Fig. 7

ASFV-induced lymphopenia and Treg increase in the coculture system. Density gradient-separated peripheral blood mononuclear cells (PBMCs) were co-cultured with primary pulmonary alveolar macrophages (PAMs) infected with/without 0.1 MOI ASFV (HB-2208 strain) in medium with porcine IL-2, IL-4 added for 7 days. a, Representative contour plot of CD3⁺ TCR γδ⁺ T cells in co-culture system at 3, 5, 7 d.p.i.. b, Analysis of cell population subsets in PMBC using five-colour flow cytometry (mean ± s.e.m., Two-way ANNOVA, **P < 0.01, ***P < 0.001, n = 3). c, Absolute count of CD3⁺ TCRγδ⁺ T and apoptosis cells in PBMCs per 10⁴cells (mean ± s.e.m., Student’s t-test, ***P < 0.001, n = 3) using FITC/Annexin V, PI apoptosis assay kit at 7 d.p.i.. d, Gating strategy of CD3⁺ CD4⁺ CD25⁺ Foxp3⁺ Treg in co-culture system. e, Analysis of Treg percentage in co-culture system at 3, 5, 7 d.p.i.. f, TGF-β mRNA expression in Control and ASFV groups at 7 d.p.i (mean ± s.e.m., Student’s t-test, *P < 0.05; **P < 0.01, ***P < 0.001, ****P < 0.0001, n = 3)