Expansion of quiescent lung adenocarcinoma CD8+ T cells by MUC1-8-mer peptide-T2 cell-β2 microglobulin complexes

Abstract

Adoptive immunotherapy requires the isolation of CD8+ T cells specific for tumor-associated antigens, their expansion in vitro and their transfusion to the patient to mediate a therapeutic effect. MUC1 is an important adenocarcinoma antigen immunogenic for T cells. The MUC1-derived SAPDTRPA (MUC1-8-mer) peptide is a potent epitope recognized by CD8+ T cells in murine models. Likewise, the T2 cell line has been used as an antigen-presenting cell to activate CD8+ T cells, but so far MUC1 has not been assessed in this context. We evaluated whether the MUC1-8-mer peptide can be presented by T2 cells to expand CD25+CD8+ T cells isolated from HLA-A2+ lung adenocarcinoma patients with stage III or IV tumors. The results showed that MUC1-8-mer peptide-loaded T2 cells activated CD8+ T cells from cancer HLA-A2+ patients when anti-CD2, anti-CD28 antibodies and IL-2 were added. The percentage of CD25+CD8+ T cells was 3-fold higher than those in the non-stimulated cells (P=0.018). HLA-A2+ patient cells showed a significant difference (2.3-fold higher) in activation status than HLA-A2+ healthy control cells (P=0.04). Moreover, 77.6% of MUC1-8-mer peptide-specific CD8+ T cells proliferated following a second stimulation with MUC1-8-mer peptide-loaded T2 cells after 10 days of cell culture. There were significant differences in the percentage of basal CD25+CD8+ T cells in relation to the cancer stage; this difference disappeared after MUC1-8-mer peptide stimulation. In conclusion, expansion of CD25+CD8+ T cells by MUC1-8 peptide-loaded T2 cells plus costimulatory signals via CD2, CD28 and IL-2 can be useful in adoptive immunotherapy.

Identification of Immune-Related lncRNA Biomarkers Associated With Prognosis In LUAD

“…6). Reports have demonstrated that activation of cytotoxic T cell responses is essential for tumor cell elimination and the adoptive immunotherapy requires isolation and expansion of speci c CD8 + T cells [59,60]. Consequently, prominent and distinct associations have been To gain more insights into the functional roles of the immune-related lncRNAs, GO and pathway enrichment analyses performed in this study revealed that most of the signi cant items for each enrichment analysis were directly related to immune response (Fig.…”

Selection of a Single Domain Antibody, Specific for an HLA-Bound Epitope of the Mycobacterial Ag85B Antigen

“…Then, cells (6x10 5 ) were incubated with 80 μg of either Ag85Bp (199-207) target peptide and Esat-6p (82-90) as non-target. Twenty μg/ml of β2m (Sigma) was added according to ( 29 ). Cells were incubated for 8 h at 37°C in 5% CO 2 atmosphere, after that cells were washed twice with PBS and incubated with 10 μg of sdAb 2C ON at 4°C in agitation, and then washed 2 times, with PBS, following by incubation with anti-c-Myc Ab (Santa Cruz-/Europe) (1/100) for 1 h at RT.…”