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In vivo gene expression and immunoreactivity of Leptospira collagenase
Authors:Weena Janwitthayanan  Somboon Keelawat  Sunchai Payungporn  Alisa Lowanitchapat  Duangjai Suwancharoen  Yong Poovorawan  Chintana Chirathaworn
Institution:1. Master of Science Program in Medical Sciences, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;2. Department of Pathology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;3. Department of Biochemistry, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;4. National Institute of Animal Health, Department of Livestock Development, Bangkok, Thailand;5. Department of Pediatrics, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand;6. Department of Microbiology, Faculty of Medicine, Chulalongkorn University, Bangkok, Thailand
Abstract:Pulmonary hemorrhage is an increasing cause of death of leptospirosis patients. Bacterial collagenase has been shown to be involved in lung hemorrhage induced by various infectious agents. According to Leptospira whole genome study, colA, a gene suggested to code for bacterial collagenase has been identified. We investigated colA gene expression in lung tissues of Leptospira infected hamsters. Golden Syrian Hamsters were injected intraperitoneally with Leptospira interrogans serovar Pyrogenes. The hamsters were sacrificed on days 3, 5 and 7 post-infection and lung tissues were collected for histological examination and RNA extraction. Lung pathologies including atelectasis and hemorrhage were observed. Expression of colA gene in lung tissues was demonstrated by both RT-PCR and real time PCR. In addition, ColA protein was cloned and the purified protein could react with sera from leptospirosis patients. Leptospira ColA protein may play a role in Leptospira survival or pathogenesis in vivo. Its reaction with leptospirosis sera suggests that this protein is immunogenic and could be another candidate for vaccine development.
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