317-W.
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Generation of Monocyte-Derived Dendritic Cells (MD-DC) in Clinical GMP Conditions and their Ability to Activate CD4 and CD8 T Cells to HIV Antigens in Early HIV+ Individuals Receiving HAART
M. Lejeune*1, F. García1, C. Gil1, J. Alcami2, J. Joseph1, J. Miró1, M. Plana1, J. M. Gatell2, and T. Gallart1
1Univ. of Barcelona and 2Inst. Carlos III, Madrid, Spain
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Background: Myeloid DC activate both naïve and memory CD4+ and CD8+ T to non-infectious antigens of the exogenous pathway, and constitute the most potent cellular adjuvant for an active immunotherapy against HIV infection. MD-DC are more accessible in a clinical setting. Objectives were to optimize the generation of MD-DC using clinical GMP conditions from healthy controls (HC) and from early stage HIV-1+ patients (ES) receiving HAART and to determine the capacity to present to autologous T cells, soluble HIV antigens and whole heat-inactivated HIV (HI-HIV).
Methods: Serum-free GMP culture medium was supplemented with 1-5% of autologous serum (AS) and with 1 M of pharmaceutical AZT to obtain MD-DC in a good yield and with optimal immunophenotypic and functional characteristics to induce potent T-lymphocyte responses against different soluble antigens.
Results: The yield and immunophenotypic and functional features of MD-DC from ES-HAART (n=12) are indistinguishable from those from HC (n=25). The presence of anti-HIV IgG antibodies in AS from ES-HAART for pulsing their MD-DC with soluble HIV antigens and HI-HIV favors antigen capture and this in turn promotes MD-DC maturation as assessed by the translocation of the intracellular HLA-DR to the membrane. The addition of pharmaceutical INF-alpha during the last 16 of a 20-hour antigen pulsing period further increases the maturation of MD-DC. Autologous MD-DC from ES-HAART pulsed with recombinant soluble HIV proteins and HI-HIV induce the activation of autologous T-cells, including highly purified CD4 and CD8 T cells as demonstrated by cell growth and IFN- production. The primary activation of CD4 T cells from HC was also obtained against HIV-1 recombinant proteins, although the magnitude of the response was lower and delayed as compared to HIV-1+ individuals.
Conclusions: Functional MD-DC can be easily obtained in GMP conditions in early stage HIV+ individuals receiving HAART. They are able to induce potent anti-HIV T-cell responses in these individuals. These data support the use of MD-DC as a cellular adjuvant for a HIV therapeutic vaccine.
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