title: Three sensitive assays do not provide evidence for circulating HuD-specific T cells in the blood of patients with paraneoplastic neurological syndromes with anti-Hu antibodies.
abstract: Anti-Hu antibody-associated paraneoplastic neurological syndromes (Hu-PNSs) are severe and often precede the detection of a malignancy, usually small-cell lung cancer. In Hu-PNS, it is hypothesized that neuronal cells are destroyed by T cells targeted against HuD, a protein expressed by small-cell lung cancer cells and neurons. There is only limited evidence for the existence of HuD-specific T cells. To detect these T cells in the blood of Hu-PNS patients, we employed 3 highly sensitive assays that included T cell stimulation with dendritic cells (DCs) to specifically expand the number of any HuD-specific T cells. A total of 17 Hu-PNS patients were tested with 1 or more of the following 3 assays: (1) tetramer staining after stimulation of T cells with conventionally generated DCs (n = 9), (2) interleukin (IL)-13 enzyme-linked immunosorbent spot (ELISpot; n = 3), IL-4 and IL-5 and interferon (IFN)-γ multiplex cytokine bead array (n = 2) to assay cytokine production by T cells after stimulation with conventionally generated DCs, and (iii) IFN-γ ELISpot and tetramer staining after T cell stimulation with accelerated co-cultured DCs (n = 11). No circulating HuD-specific T cells were found. We suggest that either autoaggressive T cells in Hu-PNS are not targeted against HuD or that their numbers in the blood are too low for detection by highly sensitive techniques.
authors.affiliation: Department of Neurology, Erasmus University Medical Center, 's-Gravendijkwal 230, 3015 CE Rotterdam, the Netherlands.
authors.forename: Adriaan H C
authors.initials: AH
authors.lastname: de Jongste
authors.forename: Marieke T
authors.initials: MT
authors.lastname: de Graaf
authors.forename: Emanuela
authors.initials: E
authors.lastname: Martinuzzi
authors.forename: Patricia D M
authors.initials: PD
authors.lastname: van den Broek
authors.forename: Jaco
authors.initials: J
authors.lastname: Kraan
authors.forename: Cor H J
authors.initials: CH
authors.lastname: Lamers
authors.forename: Roberto
authors.initials: R
authors.lastname: Mallone
authors.forename: Jan W
authors.initials: JW
authors.lastname: Gratama
authors.forename: Peter A E
authors.initials: PA
authors.lastname: Sillevis Smitt
mesh_terms: D000368:Aged; D000888:Antibodies, Anti-Idiotypic / Q000097:blood*; D001681:Biological Assay; D018414:CD8-Positive T-Lymphocytes / Q000276:immunology*; D016022:Case-Control Studies; D051959:ELAV Proteins / Q000097:blood / Q000276:immunology*; D004797:Enzyme-Linked Immunosorbent Assay; D005260:Female; D006801:Humans; D007371:Interferon-gamma / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000276:immunology; D008297:Male; D008875:Middle Aged; D020361:Paraneoplastic Syndromes, Nervous System / Q000097:blood* / Q000276:immunology*; D010446:Peptide Fragments / Q000276:immunology; D016176:T-Lymphocyte Subsets / Q000276:immunology
chemical_list: D000888:Antibodies, Anti-Idiotypic; D051959:ELAV Proteins; D010446:Peptide Fragments; D007371:Interferon-gamma