Table 1 Characteristics of macrophages generated from human monocytes and alveolar macrophages Fig. 1. Morphology of macrophages, dendritic cells and

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1 Jpn. J. Med. Mycol. Vol. 38, , 1997 ISSN

Table 1 Characteristics of macrophages generated from human monocytes and alveolar macrophages Fig. 1. Morphology of macrophages, dendritic cells and multinucleated giant cells derived from human monocytes a.

2 Table 1 Characteristics of macrophages generated from human monocytes and alveolar macrophages Fig. 1. Morphology of macrophages, dendritic cells and multinucleated giant cells derived from human monocytes a. macrophages generated from monocytes by GM-CSF b. macrophages generated from monocytes by M-CSF c. dendritic cells generated from monocytes by GM-CSF plus IL-4 d. multi-nucleated giant cells generated from monocytes by M-CSF plus IL-4

3 Jpn. J. Med. Mycol. Vol. 38 (No. 3),

Table 2 Characteristics of multi-nucleated giant cells from human monocytes by M-CSF+IL-4 Fig. 2. Expression of c-fms mrna in dendritic cells generated from monocytes by GM-CSF+IL-4.

4 Table 2 Characteristics of multi-nucleated giant cells from human monocytes by M-CSF+IL-4 Fig. 2. Expression of c-fms mrna in dendritic cells generated from monocytes by GM-CSF+IL-4. Total RNAs (10/1 g/lane) extracted from TNF- a -treated DC (lane 1), TNF- a -non-treated DC (lane 2), GM-CSFinduced macrophages (lane 3) or M-CSF-induced macrophages (lane 4) were subjected to agarose gel electrophoresis and then transferred to nylon membrane and hybridized with a labeled cdna probe of c-fms. Rehybridization was carried out wyth probes specific for Ac-LDLR or for actin.

5 J Exp Med 180: , ) Yaung DA, Lowe LD, Clark SC: Comparison of the effects of IL-3, granulocyte-macrophage colony-stimulat- ing factor, and macrophage colony-stimulating factor in supporting monocyte differentiation in culture. Analysis of macrophage antibody-dependent cellular cytotoxicity. J Immunol 145: , ) Matsuda S, Akagawa K, Honda M, Yokota Y, Takebe Y, Takemori T: Suppression of HIV replication in human monocyte-derived macrophages induced by granulocyte/macrophage colony-stimulating factor. AIDS Research and Human Retroviruses 11: , ) Doranz BJ, Rucker J, Yi Y, et al: A dual-tropic primary 1) De Nichilo MO, Burns GF: Granulocyte-macrophage and macrophage colony-stimulating factors differentially regulate a v integrin expression on cultured human macrophages. Proc Natl Acad Sci USA 90: , ) Akagawa KS, Kamoshita K, Tokunaga T: Effects of granulocyte-macrophage colony-stimulating factor and colony-stimulating factor-1 on the proliferation and differentiation of murine alveolar macrophages. J Immunol 141: , ) Dranoff G, Crawford AD, Sadelain M, et al: Involvement of granulocyte-macrophage colony-stimulating factor in pulmonary homeostasis. Science 264: , ) Stanley ER, Lieschke GJ, Grail D, et al: Granulocyte/ macrophage colony-stimulating factor-deficient mice show no major perturbation of hematopoiesis but develop a characteristic pulmonary pathology. Proc Natl Acad Sci USA 91: , ) Huffman JA, Hull WN, Dranoff R, et al: Pulmonary epithelial cell expression of GM-CSF corrects the alveolar proteinosis in GM-CSF-deficient Invest 97: , mice. J Clin 6) Nakata K, Akagawa KS, Fukayama M, Hayashi Y, Kadokura M, Tokunaga T: Granulocyte-macrophage colony-stimulating factor promotes the proliferation of human alveolar macrophages in vitro. J Immunol 147: , ) Wang AVT, Scholl PR, Geha RS: Phusical and functional association of the high affinity immunoglobulin G receptor (Fc y R I ) with the kinases Hck and Lyn. HIV-1 isolate that uses fusin and the beta-chemokine receptors CKR-5, CKR-3, and CKR-2b as fusion cofactors. Cell 85: , ) Pugin J, Heumann D, Tomasz A, et al: CD 14 is a pattern recognition receptor. Immunity 1: , ) Beekhuizen H, Blokland I, Corsel-van Tilburg AJ, Koning F, van Forth R: CD 14 contributes to the adherence of human monocytes to cytokine-stimulated endothelial cells. J Immunol 147: , ) Caux C, Dezutter-Dambuyant C, Schmitt D, et al: GM-CSF and TNF-a cooperate in the generation of dendritic Langerhans cells. Nature 360: , ) Akagawa KS, Takasuka N, Nozaki Y, Komuro I, Miyuki A, Ueda M, Naito M, Takahashi K: Generation of CD 1 + Re 1 B + Dendritic Cells and TRAP-positive Osteoclast-like Multi-nucleated Giant Cells from Human Monocytes. BLOOD 88: , ) Burkly L, Hession C, Ogata L, Reilly C, Marconi LA, Olson D, Tizaard R, Cate R, Lo D: Expression of re1b is required for the development of thymic medulla and dendritic cells. Nature 373: , ) Weih F, Carrasco D, Durham SK, Barton DS, Rizzo SA, Ryseck P-P, Lira SA, Bravo R: Multiorgan inflammation and hematopoietic abnormalities in mice with a targeted disruption of relb, a member of the NF-kB/rel family. Cell 80: , ) Yoshida H, Hayashi S, Kunisada T, et al: The murine mutation osteoetrosis is in the coding region of the macrophage colony stimulating factor gene. Nature 345: , ) Hashimoto S, Yamada M, Motoyosi K, Akagawa KS: Enhancement of macrophage-colony-stimulating factorinduced growth and differentiation of human monocytes by interleukin-10. BLOOD 89: , 1997.

6 Generation of Two Phenotypically Distinct Types of Macrophages, CD 1+Dendritic Cells and TRAP+Osteoclast-like Multi-nucleated Giant Cells from Human Monocytes Kiyoko S. Akagawa Department of Immunology, National Institute of Infectious Diseases, , Toyama, Shinjuku-ku, Tokyo 162, Japan We demonstrated that granulocyte-macrophage colony-stimulating factor (GM-CSF) and macrophage- CSF (M-csF) stimulate the differentiation of CD14+human monocytes into two phenotypically distinct types of macrophages, based on morphorogy, cell surface antigen expression and functions including phagocytosis, production of reactive oxygen intermediate, antigen presenting activity and sensitivity to HIV infection. In vivo, however, not only CSF but also many other cytokines are produced under various conditions, and those cytokines may modulate the differentiation of monocytes by CSFs. We demonstrated that CD 14+human monocytes can differentiate into CD1+re1B+dendritic cells (DC) by combination of GM-CSF plus interleukin-4 (IL-4), and that they differentiate into tartrate-resistant acid phosphatase (TRAP)-positive osteoclast-like multinucleated giant cells (MGC) by combination of M-CSF plus IL-4. However, the monocyte-derived DC were not terminally differentiated cells; they could still convert to macrophages in response to M-CSF. Tumor necrosis factor- a (TNF-a) stimulated the terminal differentiation of the DC by down-regulating the expression of the M-CSF receptor, c-fms mrna and aborting the potential to convert to macrophages. Taken together, these results provide a new aspect to our knowledge of monocyte differentiation, providing evidence that human monocytes are flexible in their differentiation potential, and that they are precursors not only of macrophages but also of CD1+re1B+DC and TRAP-positive MGC. Such a diverse pathway of monocyte differentiation may constitute one of the basic mechanisms of immune regulation.

日本消化器外科学会雑誌第29巻第9号

日本消化器外科学会雑誌第29巻第9号 Table 1 Oligonucleotide primers used for RT-PCR and internal probes used for Southern blot hybridization Cytokine Primer Sequence (5'-3') 5' 3' Internal probe s' 3' Internal probe 5' 3' Internal probe