Pariko Yorozu1, Asuka Furukawa2, Keisuke Uchida3, Takumi Akashi4, Tomoya Kakegawa5, Tomohisa Ogawa6, Junko Minami7, Yoshimi Suzuki8, Nobuyasu Awano9, Haruhiko Furusawa10, Yasunari Miyazaki11, Naohiko Inase12, Yoshinobu Eishi13. 1. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: p.yorozu.pth1@tmd.ac.jp. 2. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: askapth1@tmd.ac.jp. 3. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: uchida.path@tmd.ac.jp. 4. Division of Surgical Pathology, Tokyo Medical and Dental University Hospital, Tokyo 113-8510, Japan. Electronic address: akashi.path@tmd.ac.jp. 5. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: kakepth1@tmd.ac.jp. 6. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: ogawpth1@tmd.ac.jp. 7. Department of Clinical Engineering, School of Health Sciences, Tokyo University of Technology, Tokyo 144-8650, Japan. Electronic address: jminami@stf.teu.ac.jp. 8. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: y.suzuki.pth1@tmd.ac.jp. 9. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan; Department of Respiratory Medicine, Japanese Red Cross Medical Center, Tokyo 150-8935, Japan. Electronic address: awanobu0606@hotmail.co.jp. 10. Department of Integrated Pulmonology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: hfurusawa.pulm@tmd.ac.jp. 11. Department of Integrated Pulmonology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: miyazaki.pilm@tmd.ac.jp. 12. Department of Integrated Pulmonology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan. Electronic address: ninase.pulm@tmd.ac.jp. 13. Department of Human Pathology, Tokyo Medical and Dental University Graduate School, Tokyo 113-8510, Japan; Division of Surgical Pathology, Tokyo Medical and Dental University Hospital, Tokyo 113-8510, Japan. Electronic address: eishi.path@tmd.ac.jp.
Abstract
BACKGROUND: Propionibacterium acnes is one of the most commonly implicated etiologic agents of sarcoidosis. We screened antigenic proteins from this indigenous bacterium that increase Th1 responses in sarcoidosis patients. METHODS: Antigenic bacterial proteins were screened by probing western blots of P. acnes whole cell lysates with blood plasma samples from 52 sarcoidosis patients and 34 healthy volunteers. Soluble protein antigens from the bands most frequently detected on blotting membranes were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Recombinant proteins were prepared from DNA sequences of the proteins identified by MALDI-TOF/MS and analyzed by immunologic assays. RESULTS: MALDI-TOF/MS analysis identified propionyl-CoA carboxylase subunit beta, arginine deiminase (ADI), catalase (KAT), and UDP-N-acetylglucosamine pyrophosphorylase (UAP). Successfully prepared recombinant proteins from ADI, KAT, and UAP provoked humoral and cellular immune responses in mice immunized with P. acnes when measured by enzyme-linked immunosorbent assay for serum antibodies and enzyme-linked immunospot assay for interferon (IFN)-γ-secreting cells (ELISPOT IFN-γ assay) with lymph node cells. Plasma IgG and IgA titers to KAT and UAP were significantly higher in sarcoidosis patients than in healthy volunteers. When Th1 immune responses to ADI, KAT, and UAP were measured by ELISPOT IFN-γ assay with peripheral blood mononuclear cells from 12 sarcoidosis patients, 13 other pneumonitis patients, and 11 healthy volunteers, only the KAT protein provoked a significantly higher response in sarcoidosis patients (p=0.0032). CONCLUSION: These results suggest that P. acnes KAT is an antigen that provokes allergic Th1 immune responses in sarcoidosis patients.
BACKGROUND:Propionibacterium acnes is one of the most commonly implicated etiologic agents of sarcoidosis. We screened antigenic proteins from this indigenous bacterium that increase Th1 responses in sarcoidosispatients. METHODS: Antigenic bacterial proteins were screened by probing western blots of P. acnes whole cell lysates with blood plasma samples from 52 sarcoidosispatients and 34 healthy volunteers. Soluble protein antigens from the bands most frequently detected on blotting membranes were analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Recombinant proteins were prepared from DNA sequences of the proteins identified by MALDI-TOF/MS and analyzed by immunologic assays. RESULTS: MALDI-TOF/MS analysis identified propionyl-CoA carboxylase subunit beta, arginine deiminase (ADI), catalase (KAT), and UDP-N-acetylglucosamine pyrophosphorylase (UAP). Successfully prepared recombinant proteins from ADI, KAT, and UAP provoked humoral and cellular immune responses in mice immunized with P. acnes when measured by enzyme-linked immunosorbent assay for serum antibodies and enzyme-linked immunospot assay for interferon (IFN)-γ-secreting cells (ELISPOT IFN-γ assay) with lymph node cells. Plasma IgG and IgA titers to KAT and UAP were significantly higher in sarcoidosispatients than in healthy volunteers. When Th1 immune responses to ADI, KAT, and UAP were measured by ELISPOT IFN-γ assay with peripheral blood mononuclear cells from 12 sarcoidosispatients, 13 other pneumonitispatients, and 11 healthy volunteers, only the KAT protein provoked a significantly higher response in sarcoidosispatients (p=0.0032). CONCLUSION: These results suggest that P. acnesKAT is an antigen that provokes allergic Th1 immune responses in sarcoidosispatients.
Authors: Kristian Stødkilde; Jakob Toudahl Nielsen; Steen Vang Petersen; Bernhard Paetzold; Holger Brüggemann; Frans A A Mulder; Christian Brix Folsted Andersen Journal: Front Cell Infect Microbiol Date: 2022-02-11 Impact factor: 5.293