Title of Research:
21-2-01 Development of evaluation method of inflammatory particles based on alveolar
macrophage function.
Principal Investigator:
Etsushi Kuroda, PhD (Professor and Chairman, Hyogo Medical University, School of Medicine,
Department of Immunology)
1-1 Mukogawa-cho, Nishinomiya, Hyogo 663-8501, Japan
tel: +81-798-45-6574, e-mail: kuroetu@hyo-med.ac.jp
Collaborators:
Koubun Yasuda (Hyogo Medical University)
Kazufumi Matsushita (Hyogo Medical University)
Masakiyo Nakahira (Hyogo Medical University)
Takumi Adachi (Hyogo Medical University)
Hinata Inoue (Hyogo Medical University)Yasuo Morimoto (University of Occupational and Environmental Health)
Hiroto Izumi (University of Occupational and Environmental Health)
Yoshitaka Shirasaki (The University of Tokyo, Faculty of Pharmaceutical Siences)
Summary of Research:
The number of patients with allergic diseases have increased in developed countries, and it issuggested that particulate pollution such as PM2.5 might be one of factors of exacerbation ofallergic inflammation. These particulates function as adjuvant and induce allergic immune
responses. In general, inhaled particles are thought to be engulfed by alveolar macrophages andthen excreted. Therefore, in this study we aim to develop methods of evaluation of inflammatory
particles focusing on alveolar macrophage functions in response to particles. In addition,
animal-free toxicity testing is recommended in these days, so we also aim to develop in vitro
evaluation methods using alveolar macrophage cell lines which is useful tools for detecting
inflammatory particles.
Freshly isolated alveolar macrophages were immortalized by SV40 large T gene transfer using
lentiviral vector system, and then cultured with GM-CSF and PPAR-γ agonist to maintain theircharacteristics of alveolar macrophage. After screening of obtained cells, and we establishedalveolar macrophage cell line, ALV-3. The same as primary alveolar macrophages, ALV-3 cells
responded to inflammatory particles and induced cell death and subsequently IL-1α release.
Furthermore, ALV-3 cells expressed Siglec F and CD11c antigens, which are unique antigens foralveolar macrophages, on their surface. MH-S cells, which are commercially available alveolar
macrophage cell line, did not expressed Siglec F antigen and did not induce IL-1α release inresponse to inflammatory particles. These results suggest that ALV-3 cells might be useful cell line
to evaluate inflammatory particles, as an alternative in vitro method to animal testing. Now we arefurther investigating the characteristics of ALV-3 cells, as alternative to primary alveolar
macrophages.
Timeline:
March 1, 2022－February 28, 2023
Topics:Research meeting of LRI by JCIA, oral, “Development of evaluation method of inflammatory
particles based on alveolar macrophage function”, online, August 26, 2022.
Publications:
Kuroda Etsushi “Analysis of alveolar macrophage functions in response to fine particles" The127th Annual meeting of the Japanese Association of Anatomists, Online Symposium, 2022,March 28.
Kuroda Etsushi “DAMPs release and allergic airway inflammation caused by inhaled fine particles”
SOT 61st Annual Meeting, Online Symposium, 2022, March 29.