MOESM8 of Enhancing the toolbox to study IL-17A in cattle and sheep

Additional file 8. Neutralisation of recombinant bovine and ovine IL-17A activity on ovine cells by commercial mab. Ovine ST-6 cells were set up as described in “Bulk recombinant cytokine production and functional determination of recombinant bovine and ovine IL-17A section” but using 96 well flat bottom plates. The following treatments were pre-incubated at 37 °C for 2 h in a water bath: IMDM culture medium only (unstimulated cells), rovIL-17A (50 ng/mL), rovIL-17A (50 ng/mL) + MT504 monoclonal antibody (mab, 1 μg/mL), rbovIL-17A (50 ng/mL) and rbovIL-17A (50 ng/mL) + MT504 mab (1 μg/mL). The treatments were then added to the ovine cells for 24 h and harvested and assayed as previously described. The X-axis displays the neutralisation bioassay treatments and the Y-axis shows levels of CXCL8 in pg/mL. Data are the arithmetic mean of three technical replicate samples from one representative experiment of two. The percentage neutralisation values displayed on the graph have been calculated by firstly deducting the unstimulated (IMDM culture medium control) value from all other treatment values. The value for rbovIL-17A neutralisation with MT504 mab was calculated by: 100 minus [(rbovIL-17A/MT504 mab minus background value) divided by (rbovIL-17A minus background value) multiplied by 100]. Percentage neutralisation for rovIL-17A was calculated by substituting rbovIL-17A for rovIL-17A values into equation above.