HIV-infection resistance in PMBC-derived dendritic cells
modiﬁed with recombinant virus
Received: 25 August 2011 / Accepted: 29 November 2011 / Published online: 11 December 2011
Ó Springer-Verlag 2011
Abstract This study aimed to identify the characteristics
of recombinant-adenovirus-modiﬁed PBMC-derived den-
dritic cells and their resistance to HIV-1 infection by
integrating the CCR5D32, CCR5siRNA, HIV-1 pol and
HIV-1 int genes into a recombinant adenovirus vector
using the AdEasy system. Dendritic cells (DCs) were iso-
lated from human PBMCs from blood of healthy donors.
The expression of CCR5D32, 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 b-galactosidase
assay. Furthermore, T-cell immunity induced by the
recombinant adenovirus was measured by ELISPOT assay.
After the cells were modiﬁed by Ad-R5D32siRNA, the
expression of CCR5D32 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.
Human immunodeﬁciency virus (HIV) continues to be a
global health threat, as an estimated 60 million people cur-
rently live with the virus. About 2.5 million people become
newly infected with HIV-1 and 2.1 million people die of
AIDS-related diseases every year . As a retrovirus,
HIV-1 can infect two kinds of cells, CD4
T cells and
macrophages, leading to progressive depletion of CD4
cells. HIV-1 enters target cells by a three-step process .
Primary infections are established by macrophage-tropic
HIV-1 using CCR5 as the coreceptor, whereas X4-tropic
viruses that predominate late in infection use CXCR4 as the
major coreceptor [3, 26].
Due to the lack of effective vaccines and the develop-
ment of resistance to current drug therapies, alternative
treatment strategies need to be developed. The key to a
permanent treatment of chronic HIV infection is the ability
to elicit potent host resistance to viral infection and to
restore immune functions. Intracellular immunization
C. Xia, P. Zhu and Y. Cai contributed equally for this work.
C. Xia Á Z. Mei Á H. Huang Á P. Yan (&)
Department of Pharmacy, The Third Afﬁliated Hospital,
Medical University of Guangzhou, Guangzhou 510150, China
Guangdong Cardiovascular Institute, Guangdong General
Hospital, Guangdong Academy of Medical Sciences,
Guangzhou 510080, China
Department of Dermatology and Rheumatology,
Foshan Maternity and Child Health Care Hospital,
Foshan 528000, China
Department of Radiology, The First Afﬁliated Hospital,
Medical University of Guangzhou, Guangzhou 515120, China
D. Luo (&)
Institute of Traditional Medicine of University of South China,
Chenzhou No.1 People’s Hospital, Chenzhou 423000,
Arch Virol (2012) 157:413–421