Abstract Book 2010 - CIMT Annual Meeting

L124 Zhang | Immune monitoring
Goal of ELISPOT proficiency accomplished: ELISPOT assays provide
reproducible results among different laboratories for T-cell
immune monitoring — even in hands of ELISPOT novices
W. Zhang 1 , R. Caspell 1 , A. Y. Karulin 1 , M. Ahmad 2 , N. Haicheur 3 , A. Abdelsalam 4 , K.
Johannesen 5 , V. Vignard 6 , P. Dudzik 7 , K. Georgakopoulou 8 , A. Mihaylova 9 , K. Silina 10 , N
Aptsiauri 11 , V. Adams 12 , P. V. Lehmann 1,13 , and S. McArdle 2
1 Cellular Technology Ltd., Shaker Hts. Ohio, USA
2 Nottingham Trent University, Nottingham, UK
3 Hôpital Européen Georges Pompidou, Paris, France
4 University of Pittsburgh, Pittsburgh, PA, USA
5
Norwegian Radium Institute, Oslo,Norway
6
INSERM, Nantes, France
7
Jagiellonian University Medical College, Krakow, Poland
8 Immunology Center, St. Savas Cancer Hospital, Athens, Greece
9
UMBAL “Alexandrovska”, Sofia, Bulgaria
10
Biomedical Research Study Centre, Riga, Latvia
11
Hospital Universitario Virgen De la Nieves, Granada, Spain
12
Onyvax Ltd, London, UK
13
Department of Pathology, Case Western Reserve University, Cleveland OH, USA
72
T cell monitoring remains challenging in tumor
vaccine trials due to the necessity of testing live
cells in functional assays, and the low frequencies
of tumor antigen-specific T cells. Recent multi-center
initiatives that aimed at harmonizing T
cell assays have drawn attention to alarming- 30-,
20- and 150- fold inter-laboratory variations in test
results for ELISPOT, ICS and tetramers, respectively
(Immunity, 2009, 31: 527-528). The authors concluded:
“The high degree in variability makes the
comparison between any two labs become a game
of chance”. Puzzled and alarmed by this message,
we undertook a similar effort for ELISPOT together
with a European consortium, NEUCAPs. For
our study, we required that all participants from
the eleven reporting labs follow the same detailed
protocol using one uniform platform, and that the
study participants had never previously conducted
an ELISPOT assay (the results of their first attempt
were recorded for the study). While three of the
labs failed with the basic logistics of the trial, eight
detected the peptide-specific CD8+ T-cells in frequencies
approximating the values established by
the Reference Laboratory. These results show that
ELISPOT can produce comparable, reliable data
(even from untrained personnel) if a standardized
platform for the assay and data analysis is followed.
Since ELISPOT assays have been qualified
and validated for regulated studies, they are ideal
candidates for robust and reproducible monitoring
of Tcell immunity in tumor vaccine trials.