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elispot-enzyme-linked-immunospot

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Journal Articles
Journal: Diabetes
Diabetes 2001;50(11):2459–2463
Published: 01 November 2001
... 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...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2003;52(11):2647–2651
Published: 01 November 2003
... healthy, nondiabetic control subjects. PBMCs were screened for peptide recognition using interferon-γ enzyme-linked immunospot (ELISpot) assays. Of the nine patients with recent-onset type 1 diabetes, six had ELISpot responses to preproIAPP 5-13 that were >3 SDs above the mean of the nondiabetic...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(4):1229–1236
Published: 22 March 2011
... in NOD.β2mnull.HHD mice. This result was supported by enzyme-linked immunospot (ELISPOT) analyses indicating combined INS/IGRP-SPs diminished HLA-A2.1–restricted IGRP but not INS autoreactive CD8+ T-cell responses in NOD.β2mnull.HHD mice. CONCLUSIONS These data support the potential...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(11):3068–3074
Published: 01 November 2006
..., British Columbia V6H 3V4, Canada. E-mail: roo@interchange.ubc.ca 25 7 2006 13 1 2006 DIABETES 2006 BIMAS, BioInformatics and Molecular Analysis Section CMV, cytomegalovirus CTL, cytotoxic T-lymphocyte EBV, Epstein-Barr virus ELISpot, enzyme-linked immunospot FITC...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(2):265–270
Published: 01 February 2002
... measured by interferon-γ (IFN-γ) enzyme-linked immunospot (ELISpot) assay using peripheral blood leukocytes obtained from the recipient and donor animals. An equal number of irradiated stimulators (donor leukocytes) and responders (recipient leukocytes) were added to ester cellulose bottom plates...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(11):3061–3067
Published: 01 November 2006
... Ab, antibody BIMAS, BioInformatics and Molecular Analysis Section CTL, cytotoxic T-lymphocyte ELISpot, enzyme-linked immunospot GFAP, glial fibrillary acidic protein IA-2, insulinoma-associated antigen 2 IGRP, islet-specific glucose-6-phosphatase catalytic subunit–related protein ppIAPP...
Journal Articles
Journal: Diabetes
Diabetes 2007;56(3):613–621
Published: 01 March 2007
... Station, TX). P < 0.05 was considered to be of statistical significance. Ab, antibody ELISpot, enzyme-linked immunospot IFN, interferon IGRP, islet glucose-6-phosphatase catalytic subunit–related protein ISL8Spot, islet-specific CD8+ T-cell interferon-γ enzyme-linked...
Journal Articles
Journal: Diabetes
Diabetes 2007;56(10):2551–2560
Published: 01 October 2007
... succinimidyl ester ELISPOT, enzyme-linked immunospot IFN, interferon IGRP, islet-specific glucose-6-phosphatase catalytic subunit-related protein MHC, major histocompatibility complex Type 1 diabetes is an autoimmune disease characterized by T-cell–mediated elimination of insulin-producing pancreatic...
Journal Articles
Journal: Diabetes
Diabetes 2004;53(9):2338–2345
Published: 01 September 2004
..., allophycocyanin-conjugated streptavidin ELISPOT, enzyme-linked immunospot IFN, interferon IL, interleukin mHA, minor histocompatibility antigen MHC, major histocompatibility complex PLNC, pancreatic lymph node cell Type 1 diabetes is a T-cell–mediated autoimmune disease that causes the destruction...
Meeting Abstracts
Journal: Diabetes
Diabetes 2000;49(9):1459–1467
Published: 01 September 2000
... preventive assay; ELISPOT, enzyme-linked immunospot; EMC-D, D variant of immunities lasted longer than 10 months in the immunized encephalomyocarditis; OADC, oleic acid, albumin, dextrose, catalase; PBS, mice. These long-lasting immunities are likely to be associ- phosphate-buffered saline; PCR, polymerase...
Journal Articles
Journal: Diabetes
Diabetes 2006;55(9):2588–2594
Published: 01 September 2006
... cut into strips based on molecular weight and then solubilized, using carbonate/bicarbonate buffers. The resulting nitrocellulose particles were then used to stimulate PBMCs in vitro ( 20 ). DASP, Diabetes Autoantibody Standardization Program ELISpot, enzyme-linked immunospot IA2, insulinoma...
Journal Articles
Journal: Diabetes
Diabetes 2002;51(5):1375–1382
Published: 01 May 2002
... because of limitations of the methods to detect and isolate a small number of antigen-specific T-cells in the peripheral blood. New techniques, such as the use of multimeric peptide-MHC complexes and enzyme-linked immunospot (ELISpot) assay, have enabled the detection of autoreactive T-cells at a very...
Journal Articles
Journal: Diabetes
Diabetes 2012;61(10):2546–2555
Published: 13 September 2012
... anterioris muscle, followed 5 and 19 days later by 50 µg of GAD or noncoding plasmid ( 13 ). After 2 weeks, splenocytes were collected and plated (5 × 105 per well) on enzyme-linked immunospot (ELISpot) polyvinylidene fluoride plates (Millipore) coated with anti–interferon-γ (IFN-γ) antibodies...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2009;58(6):1302–1311
Published: 16 March 2009
...) for TGF-β1 and enzyme-linked immunospot (ELISpot) kits for IFN-γ, IL-4, and IL-17 were purchased from Ebioscience. At 1 month after the completion of treatment, animals were euthanized, and peripheral blood mononuclear cells were purified from spleen or pancreatic lymph nodes using Lympholyte-M (Cedarlane...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2012;61(4):857–865
Published: 14 March 2012
...://creativecommons.org/licenses/by-nc-nd/3.0/ for details. IFN-γ enzyme-linked immunospot (ELISPOT) assay was performed according to the manufacturer’s instructions (BD Biosciences). Spleen cells, harvested from retrogenic mice (7 × 105 cells/well), were incubated in the presence or absence of 100 μg/mL...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2017;66(8):2220–2229
Published: 10 April 2017
..., CD3+, CD4+, and TCR+ cells. Levels of CellTrace Violet fluorescence were used to determine cellular proliferation. Enzyme-Linked ImmunoSpot (ELISPOT) assays in 96-well plates (S2EM004M99; Fisher Scientific) were performed according the manufacturer’s guidelines...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2011;60(8):2112–2119
Published: 18 July 2011
..., was obtained from Sanofi Pasteur (Berkshire, U.K.) and used at 1 μL/mL as a positive control. Detection of IFN-γ production by CD4+ T-cells in response to peptide stimulation was carried out using an enzyme-linked immunospot (ELISPOT) assay that has significant discriminative ability for type 1...
Includes: Supplementary data
Journal Articles
Journal: Diabetes
Diabetes 2006;55(7):1978–1984
Published: 01 July 2006
.... Interferon (IFN)-γ and interleukin-4 ELISPOT (enzyme-linked immunosorbent spot analysis) kits (BD Biosciences, San Diego, CA) were used for splenocyte analysis. Spleen cells were prepared by homogenization in red blood cell lysis buffer and cultured at 2–4 × 105 cells/well in triplicate. Cells...
Journal Articles
Journal: Diabetes
Diabetes 2009;58(2):394–402
Published: 01 February 2009
... a mass range of 500–3,200 Da and recorded with a Voyager-DE-Pro mass spectrometer (PerSeptive Biosystems, Framingham, MA). Monoisotopic masses were calculated, and peptides corresponding to computed masses were identified ( 23 ). IFN-γ enzyme-linked immunospot (ELISpot) was performed as previously...
Journal Articles
Journal: Diabetes
Diabetes 2011;60(3):857–866
Published: 21 February 2011
... identically as that performed for the CTL assays. ELISpots were performed in 96-well flat-bottom MAIP S4510 plates (Millipore) using a human IFN-γ ELISpot kit from Mabtech. Immunospot plates were coated with 15 μg/mL capture anti-human IFN-γ mAbs (clone 1-D1 K) by overnight incubation at 4°C. A total of 5,000...