vitro Activity but Does Not Enhance Suppression of HIV Infection In Vivo

Cited by 82 publications

References 77 publications

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Novel CD4-Based Bispecific Chimeric Antigen Receptor Designed for Enhanced Anti-HIV Potency and Absence of HIV Entry Receptor Activity

Cell-based gene therapy against HIV

Contact Info

Product

Resources

About

Expression of Chimeric Receptor CD4ζby Natural Killer Cells Derived from Human Pluripotent Stem Cells Improves In Vitro Activity but Does Not Enhance Suppression of HIV Infection In Vivo

Cited by 82 publications

References 77 publications

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 + T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 + T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Novel CD4-Based Bispecific Chimeric Antigen Receptor Designed for Enhanced Anti-HIV Potency and Absence of HIV Entry Receptor Activity

Cell-based gene therapy against HIV

Contact Info

Product

Resources

About

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy

Chimeric Antigen Receptor T Cells Guided by the Single-Chain Fv of a Broadly Neutralizing Antibody Specifically and Effectively Eradicate Virus Reactivated from Latency in CD4 ⁺ T Lymphocytes Isolated from HIV-1-Infected Individuals Receiving Suppressive Combined Antiretroviral Therapy