Induction of unique macrophage subset by simultaneous stimulation with LPS and IL-4

Kei Ishida; Takahiro Nagatake; Azusa Saika; Soichiro Kawai; Eri Node; Koji Hosomi; Jun Kunisawa

doi:10.3389/fimmu.2023.1111729

Induction of unique macrophage subset by simultaneous stimulation with LPS and IL-4

Front Immunol. 2023 Apr 21:14:1111729. doi: 10.3389/fimmu.2023.1111729. eCollection 2023.

Authors

Kei Ishida^{1

2}, Takahiro Nagatake^{1

3}, Azusa Saika¹, Soichiro Kawai¹, Eri Node¹, Koji Hosomi¹, Jun Kunisawa^{1

2

4

5

6

7

8

9

10}

Affiliations

¹ Laboratory of Vaccine Materials and Laboratory of Gut Environmental Health, Microbial Research Center for Health and Medicine, National Institutes of Biomedical Innovation, Health and Nutrition (NIBIOHN), Ibaraki, Osaka, Japan.
² Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan.
³ Laboratory of Functional Anatomy, Department of Life Sciences, School of Agriculture, Meiji University, Kawasaki, Kanagawa, Japan.
⁴ Graduate School of Medicine, Osaka University, Suita, Osaka, Japan.
⁵ Graduate School of Dentistry, Osaka University, Suita, Osaka, Japan.
⁶ Graduate School of Science, Osaka University, Toyonaka, Osaka, Japan.
⁷ International Vaccine Design Center, Institute of Medical Science, The University of Tokyo, Minato, Tokyo, Japan.
⁸ Department of Microbiology and Immunology, Graduate School of Medicine, Kobe University, Kobe, Hyogo, Japan.
⁹ Research Organization for Nano and Life Innovation, Waseda University, Shinjuku, Tokyo, Japan.
¹⁰ Graduate School of Biomedical and Health Sciences, Hiroshima University, Hiroshima City, Hiroshima, Japan.

Abstract

Macrophages manifest as various subtypes that play diverse and important roles in immunosurveillance and the maintenance of immunological homeostasis in various tissues. Many in vitro studies divide macrophages into two broad groups: M1 macrophages induced by lipopolysaccharide (LPS), and M2 macrophages induced by interleukin 4 (IL-4). However, considering the complex and diverse microenvironment in vivo, the concept of M1 and M2 is not enough to explain diversity of macrophages. In this study, we analyzed the functions of macrophages induced by simultaneous stimulation with LPS and IL-4 (termed LPS/IL-4-induced macrophages). LPS/IL-4-induced macrophages were a homogeneous population showing a mixture of the characteristics of M1 and M2 macrophages. In LPS/IL-4-induced macrophages, expression of cell-surface M1 markers (I-A^b) was higher than in M1 macrophages, but lower expression of iNOS, and expression of M1-associated genes (Tnfα and Il12p40) were decreased in comparison to expression in M1 macrophages. Conversely, expression of the cell-surface M2 marker CD206 was lower on LPS/IL-4-induced macrophages than on M2 macrophages and expression of M2-associated genes (Arg1, Chi3l3, and Fizz1) varied, with Arg1 being greater than, Fizz1 being lower than, and Chi3l3 being comparable to that in M2 macrophages. Glycolysis-dependent phagocytic activity of LPS/IL-4-induced macrophages was strongly enhanced as was that of M1 macrophages; however, the energy metabolism of LPS/IL-4-induced macrophages, such as activation state of glycolytic and oxidative phosphorylation, was quite different from that of M1 or M2 macrophages. These results indicate that the macrophages induced by LPS and IL-4 had unique properties.

Keywords: LPS and IL-4; M1 macrophage; M2 macrophage; energy metabolism; macrophages.

MeSH terms

Interleukin-4* / metabolism
Lipopolysaccharides* / metabolism
Lipopolysaccharides* / pharmacology
Macrophages / metabolism

Substances

Lipopolysaccharides
Interleukin-4

Grants and funding

This work was supported by the Ministry of Education, Culture, Sports, Science and Technology of Japan (MEXT)/Japan Society for the Promotion of Science KAKENHI (grant numbers 21K20769 to AS; 19K07617 to TN; 22K15004 to KH; and 21H02757 to JK); the Japan Agency for Medical Research and Development (AMED; grant numbers 22ae0121035s012 to KH; 223fa827004h0001 to TK; and 22fk0108145h0003, 22ae0121042h0002, 22ae0121035s0102 and 223fa727001h0001 to JK); the Ministry of Health and Welfare of Japan and Public/Private R&D Investment Strategic Expansion Program: PRISM (grant number 20AC5004 to JK); the Cross-ministerial Strategic Innovation Promotion Program (SIP) (grant number 18087292 to JK); the Grant for the Joint Research Project of the Institute of Medical Science, the University of Tokyo (to JK); the Ono Medical Research Foundation (to JK); and the Canon Foundation (to JK).