Fig. 4: Analysis of cytokine (IFNγ, IL6, IL1β, IL4, IL18, TNFα, and IL12) and chemokine (CXCLi2) mRNA levels in splenocytes upon stimulation with CD83 scFv, rH9HA, and rH9HA-CD83 scFv. | npj Vaccines

Fig. 4: Analysis of cytokine (IFNγ, IL6, IL1β, IL4, IL18, TNFα, and IL12) and chemokine (CXCLi2) mRNA levels in splenocytes upon stimulation with CD83 scFv, rH9HA, and rH9HA-CD83 scFv.

From: Selectively targeting haemagglutinin antigen to chicken CD83 receptor induces faster and stronger immunity against avian influenza

Fig. 4

a Cytokines and chemokine mRNA levels in splenocytes analysed using qRT-PCR. b IFNγ protein level analysed using ELISA. For a: splenocytes were isolated from the spleen of 3-week-old SPF chickens using Histopaque 1083 and stimulated with 10 μg of CD83 scFv/rH9HA/rH9HA-CD83 scFv for 5, 22, and 30 h in vitro. Stimulated splenocytes were harvested for RNA extraction and expression levels of the respective cytokines and chemokine were measured by qRT-PCR. Data were calculated using 2−∆∆CT approach (n-fold change compared to the media only control group) and reported as values normalised to the expression level of a housekeeping gene RPLPO1. For b: supernatants from the stimulated splenocytes were analysed for the presence of IFNγ by ELISA. Data are represented as mean ± SD and analysed by one-way ANOVA followed by Tukey’s multiple comparison test. ***p < 0.001 **p < 0.01 *p < 0.05. The data represent three independent experiments.

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