In:
Cytometry Part A, Wiley, Vol. 73A, No. 11 ( 2008-11), p. 1079-1085
Abstract:
To detect HuD‐specific T cells in patients with Hu‐antibody associated paraneoplastic neurological syndromes (Hu‐PNS), we used short‐term stimulation assays with HuD protein spanning peptide pools (PSPP) with purities of at least 70% and found reproducible false‐positive CD8+ T‐cell responses in three of 127 individuals (two healthy controls and one Hu‐PNS patient), which all shared HLA‐A*2402 and HLA‐B*1801. After testing the 15‐mer peptides of the HuD antigen separately, we discovered that the same three 15‐mers yielded the CD8+ T cell response in those three individuals. This highly unusual result could not be reproduced when using new batches of peptides with a higher level of purity ( 〉 82% and 〉 95%). Therefore, we assumed this response was not directed against the HuD peptides and analyzed the HuD 15‐mers by Fourier transform ion cyclotron resonance (FT‐ICR) tandem mass spectrometry (MS/MS), which showed the presence of a cytomegalovirus (CMV)‐encoded peptide (AIAEESDEEEAIVAY) as a contaminant. The three responding individuals all were CMV‐seropositive and the contaminating peptide appeared to fit in the binding groove of HLA‐B*18. Our data reveal that synthetic PSPP may contain immunogenic contaminations which may cause false positive results in T‐cell stimulation assays. © 2008 International Society for Advancement of Cytometry
Type of Medium:
Online Resource
ISSN:
1552-4922
,
1552-4930
DOI:
10.1002/cyto.a.v73a:11
DOI:
10.1002/cyto.a.20636
Language:
English
Publisher:
Wiley
Publication Date:
2008
detail.hit.zdb_id:
2180639-1
SSG:
12
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