FIG. 2.
Transgenic parasites express murine MCP-1. (A) Total RNA was isolated from wild-type and transgenic MCP-1 parasites during promastigote development. Murine MCP-1/JE was amplified from the cDNA of transgenic L. major MCP-1 (Lm-Tg) and transgenic L. amazonensis MCP-1 (La-Tg) parasites but not from the cDNA of wild-type L. major (Lm-WT) or wild-type L. amazonensis (La-WT). Axenically grown L. amazonensis MCP-1 and wild-type L. amazonensis amastigotes were also analyzed. MCP-1/JE was amplified from transgenic amastigotes but not from the wild type. The gene gp63 was amplified from the cDNA as a loading control. (B) A total of 5 × 106 transgenic L. major, wild-type L. major, transgenic L. amazonensis, or wild-type L. amazonensis parasites was added to an ELISA plate coated with monoclonal goat anti-mouse MCP-1 antibody. After 24 h, the levels of MCP-1 production were determined. (C) MCP-1 production by transgenic L. major MCP-1 parasites grown in culture (Tg-C, □) was determined by ELISA. MCP-1 production by transgenic parasites isolated from two different mice infected 4 weeks prior (Tg-M1 and Tg-M2, ▨) or wild-type L. major parasites (WT-M3, ▪) is also shown. **, P < 0.01.