Generation in vivo of peptide-specific cytotoxic T cells and presence of regulatory T cells during vaccination with hTERT (class I and II) peptide-pulsed DCs.
BACKGROUND
Optimal techniques for DC generation for immunotherapy in cancer are yet to be established. Study aims were to evaluate: (i) DC activation/maturation milieu (TNF-alpha +/- IFN-alpha) and its effects on CD8+ hTERT-specific T cell responses to class I epitopes (p540 or p865), (ii) CD8+ hTERT-specific T cell responses elicited by vaccination with class I alone or both class I and II epitope (p766 and p672)-pulsed DCs, prepared without IFN-alpha, (iii) association between circulating T regulatory cells (Tregs) and clinical responses.


METHODS
Autologous DCs were generated from 10 patients (HLA-0201) with advanced cancer by culturing CD14+ blood monocytes in the presence of GM-CSF and IL-4 supplemented with TNF-alpha [DCT] or TNF-alpha and IFN-alpha [DCTI]. The capacity of the DCs to induce functional CD8+ T cell responses to hTERT HLA-0201 restricted nonapeptides was assessed by MHC tetramer binding and peptide-specific cytotoxicity. Each DC preparation (DCT or DCTI) was pulsed with only one type of hTERT peptide (p540 or p865) and both preparations were injected into separate lymph node draining regions every 2-3 weeks. This vaccination design enabled comparison of efficacy between DCT and DCTI in generating hTERT peptide specific CD8+ T cells and comparison of class I hTERT peptide (p540 or p865)-loaded DCT with or without class II cognate help (p766 and p672) in 6 patients. T regulatory cells were evaluated in 8 patients.


RESULTS
(i) DCTIs and DCTs, pulsed with hTERT peptides, were comparable (p = 0.45, t-test) in inducing peptide-specific CD8+ T cell responses. (ii) Class II cognate help, significantly enhanced (p < 0.05, t-test) peptide-specific CD8+T cell responses, compared with class I pulsed DCs alone. (iii) Clinical responders had significantly lower (p < 0.05, Mann-Whitney U test) T regs, compared with non-responders. 4/16 patients experienced partial but transient clinical responses during vaccination. Vaccination was well tolerated with minimal toxicity.


CONCLUSIONS
Addition of IFN-alpha to ex vivo monocyte-derived DCs, did not significantly enhance peptide-specific T cell responses in vivo, compared with TNF-alpha alone. Class II cognate help significantly augments peptide-specific T cell responses. Clinically favourable responses were seen in patients with low levels of circulating T regs.
Aloysius
Mark M
MM
Section of Surgery, Biomedical Research Unit, Nottingham Digestive Diseases Centre, University of Nottingham, UK. mark.aloysius@nottingham.ac.uk
Mc Kechnie
Alastair J
AJ
Robins
Richard A
RA
Verma
Chandan
C
Eremin
Jennifer M
JM
Farzaneh
Farzin
F
Habib
Nagy A
NA
Bhalla
Joti
J
Hardwick
Nicola R
NR
Satthaporn
Sukchai
S
Sreenivasan
Thiagarajan
T
El-Sheemy
Mohammed
M
Eremin
Oleg
O
D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D019496:Cancer Vaccines / Q000276:immunology* / Q000627:therapeutic use; D003602:Cytotoxicity, Immunologic / Q000187:drug effects / Q000276:immunology; D015232:Dinoprostone / Q000627:therapeutic use; D006681:HLA-D Antigens / Q000276:immunology*; D015395:Histocompatibility Antigens Class I / Q000276:immunology*; D006801:Humans; D016898:Interferon-alpha / Q000627:therapeutic use; D007375:Interleukin-1 / Q000627:therapeutic use; D015850:Interleukin-6 / Q000627:therapeutic use; D008198:Lymph Nodes / Q000276:immunology; D008264:Macrophages / Q000276:immunology; D009369:Neoplasms / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D013602:T-Lymphocytes, Cytotoxic / Q000276:immunology*; D050378:T-Lymphocytes, Regulatory / Q000276:immunology*; D019098:Telomerase / Q000276:immunology; D014409:Tumor Necrosis Factor-alpha / Q000627:therapeutic use
D019496:Cancer Vaccines; D006681:HLA-D Antigens; D015395:Histocompatibility Antigens Class I; D016898:Interferon-alpha; D007375:Interleukin-1; D015850:Interleukin-6; D010446:Peptide Fragments; D014409:Tumor Necrosis Factor-alpha; D019098:Telomerase; D015232:Dinoprostone