Figure 7. Pathogenic versus non-pathogenic recognition of TLR4 mediated proinflammatory cytokine CXCL2 production by Acanthamoeba in Chinese hamster.

Chinese hamsters (n  =  6/group) were infected with either pathogenic isolate of Acanthamoeba trophozoites (A. castellanii or A. culbertsoni) or non-pathogenic isolate of Acanthamoeba trophozoites (A. castellanii Neff or A. astronyxis)-laden contact lenses as described earlier [7]. Lenses were removed 1, 3, and 7 days post-infection. (7A) Corneas were scored for severity of disease for the period indicated. Each graph line represents the mean±SEM of severity of disease of 6 animals for the observed time points. The results shown are representative of three separate experiments. Asterisk indicates P value < 0.05 by the Mann-Whitney’s U test. (7B and 7C) Infected and uninfected-control corneas were dissected and homogenized. Total RNA was collected and RT-PCR was performed to examine the mRNA expression of TLR4 and CXCL2. Quantification was calculated by densitometry of bands relative to GAPDH. Densitometry of mRNA bands were quantified by three independent scanned presented as mean±SEM. (7D) CXCL2 protein production from dissected and homogenized infected and uninfected-control corneal lysates was quantified using ELISA. The data are mean±SEM of three independent experiments. Asterisk indicates P value < 0.05 by unpaired Student’s t-test.