Lysozyme M deficiency leads to an increased susceptibility to Streptococcus pneumoniae-induced otitis media.
BACKGROUND
Lysozyme is an antimicrobial innate immune molecule degrading peptidoglycan of the bacterial cell wall. Lysozyme shows the ubiquitous expression in wide varieties of species and tissues including the tubotympanum of mammals. We aim to investigate the effects of lysozyme depletion on pneumococcal clearance from the middle ear cavity.


METHODS
Immunohistochemistry was performed to localize lysozyme in the Eustachian tube. Lysozyme expression was compared between the wild type and the lysozyme M-/- mice using real time quantitative RT-PCR and western blotting. Muramidase activity and bactericidal activity of lysozyme was measured using a lysoplate radial diffusion assay and a liquid broth assay, respectively. To determine if depletion of lysozyme M increases a susceptibility to pneumococal otitis media, 50 CFU of S. pneumoniae 6B were transtympanically inoculated to the middle ear and viable bacteria were counted at day 3 and 7 with clinical grading of middle ear inflammation.


RESULTS
Immunolabeling revealed that localization of lysozyme M and lysozyme P is specific to some/particular cell types of the Eustachian tube. Lysozyme P of lysozyme M-/- mice was mainly expressed in the submucosal gland but not in the tubal epithelium. Although lysozyme M-/- mice showed compensatory up-regulation of lysozyme P, lysozyme M depletion resulted in a decrease in both muramidase and antimicrobial activities. Deficiency in lysozyme M led to an increased susceptibility to middle ear infection with S. pneumoniae 6B and resulted in severe middle ear inflammation, compared to wild type mice.


CONCLUSIONS
The results suggest that lysozyme M plays an important role in protecting the middle ear from invading pathogens, particularly in the early phase. We suggest a possibility of the exogenous lysozyme as an adjuvant therapeutic agent for otitis media, but further studies are necessary.
Gonda Department of Cell and Molecular Biology, House Ear Institute, Los Angeles, CA, USA. Jasmd513@aol.com
Jun
J
Shimada
Sung K
SK
Moon
Haa-Yung
HY
Lee
Tamotsu
T
Takeshita
Huiqi
H
Pan
Jeong-Im
JI
Woo
Robert
R
Gellibolian
Noboru
N
Yamanaka
David J
DJ
Lim
D000818:Animals; D004198:Disease Susceptibility / Q000382:microbiology; D005064:Eustachian Tube / Q000382:microbiology; D020869:Gene Expression Profiling; D005786:Gene Expression Regulation; D051379:Mice; D008810:Mice, Inbred C57BL; D008826:Microbial Sensitivity Tests; D009113:Muramidase / Q000172:deficiency* / Q000235:genetics / Q000494:pharmacology; D010033:Otitis Media / Q000235:genetics* / Q000382:microbiology; D011008:Pneumococcal Infections / Q000235:genetics* / Q000382:microbiology; D012313:RNA / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013296:Streptococcus pneumoniae / Q000187:drug effects
D012313:RNA; D009113:Muramidase; C472450:lysozyme M, mouse; C472773:lysozyme P, mouse