Report from the 1st International NOD Mouse T-Cell Workshop and the follow-up mini-workshop

A workshop on autoreactive T-cell responses in NOD mice was held to optimiz e autoreactive T-cell detection methodologies. Using different proliferatio n assay protocols, 1 of the 11 participating laboratories detected spontane ous T-cell responses to GAD(524-543) and insulin(9-23) in their NOD mice. T wo other laboratories were able to detect autoreactive responses when using enzyme-linked immunospot assay (ELISPOT) and enzyme-linked immunosorbent a ssay (ELISA) analysis of cytokines in culture supernatants, suggesting that these assays provided greater sensitivity. To address the divergent findin gs, a follow-up mini-workshop tested NOD mice from four different colonies side-by-side for T-cell proliferative responses to an expanded panel of aut oantigens, using the protocol that had enabled detection of responses in th e 1st International NOD Mouse T-Cell Workshop. Under these assay conditions , 16 of 16 NOD mice displayed proliferative responses to whole GAD65, 13 of 16 to GAD(524-543), 9 of 16 to GAD(217-236), 7 of 16 to insulin(9-23), and 5 of 16 to HSP277. Thus, spontaneous proliferative T-cell responses can be consistently detected to some P-cell autoantigens and peptides thereof. Ov erall, the results suggest that more sensitive assays (e.g., ELISPOT, ELISA analysis of cytokines in supernatants, or tetramer staining) may be prefer red for the detection of autoreactive T-cells.