24 results on '"Hua, Zheng"'
Search Results
2. Metagenomic Discovery of “ Candidatus Parvarchaeales”-Related Lineages Sheds Light on Adaptation and Diversification from Neutral-Thermal to Acidic-Mesothermal Environments
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Rao, Yang-Zhi, primary, Li, Yu-Xian, additional, Li, Ze-Wei, additional, Qu, Yan-Ni, additional, Qi, Yan-Ling, additional, Jiao, Jian-Yu, additional, Shu, Wen-Sheng, additional, Hua, Zheng-Shuang, additional, and Li, Wen-Jun, additional
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- 2023
- Full Text
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3. Pathogenicity and Structural Basis of Zika Variants with Glycan Loop Deletions in the Envelope Protein
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Meng-Li Cheng, Yun-Xiang Yang, Zhong-Yu Liu, Dan Wen, Pan Yang, Xing-Yao Huang, Hao-Long Dong, Yan-Peng Xu, Xiao-Feng Li, Yong-Qiang Deng, Qing Ye, Ling Zhu, Juan Li, Andrew D. Davidson, Ai-Hua Zheng, Wei-Feng Shi, Hui Zhao, Xiang-Xi Wang, and Cheng-Feng Qin
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Virulence ,Zika Virus Infection ,Immunology ,Zika Virus ,Virus Replication ,Microbiology ,Mice ,Disease Models, Animal ,Viral Envelope Proteins ,Polysaccharides ,Virology ,Insect Science ,Pathogenesis and Immunity ,Animals - Abstract
The glycan loop of Zika virus (ZIKV) envelope protein (E) contains the glycosylation site and has been well documented to be important for viral pathogenesis and transmission. In the present study, we report that deletions in the E glycan loop, which were recorded in African ZIKV strains previously, have re-emerged in their contemporary Asian lineages. Here, we generated recombinant ZIKV containing specific deletions in the E glycan loop by reverse genetics. Extensive in vitro and in vivo characterization of these deletion mutants demonstrated an attenuated phenotype in an adult A129 mouse model and reduced oral infections in mosquitoes. Surprisingly, these glycan loop deletion mutants exhibited an enhanced neurovirulence phenotype, and resulted in a more severe microcephalic brain in neonatal mouse models. Crystal structures of the ZIKV E protein and a deletion mutant at 2.5 and 2.6 Å, respectively, revealed that deletion of the glycan loop induces encephalitic flavivirus-like conformational alterations, including the appearance of perforations on the surface and a clear change in the topology of the loops. Overall, our results demonstrate that the E glycan loop deletions represent neonatal mouse neurovirulence markers of ZIKV. IMPORTANCE Zika virus (ZIKV) has been identified as a cause of microcephaly and acquired evolutionary mutations since its discovery. Previously deletions in the E glycan loop were recorded in African ZIKV strains, which have re-emerged in the contemporary Asian lineages recently. The glycan loop deletion mutants are not glycosylated, which are attenuated in adult A129 mouse model and reduced oral infections in mosquitoes. More importantly, the glycan loop deletion mutants induce an encephalitic flavivirus-like conformational alteration in the E homodimer, resulting in a significant enhancement of neonatal mouse neurovirulence. This study underscores the critical role of glycan loop deletion mutants in ZIKV pathogenesis, highlighting a need for global virological surveillance for such ZIKV variants.
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- 2022
4. Deciphering Symbiotic Interactions of “ Candidatus Aenigmarchaeota” with Inferred Horizontal Gene Transfers and Co-occurrence Networks
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Li, Yu-Xian, primary, Rao, Yang-Zhi, additional, Qi, Yan-Ling, additional, Qu, Yan-Ni, additional, Chen, Ya-Ting, additional, Jiao, Jian-Yu, additional, Shu, Wen-Sheng, additional, Jiang, Hongchen, additional, Hedlund, Brian P., additional, Hua, Zheng-Shuang, additional, and Li, Wen-Jun, additional
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- 2021
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5. Comparative Genomics Reveals Thermal Adaptation and a High Metabolic Diversity in “ Candidatus Bathyarchaeia”
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Qi, Yan-Ling, primary, Evans, Paul N., additional, Li, Yu-Xian, additional, Rao, Yang-Zhi, additional, Qu, Yan-Ni, additional, Tan, Sha, additional, Jiao, Jian-Yu, additional, Chen, Ya-Ting, additional, Hedlund, Brian P., additional, Shu, Wen-Sheng, additional, Hua, Zheng-Shuang, additional, and Li, Wen-Jun, additional
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- 2021
- Full Text
- View/download PDF
6. Emtricitabine-Triphosphate in Dried Blood Spots as a Marker of Recent Dosing
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Robert M. Grant, Kayla Campbell, Jose R Castillo-Mancilla, Lane R. Bushman, Sybil Hosek, Stacey S Coleman, Samantha MaWhinney, Craig M. Wilson, Edward M. Gardner, David V. Glidden, Sharon M Seifert, Peter L. Anderson, Jia-Hua Zheng, Kevin McAllister, and Albert Y. Liu
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Adult ,Male ,0301 basic medicine ,medicine.medical_specialty ,Adolescent ,Anti-HIV Agents ,Clinical Trials and Supportive Activities ,Urology ,HIV Infections ,Emtricitabine ,Microbiology ,Drug Administration Schedule ,Medication Adherence ,law.invention ,03 medical and health sciences ,0302 clinical medicine ,Pharmacokinetics ,Randomized controlled trial ,Clinical Research ,law ,medicine ,Humans ,Pharmacology (medical) ,Prospective Studies ,030212 general & internal medicine ,Dosing ,Tenofovir ,Dried blood ,Prospective cohort study ,Aged ,Pharmacology ,business.industry ,HIV ,Pharmacology and Pharmaceutical Sciences ,Middle Aged ,030112 virology ,Confidence interval ,Clinical trial ,Infectious Diseases ,Medical Microbiology ,Case-Control Studies ,HIV/AIDS ,Female ,Dried Blood Spot Testing ,business ,Half-Life ,medicine.drug - Abstract
New objective measures of antiretroviral adherence are needed. We determined if emtricitabine triphosphate (FTC-TP) in dried blood spots (DBS) can be used as a marker of recent dosing with tenofovir disoproxil fumarate-emtricitabine (TDF-FTC). The half-life of FTC-TP was estimated in DBS samples obtained from an intensive pharmacokinetic (PK) study of coformulated TDF-FTC in HIV-negative and HIV-infected participants. The concordance of quantifiable FTC-TP in DBS with tenofovir (TFV)/FTC in plasma was evaluated by utilizing paired plasma-DBS samples from participants enrolled in 2 large preexposure prophylaxis (PrEP) open-label trials. The time to FTC-TP nondetectability after TDF-FTC dosing was evaluated utilizing DBS from HIV-negative participants enrolled in a directly observed therapy study of variable adherence to TDF-FTC. The mean (95% confidence interval [CI]) terminal half-life of FTC-TP in the PK study was 35 (23 to 47) h. A total of 143/163 (88%) samples obtained 0 to 48 h post-TDF-FTC dose had quantifiable FTC-TP in DBS, compared with 2/93 (2%) and 0/87 (0%) obtained >48 and >96 h postdose. In 746 paired plasma-DBS samples from 445 participants enrolled in PrEP trials, when both TFV/FTC in plasma were below the limit of quantification, FTC-TP was as well in 98.9% of the samples, and when either TFV or FTC in plasma was quantifiable, FTC-TP was as well in 90.5% of the samples. The half-life of FTC-TP in DBS is short relative to that of TFV-diphosphate (TFV-DP), making it a surrogate for TFV-FTC detection in plasma. FTC-TP can be quantified in DBS simultaneously with TFV-DP, which quantifies cumulative adherence to TDF-FTC. (The clinical trials discussed in this article have been registered at ClinicalTrials.gov under identifiers NCT01040091, NCT02022657, NCT00458393, NCT01772823, and NCT02012621.)
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- 2016
7. Analysis of the Endogenous Deoxynucleoside Triphosphate Pool in HIV-Positive and -Negative Individuals Receiving Tenofovir-Emtricitabine
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Lane R. Bushman, Jia-Hua Zheng, Sharon M Seifert, Peter L. Anderson, Kevin B. McAllister, Jose R Castillo-Mancilla, Xinhui Chen, and Samantha MaWhinney
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Adult ,Male ,0301 basic medicine ,Tenofovir ,Anti-HIV Agents ,030106 microbiology ,Cellular homeostasis ,HIV Infections ,Endogeny ,Pharmacology ,Emtricitabine ,Peripheral blood mononuclear cell ,03 medical and health sciences ,Deoxyadenine Nucleotides ,Humans ,Thymine Nucleotides ,Medicine ,heterocyclic compounds ,Pharmacology (medical) ,Clinical significance ,Deoxynucleoside triphosphate ,business.industry ,Case-control study ,Deoxyguanine Nucleotides ,030104 developmental biology ,Infectious Diseases ,Case-Control Studies ,Deoxycytosine Nucleotides ,HIV-1 ,Leukocytes, Mononuclear ,Linear Models ,Female ,business ,medicine.drug - Abstract
Tenofovir (TFV) disoproxil fumarate (TDF) and emtricitabine (FTC), two nucleos(t)ide analogs (NA), are coformulated as an anti-HIV combination tablet for treatment and preexposure prophylaxis (PrEP). TDF/FTC may have effects on the deoxynucleoside triphosphate (dNTP) pool due to their similar structures and similar metabolic pathways. We carried out a comprehensive clinical study to characterize the effects of TDF/FTC on the endogenous dNTP pool, from baseline to 30 days of TDF/FTC therapy, in both treatment-naive HIV-positive and HIV-negative individuals. dATP, dCTP, dGTP, and TTP were quantified in peripheral blood mononuclear cells (PBMC) with a validated liquid chromatography-tandem mass spectrometry (LC-MS/MS) methodology. Forty individuals (19 HIV-positive) were enrolled and underwent a baseline visit and then received TDF/FTC for at least 30 days. Longitudinal measurements were analyzed using mixed-model segmented linear regression analysis. The dNTPs were reduced by 14% to 37% relative to the baseline level within 3 days in both HIV-negative and HIV-positive individuals (P≤ 0.003). These reductions persisted to various degrees at day 30. These findings indicate that dNTP pools are influenced by TDF/FTC therapy. This may alter cellular homeostasis and could increase the antiviral effect through a more favorable analog/dNTP ratio. Further work is needed to elucidate mechanisms, to evaluate the clinical significance of these findings, and to further probe differences between HIV-negative and HIV-positive individuals. (This study has been registered at ClinicalTrials.gov under identifier NCT01040091.)
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- 2016
8. Establishment of a Novel Humanized Mouse Model To Investigate In Vivo Activation and Depletion of Patient-Derived HIV Latent Reservoirs
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Nina C. Flerin, Elizabeth Connick, Rebecca M. Lynch, Maria Korom, Jian Hua Zheng, Ronald Truong, R. Brad Jones, Harris Goldstein, Talia M. Mota, Joy M. Folkvord, Ariola Bardhi, Erika Benko, and Colin Kovacs
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0303 health sciences ,education.field_of_study ,030306 microbiology ,Immunology ,Population ,Viremia ,Spleen ,In situ hybridization ,Biology ,medicine.disease ,Microbiology ,Virology ,Peripheral blood mononuclear cell ,Virus ,03 medical and health sciences ,medicine.anatomical_structure ,In vivo ,Insect Science ,Humanized mouse ,medicine ,education ,030304 developmental biology - Abstract
Curing HIV infection has been thwarted by the persistent reservoir of latently infected CD4+ T cells, which reinitiate systemic infection after antiretroviral therapy (ART) interruption. To evaluate reservoir depletion strategies, we developed a novel preclinical in vivo model consisting of immunodeficient mice intrasplenically injected with peripheral blood mononuclear cells (PBMC) from long-term ART-suppressed HIV-infected donors. In the absence of ART, these mice developed rebound viremia which, 2 weeks after PBMC injection, was 1,000-fold higher (mean = 9,229,281 HIV copies/ml) in mice injected intrasplenically than in mice injected intraperitoneally (mean = 6,838 HIV copies/ml) or intravenously (mean = 591 HIV copies/ml). One week after intrasplenic PBMC injection, in situ hybridization of the spleen demonstrated extensive disseminated HIV infection, likely initiated from in vivo-reactivated primary latently infected cells. The time to viremia was delayed significantly by treatment with a broadly neutralizing antibody, 10-1074, compared to treatment with 10-1074-FcRnull, suggesting that 10-1074 mobilized Fc-mediated effector mechanisms to deplete the replication-competent reservoir. This was supported by phylogenetic analysis of Env sequences from viral-outgrowth cultures and untreated, 10-1074-treated, or 10-1074-FcRnull-treated mice. The predominant sequence cluster detected in viral-outgrowth cultures and untreated mouse plasma was significantly reduced in the plasma of 10-1074-treated mice, whereas two new clusters emerged that were not detected in viral-outgrowth cultures or plasma from untreated mice. These new clusters lacked mutations associated with 10-1074 resistance. Taken together, these data indicated that 10-1074 treatment depletes the reservoir of latently infected cells harboring replication competent HIV. Furthermore, this mouse model represents a new in vivo approach for the preclinical evaluation of new HIV cure strategies.IMPORTANCE Sustained remission of HIV infection is prevented by a persistent reservoir of latently infected cells capable of reinitiating systemic infection and viremia. To evaluate strategies to reactivate and deplete this reservoir, we developed and characterized a new humanized mouse model consisting of highly immunodeficient mice intrasplenically injected with peripheral blood mononuclear cells from long-term ART-suppressed HIV-infected donors. Reactivation and dissemination of HIV infection was visualized in the mouse spleens in parallel with the onset of viremia. The applicability of this model for evaluating reservoir depletion treatments was demonstrated by establishing, through delayed time to viremia and phylogenetic analysis of plasma virus, that treatment of these humanized mice with a broadly neutralizing antibody, 10-1074, depleted the patient-derived population of latently infected cells. This mouse model represents a new in vivo approach for the preclinical evaluation of new HIV cure strategies. ispartof: Journal of Virology vol:93 issue:6 ispartof: location:United States status: published
- Published
- 2019
9. Validation and Application of a Liquid Chromatography-Tandem Mass Spectrometry Method To Determine the Concentrations of Sofosbuvir Anabolites in Cells
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Lane R. Bushman, Peter L. Anderson, Joseph E. Rower, Jennifer J. Kiser, Jia-Hua Zheng, Leah C. Jimmerson, Ariel Hodara, and Xinhui Chen
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Quality Control ,Erythrocytes ,Tandem mass spectrometry ,Antiviral Agents ,Peripheral blood mononuclear cell ,Monocytes ,Phosphates ,chemistry.chemical_compound ,Tandem Mass Spectrometry ,Liquid chromatography–mass spectrometry ,In vivo ,Humans ,Pharmacology (medical) ,Nucleotide ,Phosphorylation ,Chromatography, High Pressure Liquid ,Pharmacology ,chemistry.chemical_classification ,Reproducibility of Results ,Prodrug ,Hepatitis C ,Uridine ,Infectious Diseases ,Enzyme ,Nonlinear Dynamics ,Biochemistry ,chemistry ,Hepatocytes ,Sofosbuvir ,Half-Life - Abstract
Sofosbuvir (SOF) is a highly efficacious and well-tolerated uridine nucleotide analog that inhibits the hepatitis C virus (HCV) NS5B polymerase enzyme. SOF is administered as a prodrug, which undergoes intracellular phosphorylation by host enzymes to a monophosphate, diphosphate, and finally a pharmacologically active triphosphate. In order to fully understand the clinical pharmacology of SOF, there is a great need to determine the intracellular phosphate concentrations of the drug. We describe the validation and utilization of a method to characterize SOF's disposition into various in vivo cell types, including hepatocytes, peripheral blood mononuclear cells (PBMC), and red blood cells (RBC). Standard bioanalytical validation criteria were applied to lysed cellular matrices, with a validated linear range of 50 to 50,000 fmol/sample for each phosphate moiety. The assay was utilized to collect the first data demonstrating concentrations of phosphorylated anabolites formed in PBMC, hepatocytes, and RBC in vivo during SOF therapy. Median concentrations in PBMC were 220 (range, 51.5 to 846), 70.2 (range, 25.8 to 275), and 859 (range, 54.5 to 6,756) fmol/10 6 cells in the monophosphate, diphosphate, and triphosphate fractions, respectively. In contrast, RBC triphosphate concentrations were much lower than those of PBMC, as the median concentration was 2.91 (range, 1.14 to 10.4) fmol/10 6 cells. The PBMC triphosphate half-life was estimated at 26 h using noncompartmental approaches, while nonlinear mixed-effect modeling was used to estimate a 69 h half-life for this moiety in RBC. The validated method and the data it generates provide novel insight into the cellular disposition of SOF and its phosphorylated anabolites in vivo .
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- 2015
10. The Vaginal Acquisition and Dissemination of HIV-1 Infection in a Novel Transgenic Mouse Model Is Facilitated by Coinfection with Herpes Simplex Virus 2 and Is Inhibited by Microbicide Treatment
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John C. Kappes, Christina Ochsenbauer, Kieran Seay, Betsy C. Herold, Patrick F. Kiser, Nazanin Khajoueinejad, Jian Hua Zheng, and Harris Goldstein
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Male ,Genetically modified mouse ,Herpesvirus 2, Human ,Transgene ,Immunology ,HIV Infections ,Mice, Transgenic ,Biology ,medicine.disease_cause ,Antiviral Agents ,Microbiology ,Virus ,Mice ,Virology ,Microbicide ,CAPRISA 004 ,medicine ,Animals ,Humans ,Herpes Genitalis ,Coinfection ,virus diseases ,Prodrug ,medicine.disease ,Disease Models, Animal ,Herpes simplex virus ,Insect Science ,Vagina ,HIV-1 ,Vaginal Creams, Foams, and Jellies ,Pathogenesis and Immunity ,Female - Abstract
Epidemiological studies have demonstrated that herpes simplex virus 2 (HSV-2) infection significantly increases the risk of HIV-1 acquisition, thereby contributing to the expanding HIV-1 epidemic. To investigate whether HSV-2 infection directly facilitates mucosal HIV-1 acquisition, we used our transgenic hCD4/R5/cT1 mouse model which circumvents major entry and transcription blocks preventing murine HIV-1 infection by targeting transgenic expression of human CD4, CCR5, and cyclin T1 genes to CD4 + T cells and myeloid-committed cells. Productive infection of mucosal leukocytes, predominantly CD4 + T cells, was detected in all hCD4/R5/cT1 mice intravaginally challenged with an HIV-1 infectious molecular clone, HIV-Du151.2 env -NLuc, which expresses an env gene (C.Du151.2) cloned from an acute heterosexually infected woman and a NanoLuc luciferase reporter gene. Lower genital tract HIV-1 infection after HIV-Du151.2 env -NLuc intravaginal challenge was increased ∼4-fold in hCD4/R5/cT1 mice coinfected with HSV-2. Furthermore, HIV-1 dissemination to draining lymph nodes was detected only in HSV-2-coinfected mice. HSV-2 infection stimulated local infiltration and activation of CD4 + T cells and dendritic cells, likely contributing to the enhanced HIV-1 infection and dissemination in HSV-2-coinfected mice. We then used this model to demonstrate that a novel gel containing tenofovir disoproxil fumarate (TDF), the more potent prodrug of tenofovir (TFV), but not the TFV microbicide gel utilized in the recent CAPRISA 004, VOICE (Vaginal and Oral Interventions to Control the Epidemic), and FACTS 001 clinical trials, was effective as preexposure prophylaxis (PrEP) to completely prevent vaginal HIV-1 infection in almost half of HSV-2-coinfected mice. These results also support utilization of hCD4/R5/cT1 mice as a highly reproducible immunocompetent preclinical model to evaluate HIV-1 acquisition across the female genital tract. IMPORTANCE Multiple epidemiological studies have reported that genital herpes simplex virus 2 (HSV-2) infection increases the risk of HIV-1 sexual acquisition by severalfold. Understanding the underlying mechanisms by which HSV-2 facilitates HIV-1 infection and optimizing the efficacy of therapies to inhibit HIV-1 infection during HSV-2 coinfection should contribute to reducing HIV-1 transmission. Using our novel transgenic hCD4/R5/cT1 mouse model infectible with HIV-1, we demonstrated that HSV-2 infection enhances vaginal transmission and dissemination of HIV-1 infection while stimulating recruitment and activation of CD4 + T cells and dendritic cells in the lower genital tract. HIV acquisition by hCD4/R5/cT1 mice vaginally coinfected with HSV-2 could be completely prevented in almost half the mice by preexposure prophylaxis (PrEP) with a novel gel containing tenofovir disoproxil fumarate (TDF), the tenofovir prodrug, but not with the tenofovir microbicide gel utilized in CAPRISA-004, VOICE, and FACTS-001 clinical trials. The hCD4/R5/cT1 mice represent a new preclinical mouse model to evaluate vaginal HIV-1 acquisition.
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- 2015
11. Potent In Vivo NK Cell-Mediated Elimination of HIV-1-Infected Cells Mobilized by a gp120-Bispecific and Hexavalent Broadly Neutralizing Fusion Protein
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Wei Li, Christina Ochsenbauer, Yanling Wu, Dimiter S. Dimitrov, Ariola Bardhi, Harris Goldstein, John C. Kappes, Emily K. Jeng, Zhongyu Zhu, Weizao Chen, Jian Hua Zheng, Hing C. Wong, Tianlei Ying, and Jennifer Jones
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0301 basic medicine ,CD4 antigen ,Immunology ,Simian Acquired Immunodeficiency Syndrome ,Cellular Response to Infection ,HIV Infections ,Biology ,HIV Envelope Protein gp120 ,Microbiology ,03 medical and health sciences ,chemistry.chemical_compound ,Mice ,Immune system ,In vivo ,Virology ,Virus latency ,Antibodies, Bispecific ,medicine ,Animals ,Antibody-dependent cell-mediated cytotoxicity ,Antibody-Dependent Cell Cytotoxicity ,virus diseases ,medicine.disease ,Fusion protein ,Antibodies, Neutralizing ,Macaca mulatta ,Recombinant Proteins ,Virus Latency ,Killer Cells, Natural ,Disease Models, Animal ,030104 developmental biology ,chemistry ,Insect Science ,Humanized mouse ,CD4 Antigens ,biology.protein ,HIV-1 ,Leukocytes, Mononuclear ,Simian Immunodeficiency Virus ,Antibody - Abstract
Antibodies bound to human immunodeficiency virus type 1 (HIV-1) envelope protein expressed by infected cells mobilize antibody-dependent cellular cytotoxicity (ADCC) to eliminate the HIV-1-infected cells and thereby suppress HIV-1 infection and delay disease progression. Studies treating HIV-1-infected individuals with latency reactivation agents to reduce their latent HIV-1 reservoirs indicated that their HIV-1-specific immune responses were insufficient to effectively eliminate the reactivated latent HIV-1-infected T cells. Mobilization of ADCC may facilitate elimination of reactivated latent HIV-1-infected cells to deplete the HIV-1 reservoir and contribute to a functional HIV-1 cure. The most effective antibodies for controlling and eradicating HIV-1 infection would likely have the dual capacities of potently neutralizing a broad range of HIV-1 isolates and effectively mobilizing HIV-1-specific ADCC to eliminate HIV-1-infected cells. For this purpose, we constructed LSEVh-LS-F, a broadly neutralizing, defucosylated hexavalent fusion protein specific for both the CD4 and coreceptor gp120-binding sites. LSEVh-LS-F potently inhibited in vivo HIV-1 and simian-human immunodeficiency virus (SHIV) infection in humanized mouse and macaque models, respectively, including in vivo neutralization of HIV-1 strains resistant to the broadly neutralizing antibodies VRC01 and 3BNC117. We developed a novel humanized mouse model to evaluate in vivo human NK cell-mediated elimination of HIV-1-infected cells by ADCC and utilized it to demonstrate that LSEVh-LS-F rapidly mobilized NK cells to eliminate >80% of HIV-1-infected cells in vivo 1 day after its administration. The capacity of LSEVh-LS-F to eliminate HIV-1-infected cells via ADCC combined with its broad neutralization activity supports its potential use as an immunotherapeutic agent to eliminate reactivated latent cells and deplete the HIV-1 reservoir. IMPORTANCE Mobilization of antibody-dependent cellular cytotoxicity (ADCC) to eliminate reactivated latent HIV-1-infected cells is a strategy which may contribute to depleting the HIV-1 reservoir and achieving a functional HIV-1 cure. To more effectively mobilize ADCC, we designed and constructed LSEVh-LS-F, a broadly neutralizing, defucosylated hexavalent fusion protein specific for both the CD4 and coreceptor gp120-binding sites. LSEVh-LS-F potently inhibited in vivo HIV-1 and SHIV infection in humanized mouse and macaque models, respectively, including in vivo neutralization of an HIV-1 strain resistant to the broadly neutralizing antibodies VRC01 and 3BNC117. Using a novel humanized mouse model, we demonstrated that LSEVh-LS-F rapidly mobilized NK cells to eliminate >80% of HIV-1-infected cells in vivo 1 day after its administration. The capacity of LSEVh-LS-F to eliminate HIV-1-infected cells via ADCC combined with its broad neutralization activity supports its potential use as an immunotherapeutic agent to eliminate reactivated latent cells and deplete the HIV-1 reservoir.
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- 2017
12. In Vivo Activation of Human NK Cells by Treatment with an Interleukin-15 Superagonist Potently Inhibits Acute In Vivo HIV-1 Infection in Humanized Mice
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John C. Kappes, Candice Church, Kieran Seay, Jian Hua Zheng, Harris Goldstein, Hing C. Wong, Bai Liu, Kathryn Deneroff, Christina Ochsenbauer, and Emily K. Jeng
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Interleukin-15 ,biology ,Immunology ,Degranulation ,HIV Infections ,Mice, SCID ,Lymphocyte Activation ,Microbiology ,Killer Cells, Natural ,Granzyme B ,Disease Models, Animal ,Mice ,Interleukin 21 ,Perforin ,Interleukin 15 ,In vivo ,Virology ,Insect Science ,biology.protein ,Interleukin 12 ,Animals ,Humans ,Pathogenesis and Immunity ,Cytotoxic T cell - Abstract
Natural killer (NK) cells with anti-HIV-1 activity may inhibit HIV-1 replication and dissemination during acute HIV-1 infection. We hypothesized that the capacity of NK cells to suppress acute in vivo HIV-1 infection would be augmented by activating them via treatment with an interleukin-15 (IL-15) superagonist, IL-15 bound to soluble IL-15Rα, an approach that potentiates human NK cell-mediated killing of tumor cells. In vitro stimulation of human NK cells with a recombinant IL-15 superagonist significantly induced their expression of the cytotoxic effector molecules granzyme B and perforin; their degranulation upon exposure to K562 cells, as indicated by cell surface expression of CD107a; and their capacity to lyse K562 cells and HIV-1-infected T cells. The impact of IL-15 superagonist-induced activation of human NK cells on acute in vivo HIV-1 infection was investigated by using hu-spl-PBMC-NSG mice, NOD-SCID-IL2rγ −/− (NSG) mice intrasplenically injected with human peripheral blood mononuclear cells (PBMCs) which develop productive in vivo infection after intrasplenic inoculation with HIV-1. IL-15 superagonist treatment potently inhibited acute HIV-1 infection in hu-spl-PBMC-NSG mice even when delayed until 3 days after intrasplenic HIV-1 inoculation. Removal of NK cells from human PBMCs prior to intrasplenic injection into NSG mice completely abrogated IL-15 superagonist-mediated suppression of in vivo HIV-1 infection. Thus, the in vivo activation of NK cells, integral mediators of the innate immune response, by treatment with an IL-15 superagonist increases their anti-HIV activity and enables them to potently suppress acute in vivo HIV-1 infection. These results indicate that in vivo activation of NK cells may represent a new immunotherapeutic approach to suppress acute HIV-1 infection. IMPORTANCE Epidemiological studies have indicated that NK cells contribute to the control of HIV-1 infection, and in vitro studies have demonstrated that NK cells can selectively kill HIV-1-infected cells. We demonstrated that in vivo activation of NK cells by treatment with an IL-15 superagonist that potently stimulates the antitumor activity of NK cells markedly inhibited acute HIV-1 infection in humanized mice, even when activation of NK cells by IL-15 superagonist treatment is delayed until 3 days after HIV-1 inoculation. NK cell depletion from PBMCs prior to their intrasplenic injection abrogated the suppression of in vivo HIV-1 infection observed in humanized mice treated with the IL-15 superagonist, demonstrating that activated human NK cells were mediating IL-15 superagonist-induced inhibition of acute HIV-1 infection. Thus, in vivo immunostimulation of NK cells, a promising therapeutic approach for cancer therapy, may represent a new treatment modality for HIV-1-infected individuals, particularly in the earliest stages of infection.
- Published
- 2015
13. Granzyme-Mediated Regulation of Host Defense in the Liver in Experimental Leishmania donovani Infection
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Marisa Mitchell-Flack, Xiaojing Ma, Henry W. Murray, and Hua Zheng
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T cell ,Immunology ,Leishmania donovani ,Biology ,Microbiology ,Granzymes ,GZMB ,Interferon-gamma ,Mice ,medicine ,Animals ,Inflammation ,Mice, Knockout ,Mice, Inbred BALB C ,Granuloma ,Perforin ,Effector ,Macrophages ,Intracellular parasite ,Interleukin-18 ,Macrophage Activation ,Th1 Cells ,biology.organism_classification ,Virology ,Immunity, Innate ,Interleukin-10 ,Mice, Inbred C57BL ,Infectious Diseases ,medicine.anatomical_structure ,Liver ,Granzyme ,biology.protein ,Leishmaniasis, Visceral ,Parasitology ,Fungal and Parasitic Infections ,CD8 ,T-Lymphocytes, Cytotoxic - Abstract
In the livers of susceptible C57BL/6 (B6) mice infected with Leishmania donovani , CD8 + T cell mechanisms are required for granuloma assembly, macrophage activation, intracellular parasite killing, and self-cure. Since gene expression of perforin and granzymes A and B (GzmA and GzmB), cytolytic proteins linked to CD8 + cell effector function, was enhanced in infected liver tissue, B6 mice deficient in these granular proteins were used to gauge host defense roles. Neither perforin nor GzmA was required; however, mice deficient in GzmB (GzmB −/− , GzmB cluster −/− , and GzmA×B cluster double knockout [DKO] mice) showed both delayed granuloma assembly and initially impaired control of parasite replication. Since these two defects in B6 mice were limited to early-stage infection, innately resistant 129/Sv mice were also tested. In this genetic setting, expression of both innate and subsequent T (Th1) cell-dependent acquired resistance, including the self-cure phenotype, was entirely derailed in GzmA×B cluster DKO mice. These results, in susceptible B6 mice for GzmB and in resistant 129/Sv mice for GzmA and/or the GzmB cluster, point to granzyme-mediated host defense regulation in the liver in experimental visceral leishmaniasis.
- Published
- 2015
14. T-Cell Receptor (TCR) Clonotype-Specific Differences in Inhibitory Activity of HIV-1 Cytotoxic T-Cell Clones Is Not Mediated by TCR Alone
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Huabiao Chen, Jian Hua Zheng, Nina C. Flerin, Zaza M. Ndhlovu, Evan W. Newell, Pedro A. Lamothe, Justin Fang, Tynisha D. Glover, Mark M. Davis, Bruce D. Walker, and Harris Goldstein
- Subjects
0301 basic medicine ,medicine.medical_treatment ,Immunology ,Receptors, Antigen, T-Cell ,Epitopes, T-Lymphocyte ,Gene Expression ,Cellular Response to Infection ,Major histocompatibility complex ,Microbiology ,Jurkat cells ,Epitope ,03 medical and health sciences ,0302 clinical medicine ,Virology ,medicine ,Humans ,Cytotoxic T cell ,Cloning, Molecular ,Cells, Cultured ,biology ,T-cell receptor ,Molecular biology ,030104 developmental biology ,Cytokine ,Insect Science ,Chemokine secretion ,HIV-1 ,biology.protein ,CD8 ,T-Lymphocytes, Cytotoxic ,030215 immunology - Abstract
Functional analysis of T-cell responses in HIV-infected individuals has indicated that virus-specific CD8 + T cells with superior antiviral efficacy are well represented in HIV-1 controllers but are rare or absent in HIV-1 progressors. To define the role of individual T-cell receptor (TCR) clonotypes in differential antiviral CD8 + T-cell function, we performed detailed functional and mass cytometric cluster analysis of multiple CD8 + T-cell clones recognizing the identical HLA-B*2705-restricted HIV-1 epitope KK10 (KRWIILGLNK). Effective and ineffective CD8 + T-cell clones segregated based on responses to HIV-1-infected and peptide-loaded target cells. Following cognate peptide stimulation, effective HIV-specific clones displayed significantly more rapid TCR signal propagation, more efficient initial lytic granule release, and more sustained nonlytic cytokine and chemokine secretion than ineffective clones. To evaluate the TCR clonotype contribution to CD8 + T-cell function, we cloned the TCR α and β chain genes from one effective and two ineffective CD8 + T-cell clones from an elite controller into TCR-expressing lentivectors. We show that Jurkat/MA cells and primary CD8 + T cells transduced with lentivirus expressing TCR from one of the ineffective clones exhibited a level of activation by cognate peptide and inhibition of in vitro HIV-1 infection, respectively, that were comparable to those of the effective clonotype. Taken together, these data suggest that the potent antiviral capacity of some HIV-specific CD8 + T cells is a consequence of factors in addition to TCR sequence that modulate functionality and contribute to the increased antiviral capacity of HIV-specific CD8 + T cells in elite controllers to inhibit HIV infection. IMPORTANCE The greater ex vivo antiviral inhibitory activity of CD8 + T cells from elite controllers than from HIV-1 progressors supports the crucial role of effective HIV-specific CD8 + T cells in controlling HIV-1 replication. The contribution of TCR clonotype to inhibitory potency was investigated by delineating the responsiveness of effective and ineffective CD8 + T-cell clones recognizing the identical HLA-B*2705-restricted HIV-1 Gag-derived peptide, KK10 (KRWIILGLNK). KK10-stimulated “effective” CD8 + T-cell clones displayed significantly more rapid TCR signal propagation, more efficient initial lytic granule release, and more sustained cytokine and chemokine secretion than “ineffective” CD8 + T-cell clones. However, TCRs cloned from an effective and one of two ineffective clones conferred upon primary CD8 + T cells the equivalent potent capacity to inhibit HIV-1 infection. Taken together, these data suggest that other factors aside from intrinsic TCR-peptide-major histocompatibility complex (TCR-peptide-MHC) reactivity can contribute to the potent antiviral capacity of some HIV-specific CD8 + T-cell clones.
- Published
- 2017
15. Inhibition of In Vivo HIV Infection in Humanized Mice by Gene Therapy of Human Hematopoietic Stem Cells with a Lentiviral Vector Encoding a Broadly Neutralizing Anti-HIV Antibody
- Author
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Jian Hua Zheng, Aviva Joseph, Renate Kunert, Gabriela Stiegler, Ken Chen, Monica Dutta, Harris Goldstein, Cindy Chen, and Antonia Follenzi
- Subjects
Genetic enhancement ,Genetic Vectors ,Immunology ,HIV Infections ,Mice, SCID ,Antibodies, Viral ,Microbiology ,Viral vector ,Mice ,Immune system ,Mice, Inbred NOD ,Virology ,Animals ,Humans ,Neutralizing antibody ,biology ,Lentivirus ,virus diseases ,Genetic Therapy ,Viral Load ,Hematopoietic Stem Cells ,biology.organism_classification ,Antibodies, Neutralizing ,Transplantation ,Insect Science ,HIV-1 ,Lentivirus Infections ,biology.protein ,Pathogenesis and Immunity ,Immunotherapy ,Stem cell ,Antibody ,Spleen - Abstract
Due to the inherent immune evasion properties of the HIV envelope, broadly neutralizing HIV-specific antibodies capable of suppressing HIV infection are rarely produced by infected individuals. We examined the feasibility of utilizing genetic engineering to circumvent the restricted capacity of individuals to endogenously produce broadly neutralizing HIV-specific antibodies. We constructed a single lentiviral vector that encoded the heavy and light chains of 2G12, a broadly neutralizing anti-HIV human antibody, and that efficiently transduced and directed primary human B cells to secrete 2G12. To evaluate the capacity of this approach to provide protection from in vivo HIV infection, we used the humanized NOD/SCID/γ c null mouse model, which becomes populated with human B cells, T cells, and macrophages after transplantation with human hematopoietic stem cells (hu-HSC) and develops in vivo infection after inoculation with HIV. The plasma of the irradiated NOD/SCID/γ c null mice transplanted with hu-HSC transduced with the 2G12-encoding lentivirus contained 2G12 antibody, likely secreted by progeny human lymphoid and/or myeloid cells. After intraperitoneal inoculation with high-titer HIV-1 JR-CSF , mice engrafted with 2G12-transduced hu-HSC displayed marked inhibition of in vivo HIV infection as manifested by a profound 70-fold reduction in plasma HIV RNA levels and an almost 200-fold reduction in HIV-infected human cell numbers in mouse spleens, compared to control hu-HSC-transplanted NOD/SCID/γ c null mice inoculated with equivalent high-titer HIV-1 JR-CSF . These results support the potential efficacy of this new gene therapy approach of using lentiviral vectors encoding a mixture of broadly neutralizing HIV antibodies for the treatment of HIV infection, particularly infection with multiple-drug-resistant isolates.
- Published
- 2010
16. Increased In Vivo Activation of Microglia and Astrocytes in the Brains of Mice Transgenic for an Infectious R5 Human Immunodeficiency Virus Type 1 Provirus and for CD4-Specific Expression of Human Cyclin T1 in Response to Stimulation by Lipopolysaccharides
- Author
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Jinglin Sun, Mengliang Zhao, Jian Hua Zheng, Sunhee Lee, and Harris Goldstein
- Subjects
Lipopolysaccharides ,Chemokine ,Lipopolysaccharide ,Transgene ,Immunology ,HIV Infections ,Mice, Transgenic ,Microbiology ,Proinflammatory cytokine ,Mice ,chemistry.chemical_compound ,Proviruses ,In vivo ,Cyclins ,Virology ,medicine ,Animals ,Humans ,Chemokine CCL2 ,biology ,Microglia ,Cyclin T ,Brain ,Granulocyte-Macrophage Colony-Stimulating Factor ,Provirus ,medicine.anatomical_structure ,chemistry ,Astrocytes ,Insect Science ,CD4 Antigens ,HIV-1 ,biology.protein ,Pathogenesis and Immunity ,Astrocyte - Abstract
Inflammatory mediators and viral products produced by human immunodeficiency virus (HIV)-infected microglia and astrocytes perturb the function and viability of adjacent uninfected neuronal and glial cells and contribute to the pathogenesis of HIV-associated neurocognitive disorders (HAND). In vivo exposure to lipopolysaccharide (LPS) activates parenchymal microglia and astrocytes and induces cytokine and chemokine production in the brain. HIV-infected individuals display increased circulating LPS levels due to microbial translocation across a compromised mucosa barrier. We hypothesized that HIV-infected microglia and astrocytes display increased sensitivity to the proinflammatory effects of LPS, and this combines with the increased levels of systemic LPS in HIV-infected individuals to contribute to the development of HAND. To examine this possibility, we determined the in vivo responsiveness of HIV-infected microglia and astrocytes to LPS using our mouse model, JR-CSF/human cyclin T1 (JR-CSF/hu-cycT1) mice, which are transgenic for both an integrated full-length infectious HIV type 1 (HIV-1) provirus derived from the primary R5-tropic clinical isolate HIV-1JR-CSFregulated by the endogenous HIV-1 long terminal repeat and the hu-cycT1 gene under the control of a CD4 promoter. In the current report, we demonstrated that in vivo-administered LPS more potently activated JR-CSF/hu-cycT1 mouse microglia and astrocytes and induced a significantly higher degree of monocyte chemoattractant protein production by JR-CSF/hu-cycT1 astrocytes compared to that of the in vivo LPS response of control littermate mouse microglia and astrocytes. These results indicate that HIV infection increases the sensitivity of microglia and astrocytes to inflammatory stimulation and support the use of these mice as a model to investigate various aspects of the in vivo mechanism of HIV-induced neuronal dysfunction.
- Published
- 2008
17. Correlating Microbial Diversity Patterns with Geochemistry in an Extreme and Heterogeneous Environment of Mine Tailings
- Author
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Liu, Jun, primary, Hua, Zheng-Shuang, additional, Chen, Lin-Xing, additional, Kuang, Jia-Liang, additional, Li, Sheng-Jin, additional, Shu, Wen-Sheng, additional, and Huang, Li-Nan, additional
- Published
- 2014
- Full Text
- View/download PDF
18. Establishment of a Novel Humanized Mouse Model To Investigate In Vivo Activation and Depletion of Patient-Derived HIV Latent Reservoirs.
- Author
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Flerin, Nina C., Bardhi, Ariola, Jian Hua Zheng, Korom, Maria, Folkvord, Joy, Kovacs, Colin, Benko, Erika, Truong, Ronald, Mota, Talia, Connick, Elizabeth, Jones, R. Brad, Lynch, Rebecca M., and Goldsteina, Harris
- Subjects
- *
HIV-positive women , *VIRAL antibodies , *RESERVOIRS , *HIV , *HIV infections - Abstract
Curing HIV infection has been thwarted by the persistent reservoir of latently infected CD4+ T cells, which reinitiate systemic infection after antiretroviral therapy (ART) interruption. To evaluate reservoir depletion strategies, we developed a novel preclinical in vivo model consisting of immunodeficient mice intrasplenically injected with peripheral blood mononuclear cells (PBMC) from long-term ART-suppressed HIV-infected donors. In the absence of ART, these mice developed rebound viremia which, 2 weeks after PBMC injection, was 1,000-fold higher (mean=9,229,281 HIV copies/ml) in mice injected intrasplenically than in mice injected intraperitoneally (mean=6,838 HIV copies/ml) or intravenously (mean=591 HIV copies/ml). One week after intrasplenic PBMC injection, in situ hybridization of the spleen demonstrated extensive disseminated HIV infection, likely initiated from in vivo-reactivated primary latently infected cells. The time to viremia was delayed significantly by treatment with a broadly neutralizing antibody, 10-1074, compared to treatment with 10-1074-FcRnull, suggesting that 10-1074 mobilized Fc-mediated effector mechanisms to deplete the replication-competent reservoir. This was supported by phylogenetic analysis of Env sequences from viral-outgrowth cultures and untreated, 10-1074-treated, or 10-1074-FcRnull-treated mice. The predominant sequence cluster detected in viral-outgrowth cultures and untreated mouse plasma was significantly reduced in the plasma of 10-1074-treated mice, whereas two new clusters emerged that were not detected in viral-outgrowth cultures or plasma from untreated mice. These new clusters lacked mutations associated with 10-1074 resistance. Taken together, these data indicated that 10-1074 treatment depletes the reservoir of latently infected cells harboring replication competent HIV. Furthermore, this mouse model represents a new in vivo approach for the preclinical evaluation of new HIV cure strategies. [ABSTRACT FROM AUTHOR]
- Published
- 2019
- Full Text
- View/download PDF
19. Inhibition of In Vivo HIV Infection in Humanized Mice by Gene Therapy of Human Hematopoietic Stem Cells with a Lentiviral Vector Encoding a Broadly Neutralizing Anti-HIV Antibody.
- Author
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Joseph, Aviva, Jian Hua Zheng, Ken Chen, Dutta, Monica, Chen, Cindy, Stiegler, Gabriela, Kunert, Renate, Follenzi, Antonia, and Goldstein, Harris
- Subjects
- *
HIV , *IMMUNOGLOBULINS , *GENETIC engineering , *HIV infections , *B cells - Abstract
Due to the inherent immune evasion properties of the HIV envelope, broadly neutralizing HIV-specific antibodies capable of suppressing HIV infection are rarely produced by infected individuals. We examined the feasibility of utilizing genetic engineering to circumvent the restricted capacity of individuals to endogenously produce broadly neutralizing HIV-specific antibodies. We constructed a single lentiviral vector that encoded the heavy and light chains of 2G12, a broadly neutralizing anti-HIV human antibody, and that efficiently transduced and directed primary human B cells to secrete 2G12. To evaluate the capacity of this approach to provide protection from in vivo HIV infection, we used the humanized NOD/SCID/ γcnull mouse model, which becomes populated with human B cells, T cells, and macrophages after transplantation with human hematopoietic stem cells (hu-HSC) and develops in vivo infection after inoculation with HIV. The plasma of the irradiated NOD/SCID/ γcnull mice transplanted with hu-HSC transduced with the 2G12-encoding lentivirus contained 2G12 antibody, likely secreted by progeny human lymphoid and/or myeloid cells. After intraperitoneal inoculation with high-titer HIV-1JR-CSF, mice engrafted with 2G12-transduced hu-HSC displayed marked inhibition of in vivo HIV infection as manifested by a profound 70-fold reduction in plasma HIV RNA levels and an almost 200-fold reduction in HIV-infected human cell numbers in mouse spleens, compared to control hu-HSC-transplanted NOD/SCID/γcnull mice inoculated with equivalent high-titer HIV-1JR-CSF. These results support the potential efficacy of this new gene therapy approach of using lentiviral vectors encoding a mixture of broadly neutralizing HIV antibodies for the treatment of HIV infection, particularly infection with multiple-drug-resistant isolates. [ABSTRACT FROM AUTHOR]
- Published
- 2010
- Full Text
- View/download PDF
20. Lentiviral Vectors Encoding Human Immunodeficiency Virus Type 1 (HIV-1)-Specific T-Cell Receptor Genes Efficiently Convert Peripheral Blood CD8 T Lymphocytes into Cytotoxic T Lymphocytes with Potent In Vitro and In Vivo HIV-1-Specific Inhibitory Activity.
- Author
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Joseph, Aviva, Jian Hua Zheng, Follenzi, Antonia, DiLorenzo, Teresa, Sango, Kaori, Hyman, Jaime, Ken Chen, Piechocka-Trocha, Alicja, Brander, Christian, Hooijberg, Erik, Vignali, Dario A., Walker, Bruce D., and Goldstein, Harris
- Subjects
- *
HIV , *LENTIVIRUSES , *T cell receptors , *GENES , *LYMPHOCYTES , *BLOOD vessels - Abstract
The human immunodeficiency virus type 1 (HIV-1)-specific CD8 cytotoxic T-lymphocyte (CTL) response plays a critical role in controlling HIV-1 replication. Augmenting this response should enhance control of HIV-1 replication and stabilize or improve the clinical course of the disease. Although cytomegalovirus (CMV) or Epstein-Barr virus (EBV) infection in immunocompromised patients can be treated by adoptive transfer of ex vivo-expanded CMV- or EBV-specific CTLs, adoptive transfer of ex vivo-expanded, autologous HIV-1-specific CTLs had minimal effects on HIV-1 replication, likely a consequence of the inherently compromised qualitative function of HIV-1-specific CTLs derived from HIV-1-infected individuals. We hypothesized that this limitation could be circumvented by using as an alternative source of HIV-1-specific CTLs, autologous peripheral CD8+ T lymphocytes whose antigen specificity is redirected by transduction with lentiviral vectors encoding HIV-1-specific T-cell receptor (TCR) αand ß chains, an approach used successfully in cancer therapy. To efficiently convert peripheral CD8 lymphocytes into HIV-1-specific CTLs that potently suppress in vivo HIV-1 replication, we constructed lentiviral vectors encoding the HIV-1-specific TCR αand TCR ß chains cloned from a CTL clone specific for an HIV Gag epitope, SL9, as a single transcript linked with a self-cleaving peptide. We demonstrated that transduction with this lentiviral vector efficiently converted primary human CD8 lymphocytes into HIV-1-specific CTLs with potent in vitro and in vivo HIV-1-specific activity. Using lentiviral vectors encoding an HIV-1-specific TCR to transform peripheral CD8 lymphocytes into HIV-1-specific CTLs with defined specificities represents a new immunotherapeutic approach to augment the HIV-1-specific immunity of infected patients. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
21. In Vivo Activation of Human NK Cells by Treatment with an Interleukin-15 Superagonist Potently Inhibits Acute In Vivo HIV-1 Infection in Humanized Mice.
- Author
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Seay, Kieran, Church, Candice, Jian Hua Zheng, Deneroff, Kathryn, Ochsenbauer, Christina, Kappes, John C., Bai Liu, Jeng, Emily K., Wong, Hing C., and Goldsteina, Harris
- Subjects
- *
KILLER cells , *HIV infections , *THERAPEUTICS , *INTERLEUKIN-15 , *GENE expression , *LABORATORY mice - Abstract
Natural killer (NK) cells with anti-HIV-1 activity may inhibit HIV-1 replication and dissemination during acute HIV-1 infection. We hypothesized that the capacity of NK cells to suppress acute in vivo HIV-1 infection would be augmented by activating them via treatment with an interleukin-15 (IL-15) superagonist, IL-15 bound to soluble IL-15Ra, an approach that potentiates human NK cell-mediated killing of tumor cells. In vitro stimulation of human NK cells with a recombinant IL-15 superagonist significantly induced their expression of the cytotoxic effector molecules granzyme B and perforin; their degranulation upon exposure to K562 cells, as indicated by cell surface expression of CD107a; and their capacity to lyse K562 cells and HIV-1-infected T cells. The impact of IL-15 superagonist-induced activation of human NK cells on acute in vivo HIV-1 infection was investigated by using hu-spl-PBMC-NSG mice, NOD-SCID-IL2rγ-/- (NSG) mice intrasplenically injected with human peripheral blood mononuclear cells (PBMCs) which develop productive in vivo infection after intrasplenic inoculation with HIV-1. IL-15 superagonist treatment potently inhibited acute HIV-1 infection in hu-spl-PBMC-NSG mice even when delayed until 3 days after intrasplenic HIV-1 inoculation. Removal of NK cells from human PBMCs prior to intrasplenic injection into NSG mice completely abrogated IL-15 superagonist-mediated suppression of in vivo HIV-1 infection. Thus, the in vivo activation of NK cells, integral mediators of the innate immune response, by treatment with an IL-15 superagonist increases their anti-HIV activity and enables them to potently suppress acute in vivo HIV-1 infection. These results indicate that in vivo activation of NK cells may represent a new immunotherapeutic approach to suppress acute HIV-1 infection. [ABSTRACT FROM AUTHOR]
- Published
- 2015
- Full Text
- View/download PDF
22. Potent In Vivo NK Cell-Mediated Elimination of HIV-1-Infected Cells Mobilized by a gp120-Bispecific and Hexavalent Broadly Neutralizing Fusion Protein.
- Author
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Bardhi, Ariola, Yanling Wu, Weizao Chen, Wei Li, Zhongyu Zhu, Jian Hua Zheng, Hing Wong, Emily Jeng, Jones, Jennifer, Ochsenbauer, Christina, Kappes, John C., Dimitrov, Dimiter S., Tianlei Ying, and Goldstein, Harris
- Subjects
- *
ANTIBODY-dependent cell cytotoxicity , *HIV prevention , *CHIMERIC proteins , *IMMUNOGLOBULINS , *IMMUNE response ,PREVENTION of disease progression - Abstract
Antibodies bound to human immunodeficiency virus type 1 (HIV-1) envelope protein expressed by infected cells mobilize antibody-dependent cellular cytotoxicity (ADCC) to eliminate the HIV-1-infected cells and thereby suppress HIV-1 infection and delay disease progression. Studies treating HIV-1-infected individuals with latency reactivation agents to reduce their latent HIV-1 reservoirs indicated that their HIV-1-specific immune responses were insufficient to effectively eliminate the reactivated latent HIV-1-infected T cells. Mobilization of ADCC may facilitate elimination of reactivated latent HIV-1-infected cells to deplete the HIV-1 reservoir and contribute to a functional HIV-1 cure. The most effective antibodies for controlling and eradicating HIV-1 infection would likely have the dual capacities of potently neutralizing a broad range of HIV-1 isolates and effectively mobilizing HIV-1-specific ADCC to eliminate HIV-1-infected cells. For this purpose, we constructed LSEVh-LS-F, a broadly neutralizing, defucosylated hexavalent fusion protein specific for both the CD4 and coreceptor gp120-binding sites. LSEVh-LS-F potently inhibited in vivo HIV-1 and simian-human immunodeficiency virus (SHIV) infection in humanized mouse and macaque models, respectively, including in vivo neutralization of HIV-1 strains resistant to the broadly neutralizing antibodies VRC01 and 3BNC117. We developed a novel humanized mouse model to evaluate in vivo human NK cell-mediated elimination of HIV-1-infected cells by ADCC and utilized it to demonstrate that LSEVh-LS-F rapidly mobilized NK cells to eliminate >80% of HIV-1-infected cells in vivo 1 day after its administration. The capacity of LSEVh-LS-F to eliminate HIV-1-infected cells via ADCC combined with its broad neutralization activity supports its potential use as an immunotherapeutic agent to eliminate reactivated latent cells and deplete the HIV-1 reservoir. IMPORTANCE Mobilization of antibody-dependent cellular cytotoxicity (ADCC) to eliminate reactivated latent HIV-1-infected cells is a strategy which may contribute to depleting the HIV-1 reservoir and achieving a functional HIV-1 cure. To more effectively mobilize ADCC, we designed and constructed LSEVh-LS-F, a broadly neutralizing, defucosylated hexavalent fusion protein specific for both the CD4 and coreceptor gp120-binding sites. LSEVh-LS-F potently inhibited in vivo HIV-1 and SHIV infection in humanized mouse and macaque models, respectively, including in vivo neutralization of an HIV-1 strain resistant to the broadly neutralizing antibodies VRC01 and 3BNC117. Using a novel humanized mouse model, we demonstrated that LSEVh-LS-F rapidly mobilized NK cells to eliminate >80% of HIV-1-infected cells in vivo 1 day after its administration. The capacity of LSEVh-LS-F to eliminate HIV-1-infected cells via ADCC combined with its broad neutralization activity supports its potential use as an immunotherapeutic agent to eliminate reactivated latent cells and deplete the HIV-1 reservoir. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
23. T-Cell Receptor (TCR) Clonotype-Specific Differences in Inhibitory Activity of HIV-1 Cytotoxic T-Cell Clones Is Not Mediated by TCR Alone.
- Author
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Flerin, Nina C., Huabiao Chen, Glover, Tynisha D., Lamothe, Pedro A., Jian Hua Zheng, Fang, Justin W., Ndhlovu, Zaza M., Newell, Evan W., Davis, Mark M., Walker, Bruce D., and Goldstein, Harris
- Subjects
- *
T cells , *CELL receptors , *FUNCTIONAL analysis , *HIV-positive persons , *CELL physiology - Abstract
Functional analysis of T-cell responses in HIV-infected individuals has indicated that virus-specific CD8+ T cells with superior antiviral efficacy are well represented in HIV-1 controllers but are rare or absent in HIV-1 progressors. To define the role of individual T-cell receptor (TCR) clonotypes in differential antiviral CD8+ T-cell function, we performed detailed functional and mass cytometric cluster analysis of multiple CD8+ T-cell clones recognizing the identical HLA-B+2705-restricted HIV-1 epitope KK10 (KRWIILGLNK). Effective and ineffective CD8+ T-cell clones segregated based on responses to HIV-1-infected and peptide-loaded target cells. Following cognate peptide stimulation, effective HIV-specific clones displayed significantly more rapid TCR signal propagation, more efficient initial lytic granule release, and more sustained nonlytic cytokine and chemokine secretion than ineffective clones. To evaluate the TCR clonotype contribution to CD8+ T-cell function, we cloned the TCR α and β chain genes from one effective and two ineffective CD8+ T-cell clones from an elite controller into TCR-expressing lentivectors. We show that Jurkat/MA cells and primary CD8+ T cells transduced with lentivirus expressing TCR from one of the ineffective clones exhibited a level of activation by cognate peptide and inhibition of in vitro HIV-1 infection, respectively, that were comparable to those of the effective clonotype. Taken together, these data suggest that the potent antiviral capacity of some HIV-specific CD8+ T cells is a consequence of factors in addition to TCR sequence that modulate functionality and contribute to the increased antiviral capacity of HIV-specific CD8+ T cells in elite controllers to inhibit HIV infection. IMPORTANCE The greater ex vivo antiviral inhibitory activity of CD8++ T cells from elite controllers than from HIV-1 progressors supports the crucial role of effective HIV-specific CD8+ T cells in controlling HIV-1 replication. The contribution of TCR clonotype to inhibitory potency was investigated by delineating the responsiveness of effective and ineffective CD8+ T-cell clones recognizing the identical HLA-B*2705-restricted HIV-1 Gag-derived peptide, KK10 (KRWIILGLNK). KK10-stimulated "effective" CD8+ T-cell clones displayed significantly more rapid TCR signal propagation, more efficient initial lytic granule release, and more sustained cytokine and chemokine secretion than "ineffective" CD8+ T-cell clones. However, TCRs cloned from an effective and one of two ineffective clones conferred upon primary CD8+ T cells the equivalent potent capacity to inhibit HIV-1 infection. Taken together, these data suggest that other factors aside from intrinsic TCR-peptide-major histocompatibility complex (TCR-peptide-MHC) reactivity can contribute to the potent antiviral capacity of some HIV-specific CD8+ T-cell clones. [ABSTRACT FROM AUTHOR]
- Published
- 2017
- Full Text
- View/download PDF
24. CD4-Specific Transgenic Expression of Human Cyclin T1 Markedly Increases Human Immunodeficiency Virus Type 1 (HIV-1) Production by CD4+ T Lymphocytes and Myeloid Cells in Mice Transgenic for a Provirus Encoding a Monocyte-Tropic HIV-1 Isolate.
- Author
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Jinglin Sun, Soos, Timothy, Kewalramani, Vineet N., Osiecki, Kristin, Jian Hua Zheng, Falkin, Laurie, Santambrogio, Laura, Littman, Dan R., and Goidstein, Harris
- Subjects
- *
CD4 antigen , *CD antigens , *VIRAL receptors , *TRANSGENE expression , *HIV , *TRANSGENIC mice , *LABORATORY mice - Abstract
Human immunodeficiency virus type 1 (HIV-1)-encoded Tat provides transcriptional activation critical for efficient HIV-1 replication by interacting with cyclin T1 and recruiting P-TEFb to efficiently elongate the nascent HIV transcript. Tat-mediated transcriptional activation in mice is precluded by species-specific structural differences that prevent Tat interaction with mouse cyclin T1 and severely compromise HIV-I replication in mouse cells. We investigated whether transgenic mice expressing human cyclin T1 under the control of a murine CD4 promoter/enhancer cassette that directs gene expression to CD4+ T lymphocytes and monocytes/macrophages (hu-cycT1 mice) would display Tat responsiveness in their CD4-expressing mouse cells and selectively increase HIV-1 production in this cellular population, which is infected primarily in HIV-1-positive individuals. To this end, we crossed hu-cycT1 mice with JR-CSF transgenic mice carrying the full-length HIV-1JR-CSF provirus under the control of the endogenous HIV-1 long terminal repeat and demonstrated that human cyclin T1 expression is sufficient to support Tat-mediated transactivation in primary mouse CD4 T lymphocytes and monocytes/macrophages and increases in vitro and in vivo HIV-1 production by these stimulated cells. Increased HIV-1 production by CD4+ T lymphocytes was paralleled with their specific depletion in the peripheral blood of the JR-CSF/hu-cycT1 mice, which increased over time. In addition, increased HIV-1 transgene expression due to human cyclin T1 expression was associated with increased lipopolysaccharide-stimulated monocyte chemoattractant protein 1 production by JR-CSF mouse monocytes/ macrophages in vitro. Therefore, the JR-CSF/hu-cycT1 mice should provide an improved mouse system for investigating the pathogenesis of various aspects of HIV-1-mediated disease and the efficacies of therapeutic interventions. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
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