Figure 5. Functional comparison for rCRT monomers and oligomers.

Samples of rCRT/18-412, OrCRT, MrCRT-60, MrCRT-46, nCRT and rEGFP were titrated against freshly prepared mouse peritoneal macrophages in 96-well plates. After 24 h incubation, concentrations of TNF-α(A) and NO₂ ⁻ (B) in the culture supernatant were determined using ELISA kit or Griess reagent, respectively. For immunogenicity test, groups of C57/BL6 mice (5 per group) were s.c. injected with 100 µg of rCRT/18-412, OrCRT, MrCRT-60 kDa, MrCRT-46 kDa or rEGFP (in 100 µl PBS/mouse) and boosted with 50 µg of the antigen preparations a fortnight later. The mice were bled 10 days thereafter and their sera were assayed, in triplicate wells, in ELISAs based on OrCRT (C), MrCRT-60 kDa (D), MrCRT-46 kDa (E) or rEGFP (F). The detection Ab was HRP-conjugated goat-anti-mouse IgG with OPD as substrate, and the results are expressed as mean OD492 nm±SD. These are representatives of 3 independent experiments.