scholarly journals Determination of autoantibodies in dogs with diabetes mellitus

2021 ◽  
pp. 2694-2698
Author(s):  
Franco González-Villar ◽  
Francisco Pérez-Bravo
Keyword(s):  
Diabetes Mellitus ◽  
Detection System ◽  
Tyrosine Phosphatase ◽  
Normal Weight ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acid Decarboxylase ◽  
Insulin Deficiency ◽  
Insulin Dependent ◽  
Islet Antigen ◽  
Type 1 Dm

Background and Aim: The classification of diabetes mellitus (DM) in dogs has been controversial as currently canine insulin-dependent DM is classified together with absolute insulin deficiency, non-insulin-dependent DM, and relative insulin deficiency. Studies on human autoantibodies evaluated in canines with DM, such as anti-glutamic acid decarboxylase (GAD65), anti-islet antigen 2 (IA2), and anti-zinc transporter isoform 8 (ZnT8), have been inconclusive. Thus, this study was designed to establish the serological profile of anti-GAD65, anti-IA2, and anti-ZnT8 antibodies in a group of dogs with and without DM. Materials and Methods: Sixty-one dogs, including 31 patients with DM (with and without insulin treatment) and 30 patients without DM (normal weight and obese), were included for determining autoantibodies using a human enzyme-linked immunosorbent assay (ELISA) detection system for type 1 DM. Results: This study found the presence of anti-IA2 antibodies in 58% of the sample (18/31 patients with DM); however, the presence of anti-GAD65 was not detected, and anti-ZnT8 was found in 3 (9.6%) patients with DM. Conclusion: This study showed a higher positive frequency of anti-IA2 antibodies in a sample of canine with DM, indicating that alterations in the signaling vesicle tyrosine phosphatase 2 lead to lower insulin release and thus to an increase in patients' glycemia. These preliminary results should be taken with caution and corroborated by a canine-specific assay when an ELISA is available for such determination.

scholarly journals Fusion Proteins for Combined Analysis of Autoantibodies to the 65-kDa Isoform of Glutamic Acid Decarboxylase and Islet Antigen-2 in Insulin-dependent Diabetes Mellitus

2001 ◽  
Vol 47 (5) ◽  
pp. 926-934 ◽  
Author(s):  
Mathias Rickert ◽  
Jochen Seissler ◽  
Werner Dangel ◽  
Helga Lorenz ◽  
Wiltrud Richter
Keyword(s):  
Diabetes Mellitus ◽  
Fusion Protein ◽  
Cost Effective ◽  
Insulin Dependent Diabetes Mellitus ◽  
Insulin Dependent Diabetes ◽  
Dependent Diabetes Mellitus ◽  
Acid Decarboxylase ◽  
Immune Reactivity ◽  
Insulin Dependent ◽  
Islet Antigen

Abstract Background: Prediction, risk assessment, and diagnosis of autoimmune diseases often rely on detection of autoantibodies directed to multiple target antigens, such as the 65-kDa isoform of glutamic acid decarboxylase (GAD65-abs) and the tyrosine phosphatase-like protein islet antigen-2 (IA2-abs), the two major subspecificities of islet cell antibodies (ICAs) associated with insulin-dependent diabetes mellitus. We hypothesized that a combination of autoantigens in a fusion protein unifying the important immunodominant epitopes could provide an efficient target for cost-effective, one-step screening of sera. Methods: Chimeric proteins composed of GAD65 and IA2 residues were constructed, analyzed for their immune reactivity with monoclonal antibodies and sera, and used in a diagnostic assay with 35S-labeled protein as antigen. Results: Length and order of GAD65 and IA2 sequences were critical for conservation of the conformational epitopes in the fusion protein. Among four chimera tested, only IA2(606–979)/GAD65(1–585) retained wild-type-like folding of GAD65 and IA2 domains and yielded a stable protein after baculovirus expression. Reactivity of GAD65 antibody- and IA2 antibody-positive sera from patients newly diagnosed with insulin-dependent diabetes mellitus, from ICA-positive prediabetics, and from ICA-positive first-degree relatives demonstrated conservation of the relevant autoreactive epitopes. The assay based on the in vitro translated fusion antigen had a sensitivity and specificity identical to those for detection of GAD65- and IA2-abs based on the two separate GAD65 and IA2 proteins. Conclusions: Autoantigens such as GAD65 and IA2 can be combined successfully in a fusion protein of similar immune reactivity. This allows simultaneous detection of GAD65- and IA2-abs in a one-step screening assay and cost-effective identification of positive individuals at risk of diabetes or at onset of disease.

scholarly journals Comparison of Measurements of Autoantibodies to Glutamic Acid Decarboxylase and Islet Antigen-2 in Whole Blood Eluates from Dried Blood Spots Using the RSR-Enzyme Linked Immunosorbent Assay Kits and In-House Radioimmunoassays

2010 ◽  
Vol 2010 ◽  
pp. 1-6 ◽  
Author(s):  
Anders Persson ◽  
Charlotte Becker ◽  
Ida Hansson ◽  
Anita Nilsson ◽  
Carina Törn
Keyword(s):  
Glutamic Acid ◽  
Glutamic Acid Decarboxylase ◽  
Dried Blood Spots ◽  
Enzyme Linked Immunosorbent Assay ◽  
Acid Decarboxylase ◽  
Blood Spots ◽  
And Control ◽  
Islet Antigen ◽  
Prozone Effect

To evaluate the performance of dried blood spots (DBSs) with subsequent analyses of glutamic acid decarboxylase (GADA) and islet antigen-2 (IA-2A) with the RSR-ELISAs, we selected 80 children newly diagnosed with type 1 diabetes and 120 healthy women. DBSs from patients and controls were used for RSR-ELISAs while patients samples were analysed also with in-house RIAs. The RSR-ELISA-GADA performed well with a specificity of 100%, albeit sensitivity (46%) was lower compared to in RIA (56%;P=.008). No prozone effect was observed after dilution of discrepant samples. RSR-ELISA-IA-2A achieved specificity of 69% and sensitivity was lower (59%) compared with RIA (66%;P<.001). Negative or low positive patients and control samples in the RSR-ELISA-IA-2A increased after dilution. Eluates from DBS can readily be used to analyse GADA with the RSR-ELISA, even if low levels of autoantibodies were not detected. Some factor could disturb RSR-ELISA-IA-2A analyses.

Autoimmune thyroid disease in patients with anti-GAD positive type 1 diabetes mellitus

Open Medicine ◽  
2009 ◽  
Vol 4 (4) ◽  
pp. 415-422
Author(s):  
Kamile Gul ◽  
Ihsan Ustun ◽  
Yusuf Aydin ◽  
Dilek Berker ◽  
Halil Erol ◽  
...  
Keyword(s):  
Diabetes Mellitus ◽  
Type 1 Diabetes ◽  
Type 1 Diabetes Mellitus ◽  
Statistical Significance ◽  
Control Group ◽  
Acid Decarboxylase ◽  
Thyroid Peroxidase ◽  
The Difference ◽  
Type 1 Dm

AbstractThe aim of the study was to determine the frequency and titers of anti-thyroid peroxidase (Anti-TPO), anti-thyroglobulin (Anti-TG), and anti-glutamic acid decarboxylase (Anti-GAD) antibodies in Turkish patients with type 1 diabetes mellitus (DM), and to compare the frequency of anti-TPO and anti-TG titers in the presence or absence of anti-GAD. A total of 104 patients including 56 males and 48 females with type 1 DM and their age-, gender-, and body mass index-matched control group, including 31 males and 27 females, 58 cases in total with an age range of 15-50 years, were recruited into this study. In patients with type 1 DM, positive anti-GAD was detected in 30.8% (n=32). In patients with positive anti-GAD, rate of positive anti-TPO was 37.5%; however, in patients with negative anti-GAD, the rate of positive anti-TPO was 9.7% and the difference was statistically significant (p=0.001). In patients with positive anti-GAD, the rate of positive anti-TG was 18.8%. In patients with negative anti-GAD, the rate of positive anti-TG was 2.8%, and the difference between them was statistically significant (p=0.005). In patients with positive and negative anti-GAD, rates of both positive anti-TPO and anti-TG were 15.6% and 1.4%, respectively, with the difference showing statistical significance (p=0.004). Thyroid autoimmunity in type 1 DM patients with positive anti-GAD was apparently higher; therefore, these patients should be followed more frequently and carefully.

scholarly journals Immunological aspects of bacterial keratites in patients with diabetes mellitus

2021 ◽  
Vol 9 (2) ◽  
pp. 6-9
Author(s):  
O.V. Zavoloka ◽  
P.A. Bezditko ◽  
L.P. Abramova ◽  
V.O. Vekshyn
Keyword(s):  
Diabetes Mellitus ◽  
Anterior Segment ◽  
Enzyme Linked Immunosorbent Assay ◽  
Bacterial Keratitis ◽  
Healthy Individuals ◽  
Lacrimal Fluid ◽  
Cytokine Balance ◽  
Patients With Diabetes ◽  
Contralateral Eye ◽  
Type 1 Dm

Background. The purpose was to analyze the cytokine balance of lacrimal fluid in patients with bacterial keratitis and diabetes mellitus (DM) at the first visit and to identify the immunological aspects of the disease. Materials and methods. The analysis of pro- and anti-inflammatory cytokine concentration in the lacrimal fluid was performed in 17 patients with type 1 DM and bacterial keratitis and 15 nondiabetic patients with bacterial keratitis at the first visit. Data from 14 healthy individuals were used for comparison. In addition to standard ones, ophthalmic examination methods included bacteriological examination, fluorescein test, anterior segment optical coherence tomography, non-contact corneal aesthesiometry. The levels of interleukin (IL) 1β, IL-6 and IL-10 in the lacrimal fluid of the sick and the contralateral eye were determined by a quantitative colorimetric enzyme-linked immunosorbent assay using ELISA kits. Results. In DM patients with bacterial keratitis, the concentration of IL-1β and IL-6 in the lacrimal fluid of the sick eye exceeded that in healthy individuals (p < 0.05) and did not differ significantly from nondiabetic patients with bacterial keratitis (p > 0.05). In the lacrimal fluid of the contralateral eye of DM patients with bacterial keratitis, the level of IL-1β and IL-6 exceeded the corresponding indicators of nondiabetic patients with bacterial keratitis and healthy individuals (p < 0.05). The concentration of IL-10 in the lacrimal fluid of the contralateral eye in DM patients with bacterial keratitis exceeded that in healthy individuals (p < 0.05) and did not significantly differ from those in nondiabetic patients with bacterial keratitis (p > 0.05). Conclusions. DM patients with bacterial keratitis have immunological features of the disease.

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