Líffræðifélag Íslands - biologia.is
Líffræðiráðstefnan 2023
Höfundar / Authors: Sara Björk Stefánsdóttir1, Vilhjálmur Svansson1, Siggeir Fannar Brynjólfsson2, Sigurbjörg Þorsteinsdóttir1, Sigríður Jónsdóttir1
Starfsvettvangur / Affiliations: 1. Institute for Experimental Pathology, Biomedical Center, University of Iceland, Keldur, Reykjavik, Iceland. ; 2. Department of Immunology, Landspitali, The National University Hospital of Iceland, Reykjavik, Iceland.
Kynnir / Presenter: Hafdís María Pétursdóttir
Introduction: Insect bite hypersensitivity (IBH) in horses is an IgE mediated allergy caused by bites of Culicoides spp. It does not affect horses in Iceland due to the absence of the causative Culicoides. Currently, there's no effective treatment except avoiding Culicoides bites. We are developing both preventive and curative treatment for IBH. It is crucial to measure the cytokines following the treatment for the evaluation of the T-cell response. The aim is to determine an effective method to evaluate the cytokine response following a vaccination.
Methods: Two vaccinated horses were boosted with a pool of allergens and their PBMC stimulated in vitro with the same allergens for cytokine (IFNg, IL-4, IL-10) measurements by qPCR and Luminex. For Luminex, 500.000 and 800.000 cells were stimulated for 48h, 72h and 96h. For qPCR 3x, 5x and 8x106 cells were stimulated for 12h, 18h and 24h.
Results: For Luminex; both 500.000 and 800.000 cells, and all time points, showed increase in IFNg and IL-10 when stimulated with the vaccine pool as compared to the control pool. The best results were obtained after stimulation of 800.000 cells for 96h. Primers, probes and the parameters of the qPCR were optimized. However, due to a great difference between the horses this approach could not be fully evaluated.
Conclusion: For detection of cytokine response following a stimulation with vaccine allergens Luminex showed a promising result, and this must be confirmed on more horses.