Volume 1 Supplement 1
Optimization of ex vivo expansion of HER2 specific polyfunctional Th1/Th17 cells from HER2 vaccine primed PBMC
© Dang et al; licensee BioMed Central Ltd. 2013
Published: 7 November 2013
Adoptive transfer of ex vivo expanded neu specific polyfunctional T-cells secreting TNF-alpha (α), IFN-gamma (γ), and IL-17 (Th1/Th17) cells into tumor bearing mice can result in complete resolution of disease as compared to the use of neu specific Th1 (Lai et al 2009). Murine antigen specific Th1/Th17 cells could be readily expanded with IL-2 and IL-21 in culture, however, the use of these cytokines resulted in successful expansion of human tumor antigen specific T-cells in only a minority of patients. We sought to identify ex vivo culture conditions that would be suitable for the clinical expansion of polyfunctional HER2 specific Th1/Th17 for therapeutic infusion. PBMC, derived from the aphaeresis of patients previously immunized with a HER2 vaccine, were stimulated with HER2 peptides in the presence of different cytokines to polarize Th17 cells, and then cultured with different T-cell growth factors on Day4/8, and subsequently expanded with CD3/CD28 beads on Day 12 and IL-2 for 12 days. We found that IL-1beta (β)/IL-6 generated higher number of IL-17 secreting CD4 cells before CD3/CD28 activation. Other cytokine combinations, including IL-1β/IL-6/IL-21, IL-1β/IL-6/anti-TGFβ antibody, and IL-21 alone, failed to further increase IL-17 cells. A low dose of IL-2 alone added in the culture on Day 4/8, following HER2 peptide and IL-1β/IL-6, generated a higher number of antigen specific IL-17 secreting cells than the combinations of IL-2/IL-7 and IL-2/IL-7/IL-15. In addition, exposure to IL1-β/IL-6 at the time of antigen stimulation was superior to the cytokines added on Day 4/8. Flow cytometric studies of the T-cells generated showed the generation of a Th1/Th17 phenotype, including dual secreting IL-17 and TNF-α, IL-17 and IFN-γ, and triple secreting IL-17, IFN-γ and TNF-α. These data demonstrate a streamlined methodology, easily adaptable to the clinic, for the generation of tumor specific polyfunctional T-cells for therapeutic infusion.
This article is published under license to BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.