Table 1.
Genes and primer sequences used for real-time PCR.
Table 2.
Physical properties of PLGA NPs encapsulating different classes of TLR ligands.
Fig 1.
Nitric oxide (NO) production from chicken macrophages (six replicates/group) stimulated with encapsulated and soluble forms of three TLR ligands, measured by Griess reagent system.
(A). LPS; (B). Pam3CSK4, and (C). CpG ODN. Significance (P <0.05) between delivery systems (encapsulated and soluble TLR ligands) within a treatment dose was determined. P-LPSHi—encapsulated high dose LPS; P-LPSlo—encapsulated low dose LPS; LPSHi—soluble high dose LPS; LPSlo—soluble low dose LPS. P-PamHi—encapsulated high dose Pam3CSK4; P-Pamlo—encapsulated low dose Pam3CSK4; PamHi—soluble high dose Pam3CSK4; Pamlo—soluble low dose Pam3CSK4. Similar designations were made for CpG ODN. * indicates significant difference.
Fig 2.
Expression profiles of IFN-γ for (A). LPS; (B). Pam3CSK4, and (C). CpG ODN.
Chicken macrophages (six replicates/group) were treated with encapsulated and soluble forms of the three TLR ligands and mean fold expression of IFN-γ in treated groups were compared to the cell culture medium treated group. Significance (P <0.05) was tested between delivery systems (encapsulated and soluble TLR ligands) within a dose and at a given time point. * indicates significant difference.
Fig 3.
Expression level of IFN-β by stimulation with CpG ODN.
Chicken macrophages (six replicates/group) were treated with encapsulated and soluble CpG ODN. Mean fold expression of IFN-β in treated groups were compared to the cell culture medium treated group. Significance (P <0.05) was tested between delivery systems within a dose and at a given time point. * indicates significant difference.
Fig 4.
IL-1β mRNA expression in chicken macrophages stimulated with (A). LPS; (B). Pam3CSK4; and (C) CpG ODN.
Chicken macrophages (six replicates/group) were treated with encapsulated and soluble forms of the three TLR ligands and mean fold expression of IL-1β in treated groups were compared to the cell culture medium treated group. Significance (P <0.05) was tested between delivery systems within a dose and at a given time point. * indicates significant difference.
Fig 5.
Quantification of IFN-γ (A, B), IL-1β (C, D) and IL-8 (E, F) expression in spleen of chickens intramuscularly injected with encapsulated and soluble LPS.
At 3, 18, 48 and 96 hr post-injection, spleens were collected and the relative expression of cytokine genes was determined. Bars represent mean fold expression of cytokines in treated chickens compared to the control chickens. Cytokine gene expressions were compared between delivery systems (encapsulated and soluble) within a dose and at a given time point. * indicates significant difference at P <0.05.