scholarly journals 25-Hydroxyvitamin D potentializes extracellular cathelicidin release from human PBMC stimulated ex vivo with either bacterial (LPS) or viral (P: IC) mimetics

Author(s):  
Sahar Al Dekwer ◽  
Nicolas Goncalves-Mendes ◽  
Rea Bingula ◽  
Guillaume Martinroche ◽  
Kassandra Lanchais ◽  
...  
2015 ◽  
Vol 61 (4) ◽  
pp. 627-635 ◽  
Author(s):  
Kazuya Omi ◽  
Tsuyoshi Ando ◽  
Takuya Sakyu ◽  
Takashi Shirakawa ◽  
Yoshiaki Uchida ◽  
...  

Abstract BACKGROUND Small molecules classified as haptens are generally measured by competitive immunoassay, which is theoretically inferior to noncompetitive sandwich immunoassay in terms of sensitivity and specificity. We created a method for developing sandwich immunoassays to measure haptens on the basis of antimetatype antibodies. METHODS We generated antimetatype monoclonal antibodies against a hapten–antibody immunocomplex using an ex vivo antibody development system, the Autonomously Diversifying Library (ADLib) system. We selected 2 haptens, estradiol (E2) and 25-hydroxyvitamin D [25(OH)D], as analytes. Sandwich immunoassays for these 2 haptens were developed by use of a 96-well microtiter plate and a fully automated chemiluminescence analyzer, and the performances of these immunoassays were investigated. RESULTS The developed assays exhibited sensitivity high enough to detect target haptens in serum samples. The limit of detection of the ELISA for E2 was 3.13 pg/mL, and that of the fully automated chemiluminescent enzyme immunoassay (CLEIA) system was 2.1 ng/mL for 25(OH)D. The cross-reactivity with immunoreactive derivatives was effectively improved compared with the competitive assay. The CVs for the sandwich ELISA for E2 were 4.2%–12.6% (intraassay) and 6.2%–21.8% (total imprecision). The CVs for the sandwich CLEIA for 25(OH)D were 1.0%–2.3% (intraassay) and 1.9%–3.5% (total imprecision). In particular, the sandwich CLEIA for 25(OH)D showed correlations of r = 0.99 with both LC-MS/MS and a commercially available 125I RIA. CONCLUSIONS Our method represents a potentially simple and practical approach for routine assays of haptens, including vitamins, hormones, drugs, and toxins.


2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Yi-Ching Tung ◽  
Tsan-Teng Ou ◽  
Wen-Chan Tsai

A uremic patient developed hypercalcemia after tuberculosis infection, and his ionized calcium levels correlated with 1,25-dihydroxyvitamin D3(1,25(OH)2D3) levels. We performed further studies to determine whether monocytes are alternative sites of 1,25(OH)2D3conversion beyond renal tubular cells. Using an ex vivo bioassay, in this study, we found that 1-αhydroxylase (CYP27B1) activity in monocytes is significantly higher in patients with active tuberculosis (TB) than in those with frequent TB contact. However, when monocytes from patients with active TB were restimulated with antigen derived fromMycobacterium tuberculosis, less 1,25(OH)2D3was observed. In contrast, the level of 1,25(OH)2D3was unchanged in those with frequent TB contact. We conclude that monocytes may be an alternative source of 1-αhydroxylase that could convert 25-hydroxyvitamin D3to the more active 1,25(OH)2D3.


2016 ◽  
Vol 22 ◽  
pp. 6
Author(s):  
Leena Kinnunen ◽  
Valma Harjutsalo ◽  
Heljä-Marja Surcel ◽  
Christel Lamberg-Allardt ◽  
Jaakko Tuomilehto ◽  
...  

2015 ◽  
Vol 63 (S 03) ◽  
Author(s):  
F.E.A. Udink ten Cate ◽  
F. Holler ◽  
T. Hannes ◽  
I. Germund ◽  
M. Emmel ◽  
...  

2015 ◽  
Author(s):  
Anna Challa ◽  
Eleni Evagelidou ◽  
Ekaterini Siomou ◽  
Alexandros Tzallas ◽  
Vasileios Giapros

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