Report From the 1st International NOD Mouse T-Cell Workshop and the Follow-Up Mini-Workshop
- Daniel L. Kaufman1,
- Roland Tisch2,
- Nora Sarvetnick3,
- Lucienne Chatenoud4,
- Leonard C. Harrison5,
- Kathryn Haskins6,
- Anthony Quinn7,
- Eli Sercarz7,
- Bhagi Singh8,
- Matthias von Herrath9,
- Dale Wegmann10,
- Li Wen11 and
- Dan Zekzer12
- 1Department of Molecular and Medical Pharmacology, University of California, Los Angeles, Los Angeles, California
- 2Department of Microbiology and Immunology, University of North Carolina, Chapel Hill, North Carolina
- 3Department of Immunology, Scripps Research Institute, La Jolla, California
- 4Institut National de la Santé et de la Récherche Medicale, Hopital Necker, Paris, France
- 5Walter and Eliza Hall Institute of Medical Research, PO Royal Melbourne Hospital, Victoria, Australia
- 6Health Science Center Department of Immunology, University of Colorado, Denver, Colorado
- 7La Jolla Institute for Allergy and Immunology, San Diego, California
- 8University of Western Ontario, Department of Microbiology, London, Ontario, Canada
- 9Department of Virology, Scripps Research Institute, La Jolla, California
- 10Barbara Davis Center for Childhood Diabetes, Denver, Colorado
- 11Department of Internal Medicine, Endocrinology, Yale University, New Haven, Connecticut
- 12Sankyo, La Jolla, California
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 β-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.
Footnotes
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Address correspondence and reprint requests to Daniel Kaufman, Department of Molecular and Medical Pharmacology, UCLA School of Medicine, Box 173517, UCLA, Los Angeles, CA. 90095-1735. E-mail: dkaufman{at}mednet.ucla.edu.
Received for publication 24 April 2001 and accepted in revised form 10 August 2001.
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.
Additional information can be found in an online appendix at http://diabetes.diabetesjournals.org.
ELISA, enzyme-linked immunosorbent assay; ELISPOT, enzyme-linked immunospot assay; SI, stimulation index; UCLA, University of California, Los Angeles.
