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    ZeptoMetrix hiv 1 p24 antigen elisa kit
    Treatment of NFκ-B inhibitors reduce <t>HIV-1</t> replication in U1 cells due to BaP exposure. U1 cells were concomitantly treated with BaP (1 µM) and NFκ-B inhibitors, IKK-16 (400 nM) ( A ), and SC-514 (10 µM) ( B ) for 3 days. After the treatment, supernatants were collected to determine the viral load using the <t>p24</t> <t>ELISA</t> assay. HIV-1 replication due to BaP (1 µM) exposure was significantly rescued by NFκ-B inhibitors, IKK-16 (400 nM) and SC-514 (10 µM). *Represents p ≤ 0.05 compared with the control group while ## represents p ≤ 0.005 compared to the BaP-treated groups. Western blots were run using the nuclear fraction proteins obtained from the BaP-exposed cells treated with IKK-16 ( C ) SC-524 ( D ) or siRNA CYP1A1 ( E ) to determine the expression of NFκ-B p65 subunits. The blots indicate that treatment with both the NFκ-B inhibitors and CYP1A1 siRNA reduced the expression of NFκ-B p65 in the nuclear fraction protein of the BaP-treated cells compared to the control. The blots presented are representative of at least three different experiments.
    Hiv 1 P24 Antigen Elisa Kit, supplied by ZeptoMetrix, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv 1 p24 antigen elisa kit/product/ZeptoMetrix
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    Price from $9.99 to $1999.99
    hiv 1 p24 antigen elisa kit - by Bioz Stars, 2021-06
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    86
    PerkinElmer alliance hiv 1 p24 antigen elisa kit
    Gp120 aptamer-siRNA 362 represses <t>HIV-1</t> by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV <t>p24</t> in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p
    Alliance Hiv 1 P24 Antigen Elisa Kit, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alliance hiv 1 p24 antigen elisa kit/product/PerkinElmer
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alliance hiv 1 p24 antigen elisa kit - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    86
    PerkinElmer hiv 1 p24 elisa kit
    L-selectin shedding is required for <t>HIV-1</t> release. a . b The inhibition of X4-tropic HIV-1 LAI infection of CEM cells by BB-94. c , d The inhibition of R5-tropic HIV-1 BAL ( c ) or X4-tropic HIV-1 LAI ( d ) infections of PBMC by BB-94 on day 6 p.i. e The expression of CD62L during HIV-1 BAL infection ( c ) from uninfected (gray area) and HIV-1 BAL infected PBMC in the absence (blue lines) or presence of 100 µM BB-94 (red). f Shedding of L-selectin in the infected and uninfected supernatants during HIV-1 BAL infection of PBMC (panel c) in the presence and absence of BB-94. g The western blot analysis of a cleaved 6kD C-terminal L-selectin fragment in the presence and absence of BB-94 during HIV-1 BAL infection ( c ) ( g ). Uninfected PBMC were treated with PMA for 30 min to induce L-selectin shedding. Lanes 1 and 2 are cell lysates from the infected samples in the presence of DMSO or BB-94. Lanes 3 and 4 are cell lysates from uninfected samples in the absence or presence of PMA. β-actin (lower panel) is used as loading control. h The effect of ADAM17-specific inhibitors TAPI-1, TAPI-2 on HIV-1 BAL infections of PBMC. i The infection of PBMC by JRFL- and SF33-pseudotyped viruses in the presence of BB-94 or DMSO. Luciferase activity was measured at day 3 p.i. j Trypsin-mediated viral release assay. The release of viral particles upon trypsin digestion from day 6 of infected PBMC in the presence and absence of BB-94 or DMSO was measured by <t>p24</t> <t>ELISA.</t> k BB-94 inhibition of cell−cell transfer-mediated HIV-1 BAL infections. TZM-BL cells were infected through coincubation with titrating amount of infected PBMC in the presence of BB-94 or DMSO. The infection of TZM-BL cells was measured by luciferase activity 48–60 h p.i. The results are from at least two independent experiments with all data included in the analysis
    Hiv 1 P24 Elisa Kit, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/hiv 1 p24 elisa kit/product/PerkinElmer
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    hiv 1 p24 elisa kit - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    86
    PerkinElmer alliance hiv 1 p24 elisa kit
    Gp120 aptamer-siRNA 362 represses <t>HIV-1</t> by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV <t>p24</t> in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p
    Alliance Hiv 1 P24 Elisa Kit, supplied by PerkinElmer, used in various techniques. Bioz Stars score: 86/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/alliance hiv 1 p24 elisa kit/product/PerkinElmer
    Average 86 stars, based on 1 article reviews
    Price from $9.99 to $1999.99
    alliance hiv 1 p24 elisa kit - by Bioz Stars, 2021-06
    86/100 stars
      Buy from Supplier

    Image Search Results


    Treatment of NFκ-B inhibitors reduce HIV-1 replication in U1 cells due to BaP exposure. U1 cells were concomitantly treated with BaP (1 µM) and NFκ-B inhibitors, IKK-16 (400 nM) ( A ), and SC-514 (10 µM) ( B ) for 3 days. After the treatment, supernatants were collected to determine the viral load using the p24 ELISA assay. HIV-1 replication due to BaP (1 µM) exposure was significantly rescued by NFκ-B inhibitors, IKK-16 (400 nM) and SC-514 (10 µM). *Represents p ≤ 0.05 compared with the control group while ## represents p ≤ 0.005 compared to the BaP-treated groups. Western blots were run using the nuclear fraction proteins obtained from the BaP-exposed cells treated with IKK-16 ( C ) SC-524 ( D ) or siRNA CYP1A1 ( E ) to determine the expression of NFκ-B p65 subunits. The blots indicate that treatment with both the NFκ-B inhibitors and CYP1A1 siRNA reduced the expression of NFκ-B p65 in the nuclear fraction protein of the BaP-treated cells compared to the control. The blots presented are representative of at least three different experiments.

    Journal: Scientific Reports

    Article Title: Benzo(a)pyrene in Cigarette Smoke Enhances HIV-1 Replication through NF-κB Activation via CYP-Mediated Oxidative Stress Pathway

    doi: 10.1038/s41598-018-28500-z

    Figure Lengend Snippet: Treatment of NFκ-B inhibitors reduce HIV-1 replication in U1 cells due to BaP exposure. U1 cells were concomitantly treated with BaP (1 µM) and NFκ-B inhibitors, IKK-16 (400 nM) ( A ), and SC-514 (10 µM) ( B ) for 3 days. After the treatment, supernatants were collected to determine the viral load using the p24 ELISA assay. HIV-1 replication due to BaP (1 µM) exposure was significantly rescued by NFκ-B inhibitors, IKK-16 (400 nM) and SC-514 (10 µM). *Represents p ≤ 0.05 compared with the control group while ## represents p ≤ 0.005 compared to the BaP-treated groups. Western blots were run using the nuclear fraction proteins obtained from the BaP-exposed cells treated with IKK-16 ( C ) SC-524 ( D ) or siRNA CYP1A1 ( E ) to determine the expression of NFκ-B p65 subunits. The blots indicate that treatment with both the NFκ-B inhibitors and CYP1A1 siRNA reduced the expression of NFκ-B p65 in the nuclear fraction protein of the BaP-treated cells compared to the control. The blots presented are representative of at least three different experiments.

    Article Snippet: We used the HIV-1 p24 Antigen ELISA kit (Zeptometrix Corporation, Buffalo, NY) for this purpose.

    Techniques: Enzyme-linked Immunosorbent Assay, Western Blot, Expressing

    Treatment of antioxidants and CYP1A1 inhibitors reduce HIV-1 replication in U1 cells due to BaP exposure. U1 cells were concomitantly treated with BaP (1 µM) and antioxidants [vitamin C (100 µM) and vitamin E (100 µM), pinostilbene (2 µM), and resveratrol (50 µM)] ( A ) or CYP1A1 inhibitor ellipticine (1 µM)] ( B ) for 3 days. Prior to BaP treatment, the CYP1A1 gene was knocked down in the U1 cells using siRNA specific to CYP1A1. ( C ) The cells were then treated with BaP (100 nM) for 3 days. After the treatment, supernatants were collected to determine the viral load using the p24 ELISA assay. HIV-1 replication significantly increased with 3-days exposure of BaP (1 µM), which was rescued by all the antioxidants (vitamin C and E, and resveratrol) as well as the CYP1A1 inhibitor, ellipticine . The knock-down of the CYP1A1 gene also rescued HIV-1 replication in BaP-exposed U1 cells. The data were obtained from the mean of at least three independent experiments. * and ** represents p ≤ 0.05 and p ≤ 0.005 compared with the control group while #,## and ### represents p ≤ 0.05, p ≤ 0.005 and p ≤ 0.0005, respectively, compared to the BaP-treated groups.

    Journal: Scientific Reports

    Article Title: Benzo(a)pyrene in Cigarette Smoke Enhances HIV-1 Replication through NF-κB Activation via CYP-Mediated Oxidative Stress Pathway

    doi: 10.1038/s41598-018-28500-z

    Figure Lengend Snippet: Treatment of antioxidants and CYP1A1 inhibitors reduce HIV-1 replication in U1 cells due to BaP exposure. U1 cells were concomitantly treated with BaP (1 µM) and antioxidants [vitamin C (100 µM) and vitamin E (100 µM), pinostilbene (2 µM), and resveratrol (50 µM)] ( A ) or CYP1A1 inhibitor ellipticine (1 µM)] ( B ) for 3 days. Prior to BaP treatment, the CYP1A1 gene was knocked down in the U1 cells using siRNA specific to CYP1A1. ( C ) The cells were then treated with BaP (100 nM) for 3 days. After the treatment, supernatants were collected to determine the viral load using the p24 ELISA assay. HIV-1 replication significantly increased with 3-days exposure of BaP (1 µM), which was rescued by all the antioxidants (vitamin C and E, and resveratrol) as well as the CYP1A1 inhibitor, ellipticine . The knock-down of the CYP1A1 gene also rescued HIV-1 replication in BaP-exposed U1 cells. The data were obtained from the mean of at least three independent experiments. * and ** represents p ≤ 0.05 and p ≤ 0.005 compared with the control group while #,## and ### represents p ≤ 0.05, p ≤ 0.005 and p ≤ 0.0005, respectively, compared to the BaP-treated groups.

    Article Snippet: We used the HIV-1 p24 Antigen ELISA kit (Zeptometrix Corporation, Buffalo, NY) for this purpose.

    Techniques: Enzyme-linked Immunosorbent Assay

    Chronic treatment of BaP induces HIV-1 replication and apoptotic DNA damage in HIV-1-infected macrophages. ( A ) The U1 cells were treated with 10 nM and 100 nM BaP for seven days. After the BaP treatment, the U1 cells were stimulated with 100 nM of Phorbol 12-myristate 13-acetate (PMA) to produce HIV-1. Supernatants were collected after two days of differentiation, which were used for the p24 ELISA assay to assess the viral load. The chronic (7 days) treatment of BaP (100 nM) significantly increased the viral replication in U1 cells, while the 10-fold lower concentration did not have any significant effect. The data is displayed as mean ± SEM (n = 6), calculated as a percentage of the control. ( B ) HIV-infected human primary macrophages were treated with BaP (100 nM) for 3 days. The supernatant was collected thereafter and used for the p24 ELISA assay to assess the viral load. The acute (3 days) treatment of BaP (100 nM) significantly increased the viral replication in HIV-1-infected primary macrophages. The data is displayed as mean ± SEM (n = 4). For calculating the viral load, we subtracted the nonspecific background reading from the actual absorbance values. Since the residual viral load varies from experiment to experiment in U1 cells, we normalized the control values for each experiment to 100% and calculated the values for the treated, as the percentage of the control. The statistical significance was calculated at *p ≤ 0.05, where *** represents p ≤ 0.0005, compared with the control group. ( C ) The apoptotic DNA damage assay was performed on the treated cells. DAPI, FAM and CR590 stained nucleus (blue), apoptotic DNA damage with DNase Type II ends (green) and Type I ends (red) respectively. A higher signal for CR590 is visible in the fluorescent images, indicating apoptotic DNA fragmentation with DNase Type I ends in the infected human primary macrophages after BaP (100 nM) exposure for 3 days. DNA fragmentation with DNase Type II ends (green) was not visible in either the control or the treated cells. Therefore, the images indicate that BaP (100 nM) induces DNA fragmentation during the early phase of apoptosis in the HIV-1-infected human primary macrophages.

    Journal: Scientific Reports

    Article Title: Benzo(a)pyrene in Cigarette Smoke Enhances HIV-1 Replication through NF-κB Activation via CYP-Mediated Oxidative Stress Pathway

    doi: 10.1038/s41598-018-28500-z

    Figure Lengend Snippet: Chronic treatment of BaP induces HIV-1 replication and apoptotic DNA damage in HIV-1-infected macrophages. ( A ) The U1 cells were treated with 10 nM and 100 nM BaP for seven days. After the BaP treatment, the U1 cells were stimulated with 100 nM of Phorbol 12-myristate 13-acetate (PMA) to produce HIV-1. Supernatants were collected after two days of differentiation, which were used for the p24 ELISA assay to assess the viral load. The chronic (7 days) treatment of BaP (100 nM) significantly increased the viral replication in U1 cells, while the 10-fold lower concentration did not have any significant effect. The data is displayed as mean ± SEM (n = 6), calculated as a percentage of the control. ( B ) HIV-infected human primary macrophages were treated with BaP (100 nM) for 3 days. The supernatant was collected thereafter and used for the p24 ELISA assay to assess the viral load. The acute (3 days) treatment of BaP (100 nM) significantly increased the viral replication in HIV-1-infected primary macrophages. The data is displayed as mean ± SEM (n = 4). For calculating the viral load, we subtracted the nonspecific background reading from the actual absorbance values. Since the residual viral load varies from experiment to experiment in U1 cells, we normalized the control values for each experiment to 100% and calculated the values for the treated, as the percentage of the control. The statistical significance was calculated at *p ≤ 0.05, where *** represents p ≤ 0.0005, compared with the control group. ( C ) The apoptotic DNA damage assay was performed on the treated cells. DAPI, FAM and CR590 stained nucleus (blue), apoptotic DNA damage with DNase Type II ends (green) and Type I ends (red) respectively. A higher signal for CR590 is visible in the fluorescent images, indicating apoptotic DNA fragmentation with DNase Type I ends in the infected human primary macrophages after BaP (100 nM) exposure for 3 days. DNA fragmentation with DNase Type II ends (green) was not visible in either the control or the treated cells. Therefore, the images indicate that BaP (100 nM) induces DNA fragmentation during the early phase of apoptosis in the HIV-1-infected human primary macrophages.

    Article Snippet: We used the HIV-1 p24 Antigen ELISA kit (Zeptometrix Corporation, Buffalo, NY) for this purpose.

    Techniques: Infection, Enzyme-linked Immunosorbent Assay, Concentration Assay, Staining

    Gp120 aptamer-siRNA 362 represses HIV-1 by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV p24 in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p

    Journal: Theranostics

    Article Title: Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1

    doi: 10.7150/thno.23085

    Figure Lengend Snippet: Gp120 aptamer-siRNA 362 represses HIV-1 by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV p24 in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p

    Article Snippet: The p24 antigen analyses were performed using a HIV-1 p24 Antigen Assay (Alliance HIV-1 p24 Antigen ELISA kit, PerkinElmer, CA) according to the manufacturer's instructions.

    Techniques: Expressing, Infection

    Aptamer-directed TGS. ( A ) A schematic is shown depicting gp120 aptamer conjugates containing the LTR-362 siRNA directed to transcriptionally silence HIV-1 LTR activity and inhibit HIV-1 expression. ( B ) The candidate gp120-aptamer siRNA chimeras and controls assessed are shown. ( C ) The effects of the various gp120-aptamer-siRNA chimeras and controls on HIV-1 expression in HIV-1-infected CCRF-CEM cells following one 800 nM treatment. A schematic depicting the experimental procedure is shown along with p24 values from days 3-10 post-aptamer treatment.

    Journal: Theranostics

    Article Title: Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1

    doi: 10.7150/thno.23085

    Figure Lengend Snippet: Aptamer-directed TGS. ( A ) A schematic is shown depicting gp120 aptamer conjugates containing the LTR-362 siRNA directed to transcriptionally silence HIV-1 LTR activity and inhibit HIV-1 expression. ( B ) The candidate gp120-aptamer siRNA chimeras and controls assessed are shown. ( C ) The effects of the various gp120-aptamer-siRNA chimeras and controls on HIV-1 expression in HIV-1-infected CCRF-CEM cells following one 800 nM treatment. A schematic depicting the experimental procedure is shown along with p24 values from days 3-10 post-aptamer treatment.

    Article Snippet: The p24 antigen analyses were performed using a HIV-1 p24 Antigen Assay (Alliance HIV-1 p24 Antigen ELISA kit, PerkinElmer, CA) according to the manufacturer's instructions.

    Techniques: Activity Assay, Expressing, Infection

    L-selectin shedding is required for HIV-1 release. a . b The inhibition of X4-tropic HIV-1 LAI infection of CEM cells by BB-94. c , d The inhibition of R5-tropic HIV-1 BAL ( c ) or X4-tropic HIV-1 LAI ( d ) infections of PBMC by BB-94 on day 6 p.i. e The expression of CD62L during HIV-1 BAL infection ( c ) from uninfected (gray area) and HIV-1 BAL infected PBMC in the absence (blue lines) or presence of 100 µM BB-94 (red). f Shedding of L-selectin in the infected and uninfected supernatants during HIV-1 BAL infection of PBMC (panel c) in the presence and absence of BB-94. g The western blot analysis of a cleaved 6kD C-terminal L-selectin fragment in the presence and absence of BB-94 during HIV-1 BAL infection ( c ) ( g ). Uninfected PBMC were treated with PMA for 30 min to induce L-selectin shedding. Lanes 1 and 2 are cell lysates from the infected samples in the presence of DMSO or BB-94. Lanes 3 and 4 are cell lysates from uninfected samples in the absence or presence of PMA. β-actin (lower panel) is used as loading control. h The effect of ADAM17-specific inhibitors TAPI-1, TAPI-2 on HIV-1 BAL infections of PBMC. i The infection of PBMC by JRFL- and SF33-pseudotyped viruses in the presence of BB-94 or DMSO. Luciferase activity was measured at day 3 p.i. j Trypsin-mediated viral release assay. The release of viral particles upon trypsin digestion from day 6 of infected PBMC in the presence and absence of BB-94 or DMSO was measured by p24 ELISA. k BB-94 inhibition of cell−cell transfer-mediated HIV-1 BAL infections. TZM-BL cells were infected through coincubation with titrating amount of infected PBMC in the presence of BB-94 or DMSO. The infection of TZM-BL cells was measured by luciferase activity 48–60 h p.i. The results are from at least two independent experiments with all data included in the analysis

    Journal: Nature Communications

    Article Title: HIV-1 targets L-selectin for adhesion and induces its shedding for viral release

    doi: 10.1038/s41467-018-05197-2

    Figure Lengend Snippet: L-selectin shedding is required for HIV-1 release. a . b The inhibition of X4-tropic HIV-1 LAI infection of CEM cells by BB-94. c , d The inhibition of R5-tropic HIV-1 BAL ( c ) or X4-tropic HIV-1 LAI ( d ) infections of PBMC by BB-94 on day 6 p.i. e The expression of CD62L during HIV-1 BAL infection ( c ) from uninfected (gray area) and HIV-1 BAL infected PBMC in the absence (blue lines) or presence of 100 µM BB-94 (red). f Shedding of L-selectin in the infected and uninfected supernatants during HIV-1 BAL infection of PBMC (panel c) in the presence and absence of BB-94. g The western blot analysis of a cleaved 6kD C-terminal L-selectin fragment in the presence and absence of BB-94 during HIV-1 BAL infection ( c ) ( g ). Uninfected PBMC were treated with PMA for 30 min to induce L-selectin shedding. Lanes 1 and 2 are cell lysates from the infected samples in the presence of DMSO or BB-94. Lanes 3 and 4 are cell lysates from uninfected samples in the absence or presence of PMA. β-actin (lower panel) is used as loading control. h The effect of ADAM17-specific inhibitors TAPI-1, TAPI-2 on HIV-1 BAL infections of PBMC. i The infection of PBMC by JRFL- and SF33-pseudotyped viruses in the presence of BB-94 or DMSO. Luciferase activity was measured at day 3 p.i. j Trypsin-mediated viral release assay. The release of viral particles upon trypsin digestion from day 6 of infected PBMC in the presence and absence of BB-94 or DMSO was measured by p24 ELISA. k BB-94 inhibition of cell−cell transfer-mediated HIV-1 BAL infections. TZM-BL cells were infected through coincubation with titrating amount of infected PBMC in the presence of BB-94 or DMSO. The infection of TZM-BL cells was measured by luciferase activity 48–60 h p.i. The results are from at least two independent experiments with all data included in the analysis

    Article Snippet: HIV-1 p24 ELISA kit was obtained from PerkinElmer Life Sciences, Inc. (Waltham, MA).

    Techniques: Inhibition, Infection, Expressing, Western Blot, Luciferase, Activity Assay, Release Assay, Enzyme-linked Immunosorbent Assay

    L-selectin facilitates HIV-1 infection of CD4 + T cells. a ). b HIV infection of CEM cell lines in the presence and absence of 1 µg/mL polybrene. c HIV-1 BAL infection of PBMC in the presence of EDTA or anti-CD4 (RPA-T4). The infection was measured by intracellular p24 + staining. d Dose-dependent HIV-1 BAL infection of PBMC in the presence of CD62L blocking antibody DREG-56 or isotype controls. Infections were measured as copy numbers of viral DNA by real-time PCR and displayed as % relative to the isotype controls. The results are from at least two independent experiments and statistics are performed without data rejections

    Journal: Nature Communications

    Article Title: HIV-1 targets L-selectin for adhesion and induces its shedding for viral release

    doi: 10.1038/s41467-018-05197-2

    Figure Lengend Snippet: L-selectin facilitates HIV-1 infection of CD4 + T cells. a ). b HIV infection of CEM cell lines in the presence and absence of 1 µg/mL polybrene. c HIV-1 BAL infection of PBMC in the presence of EDTA or anti-CD4 (RPA-T4). The infection was measured by intracellular p24 + staining. d Dose-dependent HIV-1 BAL infection of PBMC in the presence of CD62L blocking antibody DREG-56 or isotype controls. Infections were measured as copy numbers of viral DNA by real-time PCR and displayed as % relative to the isotype controls. The results are from at least two independent experiments and statistics are performed without data rejections

    Article Snippet: HIV-1 p24 ELISA kit was obtained from PerkinElmer Life Sciences, Inc. (Waltham, MA).

    Techniques: Infection, Recombinase Polymerase Amplification, Staining, Blocking Assay, Real-time Polymerase Chain Reaction

    HIV-1 infection resulted in L-selectin shedding. a PBMC were infected with HIV-1 BAL and CD4 and CD62L expression was measured on days 6 and 11 p.i. for p24 + , p24 − , and uninfected cells. The results are the representative of three individual experiments. b Histograms showing the expression of CD4, CD62L on days 6 and 11 p.i. for p24 + (solid blue), p24 − (dotted), and uninfected (shaded) cells. c CCR7 and CD27 expression as defined in b . d Downregulation of CD4 and CD62L in infected PBMC is evident from the decrease in % of CD4 + /CD62L + cells and the increase in CD4 - /CD62L − cells on day 6 and further on day 11 in p24 + compared to p24 − and uninfected populations. e The accumulation of soluble L-selectin in the supernatant of infected and uninfected samples was measured by ELISA on days 3, 6, and 7 post HIV-1 BAL A−C). f HIV-1 infection of PBMC induced downregulation of L-selectin expression. The loss of CD62L + CD4 T cells during HIV-1 BAL infection (upper panel) is accompanied by the increase in the viral infection (lower panel) at days 6 and 11 p.i. The results are from at least two independent experiments with all data included in the analysis

    Journal: Nature Communications

    Article Title: HIV-1 targets L-selectin for adhesion and induces its shedding for viral release

    doi: 10.1038/s41467-018-05197-2

    Figure Lengend Snippet: HIV-1 infection resulted in L-selectin shedding. a PBMC were infected with HIV-1 BAL and CD4 and CD62L expression was measured on days 6 and 11 p.i. for p24 + , p24 − , and uninfected cells. The results are the representative of three individual experiments. b Histograms showing the expression of CD4, CD62L on days 6 and 11 p.i. for p24 + (solid blue), p24 − (dotted), and uninfected (shaded) cells. c CCR7 and CD27 expression as defined in b . d Downregulation of CD4 and CD62L in infected PBMC is evident from the decrease in % of CD4 + /CD62L + cells and the increase in CD4 - /CD62L − cells on day 6 and further on day 11 in p24 + compared to p24 − and uninfected populations. e The accumulation of soluble L-selectin in the supernatant of infected and uninfected samples was measured by ELISA on days 3, 6, and 7 post HIV-1 BAL A−C). f HIV-1 infection of PBMC induced downregulation of L-selectin expression. The loss of CD62L + CD4 T cells during HIV-1 BAL infection (upper panel) is accompanied by the increase in the viral infection (lower panel) at days 6 and 11 p.i. The results are from at least two independent experiments with all data included in the analysis

    Article Snippet: HIV-1 p24 ELISA kit was obtained from PerkinElmer Life Sciences, Inc. (Waltham, MA).

    Techniques: Infection, Expressing, Enzyme-linked Immunosorbent Assay

    Gp120 aptamer-siRNA 362 represses HIV-1 by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV p24 in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p

    Journal: Theranostics

    Article Title: Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1

    doi: 10.7150/thno.23085

    Figure Lengend Snippet: Gp120 aptamer-siRNA 362 represses HIV-1 by transcriptional gene silencing. ( A ) A schematic depicting the experimental plan to test the various aptamer conjugates. ( B ) The effects of the various aptamer siRNA conjugates on HIV-1 expression in HIV-1-IIIB-infected CCRF-CEM cells (MOI=0.001). ( C-E ) Nuclear run-on analysis of LTR transcription in differentially treated aptamer-siRNA conjugates in ( C ) the absence of any drug and ( D ) the presence of TSA (1 µM) or ( E ) 5' AzaC (4 µM). ( F-H ) expression of HIV p24 in ( F ) untreated, TSA- or 5' AzaC-treated HIV-1-infected CCRF-CEM cultures. ( G ) Nuclear localization of the antisense strand of LTR-362-directed siRNAs following gp120 aptamer delivery in gp120 positive CHO-WT cells. ( H ) The cytoplasmic vs. nuclear distribution of siRNA Tat/rev , LTR-362 siRNA, and Scr siRNA in aptamer conjugate-treated HIV-1-infected CCRF-CEM cells. For B-F triplicate treated cells are shown with the standard deviations and (*) representing p

    Article Snippet: Virus was titrated using the Alliance HIV-1 p24 ELISA kit (PerkinElmer, Waltham, MA).

    Techniques: Expressing, Infection

    Aptamer-directed TGS. ( A ) A schematic is shown depicting gp120 aptamer conjugates containing the LTR-362 siRNA directed to transcriptionally silence HIV-1 LTR activity and inhibit HIV-1 expression. ( B ) The candidate gp120-aptamer siRNA chimeras and controls assessed are shown. ( C ) The effects of the various gp120-aptamer-siRNA chimeras and controls on HIV-1 expression in HIV-1-infected CCRF-CEM cells following one 800 nM treatment. A schematic depicting the experimental procedure is shown along with p24 values from days 3-10 post-aptamer treatment.

    Journal: Theranostics

    Article Title: Receptor-targeted aptamer-siRNA conjugate-directed transcriptional regulation of HIV-1

    doi: 10.7150/thno.23085

    Figure Lengend Snippet: Aptamer-directed TGS. ( A ) A schematic is shown depicting gp120 aptamer conjugates containing the LTR-362 siRNA directed to transcriptionally silence HIV-1 LTR activity and inhibit HIV-1 expression. ( B ) The candidate gp120-aptamer siRNA chimeras and controls assessed are shown. ( C ) The effects of the various gp120-aptamer-siRNA chimeras and controls on HIV-1 expression in HIV-1-infected CCRF-CEM cells following one 800 nM treatment. A schematic depicting the experimental procedure is shown along with p24 values from days 3-10 post-aptamer treatment.

    Article Snippet: Virus was titrated using the Alliance HIV-1 p24 ELISA kit (PerkinElmer, Waltham, MA).

    Techniques: Activity Assay, Expressing, Infection