Chitin Oligomer Innate Immune Response

Custom-sized purified chitin particles were obtained from Elicityl, ranging from chitinbiose to chitinhexaose [10 mg]. Chitin oligomer (C10-15) was generated as described previously [12 (link)]. Purified chitin particle suspensions were delivered by involuntary aspiration to isoflurane-anesthetized mice. To assess size dependent innate immune responses to purified chitin particles, mice were sacrificed 6 hours post challenge. Suspensions of 5 x 10⁷ conidia were delivered by a single involuntary aspiration. Mice were sacrificed 24 hours post-challenge with sodium pentobarbitol as described previously [17 (link)], and lungs were perfused with 10ml phosphate buffered saline (PBS) for quantitative RT-PCR (qRT-PCR) assay. Bronchoalveolar lavage was performed to collect samples to assay immune cell composition by flow cytometry. All animal procedures were approved by the Animal Care and Use Committee of Indiana State University, the host campus of IUSM-Terre Haute. All animal handling and experimental procedures were performed in accordance with the recommendations found in the Guide for the Care and Use of Laboratory Animals of the National Institutes of Health. This study was carried out in accordance with the recommendations of the PHS Policy on Humane Care and Use of Laboratory Animals. The protocol was approved by the Indiana State University Animal Care and Use Committee, the host campus of IUSM-Terre Haute. IUSM- Biosafety committee approved of the use of human cell line (approval number: SM-847-06). Indiana State University IACUC committee approved of the use of murine studies (approval number: 1507790).