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To investigate the phenotypic knockout of HIV-1 chemokine coreceptor CXCR4 and CCR5 by intrakines and its inhibitory effect on HIV-1 infection. Primary human PBLs were transduced with the recombinant vector pLNCX-R-K-S-K followed by anti-NGFR/anti-IgG-magnetic bead method selection and FCM detection. The transduced PBI.S were infected with DP1 HFV-1 virus thereafter envelope-mediated syncytium formation and p24 detection were carried out to study the blockage of HIV-1 infection by co-inactivation of CCR5 and CXCR4. pLNCX-R-K-S-K -transduced PBLs were isolated with an anti-NGFR/anti-IgG-magnetic bead method. After isolation, about 70% of the PBI.S were posi- tive for the NGFR marker. When the transduced PBLs were infected with DP1 HIV-1 virus, envelop-mediated syncytium for- mation was almost completely inhibited by pLNCX-R-K-S-K transfection. Also, p24 antigen was very low in the cultures of pLNCX-R-K-S-K transduced PBLs. pLNCX-R-K-S-K transduction inhibited the produc- tion of DP1 p24 antigen by 15%, 43% and 19% on days 4, 7 and 10 respectively. The lymphocytes with the phenotypic knockout of CCR5 and CXCR4 could protect primary human PBLs from DP1 HIV-1 virus infection.
To investigate the phenotypic knockout of HIV-1 chemokine receptor CXCR4 and CCR5 by intrakines and its inhibitory effect on HIV-1 infection. Primary human PBLs were transduced with the recombinant vector pLNCX-RKSK followed by anti-NGFR / anti-IgG-magnetic bead The selection of FCI detection. The transduced PBI-S were infected with DP1 HFV-1 virus infected envelope-mediated syncytium formation and p24 detection were carried out to study the blockage of HIV-1 infection by co-inactivation of CCR5 and CXCR4. pLNCX After isolation, about 70% of the PBI.S were posi tive for the NGFR marker. When the transduced PBLs were isolated with an anti-NGFR / anti-IgG-magnetic bead method. -1 virus, envelop-mediated syncytium for-mation was almost completely inhibited by pLNCX-RKSK transfection. Also, p24 antigen was very low in the cultures of pLNCX-RKSK transduced PBLs. PLNCX-RKSK transduction inhibited the productions of DP1 p24 antigen by 15%, 43% and 19% on days 4, 7 and 10 respectively. The lymphocytes with the phenotypic knockout of CCR5 and CXCR4 could protect primary human PBLs from DP1 HIV-1 virus infection.