title: Host response and bacterial virulence factor expression in Pseudomonas aeruginosa and Streptococcus pneumoniae corneal ulcers.
abstract: P. aeruginosa and S. pneumoniae are major bacterial causes of corneal ulcers in industrialized and in developing countries. The current study examined host innate immune responses at the site of infection, and also expression of bacterial virulence factors in clinical isolates from patients in south India. Corneal ulcer material was obtained from 49 patients with confirmed P. aeruginosa and 27 patients with S. pneumoniae, and gene expression of Toll Like Receptors (TLR), cytokines and inflammasome proteins was measured by quantitative PCR. Expression of P. aeruginosa type III secretion exotoxins and S. pneumoniae pneumolysin was detected by western blot analysis. We found that neutrophils comprised >90% cells in corneal ulcers, and that there was elevated expression of TLR2, TLR4, TLR5 and TLR9, the NLRP3 and NLRC4 inflammasomes and the ASC adaptor molecule. IL-1α IL-1β and IFN-γ expression was also elevated; however, there was no significant difference in expression of any of these genes between corneal ulcers from P. aeruginosa and S. pneumoniae infected patients. We also show that 41/49 (84%) of P. aeruginosa clinical isolates expressed ExoS and ExoT, whereas 5/49 (10%) of isolates expressed ExoS, ExoT and ExoU with only 2/49 isolates expressing ExoT and ExoU. In contrast, all 27 S. pneumoniae clinical isolates produced pneumolysin. Taken together, these findings demonstrate that ExoS/T expressing P. aeruginosa and pneumolysin expressing S. pneumoniae predominate in bacterial keratitis. While P. aeruginosa strains expressing both ExoU and ExoS are usually rare, these strains actually outnumbered strains expressing only ExoU in the current study. Further, as neutrophils are the predominant cell type in these corneal ulcers, they are the likely source of cytokines and of the increased TLR and inflammasome expression.
authors.affiliation: Dr. G. Venkatasamy Eye Research Institute, Aravind Eye Hospital, Madurai, Tamil Nadu, India.
authors.forename: Rajapandian SivaGanesa
authors.initials: RS
authors.lastname: Karthikeyan
authors.forename: Jeganathan Lakshmi
authors.initials: JL
authors.lastname: Priya
authors.forename: Sixto M
authors.initials: SM
authors.lastname: Leal
authors.forename: Jonida
authors.initials: J
authors.lastname: Toska
authors.forename: Arne
authors.initials: A
authors.lastname: Rietsch
authors.forename: Venkatesh
authors.initials: V
authors.lastname: Prajna
authors.forename: Eric
authors.initials: E
authors.lastname: Pearlman
authors.forename: Prajna
authors.initials: P
authors.lastname: Lalitha
mesh_terms: D000293:Adolescent; D000328:Adult; D000368:Aged; D058947:Bacterial Secretion Systems / Q000235:genetics; D002648:Child; D003320:Corneal Ulcer / Q000276:immunology* / Q000382:microbiology; D005260:Female; D015964:Gene Expression Regulation, Bacterial; D054884:Host-Pathogen Interactions; D006801:Humans; D007113:Immunity, Innate; D008297:Male; D008875:Middle Aged; D011550:Pseudomonas aeruginosa / Q000235:genetics* / Q000502:physiology*; D013296:Streptococcus pneumoniae / Q000235:genetics* / Q000502:physiology*; D037521:Virulence Factors / Q000235:genetics*; D055815:Young Adult
chemical_list: D058947:Bacterial Secretion Systems; D037521:Virulence Factors