Journal: Cell reports

Article Title: Slow Receptor Binding of the Noncytopathic HIV-2 UC1 Envs Is Balanced by Long-Lived Activation State and Efficient Fusion Activity

doi: 10.1016/j.celrep.2020.107749

Figure Lengend Snippet: (A) Infection of cells expressing different receptors. HIV-2UC1, HIV-1JRFL, or A-MLV PVs, were used to infect Cf2 cells expressing CD4/CCR5, CCR5, or CD4/CXCR4 receptors. HIV-1JRFL Envs share many properties with other HIV-1 strains (Figure S1).

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-human CD4 RPA-T4 BD PharMingen Cat# 550369; RRID:AB_393640 Chemicals, Peptides, and Recombinant Proteins Effectene Transfection Reagent QIAGEN Cat# 301425 Passive Lysis Buffer Promega Cat# E1941 293fectin Invitrogen Cat# 12347019 Genticin G418 Sulfate Invitrogen Cat# 10131027 Penicillin/Streptomycin ThermoFisher Cat# 15140122 Hygromycin B Invitrogen Cat# 10687010 Maraviroc (CCR5 inhibitor) NIH AIDS Reagent Program Cat# 11580 D-Luciferin Phosphate BD Biosciences Cat# 556879 10 mM glycine (pH 2.5) GE Life Sciences Regeneration Buffer Cat# BR100356 Soluble HIV-1 JRFL gp120 Current study N/A Soluble HIV-2 UC1 gp125 Current study N/A Critical Commercial Assays HIV-1 p24 antigen capture assay Advanced BioScience Laboratories Cat# 5421 CM5 Biacore sensor chip GE Life Sciences Cat# 29104988 Deposited Data Compile, Analyze and Tally Neutralizing Antibodies Panel (CATNAP) HIV Database https://www.hiv.lanl.gov/content/index Protein Data Bank (PDB): 5CAY (HIV-2 ST ) Research Collaboratory for Structural Bioinformatics (RCSB) https://www.rcsb.org PDB: 2QAD (HIV-1 YU2 ) RCSB https://www.rcsb.org PDB: 1RZK (HIV-2 YU2 ) RCSB https://www.rcsb.org PDB: 4R2G (HIV-1 JRCSF ) RCSB https://www.rcsb.org PDB: 3LQA(HIV-1 CAP210 ) RCSB https://www.rcsb.org Experimental Models: Cell Lines HEK293T/17 cells ATCC CRL-11268 TZM-bl cells NIH AIDS Reagent Program Cat# 8129 Cf2Th CD4/CCR5 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 R5C9 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 T4×4 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Recombinant DNA p1054.TC4.1499 NIH AIDS Reagent Program Env-expression plasmid Cat# 11561 pPRB926_04.A9.4237 NIH AIDS Reagent Program Env-expression plasmid Cat# 11573 p9021_14.B2.4571 NIH AIDS Reagent Program Env-expression plasmid Cat # 11572 psPAX2 NIH AIDS Reagent Program Cat# 11348 pHIVec2.luc Laboratory of Joseph Sodroski N/A pCO-HIV-2 UC1 env Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pUC57-CO-HIV-2 UC1 env GenScript Gene synthesis pCO-HIV-1 JRFL env Previous study Mao et al., 2012 pCO-HIV-1 JRFL gp120 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-HIV-2 UC1 gp125 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-sCD4 Current study pcDNA 3-based backbone plasmid (Invitrogen) HIV-2 ROD10 Env Laboratory of Paula Cannon Env-expressing plasmid HIV-2 ROD14 Env Laboratory of Paula Cannon Env-expressing plasmid Software and Algorithms UCSF Chimera UCSF https://www.cgl.ucsf.edu/chimera/ BIAcore BIAevaluation 3.2 GE Healthcare BR-1005–97 MikroWin 2000 Lite Berthold Technologies GmbH Id.

Techniques: Infection, Expressing

Journal: Cell reports

Article Title: Slow Receptor Binding of the Noncytopathic HIV-2 UC1 Envs Is Balanced by Long-Lived Activation State and Efficient Fusion Activity

doi: 10.1016/j.celrep.2020.107749

Figure Lengend Snippet: (A) HIV Env cold sensitivity. HIV-2UC1 or the control HIV-1JRFL PVs were incubated on ice for the indicated times and then used to infect Cf2Th-CD4/CCR5 cells. (B and C) Cell-cell fusion activity. Effector cells expressing the specified Envs were incubated with TZM-bl reporter cells for the indicated times, and cell-cell fusion was monitored by measuring the firefly luciferase activity.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-human CD4 RPA-T4 BD PharMingen Cat# 550369; RRID:AB_393640 Chemicals, Peptides, and Recombinant Proteins Effectene Transfection Reagent QIAGEN Cat# 301425 Passive Lysis Buffer Promega Cat# E1941 293fectin Invitrogen Cat# 12347019 Genticin G418 Sulfate Invitrogen Cat# 10131027 Penicillin/Streptomycin ThermoFisher Cat# 15140122 Hygromycin B Invitrogen Cat# 10687010 Maraviroc (CCR5 inhibitor) NIH AIDS Reagent Program Cat# 11580 D-Luciferin Phosphate BD Biosciences Cat# 556879 10 mM glycine (pH 2.5) GE Life Sciences Regeneration Buffer Cat# BR100356 Soluble HIV-1 JRFL gp120 Current study N/A Soluble HIV-2 UC1 gp125 Current study N/A Critical Commercial Assays HIV-1 p24 antigen capture assay Advanced BioScience Laboratories Cat# 5421 CM5 Biacore sensor chip GE Life Sciences Cat# 29104988 Deposited Data Compile, Analyze and Tally Neutralizing Antibodies Panel (CATNAP) HIV Database https://www.hiv.lanl.gov/content/index Protein Data Bank (PDB): 5CAY (HIV-2 ST ) Research Collaboratory for Structural Bioinformatics (RCSB) https://www.rcsb.org PDB: 2QAD (HIV-1 YU2 ) RCSB https://www.rcsb.org PDB: 1RZK (HIV-2 YU2 ) RCSB https://www.rcsb.org PDB: 4R2G (HIV-1 JRCSF ) RCSB https://www.rcsb.org PDB: 3LQA(HIV-1 CAP210 ) RCSB https://www.rcsb.org Experimental Models: Cell Lines HEK293T/17 cells ATCC CRL-11268 TZM-bl cells NIH AIDS Reagent Program Cat# 8129 Cf2Th CD4/CCR5 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 R5C9 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 T4×4 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Recombinant DNA p1054.TC4.1499 NIH AIDS Reagent Program Env-expression plasmid Cat# 11561 pPRB926_04.A9.4237 NIH AIDS Reagent Program Env-expression plasmid Cat# 11573 p9021_14.B2.4571 NIH AIDS Reagent Program Env-expression plasmid Cat # 11572 psPAX2 NIH AIDS Reagent Program Cat# 11348 pHIVec2.luc Laboratory of Joseph Sodroski N/A pCO-HIV-2 UC1 env Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pUC57-CO-HIV-2 UC1 env GenScript Gene synthesis pCO-HIV-1 JRFL env Previous study Mao et al., 2012 pCO-HIV-1 JRFL gp120 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-HIV-2 UC1 gp125 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-sCD4 Current study pcDNA 3-based backbone plasmid (Invitrogen) HIV-2 ROD10 Env Laboratory of Paula Cannon Env-expressing plasmid HIV-2 ROD14 Env Laboratory of Paula Cannon Env-expressing plasmid Software and Algorithms UCSF Chimera UCSF https://www.cgl.ucsf.edu/chimera/ BIAcore BIAevaluation 3.2 GE Healthcare BR-1005–97 MikroWin 2000 Lite Berthold Technologies GmbH Id.

Techniques: Control, Incubation, Activity Assay, Expressing, Luciferase

Journal: Cell reports

Article Title: Slow Receptor Binding of the Noncytopathic HIV-2 UC1 Envs Is Balanced by Long-Lived Activation State and Efficient Fusion Activity

doi: 10.1016/j.celrep.2020.107749

Figure Lengend Snippet: Slow association kinetics of HIV-2UC1 Env binding to the CD4 receptor allows the CD4 to escape downregulation during HIV-1 production in infected cells. The resulting low-affinity binding to the CD4 is compensated by (1) the long-lived activation state upon CD4 binding and (2) the efficient fusion activity of HIV-2UC1 Env that mediates viral entry.

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-human CD4 RPA-T4 BD PharMingen Cat# 550369; RRID:AB_393640 Chemicals, Peptides, and Recombinant Proteins Effectene Transfection Reagent QIAGEN Cat# 301425 Passive Lysis Buffer Promega Cat# E1941 293fectin Invitrogen Cat# 12347019 Genticin G418 Sulfate Invitrogen Cat# 10131027 Penicillin/Streptomycin ThermoFisher Cat# 15140122 Hygromycin B Invitrogen Cat# 10687010 Maraviroc (CCR5 inhibitor) NIH AIDS Reagent Program Cat# 11580 D-Luciferin Phosphate BD Biosciences Cat# 556879 10 mM glycine (pH 2.5) GE Life Sciences Regeneration Buffer Cat# BR100356 Soluble HIV-1 JRFL gp120 Current study N/A Soluble HIV-2 UC1 gp125 Current study N/A Critical Commercial Assays HIV-1 p24 antigen capture assay Advanced BioScience Laboratories Cat# 5421 CM5 Biacore sensor chip GE Life Sciences Cat# 29104988 Deposited Data Compile, Analyze and Tally Neutralizing Antibodies Panel (CATNAP) HIV Database https://www.hiv.lanl.gov/content/index Protein Data Bank (PDB): 5CAY (HIV-2 ST ) Research Collaboratory for Structural Bioinformatics (RCSB) https://www.rcsb.org PDB: 2QAD (HIV-1 YU2 ) RCSB https://www.rcsb.org PDB: 1RZK (HIV-2 YU2 ) RCSB https://www.rcsb.org PDB: 4R2G (HIV-1 JRCSF ) RCSB https://www.rcsb.org PDB: 3LQA(HIV-1 CAP210 ) RCSB https://www.rcsb.org Experimental Models: Cell Lines HEK293T/17 cells ATCC CRL-11268 TZM-bl cells NIH AIDS Reagent Program Cat# 8129 Cf2Th CD4/CCR5 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 R5C9 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 T4×4 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Recombinant DNA p1054.TC4.1499 NIH AIDS Reagent Program Env-expression plasmid Cat# 11561 pPRB926_04.A9.4237 NIH AIDS Reagent Program Env-expression plasmid Cat# 11573 p9021_14.B2.4571 NIH AIDS Reagent Program Env-expression plasmid Cat # 11572 psPAX2 NIH AIDS Reagent Program Cat# 11348 pHIVec2.luc Laboratory of Joseph Sodroski N/A pCO-HIV-2 UC1 env Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pUC57-CO-HIV-2 UC1 env GenScript Gene synthesis pCO-HIV-1 JRFL env Previous study Mao et al., 2012 pCO-HIV-1 JRFL gp120 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-HIV-2 UC1 gp125 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-sCD4 Current study pcDNA 3-based backbone plasmid (Invitrogen) HIV-2 ROD10 Env Laboratory of Paula Cannon Env-expressing plasmid HIV-2 ROD14 Env Laboratory of Paula Cannon Env-expressing plasmid Software and Algorithms UCSF Chimera UCSF https://www.cgl.ucsf.edu/chimera/ BIAcore BIAevaluation 3.2 GE Healthcare BR-1005–97 MikroWin 2000 Lite Berthold Technologies GmbH Id.

Techniques: Binding Assay, Infection, Activation Assay, Activity Assay

Journal: Cell reports

Article Title: Slow Receptor Binding of the Noncytopathic HIV-2 UC1 Envs Is Balanced by Long-Lived Activation State and Efficient Fusion Activity

doi: 10.1016/j.celrep.2020.107749

Figure Lengend Snippet: KEY RESOURCES TABLE

Article Snippet: ​ REAGENT or RESOURCE SOURCE IDENTIFIER Antibodies Anti-human CD4 RPA-T4 BD PharMingen Cat# 550369; RRID:AB_393640 Chemicals, Peptides, and Recombinant Proteins Effectene Transfection Reagent QIAGEN Cat# 301425 Passive Lysis Buffer Promega Cat# E1941 293fectin Invitrogen Cat# 12347019 Genticin G418 Sulfate Invitrogen Cat# 10131027 Penicillin/Streptomycin ThermoFisher Cat# 15140122 Hygromycin B Invitrogen Cat# 10687010 Maraviroc (CCR5 inhibitor) NIH AIDS Reagent Program Cat# 11580 D-Luciferin Phosphate BD Biosciences Cat# 556879 10 mM glycine (pH 2.5) GE Life Sciences Regeneration Buffer Cat# BR100356 Soluble HIV-1 JRFL gp120 Current study N/A Soluble HIV-2 UC1 gp125 Current study N/A Critical Commercial Assays HIV-1 p24 antigen capture assay Advanced BioScience Laboratories Cat# 5421 CM5 Biacore sensor chip GE Life Sciences Cat# 29104988 Deposited Data Compile, Analyze and Tally Neutralizing Antibodies Panel (CATNAP) HIV Database https://www.hiv.lanl.gov/content/index Protein Data Bank (PDB): 5CAY (HIV-2 ST ) Research Collaboratory for Structural Bioinformatics (RCSB) https://www.rcsb.org PDB: 2QAD (HIV-1 YU2 ) RCSB https://www.rcsb.org PDB: 1RZK (HIV-2 YU2 ) RCSB https://www.rcsb.org PDB: 4R2G (HIV-1 JRCSF ) RCSB https://www.rcsb.org PDB: 3LQA(HIV-1 CAP210 ) RCSB https://www.rcsb.org Experimental Models: Cell Lines HEK293T/17 cells ATCC CRL-11268 TZM-bl cells NIH AIDS Reagent Program Cat# 8129 Cf2Th CD4/CCR5 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 R5C9 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Cf2 T4×4 cells Laboratory of Joseph Sodroski Parental Cf2Th cells are from ATCC (CRL-1430) Recombinant DNA p1054.TC4.1499 NIH AIDS Reagent Program Env-expression plasmid Cat# 11561 pPRB926_04.A9.4237 NIH AIDS Reagent Program Env-expression plasmid Cat# 11573 p9021_14.B2.4571 NIH AIDS Reagent Program Env-expression plasmid Cat # 11572 psPAX2 NIH AIDS Reagent Program Cat# 11348 pHIVec2.luc Laboratory of Joseph Sodroski N/A pCO-HIV-2 UC1 env Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pUC57-CO-HIV-2 UC1 env GenScript Gene synthesis pCO-HIV-1 JRFL env Previous study Mao et al., 2012 pCO-HIV-1 JRFL gp120 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-HIV-2 UC1 gp125 Current study pcDNA3.1/Zeo (−) backbone plasmid (Invitrogen) pCO-sCD4 Current study pcDNA 3-based backbone plasmid (Invitrogen) HIV-2 ROD10 Env Laboratory of Paula Cannon Env-expressing plasmid HIV-2 ROD14 Env Laboratory of Paula Cannon Env-expressing plasmid Software and Algorithms UCSF Chimera UCSF https://www.cgl.ucsf.edu/chimera/ BIAcore BIAevaluation 3.2 GE Healthcare BR-1005–97 MikroWin 2000 Lite Berthold Technologies GmbH Id.

Techniques: Recombinant, Transfection, Lysis, Plasmid Preparation, Software