Interleukin 2 (il 2)

The IL-10, IL-6, tumor necrosis factor-α (TNF-α), IFN-α, IFN-β, transforming growth factor-β (TGF-β), and CXCL13 enzyme linked immunosorbent assay (ELISA) kits were purchased from MyBioSource (Vancouver, DC, USA), while the IL-2, IL-17, IL-18, IL-1β, and CXCL1 ELISA kits were purchased from Cloud Clone Corp (Wuhan, China). The level of cytokines in the serum was measured according to the manufacturer’s instructions.

The blood samples were examined using commercial ELISA kits according to the manufacturer’s instructions for the following parameters: a concentration of acute phase proteins (APPs) including serum amyloid A (SAA; Tridelta Development Ltd.), haptoglobin (Hp; Tridelta Development Ltd.), alpha-2-Macroglobulin (A2M; Cusabio) and albumin (Alb, BSA; Cloud-Clone Corp.) as well as cytokine activity such as IL-1, IL-2, IL-10 and TNF-α (Cloud-Clone Corp.).
The optical densities in the microwells for APP and cytokine concentrations as well as for the antibodies to M. bovis and M. bovis antigen detection were read by an automated plate reader (Elx800 Microplate Reader, BioTek Instruments, Inc., USA) using the KC Junior programme (BioTek Instruments, Inc., USA).

The ELISAs used to detect the porcine IFNγ, IL-2, IL-12B (Cloud-Clone Corporation, Houston, TX, USA), IL-7, and IL-15 (Wuhan Fine Biotech Co., Wuhan, China) in the serum were performed according to the manufacturer’s instructions.