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

Result code in 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>

Alternative codes found

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

Result on the web

<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>

Result language

angličtina

Original language name

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

Original language description

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.

Czech name

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Czech description

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Type

J_imp - Article in a specialist periodical, which is included in the Web of Science database

CEP classification

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OECD FORD branch

40301 - Veterinary science

Project

Result was created during the realization of more than one project. More information in the Projects tab.

Continuities

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

Publication year

2019

Confidentiality

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

Name of the periodical

Veterinární Medicína

ISSN

0375-8427

e-ISSN

1805-9392

Volume of the periodical

64

Issue of the periodical within the volume

12

Country of publishing house

CZ - CZECH REPUBLIC

Number of pages

11

Pages from-to

547-557

UT code for WoS article

000504204700005

EID of the result in the Scopus database

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