Abstract Book 2010 - CIMT Annual Meeting

028 Laske | Immune monitoring
T-cell immunomonitoring in patients with renal cell carcinoma
after multi-peptide vaccination
Karoline Laske 1 , Annemarie Dröge 1 , Susan Feyerabend 2 , Jörg Hennenlotter 2 , Jens Bedke 2 , Patricia
Hrstic 1 , Stefan Stevanovic 1 , Arnulf Stenzl 2 , Cécile Gouttefangeas 1 , Hans-Georg Rammensee 1
1 Institute for Cell Biology, Department of Immunology, Eberhard Karls University Tuebingen,
Auf der Morgenstelle 15, 72076 Tuebingen, Germany
2 Department of Urology, Eberhard Karls University Tuebingen, Hoppe-Seyler-Str. 3, 72076 Tuebingen,
Germany
In this clinical trial, patients with advanced or
metastatic renal cell carcinoma after complete resection
receive a multi-peptide cocktail containing
HLA-class I and II binding peptides. The vaccine is
applied either intradermally with GM-CSF as adjuvant
or subcutaneously in Montanide ISA52 at
18 vaccination time points over 12 months or until
progression. Computer tomography imaging is performed
to evaluate progression-free survival every
3 months. HLA-A*02 positive or negative patients
receive an individual peptide cocktail containing
between 3 and 12 HLA-class I binding peptides.
These peptides are known epitopes or ligands
derived from selected tumor-associated antigens.
Furthermore, the peptide cocktail for HLA-A*02 positive
patients contains one peptide (from the influenza
virus) as recall control and one peptide (from
HBV) as priming control for the vaccination.
Additionally, 3 to 4 HLA-class II binding peptides
are included for CD4+ T-cell stimulation. Until
now, 17/40 patients have been recruited and T-cell
monitoring was performed for 9 patients. Blood
was collected at each vaccination time point, serum
and peripheral blood mononuclear cells (PBMCs)
were isolated and frozen. For T-cell monitoring,
PBMCs from different time points were thawed and
stimulated with the individual peptide cocktail for
12 days. Detection of specific T-cells was performed
by IFN-gamma Elispot, tetramer staining (for
HLA-A*02 positive patients) and intracellular cytokine
staining. So far, all 7 monitored HLA-A*02
positive patients developed a strong CD4+ T-cell
response against 2 of the 4 HLAclass II binding
epitopes (derived from carbonic anhydrase 9 and
cyclin-D1 protein) which were detected by Elispot
and further characterized by intracellular cytokine
staining. We could detect a CD8+ T-cell response
in 4 of 7 HLA-A*02 positive patients. The most frequently
recognized peptides are from the proteins
cyclin-D1 and guanylate cyclase 1.
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