Optimisation of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

Kód výsledku v IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00027162%3A_____%2F19%3AN0000179" target="_blank" >RIV/00027162:_____/19:N0000179 - isvavai.cz</a>

Nalezeny alternativní kódy

RIV/62156489:43210/19:43916913 RIV/62157124:16270/19:43877473

Výsledek na webu

<a href="https://www.agriculturejournals.cz/web/vetmed.htm?type=article&id=98_2019-VETMED" target="_blank" >https://www.agriculturejournals.cz/web/vetmed.htm?type=article&id=98_2019-VETMED</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.17221/98/2019-VETMED" target="_blank" >10.17221/98/2019-VETMED</a>

Jazyk výsledku

angličtina

Název v původním jazyce

Optimisation of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

Popis výsledku v původním jazyce

The lymphocyte proliferation assay is a valuable method used for the evaluation of the fish immune system. However, there are many variations and optimal results are not always obtained. Unification is necessary to ensure the comparability between different studies. The aim of this study was to optimise the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss). This goal included the determination of the optimal incubation length, serum type, incubation temperature, type of mitogen and its concentration, and anticoagulant. The peripheral blood and head kidney lymphocytes were isolated by density gradient centrifugation. Subsequently, the cells were incubated for 3–8 days with different mitogens (pokeweed mitogen 5, 10 and 50 μg/ml, concanavalin A 1, 10 and 20 μg/ml, phytohaemagglutinin 25, 50 and 100 μg/ml, lipopolysaccharide 1, 50 and 100 μg/ml). The use of the different serum types (foetal bovine serum, trout serum), incubation temperatures (10–20 °C) and anticoagulants (heparin, EDTA) was compared. Labelled thymidine was used to evaluate the assay. The best results were obtained after seven days of incubation at 15 °C with foetal bovine serum (FBS). The head kidney lymphocytes showed the highest proliferative response with 50 μg/ml phytohaemagglutinin. With the peripheral blood lymphocytes (heparin and EDTA), the best results were obtained with 50 μg/ml pokeweed mitogen. The highest proliferation levels were detected with heparinised blood. In conclusion, optimisation of this assay contributes to the improved assessment of the rainbow trout immune function.

Název v anglickém jazyce

Optimisation of the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss)

Popis výsledku anglicky

The lymphocyte proliferation assay is a valuable method used for the evaluation of the fish immune system. However, there are many variations and optimal results are not always obtained. Unification is necessary to ensure the comparability between different studies. The aim of this study was to optimise the lymphocyte proliferation assay in rainbow trout (Oncorhynchus mykiss). This goal included the determination of the optimal incubation length, serum type, incubation temperature, type of mitogen and its concentration, and anticoagulant. The peripheral blood and head kidney lymphocytes were isolated by density gradient centrifugation. Subsequently, the cells were incubated for 3–8 days with different mitogens (pokeweed mitogen 5, 10 and 50 μg/ml, concanavalin A 1, 10 and 20 μg/ml, phytohaemagglutinin 25, 50 and 100 μg/ml, lipopolysaccharide 1, 50 and 100 μg/ml). The use of the different serum types (foetal bovine serum, trout serum), incubation temperatures (10–20 °C) and anticoagulants (heparin, EDTA) was compared. Labelled thymidine was used to evaluate the assay. The best results were obtained after seven days of incubation at 15 °C with foetal bovine serum (FBS). The head kidney lymphocytes showed the highest proliferative response with 50 μg/ml phytohaemagglutinin. With the peripheral blood lymphocytes (heparin and EDTA), the best results were obtained with 50 μg/ml pokeweed mitogen. The highest proliferation levels were detected with heparinised blood. In conclusion, optimisation of this assay contributes to the improved assessment of the rainbow trout immune function.

Druh

J_imp - Článek v periodiku v databázi Web of Science

CEP obor

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OECD FORD obor

40301 - Veterinary science

Projekt

Výsledek vznikl pri realizaci vícero projektů. Více informací v záložce Projekty.

Návaznosti

P - Projekt vyzkumu a vyvoje financovany z verejnych zdroju (s odkazem do CEP)

Rok uplatnění

2019

Kód důvěrnosti údajů

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Název periodika

Veterinární Medicína

ISSN

0375-8427

e-ISSN

1805-9392

Svazek periodika

64

Číslo periodika v rámci svazku

12

Stát vydavatele periodika

CZ - Česká republika

Počet stran výsledku

11

Strana od-do

547-557

Kód UT WoS článku

000504204700005

EID výsledku v databázi Scopus

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