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Preparation of Tumor Antigen-loaded Mature Dendritic Cells for Immunotherapy
Published on: August 1, 2013
In vitro functional tests for evaluation of stimulating capacity of cultured human dendritic cells
T Hajdinjak1, M Jeras, M Knezević
1Tissue Typing Centre, Blood Transfusion Centre of Slovenia, Ljubljana.
Insights
Mature dendritic cells (DC) significantly accelerate mixed lymphocyte reactions (MLR) compared to macrophages. This finding enables earlier confirmation of mature DC in cell suspensions, optimizing functional testing.
Area of Science:
- Immunology
- Cell Biology
Background:
- Dendritic cells (DC) are key antigen-presenting cells.
- Evaluating DC function typically involves allogeneic mixed lymphocyte reactions (MLR).
- Current MLR assays measure cell proliferation via 3H-thymidine incorporation, usually on days 5-7.
Purpose of the Study:
- To investigate the kinetics of MLR using different stimulator cells.
- To determine if MLR time can be reduced for DC functional testing.
- To compare the stimulating capacity of mature dendritic cells versus macrophages.
Main Methods:
- In vitro cultured human dendritic cells (DC) and macrophages were used as stimulators.
- Allogeneic one-way mixed lymphocyte reaction (MLR) was performed.
- Cell proliferation was measured by 3H-thymidine incorporation over several days.
Main Results:
- Macrophage-stimulated MLR showed significant differences from controls starting on day 4.
- Mature DC-stimulated MLR demonstrated consistently higher proliferation from day 2.
- DC-initiated reactions peaked 2-3 days earlier and were over twice as intense as controls.
Conclusions:
- Mature dendritic cells provide a significantly faster and more intense proliferative response in MLR.
- This accelerated response allows for earlier detection (within 48 hours) of mature DC in cell suspensions.
- The findings optimize functional testing of dendritic cells.
Abstract:
Basic functional test for evaluation of in vitro cultured human dendritic cells (DC) is primary allogeneic one-way mixed lymphocyte reaction (MLR). In this way, one can evaluate stimulating capacity, which is a basic characteristic of DC. The proliferation of cells is measured through incorporation of 3H-thymidine. Normally proliferation is measured at days 5-7. We studied kinetics of proliferative responses initiated with different stimulating cell suspensions to evaluate differences and possibly reduce time needed to perform this test. Gradual increase in response from days 1 to 7 and a significant difference from controls (peripheral blood mononuclear cells) seen from day 4 was noted if macrophages were used as stimulators. A consistently higher proliferation, compared to controls, was always found already on day 2 when mature DC were used as stimulators. The reaction peaked 2 to 3 days earlier and was also more than two times more intense. This maximal and significantly higher response, consistently seen already after 48 hours, allows us to confirm the presence of mature DC in stimulating suspensions much earlier than previously.
