Ehrlichia secretes Etf-1 to induce autophagy and capture nutrients for its growth through RAB5 and class III phosphatidylinositol 3-kinase

Lin, Mingqun; Liu, Hongyan; Xiong, Qingming; Niu, Hua; Cheng, Zhihui; Yamamoto, Akitsugu; Rikihisa, Yasuko

The E. chaffeensis Arkansas strain 1 was cultured in THP-1 cells (ATCC, TIB-202), 122 and the A. phagocytophilum HZ strain 123 was cultured in HL-60 cells (ATCC, CCL-240) in RPMI 1640 medium (Mediatech, 10-040-CV) supplemented with 10% fetal bovine serum (Atlanta Biologicals, S12450) and 2 mM L-glutamine (Gibco, 25030). RF/6A cells (ATCC, CRL-1780) were cultured in advanced minimal essential medium (Gibco, 12492) supplemented with 10% fetal bovine serum and 2 mM L-glutamine. The method of synchronous culture of E. chaffeensis in HL-60 cells was similar to those described previously. 9, 43 Host cell-free E. chaffeensis was used to initiate infection as previously described. 9,43 HEK293 cells (ATCC, CRL-1573) and canine histiocytic leukemia DH82 cells were cultured in DMEM (Dulbecco's minimal essential medium; Mediatech, 10-013-CV) supplemented with 10% fetal bovine serum and 2 mM L-glutamine. 1 Cultures were incubated at 37 C under 5% CO 2 in a humidified atmosphere.