المؤلفون: Levine, Bruce L., Humeau, Laurent M., Boyer, Jean, MacGregor, Rob-Roy, Rebello, Tessio, Lu, Xiaobin, Binder, Gwendolyn K., Slepushkin, Vladimir, Lemiale, Franck, Mascola, John R., Bushman, Frederic D., Dropulic, Boro, June, Carl H.

المصدر: Proceedings of the National Academy of Sciences of the United States of America, 2006 Nov . 103(46), 17372-17377.

الوصول الحر: https://www.jstor.org/stable/30052439Test

المؤلفون: Wang, Gary P.^1,2, Levine, Bruce L.³, Binder, Gwendolyn K.³, Berry, Charles C.⁴, Malani, Nirav¹, McGarrity, Gary⁵, Tebas, Pablo², June, Carl H.³, Bushman, Frederic D.¹ bushman@mail.med.upenn.edu

المصدر: Molecular Therapy. May2009, Vol. 17 Issue 5, p844-850. 7p. 1 Chart, 5 Graphs.

مصطلحات موضوعية: *GENE therapy, *HIV, *PROTO-oncogenes, *CELLS, *TUMOR suppressor genes, *DNA

مستخلص: Lentiviral vector-based gene therapy has been used to target the human immunodeficiency virus (HIV) using an antisense env payload. We have analyzed lentiviral-vector integration sites from three treated individuals. We compared integration sites from the ex vivo vector-transduced CD4+ cell products to sites from cells recovered at several times after infusion. Integration sites were analyzed using 454 pyrosequencing, yielding a total of 7,782 unique integration sites from the ex vivo product and 237 unique sites from cells recovered after infusion. Integrated vector copies in both data sets were found to be strongly enriched within active genes and near epigenetic marks associated with active transcription units. Analysis of integration relative to nucleosome structure on target DNA indicated favoring of integration in outward facing DNA major grooves on the nucleosome surface. There was no indication that growth of transduced cells after infusion resulted in enrichment for integration sites near proto-oncogene 5′-ends or within tumor suppressor genes. Thus, this first look at the longitudinal evolution of cells transduced with a lentiviral vector after infusion of gene modified CD4+ cells provided no evidence for abnormal expansions of cells due to vector-mediated insertional activation of proto-oncogenes.Molecular Therapy (2009) 17 5, 844–850 doi:10.1038/mt.2009.16 [ABSTRACT FROM AUTHOR]

المؤلفون: Levine, Bruce L., Bernstein, Wendy B., Aronson, Naomi E., Schlienger, Katia, Cotte, Julio, Perfetto, Steven, Humphries, Mary J., Ratto-Kim, Silvia, Birx, Deborah L., Steffens, Carolyn, Landay, Alan, Carroll, Richard G., June, Carl H.

المصدر: Nature Medicine. Jan2002, Vol. 8 Issue 1, p47. 7p.

مصطلحات موضوعية: *HIV, *T cells, *CYTOKINES, *CELLS

مستخلص: To study the safety and feasibility of T-cell reconstitution in HIV-infected individuals, we adoptively transferred activated autologous CD4+ T cells. Polyclonal peripheral blood CD4+ cells were costimulated ex vivo and subjects were given infusions of up to 3 × 1010 activated CD4+ cells. Dose-dependent increases in CD4+ cell counts and in the CD4:CD8 ratio were observed. Sustained increases in the fraction of cytokine-secreting T cells and decreases in the percentage of CD4+CCR5+ cells were noted in vivo, suggesting enhanced function and resistance to HIV infection. The frequency of CD4+Ki-67+ cells increased whereas CD4+ T cells containing T cell?receptor rearrangement excision circles (TRECs) decreased. These findings indicate that expansion of the peripheral T-cell pool mediated the increase in CD4 counts and suggest that approaches to reconstitute CD4 helper cell activity and decrease CCR5 expression may augment natural immunity to HIV infection. [ABSTRACT FROM AUTHOR]

المؤلفون: Tebas, Pablo¹, Jadlowsky, Julie K.², Shaw, Pamela A.³, Lifeng Tian⁴, Esparza, Erin⁴, Brennan, Andrea⁴, Sukyung Kim¹, Soe Yu Naing¹, Richardson, Max W.², Vogel, Ashley N.⁴, Maldini, Colby R.², Hong Kong², Xiaojun Liu⁴, Lacey, Simon F.⁴, Bauer, Anya M.¹, Mampe, Felicity¹, Richman, Lee P.¹, Lee, Gary⁵, Ando, Dale⁵, Levine, Bruce L.⁴

المصدر: Journal of Clinical Investigation. 4/1/2021, Vol. 131 Issue 7, p1-12. 12p.

مصطلحات موضوعية: *T cells, *HIV, *ZINC-finger proteins, *VIRAL load, *DRUG abuse

الشركة/الكيان: NATIONAL Institute on Drug Abuse , NATIONAL Institute of Neurological Disorders & Stroke (U.S.) , NATIONAL Institutes of Health (U.S.)

مستخلص: BackgroundWe conducted a phase I clinical trial that infused CCR5 gene-edited CD4+ T cells to determine how these T cells can better enable HIV cure strategies.MethodsThe aim of trial was to develop RNA-based approaches to deliver zinc finger nuclease (ZFN), evaluate the effect of CCR5 gene-edited CD4+ T cells on the HIV-specific T cell response, test the ability of infused CCR5 gene-edited T cells to delay viral rebound during analytical treatment interruption, and determine whether individuals heterozygous for CCR5 Δ32 preferentially benefit. We enrolled 14 individuals living with HIV whose viral load was well controlled by antiretroviral therapy (ART). We measured the time to viral rebound after ART withdrawal, the persistence of CCR5-edited CD4+ T cells, and whether infusion of 10 billion CCR5-edited CD4+ T cells augmented the HIV-specific immune response.ResultsInfusion of the CD4+ T cells was well tolerated, with no serious adverse events. We observed a modest delay in the time to viral rebound relative to historical controls; however, 3 of the 14 individuals, 2 of whom were heterozygous for CCR5 Δ32, showed post-viral rebound control of viremia, before ultimately losing control of viral replication. Interestingly, only these individuals had substantial restoration of HIV-specific CD8+ T cell responses. We observed immune escape for 1 of these reinvigorated responses at viral recrudescence, illustrating a direct link between viral control and enhanced CD8+ T cell responses.ConclusionThese findings demonstrate how CCR5 gene-edited CD4+ T cell infusion could aid HIV cure strategies by augmenting preexisting HIV-specific immune responses.REGISTRATIONClinicalTrials.gov NCT02388594.FundingNIH funding (R01AI104400, UM1AI126620, U19AI149680, T32AI007632) was provided by the National Institute of Allergy and Infectious Diseases (NIAID), the National Institute on Drug Abuse (NIDA), the National Institute of Mental Health (NIMH), and the National Institute of Neurological Disorders and Stroke (NINDS). Sangamo Therapeutics also provided funding for these studies. [ABSTRACT FROM AUTHOR]

المؤلفون: Yu, Jianqing J.¹, Wu, Te Lang², Liszewski, Megan K.³, Dai, Jihong¹, Swiggard, William J.⁴, Baytop, Clifford¹, Frank, Ian⁵, Levine, Bruce L.¹, Yang, Wei⁶, Theodosopoulos, Theodore⁷, O'Doherty, Una¹ unao@mail.med.upenn.edu

المصدر: Virology. Sep2008, Vol. 379 Issue 1, p78-86. 9p.

مصطلحات موضوعية: *HIV-positive persons, *ANTIRETROVIRAL agents, *THERAPEUTICS, *NUCLEIC acids

مستخلص: Abstract: Many studies report the level of total viral DNA in HIV-infected patients, but few studies report the level of integrated DNA. It is important to measure integrated DNA in HIV-infected patients because the information could shed light on the effectiveness of antiretroviral therapy, especially intensified therapy, when viral loads may remain undetectable. In order to develop an integration assay for patient samples, we enhanced the sensitivity of our prior integration assay. To do this, we exploited a technique that we developed, called repetitive sampling, and optimized reaction conditions for rare event detection, rather than large dynamic range. We also designed our primers to match more conserved regions of HIV. The result is a new, sensitive, quantitative assay that allows us to measure integrated DNA in HIV-infected patients. When we applied our integration assay to patient PBMCs, we found that the use of HAART is associated with reduced levels of integrated DNA. [Copyright &y& Elsevier]

المؤلفون: Bernstein, Wendy B.^1,2,3, Cox, Josephine H.⁴, Aronson, Naomi E.^2,5, Tracy, LaRee⁶, Schlienger, Katia⁷, Ratto-Kim, Silvia⁴, Garner, Robin⁴, Cotte, Julio⁷, Zheng, Zhaohui⁷, Winestone, Lena⁷, Liebig, Caroline⁴, Galley, Lynee M.⁴, Connors, Mark⁸, Birx, Deborah L.¹, Carroll, Richard G.⁷, Levine, Bruce L.⁷ levinebl@mail.med.upenn.edu

المصدر: Clinical Immunology. Jun2004, Vol. 111 Issue 3, p262-274. 13p.

مصطلحات موضوعية: *T cell receptors, *LYMPHOCYTES, *ANTIGENS, *IMMUNITY

مستخلص: We have previously shown that adoptive transfer of in vitro CD3/CD28 activated autologous CD4+ T cells results in increased CD4 counts and CD4/CD8 ratios in HIV+ subjects. In this report, analysis of variable beta (Vβ) chain T cell receptor (TCR) repertoire showed that CD3/CD28 stimulation was able to increase polyclonality within skewed spectra types in vitro. In vivo, two of eight subjects showed increase in TCR diversity and importantly, in no subject did a highly skewed in vivo repertoire emerge. Measurement of proliferative response to alloantigen showed increases following infusions. Response to pharmacological stimulus and lectin via Interferon-γ ELISpot assay showed increases in a subset of subjects following infusions. However, interferon-γ response to HIV antigens and peptides declined concurrent with stable or diminishing latent infectious viral load in CD4+ T cells. These data provide further evidence that adoptive transfer of activated autologous CD4+ T cells can augment the immune system. [Copyright &y& Elsevier]