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. 2021 Jul 30;4:929. doi: 10.1038/s42003-021-02446-x

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© This is a U.S. Government work and not under copyright protection in the US; foreign copyright protection may apply 2021

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Fig. 4 — a Schematic representation of the experimental plan. b Kaplan–Meier survival curves of LmCen^−/⁻-immunized hamsters (Imm Chal; blue lines, n = 8) following challenge with L. donovani-infected sand flies and compared with age-matched nonimmunized-challenged group (Non Imm Chal; red lines, n = 8). c Picture of hamsters from each group after 9 months of sand fly-transmitted L. donovani challenge. d Photographs (left panel) of representative two spleen samples of both LmCen^−/⁻-immunized and nonimmunized hamsters following 9 months post challenge as well as one age-matched naive hamster is shown. Right panel showing stamp smear of respective spleens stained with H&E, black arrows indicates intracellular parasites (bar-20 μm). e, f Parasite load in the spleen (e) and liver (f) (n = 8, n = 10, and n = 5 for 3.5M, 9M, and 12M post challenge, respectively, of nonimmunized and immunized-challenged groups). Results represent (the geometric means with 95% Cl) cumulative effect of two (3.5MPC and 9MPC) and one (12MPC) independent experiment respectively (p values were determined by Mann–Whitney two-tailed test). g Liver sections are from animals at 3.5 months after challenge and stained with H&E (left panel ×20 and right panel ×100 magnification). h–j IFN-γ (h) and IL-10 (i) expression in the spleen of LmCen^−/⁻ immunized and age-matched nonimmunized hamsters (n = 8/per group) was evaluated by qPCR following 3.5 months post-L. donovani-infected sand fly challenge. The ratio of IFN-γ/IL-10 expression in the spleen was also determined (j). Results (the geometric means with 95% Cl) represent the cumulative effect of two independent experiments (p values were determined by Mann–Whitney two-tailed test). WPI weeks postimmunization, MPC months post challenge.