cemx174 5 25 m7 cells  (ATCC)

Journal: Current Research in Microbial Sciences

Article Title: ADS-J21 is a novel HIV-1 entry inhibitor targeting gp41

doi: 10.1016/j.crmicr.2024.100260

Figure Lengend Snippet: Inhibitory activity of ADS-J21 on infection by primary HIV-1 strain pseudovirus. Briefly, ADS-J21 were mixed with 50 µL of the pseudovirus about 100 TCID 50 before the mixtures were added to the U87.CD4.CCR5.CXCR4 cells, then incubated at 37 °C for 30 min. After 2 days, the cells were lysed and measured for luciferase activity as described above. ( A ) subtype A, ( B ) subtype B, ( C ) subtype C, ( D ) subtype C/D, ( E ) subtype D. Single-cycle infection experiments were performed as described in Materials and Methods. ( F ) Laboratory strains. Each sample was tested in triplicate and the experiment was repeated twice. The data from one representative experiment are presented in mean ± standard deviation.

Article Snippet: The cytotoxicity of ADS-J21 and ADS-J1 in cells (MT-2, U87.CD4.CXCR4.CCR5 and CEMx174 5.25M7 cells) was tested using the CCK-8 kit (YEASEN).

Techniques: Activity Assay, Infection, Incubation, Luciferase, Standard Deviation

Journal: Current Research in Microbial Sciences

Article Title: ADS-J21 is a novel HIV-1 entry inhibitor targeting gp41

doi: 10.1016/j.crmicr.2024.100260

Figure Lengend Snippet: In vitro cytotoxicity of ADS-J21 and ADS-J1. a

Article Snippet: The cytotoxicity of ADS-J21 and ADS-J1 in cells (MT-2, U87.CD4.CXCR4.CCR5 and CEMx174 5.25M7 cells) was tested using the CCK-8 kit (YEASEN).

Techniques: In Vitro

Journal:

Article Title: The V1, V2, and V3 Regions of the Human Immunodeficiency Virus Type 1 Envelope Differentially Affect the Viral Phenotype in an Isolate-Dependent Manner

doi: 10.1128/JVI.79.14.9069-9080.2005

Figure Lengend Snippet: CD4 binding and fusogenic potential of SF162 envelopes with deletions in the V3 loop. (A) The indicated deletions were introduced within the V3 loop of the SF162 envelope. The deleted sequences were replaced by the GAG tripeptide. (B) The binding of IgGCD4 to SF162 and SF162ΔV2 virions with and without deletions in the V3 loop was determined by monitoring the binding of IgGCD4 to virions. (C) The cell fusion potential of SF162 and SF162ΔV2 envelopes lacking elements of the V3 loop was assessed using U87-CD4-CCR5 as target cells. Δenv, pseudovirions lacking envelope were used as negative controls. The values represent the average and standard deviations from quadruplicate wells from a single experiment. Experiments were performed at least three times with similar results.

Article Snippet: CEMx174 5.25 M7 cells (expressing CD4, CCR5, and CXCR4, as well as being stably transduced with an HIV-1 long terminal repeat-green fluorescent protein and HIV-1 long terminal repeat-luciferase reporter construct) were cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 μg/ml), l -glutamine (2 mM), puromycin (0.5 μg/ml), G418 (200 μg/ml; Cellgro), and hygromycin B (200 μg/ml; Cellgro).

Techniques: Binding Assay

Journal:

Article Title: The V1, V2, and V3 Regions of the Human Immunodeficiency Virus Type 1 Envelope Differentially Affect the Viral Phenotype in an Isolate-Dependent Manner

doi: 10.1128/JVI.79.14.9069-9080.2005

Figure Lengend Snippet: Generation of the SF162ΔV1V2 virus. (A) Human PBMC were inoculated with supernatant from 293T cells that were transfected with plasmids encoding the SF162ΔV1V2 genome. Supernatant collected at day 28 post-PBMC-inoculation was used to inoculate a new batch of human PBMC. Note the difference in scale of the y-axis between the two graphs. (B) Single-round competent virions expressing the indicated envelopes were incubated with U87-CD4-CCR5 cells, as described in the Materials and Methods and cell-associated luciferase was determined 48 h later. Δenv, Virions that do not express envelope. D/N, aspartic acid at position 112 in the C1 region was replaced by asparagine. Black bars, 1 ng of p24 inoculum. White bars, 0.1 ng of p24 inoculum. Results are from one out of thee independent experiments.

Article Snippet: CEMx174 5.25 M7 cells (expressing CD4, CCR5, and CXCR4, as well as being stably transduced with an HIV-1 long terminal repeat-green fluorescent protein and HIV-1 long terminal repeat-luciferase reporter construct) were cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 μg/ml), l -glutamine (2 mM), puromycin (0.5 μg/ml), G418 (200 μg/ml; Cellgro), and hygromycin B (200 μg/ml; Cellgro).

Techniques: Transfection, Expressing, Incubation, Luciferase

Journal:

Article Title: The V1, V2, and V3 Regions of the Human Immunodeficiency Virus Type 1 Envelope Differentially Affect the Viral Phenotype in an Isolate-Dependent Manner

doi: 10.1128/JVI.79.14.9069-9080.2005

Figure Lengend Snippet: Processing and CD4 binding potential of SF128A-derived envelopes lacking the V2 loop. (A) 293T cells were transfected with DNA vectors expressing wither the SF128A or SF128AΔV2 (two clones, 5 and 9) gp160 envelopes. The presence of gp160 and gp120 in the cell-lysates or cell-supernatants was determined as described in the Materials and Methods section. (B) Single-round competent virions expressing the SF128A or SF128AΔV2 envelopes were purified through sucrose and then incubated with the indicated concentrations of IgGCD4. Virion-associated IgGCD4 was determined as described in the Materials and Methods section.

Article Snippet: CEMx174 5.25 M7 cells (expressing CD4, CCR5, and CXCR4, as well as being stably transduced with an HIV-1 long terminal repeat-green fluorescent protein and HIV-1 long terminal repeat-luciferase reporter construct) were cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 μg/ml), l -glutamine (2 mM), puromycin (0.5 μg/ml), G418 (200 μg/ml; Cellgro), and hygromycin B (200 μg/ml; Cellgro).

Techniques: Binding Assay, Derivative Assay, Transfection, Expressing, Clone Assay, Purification, Incubation

Journal:

Article Title: The V1, V2, and V3 Regions of the Human Immunodeficiency Virus Type 1 Envelope Differentially Affect the Viral Phenotype in an Isolate-Dependent Manner

doi: 10.1128/JVI.79.14.9069-9080.2005

Figure Lengend Snippet: Fusion potential of SF170 or SF128A-derived envelopes. Single-round competent virions (5 ng of p24 equivalent per well of a 96 well plate) expressing the indicated envelopes derived either from SF170 or SF128A were incubated with U87-CD4-CCR5 cells for 72 h and the cell-associated luciferase was determined. Data are the average of triplicate wells, and two independent clones each of SF170ΔV2 and SF128AΔV2 were tested in parallel with similar results.

Article Snippet: CEMx174 5.25 M7 cells (expressing CD4, CCR5, and CXCR4, as well as being stably transduced with an HIV-1 long terminal repeat-green fluorescent protein and HIV-1 long terminal repeat-luciferase reporter construct) were cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 μg/ml), l -glutamine (2 mM), puromycin (0.5 μg/ml), G418 (200 μg/ml; Cellgro), and hygromycin B (200 μg/ml; Cellgro).

Techniques: Derivative Assay, Expressing, Incubation, Luciferase, Clone Assay

Journal:

Article Title: The V1, V2, and V3 Regions of the Human Immunodeficiency Virus Type 1 Envelope Differentially Affect the Viral Phenotype in an Isolate-Dependent Manner

doi: 10.1128/JVI.79.14.9069-9080.2005

Figure Lengend Snippet: Neutralization susceptibility of SF162, SF162ΔV1, SF162ΔV2, and SF162ΔV1V2 viruses. The neutralization susceptibility of the indicated viruses against a panel of MAbs and sera was determined using luciferase-expressing single-round replication-competent viruses and U87-CD4-CCR5 cells as targets. Values indicate the average and standard deviation from quadruplicate wells. Each neutralization assay was repeated at least two independent times. (A) IgGCD4, IgG1b12, F105, and M18 bind to the CD4 binding site, while 2909 recognizes a complex epitope. (B) 447D and 391-95D bind to the V3 loop, while 17b andX5 bind to CD4 induced epitopes. (C) 2F5 and 4E10 bind to the extracellular region of gp41, while 2G12 binds to a complex carbohydrate epitope; 1418 is a non-anti-HIV MAb used as a negative control. (D) Sera from macaques C640 and A141 were isolated from animals infected with SHIVSF162P4, while HIVIG is heterologous human serum IgG.

Article Snippet: CEMx174 5.25 M7 cells (expressing CD4, CCR5, and CXCR4, as well as being stably transduced with an HIV-1 long terminal repeat-green fluorescent protein and HIV-1 long terminal repeat-luciferase reporter construct) were cultured in RPMI 1640 medium supplemented with 10% FBS, penicillin (100 U/ml), streptomycin (100 μg/ml), l -glutamine (2 mM), puromycin (0.5 μg/ml), G418 (200 μg/ml; Cellgro), and hygromycin B (200 μg/ml; Cellgro).

Techniques: Neutralization, Luciferase, Expressing, Standard Deviation, Binding Assay, Negative Control, Isolation, Infection

Journal:

Article Title: Increased Mucosal Transmission but Not Enhanced Pathogenicity of the CCR5-Tropic, Simian AIDS-Inducing Simian/Human Immunodeficiency Virus SHIV SF162P3 Maps to Envelope gp120

doi: 10.1128/JVI.77.2.989-998.2003

Figure Lengend Snippet: Relative entry of luciferase reporter viruses expressing SF162 gp120 or P3 gp120. (A) Entry of HOS.CD4.CCR5 cells. (B) Entry of CEMx174 5.25 M7 cells. Data for relative entry of HOS.CD4.CCR5 cells are the means and standard errors of six independent experiments, while those for CEMx174 5.25 M7 cells are the averages of two independent experiments.

Article Snippet: The CEMx174 5.25 M7 cell line (also a generous gift from N. Landau), expressing both CD4 and CCR5, was stably transduced with a long terminal repeat (LTR)-luciferase and LTR-green fluorescent protein cassette and was maintained in RPMI-10% FBS containing 200 μg of G418 (Life Technologies, Carlsbad, Calif.)/ml, 200 μg of hygromycin (Invitrogen, Carlsbad, Calif.)/ml, and 0.5 μg of puromycin (Sigma-Aldrich, St. Louis, Mo.)/ml.

Techniques: Luciferase, Expressing

Journal:

Article Title: Increased Mucosal Transmission but Not Enhanced Pathogenicity of the CCR5-Tropic, Simian AIDS-Inducing Simian/Human Immunodeficiency Virus SHIV SF162P3 Maps to Envelope gp120

doi: 10.1128/JVI.77.2.989-998.2003

Figure Lengend Snippet: Fusogenic capacity and susceptibility to T-20. (A) Fusion of SF162 and P3 gp120-expressing 293T cells with CEMx174 5.25 M7 cells. Results shown are the means and standard errors from three independent experiments. (B) Inhibition of luciferase reporter virus fusion with HOS.CD4.CCR5 cells by T-20. Viruses and T-20 were incubated for 1 h at 37°C before addition to cells, and inhibition relative to viruses in the absence of drug was calculated. The means and standard errors from three independent experiments are shown.

Article Snippet: The CEMx174 5.25 M7 cell line (also a generous gift from N. Landau), expressing both CD4 and CCR5, was stably transduced with a long terminal repeat (LTR)-luciferase and LTR-green fluorescent protein cassette and was maintained in RPMI-10% FBS containing 200 μg of G418 (Life Technologies, Carlsbad, Calif.)/ml, 200 μg of hygromycin (Invitrogen, Carlsbad, Calif.)/ml, and 0.5 μg of puromycin (Sigma-Aldrich, St. Louis, Mo.)/ml.

Techniques: Expressing, Inhibition, Luciferase, Incubation