Differential host response to LPS variants in amniochorion and the TLR4/MD-2 system in Macaca nemestrina

Structures of LPS variants. PgLPSweak and PgLPSantag were isolated from P. gingivalis grown under low and high haemin conditions, respectively. The structures have been elucidated using MALDI-TOF mass spectrometry. PgLPSweak and PgLPSantag are a mixture of predominantly two LPS variants as shown.

Luminex and ELISA analysis of cytokines and prostaglandins in amniochorion explant supernatant 24-h after LPS stimulation. Values are means ± SEMs of N = 6 independent experiments. Values were log-transformed and analyzed using ANOVA with a Bonferroni correction applied to pairwise comparisons.

Quantitative RT-PCR analysis of gene expression in amniochorion explants 24-h after LPS stimulation. Values are means ± SEMs of N = 6 independent experiments. Analysis by ANOVA with a Bonferroni correction yielded no significant differences.

Recombinant TLR4 signaling system to compare human and M. nemestrina TLR4 activation in HEK293 cells. Data are expressed as the fold-increase in relative light units (which represents the ratio of NF-κB-luciferase to β-actin Renilla-luciferase expression) relative to that of a no-stimulation control. There were no significant differences between human and M. nemestrina activation of the reporter assay by Wilcoxon rank sum with Bonferroni correction.

Fig. S1. TLR4 cDNA sequence for M. nemestrina aligned with sequence for human and M. mulatta.

Fig. S2. TLR4 protein sequence for M. nemestrina aligned with sequence for human and M. mulatta.