Figure 2.

Leishmania major matrix-metalloprotease, glycoprotein-63, is necessary and sufficient to cleave CXCL10. (A) Zinc chelation prevents CXCL10 suppression. Concentration of human recombinant CXCL10 was measured by ELISA after incubation for 12 h with filtered conditioned media from L. major WT promastigote culture and addition of the zinc-chelator 1,10-phenanthroline (n = 8 from 4 experiments). (B) gp63 is required for L. major CXCL10 suppression. Human recombinant CXCL10 concentrations were measured by ELISA after 12 h incubation with conditioned media from L. major WT, Δgp63, or Δgp63+1 (n = 6 from 3 experiments). (C) GP63 expressed and secreted by HEK293Ts is sufficient for CXCL10 suppression. Human recombinant CXCL10 concentrations were measured by ELISA after 12 h incubation with culture supernatant from HEK293Ts transfected with pCDNA3.1-gp63^WT or pCDNA3.1-gp63^E285A (n = 6 from 3 experiments). Concentration is represented as fold change relative to supernatants from YFP transfection control. P-values calculated by one-way ANOVA with Tukey's post-hoc test. Error bars represent standard error of the mean.