Quantitative single-cell analysis of Leishmania major amastigote differentiation demonstrates variably extended expression of the lipophosphoglycan (LPG) virulence factor in different host cell types
Fig 6
LPG-retaining parasites within DCs 72 hours post-infection have not undergone DNA synthesis.
(A, B) Parasites were cultured in the presence of thymidine analogue (EdU) for 72 hours following infection of BMDC. Cells were stained to detect EdU-incorporation (green), LPG (red) and parasite nuclei (blue). (A) Representative image of parasites within DCs stained as above. Arrows, parasites showing LPG-positivity. Scale bar, 5 μm. (B) Percent of parasites within DCs 72 hours post-infection that are positive for EdU-incorporation. Data shown include the percent of total parasites that are EdU+, as well as the EdU-positivity of parasites that are either LPG-negative or LPG-positive. Data, means ± S.E., n = 3 experiments, ****, P < 0.0001 (Chi-square; N = 1185 parasites). (C) Comparison of L. major amastigogenesis marker transitions, LPG loss, and cell cycle re-entry following infection of PEM, BMM, and BMDC. Overall, the acquisition of early amastigote characteristics is the same in the three host cell types, but LPG loss and DNA replication are reduced or substantially delayed following infection of BMM and BMDC.