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HKL centrifugation on gradient yielded an interface, between 34% and 51% of Percoll, enriched in viable macrophages which were stabilized in culture.
HKL cells stabilized in culture for 5 days, hereinafter macrophages, were induced to generate iNOS in response to 1 mg [mL.sup.-1] of Zymosan A.
From head kidney was obtained a phase of leukocytes (HKL) enriched in macrophages, which were grown successfully.
Our results confirm the ability of HKL to respond to the challenge with Zymosan A, in all the tested assays, both at enzyme level as in ELISAs.
This parameter has been previously established and the test conditions of rainbow trout HKL, determined (Chettri et al., 2010).
The main objective of this work focused on establishing an analytical model of HKL response to Zymosan A, as a reference to test immunostimulants in salmonids, considering a high-capacity screening system.
Our results confirm that increased transcriptional activity, previously demonstrated, that occurs in trout HKL in response to Zymosan A (Chettri et al., 2011), also causes increased secretion of pro-inflammatory cytokines TNF-[alpha], IL-1[beta] and IL-6.
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