Uc7 13d5

5 × 10⁴ sorted CXCR5⁻ γδ T cells from a pool of spleen and lymph nodes of 15 naïve WT mice were cultured for 48 h at 37 °C in the presence of medium only (unstimulated); anti-TCRγδ monoclonal antibody (UC7-13D5; Biolegend; 2 μg ml⁻¹); anti-TCRγδ (2 μg ml⁻¹) + anti-CD28 (37.52; BD Biosciences; 5 μg ml⁻¹) monoclonal antibodies; TLR1/2 agonist (Pam3CSK4 (Invivogen), 500 ng ml⁻¹); anti-TCRγδ (2 μg ml⁻¹) + TLR1/2 agonist (500 ng ml⁻¹); M. tuberculosis (Difco, 10 μg ml⁻¹); anti-TCRγδ (2 μg ml⁻¹) + MT (10 μg ml⁻¹); TLR1/2 antagonist (CU-CPT22 (Millipore Sigma), 0.5 μM); TLR1/2 agonist (500 ng ml⁻¹) + TLR1/2 antagonist (TLR1/2 ant, 0.5 μM) or MT (10 μg ml⁻¹) + TLR1/2 antagonist (0.5 μM). CXCR5 expression on γδ T cells was analyzed by flow cytometry.
For in vivo study of CXCR5 expression on γδ T cells, 500 μg per mouse of anti-TCRγδ monoclonal antibody (UC7-13D5; Biolegend) was administered i.p. and 24 h later CXCR5 expression on γδ T cells was analyzed by flow cytometry.