Report From the 1st International NOD Mouse T-Cell Workshop and the Follow-Up Mini-Workshop

Daniel L Kaufman,Roland Tisch,Eli Sercarz,Dale Wegmann,Nora Sarvetnick,Dan Zekzer,Bhagi Singh,Kathryn Haskins,Lucienne Chatenoud,Matthias Von Herrath,Leonard C Harrison,Anthony Quinn,Li Wen

doi:10.2337/diabetes.50.11.2459

Abstract

A workshop on autoreactive T-cell responses in NOD mice was held to optimize autoreactive T-cell detection methodologies. Using different proliferation assay protocols, 1 of the 11 participating laboratories detected spontaneous T-cell responses to GAD(524-543) and insulin(9-23) in their NOD mice. Two other laboratories were able to detect autoreactive responses when using enzyme-linked immunospot assay (ELISPOT) and enzyme-linked immunosorbent assay (ELISA) analysis of cytokines in culture supernatants, suggesting that these assays provided greater sensitivity. To address the divergent findings, 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 autoantigens, using the protocol that had enabled detection of responses in the 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 beta-cell autoantigens and peptides thereof. Overall, the results suggest that more sensitive assays (e.g., ELISPOT, ELISA analysis of cytokines in supernatants, or tetramer staining) may be preferred for the detection of autoreactive T-cells.

Highlights

A workshop on autoreactive T-cell responses in NOD mice was held to optimize autoreactive T-cell detection methodologies
D.L.K. is on the advisory board of Diamyd (Stockholm), which is involved in GAD-based therapeutics, and CTL (Cleveland, OH), which is involved in T-cell detection equipment
The use of synthetic peptides would circumvent issues concerning the purity of the antigen

Summary

Introduction

A workshop on autoreactive T-cell responses in NOD mice was held to optimize autoreactive T-cell detection methodologies. Laboratories that can detect proliferative responses to whole ␤-cell autoantigens directly ex vivo often do not detect responses to peptides that were identified as containing autoantigen target determinants in the NOD mice of another colony. Only 3 of the 11 participating laboratories could detect T-cell responses to either insulin(9-23) or GAD(524-543) in their NOD mice (Table 1).

Results

Conclusion