2014
Chemiluminescent determination of reactive oxygen species using Pholasin luminophore in birds and insect phagocytes
VOJTEK, Libor, Pavel DOBEŠ, Lucie PROKOPOVÁ, Jitka VINKLEROVA, Michal VINKLER et. al.Základní údaje
Originální název
Chemiluminescent determination of reactive oxygen species using Pholasin luminophore in birds and insect phagocytes
Autoři
VOJTEK, Libor (203 Česká republika, domácí), Pavel DOBEŠ (203 Česká republika, domácí), Lucie PROKOPOVÁ (203 Česká republika, domácí), Jitka VINKLEROVA (203 Česká republika), Michal VINKLER (203 Česká republika) a Pavel HYRŠL (203 Česká republika, garant, domácí)
Vydání
18th International Symposium on Bioluminescence and Chemiluminescence, 2014
Další údaje
Jazyk
angličtina
Typ výsledku
Konferenční abstrakt
Obor
30102 Immunology
Stát vydavatele
Velká Británie a Severní Irsko
Utajení
není předmětem státního či obchodního tajemství
Impakt faktor
Impact factor: 1.518
Kód RIV
RIV/00216224:14310/14:00073254
Organizační jednotka
Přírodovědecká fakulta
ISSN
Klíčová slova česky
Chemiluminiscence; fagocyty; Pholasin; reaktivní kyslíkové metabolity
Klíčová slova anglicky
Chemiluminiscence; phagocytes; Pholasin; reactive oxygen species
Příznaky
Mezinárodní význam, Recenzováno
Změněno: 31. 3. 2015 14:31, doc. RNDr. Pavel Hyršl, Ph.D.
Anotace
V originále
Phagocytosis is one of the most important innate immunity mechanisms which prevents organism against pathogens overcoming the natural barriers. There are several methods widely used for evaluation of phagocytosis efficiency. One of them is to measure the level of reactive oxygen species (ROS) produced by phagocytes - respiratory burst of phagocytes. ROS production can be elicited by addition of activators such as lipopolysaccharide (LPS), zymosan, starch particles, phorbol-12-miristate-13-acetate (PMA) or N-formyl-methionyl-leucyl-phenylalanin (fMLP). Produced ROS cause oxidation of luminophore which subsequently emits the energy in form of light measured by luminometers. Luminol is the most widely used luminophore. On the other hand evaluation of respiratory burst using this method on birds or insect samples is not possible due to a low production of ROS and low sensitivity of luminol. In birds is production limited by absence of myeloperoxidase, enzyme responsible for the main production of ROS and hypochlorous acid. Purpose of this work was to find more sensitive luminophore and optimize chemiluminescent (CL) measurement of oxidative burst for birds' and insects' samples. As more sensitive (with app. thirty times higher luminescent signal than luminol) was found the luminophore Pholasin - photoprotein extracted from bioluminescent mollusc Pholas dactylus. For CL measurement of oxidative burst in birds we combined this luminophore with Salmonella enterica and Escherichia coli LPS; an ideal activators that give fast and stable enhancement of ROS production (Fig. 1). Data acquired by this assay can be subsequently compared to results of other experiments by evaluation of peak of the reaction (maximum intensity of respiratory burst in counts per second, CPS) or integral of the reaction. So far there are four studies of heterophil respiratory burst determination in birds' immunology with the use of Pholasin and the first study on great tits (Parus major).
Návaznosti
| CZ.1.07/2.3.00/30.0009, interní kód MU (Kód CEP: EE2.3.30.0009) |
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| QJ1210047, projekt VaV |
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