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Role of glycosaminoglycans in infectious disease - PubMed

Role of glycosaminoglycans in infectious disease

Akiko Jinno et al. Methods Mol Biol. 2015.

Abstract

Glycosaminoglycans (GAGs) have been shown to bind to a wide variety of microbial pathogens, including viruses, bacteria, parasites, and fungi in vitro. GAGs are thought to promote pathogenesis by facilitating pathogen attachment, invasion, or evasion of host defense mechanisms. However, the role of GAGs in infectious disease has not been extensively studied in vivo and therefore their pathophysiological significance and functions are largely unknown. Here we describe methods to directly investigate the role of GAGs in infections in vivo using mouse models of bacterial lung and corneal infection. The overall experimental strategy is to establish the importance and specificity of GAGs, define the essential structural features of GAGs, and identify a biological activity of GAGs that promotes pathogenesis.

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Figures

Fig. 1
Fig. 1

Syndecan-1 promotes S. aureus corneal infection in an HS-dependent manner. (a) Corneas of anesthetized Wt and Sdc1−/− mice on the BALB/c background were scratched with a 29 G needle and infected topically with 1 × 109 cfu of S. aureus strain 8325-4. The corneal bacterial burden was quantified at 10 h postinfection. Data shown are mean ± S.E. (n=9 in Wt and n=6 in Sdc1−/− group). (b) Scarified Wt and Sdc1−/− corneas were infected with 1 × 109 cfu of 8325-4 with or without 200 ng of HS or heparin (HP), or 500 ng of CS-A (CS) or heparosan (H), and the corneal bacterial burden was quantified at 10 h postinfection. Data shown are mean ± S.E. (n=11 in PBS, n=10 in HS, n=7 in HP, n=4 in CS, and n=5 in H group). (c) Paraffin-embedded eye sections of infected Wt and Sdc1−/− mice were Gram stained (arrowhead indicates injured areas). Note the increased number of Gram-positive cocci in Wt cornea infected with S. aureus only and Sdc1−/− cornea co-infected with S. aureus and HS compared to Sdc1−/− cornea infected with S. aureus only

Fig. 2
Fig. 2

HS specifically inhibits the killing of P. aeruginosa by LL-37. (a) P. aeruginosa (103 cfu) was incubated with LL-37 (3 μg/ml) in 30 μl PBS for 2 h at 37 °C in the absence or presence of increasing doses of CS-A, CS-C, DS, or HS. Bacterial killing was determined by plating out serial dilutions. Data shown are mean ± S.E. (n=4 in each group). Note the significantly increased inhibitory activity of HS at doses ≥15 μg/ml compared to other GAGs. (b) P. aeruginosa was incubated with LL-37 (3 μg/ml) (LL-37 group), preincubated with HS (20 μg/ml) for 30 min, washed free of HS, and then incubated with LL-37 (Pre-HS group), or co-incubated with LL-37 and HS (Co-HS group) for 2 h at 37 °C in a microfuge tube. Bacterial killing was determined by plating out serial dilutions. Data shown are mean ± S.E. (n=4 in each group). This experiment shows that HS does not inhibit LL-37 activity by binding to the bacteria, but rather by directly binding to LL-37 and inhibiting its antibacterial activity

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