Quantification of Immune Mediator Levels in Human Mucosal Lining Fluid

Published: January 31, 2024

Abstract

Source: Wolsk, H. M. et al., Noninvasive Sampling of Mucosal Lining Fluid for the Quantification of In Vivo Upper Airway Immune-mediator Levels. J. Vis. Exp. (2017)

This video demonstrates the quantification of immune mediator levels in neonatal human nasal mucosal lining fluid extract. An electrochemiluminescence-based multiplex assay system measures these mediators by evaluating the intensity of emitted light when they are excited.

Protocol

All procedures involving sample collection have been performed in accordance with the institute's IRB guidelines. 1. Experimental Setup Use sheets of filter paper (fibrous hydroxylated-polyester sheets, see the Table of Materials). Cut out strips 3 x 15 mm in size for a 1-month-old child and in an L-shape for older children and adults (5 x 20 x 10 mm (short arm of the L)). Insert one piece…

Divulgations

The authors have nothing to disclose.

Materials

Fibrous hydroxylatedpolyester sheets Accuwik Ultra  SPR0730 Filter paper
Milliplex Assay Buffer  Millipore L-AB Buffer
low-protein binding storage plates  Thermo Scientific CLS8161 Plates
Protease Inhibitor Roche 11873580001 Complete EDTA-free Protease Inhibitor Cocktail
Reader of multi-spot plates Mesoscale NA Sector imager 6000
Assays  Mesoscale Human 10-plex TH1/TH2 cytokine assay and 9-plex chemokine assay, and singleplex IL-17A, TGF-β1and TSLP. A description can be found online on www.mesoscale.com

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Quantification of Immune Mediator Levels in Human Mucosal Lining Fluid. J. Vis. Exp. (Pending Publication), e21898, doi: (2024).

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