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Macrophage depletion using clodronate liposomes reveals latent dysfunction of the hematopoietic microenvironment associated with persistently imbalanced M1/M2 macrophage polarization in a mouse model of hemophagocytic lymphohistiocytosis

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Abstract

Hemophagocytic lymphohistiocytosis (HLH), a hyperinflammatory syndrome, is caused by the incessant activation of lymphocytes and macrophages, resulting in damage to organs, including hematopoietic organs. Recently, we demonstrated that repeated lipopolysaccharide (LPS) treatment induces HLH-like features in senescence-accelerated (SAMP1/TA-1) mice but not in senescence-resistant control (SAMR1) mice. Hematopoietic failure in LPS-treated SAMP1/TA-1 mice was attributed to hematopoietic microenvironment dysfunction, concomitant with severely imbalanced M1 and M2 macrophage polarization. Macrophages are a major component of the bone marrow (BM) hematopoietic microenvironment. Clodronate liposomes are useful tools for in vivo macrophage depletion. In this study, we depleted macrophages using clodronate liposomes to determine their role in the hematopoietic microenvironment in SAMP1/TA-1 and SAMR1 mice. Under clodronate liposome treatment, the response between SAMR1 and SAMP1/TA-1 mice differed as follows: (1) increase in the number of activated M1 and M2 macrophages derived from newly generated macrophages and M2-dominant and imbalanced M1 and M2 macrophage polarization in the BM and spleen; (2) severe anemia and thrombocytopenia; (3) high mortality rate; (4) decrease in erythroid progenitors and B cell progenitors in the BM; and (5) decrease in the mRNA expression of erythroid-positive regulators such as erythropoietin and increase in that of erythroid- and B lymphoid-negative regulators such as interferon-γ in the BM. Depletion of residual macrophages in SAMP1/TA-1 mice impaired hematopoietic homeostasis, particularly erythropoiesis and B lymphopoiesis, owing to functional impairment of the hematopoietic microenvironment accompanied by persistently imbalanced M1/M2 polarization. Thus, macrophages play a vital role in regulating the hematopoietic microenvironment to maintain homeostasis.

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Funding

This study was supported by the Japan Society for the Promotion of Science KAKENHI (grant number JP18K06846).

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Authors and Affiliations

  1. Division of Hematology and Oncology, Department of Internal Medicine, Nihon University School of Medicine, 30-1 Ohyaguchi-kami-cho, Itabashi-ku, Tokyo, 173-8610, Japan

    Takashi Koike, Katsuhiro Miura, Yoshihiro Hatta & Hideki Nakamura

  2. Center for Biological Safety and Research, National Institute of Health Sciences, Kawasaki, Japan

    Yoko Hirabayashi

  3. Division of Anatomical Science, Department of Functional Morphology, Nihon University School of Medicine, Tokyo, Japan

    Miyuki Yuda, Tomonori Harada, Shuichi Hirai, Isao Tsuboi & Shin Aizawa

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  1. Takashi Koike
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Contributions

Study design, data analysis, interpretation, and manuscript review: Takashi Koike, Katsuhiro Miura, Yoshihiro Hatta, Hideki Nakamura, Yoko Hirabayashi, Miyuki Yuda, Tomonori Harada, Shuichi Hirai, Isao Tsuboi, and Shin Aizawa. Experimentation: Takashi Koike, Yoshihiro Hatta, Miyuki Yuda, Tomonori Harada, Isao Tsuboi, and Shin Aizawa. Manuscript writing: Takashi Koike, Katsuhiro Miura, Yoshihiro Hatta, Hideki Nakamura, Yoko Hirabayashi, Tomonori Harada, Shuichi Hirai, Isao Tsuboi, and Shin Aizawa.

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Correspondence to Katsuhiro Miura.

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This study was reviewed and approved by the Animal Care and Use Committee of Nihon University (experimental code AP19MED019-2, AP21MED026-1, and AP19Med050-3).

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Koike, T., Miura, K., Hatta, Y. et al. Macrophage depletion using clodronate liposomes reveals latent dysfunction of the hematopoietic microenvironment associated with persistently imbalanced M1/M2 macrophage polarization in a mouse model of hemophagocytic lymphohistiocytosis. Ann Hematol 102, 3311–3323 (2023). https://doi.org/10.1007/s00277-023-05425-w

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