Recombinant Human Anti-HIV-1 Antibody scFv Fragment (N12-i15) (CAT#: PABX-096-S (P))

Figure 1 Representative experiments of Western blot show the reactive intensity of nAbs and non-nAbs with HIV-1 Env in the cellular lysates of ACH-2 cells 24 h after stimulation. PMA, PMA (10 ng/ml) plus ionomycin (1 µg/ml).

Yang, K., Lan, J., Shepherd, N., Hu, N., Xing, Y., Byrd, D., ... & Gao, J. (2015). Blockage of CD59 function restores activities of neutralizing and nonneutralizing antibodies in triggering antibody-dependent complement-mediated lysis of HIV-1 virions and provirus-activated latently infected cells. Journal of virology, 89(18), 9393-9406.

Figure 2 Colocalization of Env, CD59 and lipid rafts on the surface of ACH-2 cells upon provirus activation.

ACH-2 cells were treated with PMA (10 ng/ml) plus ionomycin (1 µg/ml) or DMSO for 24 h. After treatment, cells are subjected to immunostaining for confocal microscopy analysis of Env, CD59, and lipid raft colocalization on the cell surface. Cells were subjected to surface staining with CTB conjugated with Alexa Fluor 488 (for lipid rafts), an individual nAbs (2G12, 2F5, and 4E10) or non-nAbs (2.2C, A32, N5-i5, and N12-i15) at 1 µg/ml (for HIV-1 Env), and BRIC229 (for hCD59). The white color in the merged panel indicates the colocalization of HIV-1 Env (red) with lipid rafts (green) and CD59 (magenta) on the surfaces of provirus-activated ACH-2 cells. Cellular DNA content and nuclei were stained with DAPI (blue). Scale bars, 5 µm.

Yang, K., Lan, J., Shepherd, N., Hu, N., Xing, Y., Byrd, D., ... & Gao, J. (2015). Blockage of CD59 function restores activities of neutralizing and nonneutralizing antibodies in triggering antibody-dependent complement-mediated lysis of HIV-1 virions and provirus-activated latently infected cells. Journal of virology, 89(18), 9393-9406.

Figure 3 Representative experiments of flow cytometry analysis show the results of ACH-2 cells expressing Env on the cell surface 24 h after stimulation.

Yang, K., Lan, J., Shepherd, N., Hu, N., Xing, Y., Byrd, D., ... & Gao, J. (2015). Blockage of CD59 function restores activities of neutralizing and nonneutralizing antibodies in triggering antibody-dependent complement-mediated lysis of HIV-1 virions and

Figure 4 Anti-Env nAbs and non-nAbs triggered ADCML of latently HIV-1-infected cells upon provirus activation.

Specific lysis of provirus-activated ACH-2 cells by anti-Env nAbs and non-nAbs. The calcein release assay was used to quantitate ADCML of latently HIV-1-infected cells by anti-HIV-1 nAbs and non-nAbs. ACH-2 or A3.01 cells treated or untreated with PMA/ionomycycin or DMSO. Cells were preincubated with BRIC229 (20 µg/ml) to block hCD59 function. An irrelevant IgG control Ab (20 µg/ml) was used in parallel as a negative control of hCD59 blockage. After preincubation, nAbs or non-nAbs were added, followed by the exposure to either complement-competent or heat-inactivated sera from three healthy blood donors. Triton X-100 was used for determining the total cytolysis. IgG + C, irrelevant Ab IgG plus
complement; BRIC229 + C, BRIC229 Ab plus complement; BRIC229 + iC, BRIC229 Ab plus inactivated complement. Statistical significance (**, P < 0.01 versus IgG plus complement or BRIC229 plus inactivated complement) is indicated by asterisks. ns, not significant.

Yang, K., Lan, J., Shepherd, N., Hu, N., Xing, Y., Byrd, D., ... & Gao, J. (2015). Blockage of CD59 function restores activities of neutralizing and nonneutralizing antibodies in triggering antibody-dependent complement-mediated lysis of HIV-1 virions and