Skip to main page content
U.S. flag

An official website of the United States government

Dot gov

The .gov means it’s official.
Federal government websites often end in .gov or .mil. Before sharing sensitive information, make sure you’re on a federal government site.

Https

The site is secure.
The https:// ensures that you are connecting to the official website and that any information you provide is encrypted and transmitted securely.

Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2009 Nov;58(11):2588-95.
doi: 10.2337/db09-0249. Epub 2009 Aug 12.

Validity and reproducibility of measurement of islet autoreactivity by T-cell assays in subjects with early type 1 diabetes

Kevan C Herold  1 , Barbara Brooks-Worrell, Jerry Palmer, H Michael Dosch, Mark Peakman, Peter Gottlieb, Helena Reijonen, Sefina Arif, Lisa M Spain, Clinton Thompson, John M Lachin; Type 1 Diabetes TrialNet Research Group
Affiliations

Validity and reproducibility of measurement of islet autoreactivity by T-cell assays in subjects with early type 1 diabetes

Kevan C Herold et al. Diabetes. 2009 Nov.

Abstract

Objective: Type 1 diabetes results from an immunemediated destruction of beta-cells, likely to be mediated by T lymphocytes, but the sensitivity, specificity, and other measures of validity of existing assays for islet autoreactive T-cells are not well established. Such assays are vital for monitoring responses to interventions that may modulate disease progression.

Research design and methods: We studied the ability of cellular assays to discriminate responses in patients with type 1 diabetes and normal control subjects in a randomized blinded study in the U.S. and U.K. We evaluated the reproducibility of these measurements overall and to individual analytes from repeat collections.

Results: Responses in the cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays could differentiate patients from control subjects with odds ratios of 21.7, 3.44, and 3.36, respectively, with sensitivity and specificity as high as 74 and 88%. The class II tetramer and U.S. ELISPOT assays performed less well. Despite the significant association of the responses with type 1 diabetes, the reproducibility of the measured responses, both overall and individual analytes, was relatively low. Positive samples from normal control subjects (i.e., false positives) were generally isolated to single assays.

Conclusions: The cellular immunoblot, U.K.-ELISPOT, and T-cell proliferation assays can distinguish responses from patients with type 1 diabetes and healthy control subjects. The limited reproducibility of the measurements overall and of responses to individual analytes may reflect the difficulty in detection of low frequency of antigen-specific T-cells or variability in their appearance in peripheral blood.

PubMed Disclaimer

Figures

FIG. 1.
FIG. 1.
The percentage of subjects with positive autoantibodies. The data show the percentage of subjects with 0–3 positive autoantibodies.
FIG. 2.
FIG. 2.
The frequency of positive analytes within T-cell assays. The number of positive analytes for each of the assays are shown for patients with type 1 diabetes and healthy control subjects. A: Combined immunoblot assay; B: U.K.-ELISPOT assay; C: TCP assay.
FIG. 3.
FIG. 3.
Sensitivity and specificity of each of the analytes used in the T-cell assays. A: Combined immunoblot assay; B: U.K.-ELISPOT assay; C: TCP assay.
See this image and copyright information in PMC

References

    1. Atkinson MA: ADA Outstanding Scientific Achievement Lecture 2004 Thirty years of investigating the autoimmune basis for type 1 diabetes: why can't we prevent or reverse this disease? Diabetes 2005;54:1253–1263 - PubMed
    1. Ellis T, Jodoin E, Ottendorfer E, Salisbury P, She JX, Schatz D, Atkinson MA: Cellular immune responses against proinsulin: no evidence for enhanced reactivity in individuals with IDDM Diabetes 1999;48:299–303 - PubMed
    1. Dosch H, Cheung RK, Karges W, Pietropaolo M, Becker DJ: Persistent T cell anergy in human type 1 diabetes J Immunol 1999;163:6933–6940 - PubMed
    1. Reijonen H, Novak EJ, Kochik S, Heninger A, Liu AW, Kwok WW, Nepom GT: Detection of GAD65-specific T-cells by major histocompatibility complex class II tetramers in type 1 diabetic patients and at-risk subjects Diabetes 2002;51:1375–1382 - PubMed
    1. Arif S, Tree TI, Astill TP, Tremble JM, Bishop AJ, Dayan CM, Roep BO, Peakman M: Autoreactive T cell responses show proinflammatory polarization in diabetes but a regulatory phenotype in health J Clin Invest 2004;113:451–463 - PMC - PubMed

MeSH terms