title: HIV-infection resistance in PMBC-derived dendritic cells modified with recombinant virus.
abstract: This study aimed to identify the characteristics of recombinant-adenovirus-modified PBMC-derived dendritic cells and their resistance to HIV-1 infection by integrating the CCR5∆32, CCR5siRNA, HIV-1 pol and HIV-1 int genes into a recombinant adenovirus vector using the AdEasy system. Dendritic cells (DCs) were isolated from human PBMCs from blood of healthy donors. The expression of CCR5∆32, CCR5, CXCR4 and HIV-1 p24 in PBMCs or modified cells was measured by western blot, p24 expression in cell lysates was measured by ELISA, and HIV-1 entry was measured by β-galactosidase assay. Furthermore, T-cell immunity induced by the recombinant adenovirus was measured by ELISPOT assay. After the cells were modified by Ad-R5∆32siRNA, the expression of CCR5∆32 increased, while the expression of CCR5 and CXCR4 decreased. There was no adverse effect of adenoviral gene transfer on DC development. CD83 expression on the surface of mature DCs did not change after gene transfer. The expression of p24 remained at low levels in modified cells when challenged by HIV-1. The modified cells showed resistance to HIV-1 infection. Results indicated that recombinant-adenovirus-modified cells demonstrated good resistance to HIV-1 infection. Modification of HSC-derived immune cells, such as DCs, may be a potent strategy to resist HIV-1 infection.
authors.affiliation: Department of Pharmacy, The Third Affiliated Hospital, Medical University of Guangzhou, Guangzhou 510150, China.
authors.forename: Cheng-Lai
authors.initials: CL
authors.lastname: Xia
authors.forename: Ping
authors.initials: P
authors.lastname: Zhu
authors.forename: Yan-Tao
authors.initials: YT
authors.lastname: Cai
authors.forename: Guang-Bin
authors.initials: GB
authors.lastname: Zhu
authors.forename: Zheng-Rong
authors.initials: ZR
authors.lastname: Mei
authors.forename: Hanhui
authors.initials: H
authors.lastname: Huang
authors.forename: Di-Xian
authors.initials: DX
authors.lastname: Luo
authors.forename: Peng-Ke
authors.initials: PK
authors.lastname: Yan
mesh_terms: D000256:Adenoviridae / Q000235:genetics*; D003713:Dendritic Cells / Q000821:virology*; D020868:Gene Silencing; D005822:Genetic Vectors; D019427:HIV Integrase / Q000096:biosynthesis / Q000235:genetics; D015497:HIV-1 / Q000472:pathogenicity*; D006801:Humans; D019713:Receptors, CCR5 / Q000096:biosynthesis / Q000235:genetics; D015728:Receptors, HIV / Q000096:biosynthesis / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000235:genetics; D053585:Virus Attachment; D014779:Virus Replication; D054302:pol Gene Products, Human Immunodeficiency Virus / Q000096:biosynthesis / Q000235:genetics
chemical_list: D019713:Receptors, CCR5; D015728:Receptors, HIV; D011994:Recombinant Proteins; D054302:pol Gene Products, Human Immunodeficiency Virus; D019427:HIV Integrase