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Abstract
A Feasibility study for detection of HIV specific cytotoxic T lymphocytes using rAAV delivery system
Zheng B.J.1, Lee S.S.2, Wong K.H.2, Chan K.C.W.2, Chan C.S.1, Yuen K.Y.1
Objectives: This study aims to develop a new technique for detection of HIV specific cytotoxic T lymphocytes (CTL). Although methods for the detection of HIV specific CTL have been established, i.e. flow cytometry, Tetramer and Elispot using specific CTL peptides, their application is still limited because: (1) the patients’ HLA-I subtypes must be determined; (2) only a few HLA-I HIV specific peptides have been identified; and (3) HLA typing and synthesis of specific peptides are expensive.
Methods: Recombinant adeno-associated virus (rAAV) expressing HIV antigens gp120, gp41 and gag were constructed. HIV antigen-specific CTLs were detected in 10 HIV patients with HLA-I subtype A2 (HLA-A2) by a standard method of flow cytometry for detection of intracellular IFN-g using HLA-A2-restricted HIV-1 CTL specific peptides (gp120 194-202, gp41 741-749 and gag 77-85) and the new technique using rAAV infected autologous B cells or dendritic cells (DC) as stimulator. The results obtained from these 2 methods were compared.
Results: Frequency of CD3+/CD8+/IFN-gamma+ cells was lower in the test using rAAV infected B cells than in the standard method using HIV specific CTL peptides. It might be due to the infection rate of rAAV was less than 3% in B cells. However, frequencies of CD3+/CD8+/IFN-gamma+ cells were similar when compared results between the tests using rAAV infected DC or purified rAAV infected B cells for the stimulation and those tested by the standard method.
Conclusions: Theoretically, this may be used for detection of HIV specific CTL, but the infection rate of rAAV in B cells should be improved. Alternately, the other viral delivery system which has higher infection rate for B cells, such as lentivirus, may be used.
This study was supported in part by AIDS Trust Fund, Hong Kong.
4th IAS Conference on HIV Pathogenesis, Treatment and Prevention
Abstract no.
TUPEA003
Suggested Citation
"ZhengB.J., et al.
A Feasibility study for detection of HIV specific cytotoxic T lymphocytes using rAAV delivery system.
Poster exhibition:
4th IAS Conference on HIV Pathogenesis, Treatment and Prevention:
Abstract no.
TUPEA003"
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