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Liver X receptor (LXR)はステロールセンサーとしてコレステロールや脂質の代謝調節に働く核内受容体である.LXRαはマクロファージ,肝細胞や脂肪細胞などに発現することが知られている.しかし個体発生におけるLXR_αの組織発現については十分な検討がなされていない.本研究では,新規に作製されたLXR_α抗体を用いて,ラットの個体発生におけるLXR_αの細胞・組織学的発現について検討した.LXR_αは,胎生11日の卵黄嚢の内胚葉層に発現し,その後,肝臓,脾臓および胸腺のマクロファージに発現した.肝原基では,LXR_α陽性マクロファージは胎生12日に出現し,その数は胎生18日に最多となった.一方,肝細胞や褐色脂肪組織は胎生後期にLXR_α陽性となり,これら細胞・組織の脂質代謝機能の成熟を示唆するものと考えられた.マクロファージコロニー刺激因子(M-CSF)存在下で培養した成熟ラット単球由来のマクロファージは細胞の成熟とともにLXR_αの発現が亢進した.以上の成績からLXR_αはマクロファージの分化や脂質代謝に関わる細胞の分化成熟に重要な役割を果たしていると考えられた.

甲医博第798号

本研究は、マウス腹腔マクロファージおよび脾細胞のサイトカイン産生能に及ぼす自由運動の影響について検討した。BALB/c系、雌マウス(12週齢)を運動群と対照群に区分し実験を行った。運動は、回転ケージを用いた自由運動を週3日、8週間実施した。腹腔マクロファージ機能において、リポ多糖刺激に対する一酸化窒素産生能およびインターロイキン(IL)-1β産生能は、対照群と比較して運動群が高い値を示し有意差が認められた。脾細胞機能において、concanavalin A (Con A) 刺激に対する脾細胞増殖反応は、運動群が有意に高い値を示した。Con A刺激に対する脾細胞のサイトカイン産生能では、運動群のIL-2およびインターフェロン-γが対照群より有意に高い値を示したが、IL-4産生能ではみられなかった。以上の結果から、8週間の自由運動は若齢マウスの腹腔マクロファージ機能および脾細胞の機能を向上させ、免疫機能増強作用のあることが示唆された。, This study was designed to evaluate the effects of voluntary running exercise on cytokine production of macrophage and lymphocyte. Twelve weeks old female BALB/c inbred mice were divided into two groups: a group has given voluntary running exercise (exercise group) and a non-exercise group (control group). Exercise consisted of spontaneous running in wheels 3 days per week for 8 weeks. The levels of nitric oxide and interleukin (IL)-1β production by lipopolysaccharide-stimulated peritoneal macrophages from the exercise group were significantly higher than those in the control group. Concanavalin A (Con A)-induced cell proliferation of the spleen in the exercise group was significantly higher than that in the control group. IL-2 and interferon-γ productions stimulated by Con A in the exercise group were significantly increased compared with the control group, but not IL-4 production. These results might suggest that 8-week voluntary running exercise effectively enhanced macrophage and lymphocyte functions in mice.

From early twentieth,it has been considered that reticuloendothelial cells play an important role in the pathogenesis of amyloidosis. In the present review I discussed the role of macrophages in the formation of amyloid fibrils. In addition I described the role of amyloid enhancing factor in amyloidosis,which exceedingly shortened a lag period of amyloid deposition during amyloid induction.

色素細胞の産生するメラニン顆粒を抗原と見立てて, 皮膚からの抗原輸送機構を検討した。その結果, 定常状態でも恒常的に抗原が所属リンパ節に輸送されていること, その際の輸送細胞はTGF-β1依存性のランゲルハンス細胞であることが示された。

To evaluate phagocytic cells in periodontal membrane under physiological condition, we used horseradish peroxidase (HRP) as a phagocytic tracer. 10-week-old male rats were injected intravenously with HRP, and sacrificed 1 hour after the injection. Upper jaws were dissected out, decalcified, and prepared for light and electron microscopic observations. Furthermore, immunohistochemical localization of macrophages in periodontal membrane was examined by using anti-rat macrophage monoclonal antibody, ED 2. The following results were obtained: 1. Most of the phagocytic cells in the periodontal membrane can be found adjacent to the blood capillaries. 2. Electron microscopic observation revealed that most of these phagocytic cells were macrophages which had microvilli, secondary lysosomes with HRP and components of lipid-like drops in cytoplasm. 3. ED2 positive cells were found adjacent to the blood vessels in the periodontal membrane. The results of this study suggest that phagocytic cells of HRP in the periodontal membrane are macrophages.

Multi-ligand, cell surface receptors for modified lipoproteins are called scavenger receptors. They include three classes；class A (type I and type II scavenger receptors and macrophage receptor with collagenous structure designated as MARCO), class B (CD36 and SR-BI), and class C (dSR-CI, Fcγ RII-B2, and CD68/macrosialin). Among these scavenger receptors, type I and II class A macrophage scavenger receptors (MSR-A) are trimeric glycoproteins which mediate the recognition and uptake of a wide range of negatively charged macromolecules and are implicated in the deposition of cholesterol in arterial walls during atherogenesis through the receptor-mediated endocytosis of chemically modified low density lipoproteins (LDL). In the early lesions of atherosclerosis, it has been confirmed that blood monocytes migrate into the intima, differentiate into macrophages, and transform into foam cells by ingesting modified LDL. In order to analyze the mechanisms of monocyte chemotaxis and transformation of monocytes into foamy macrophages, we made a model system of aortic walls. Ultrastructural observation of transmigration of monocytes into collagen gel through the endothelial cell layer was performed.

Propionibacterium acnes has been implicated as the etiologic agent of sarcoidosis since the isolation of this bacterium at a high incidence from sarcoidosis specimens in 1978. However, this possibility has not been established or accepted widely yet. We have obtained the P. acnes strains isolated in that study. Based on the idea that inflammatory cytokines are involved in granuloma formation characteristic for pathology of sarcoidosis, we have compared the cytokine-inducing ability among various strains. ［Method］ Peripheral blood mononuclear cells were obtained from sarcoidosis patients and healthy controls. Cells were stimulated with P. acnes strains in vitro, and inflammatory cytokines were determined by using cytokine-specific EIA. The effect of T-cell depletion was examined. ［Result］ Th1-type of antigen-specific response was not observed when patient cells were stimulated with P. acnes. Strains from sarcoidosis specimen showed a significantly higher activity for induction of IL-12 and IFN-γ for whole mononuclear cells and also for T cell-depleted population. ［Conclusion］ The results suggested that particular P. acnes strains capable of inducing a high level of inflammatory cytokine are involved in the pathogenesis of sarcoidosis. It was postulated that P. acnes first stimulate macrophages to produce IL-12 then in turn stimulate NK cells to produce IFN-γ which may contribute to granuloma formation.

Mice homozygous for the osteopetrosis (op) mutation are characterized by defective differentiation of osteoclasts, monocytes, and tissue macrophages due to a lack of functional macrophage colony-stimulating factor (M-CSF/CSF-1) activity. In young (4-6 week-old) op/op mice, the bone marrow cavities were filled with spongious bone. In aged (50-72-week-old) op/op mice, the bone marrow cavities were markedly reconstructed and marrow hematopoiesis was expanded. Numbers of osteoclasts and bone marrow macrophages in aged op/op mice were increased but most of the osteoclasts were mononuclear cells and showed poorly developed ruffled borders. In contrast to the marked increase in numbers of osteoclasts and macrophages in the bone marrow, the number of Kupffer cells in the liver did not increase in aged op/op mice. M-CSF administration to aged op/op mice induced numerical increases in Kupffer cells and the development of ruffled border in osteoclasts. These findings indicate that M-CSF-independent mechanisms for bone marrow hematopoiesis and macrophage and osteoclast development function in aged op/op mice.

Hemodialysis associated amyloidosis (HA) is a new type systemic amyloidosis. Gejyo elucidated precursor protein of HA was β2-microglobulin (β2-MG), which was a low molecular protein accumulating in the serum of long term hemodialysis patients, in 1985. The histological features of this amyloidosis, which are unique in fiblilar structures and histochemical characters, are clearly different from those of other types of systemic amyloidosis, AL and AA. HA shows specific aspects in clinical features, either. The incidence of various bone and articular complications, such as bone cysts, carpal tunnel syndrome and destructive spondylarthropathy, increases according to the duration of hemodialysis. These complications result from the deposition of amyloid fibrils derived from β2-MG and are suspected to be confirmed in all patients who continue the dialysis treatment more than 10 years. The affinity of amyloid deposition to bone and articular tissues supposed to be an interesting key character in the pathogenic consideration of amyloid fibril formation. We discussed clinical and histological features of HA and summarized the recent studies of pathogenesis about amyloidgenesis including in our research results.

嚥下協調障害・胃食道逆流による慢性誤嚥を呈する患者群では, 軽微な気道感染から急性呼吸窮迫症候群(acute respiratory distress syndrome: ARDS)への進展を経験することも少なくないが, その病態は充分に解明されていない。慢性誤嚥が小児ADRSの発症に関与している病態を明確にすることを目的として, 気管支肺胞洗浄液(bronchoalveolar lavage fluid: BALF)を用いた検討を行った。慢性誤嚥が基礎にある症例では, BALF分画における好中球比率の変遷は通常のARDSと同様であったが, マクロファージの比率は発症初期からBALF中に増加しており, さらにOil red O染色にて脂肪貪食像を認めた。この様な症例では, 末梢気道のレベルで肺胞マクロファージによる活発な脂肪貪食が行われ, マクロファージの活性化が惹起されていると推察された。BALF中マクロファージの脂肪貪食像は, 慢性誤嚥による肺傷害の病態解明に有用と考えられた。, Although there is ample evidence for the involvement of recurrent aspiration in the development of ARDS, the exact mechanism has not been clarified. This study aimed to know the pathophysiolosy of chronic aspiration which was involved in childhood acute lung injuries (ALI). We performed bronchoalveolar lavage (BAL) and collected bronchoalveolar lavage fluids (BALF) in ARDS patients. The BALF specimens were stained with Hansel or Oil red 0 stain to examine the BALF cytology and to identify lipid-laden alveolar macrophages. There were many lipid-laden alveolar macrophages in BALF in the acute phase of ARDS. It is considered that alveolar macrophages have been stimulated by the process of lipid phagocytosis prior to the onset of ARDS in cases of chronic aspiration. This raises the possibility that lipid-laden alveolar macrophages have a significant role in the case of ALI in children with chronic aspiration.

Article, 信州医学雑誌 45(3): 177-191(1997)

初期動脈硬化病巣においてはマクロファージが変性low density lipoprotein (LDL)を取り込んで泡沫細胞となる過程が重要と考えられている。コレステロールの細胞内輸送・代謝に関与する最も有力な細胞内脂質転送蛋白と考えられているsterol carrier protein_2 (SCP_2)は変性LDLの取り込みに伴いマクロファージが泡沫化する過程においてその含量が増加する事が明らかにされている。今回, マクロファージ泡沫細胞形成過程におけるSCP_2の遺伝子発現とその調節機構について検討を行った。まず変性LDLであるAcetylated-LDL (Ac-LDL)によりマクロファージが泡沫化しSCP_2含量の増加する際, mRNAレベルの変動をNorthern blot法で解析したところSCP_2のmRNAの増加が認められた。次にこのSCP_2の遺伝子発現が増強する機序について検討した。acyl-CoA: cholesterol acyltransferase (ACAT)阻害剤であるEAB-309の添加によりAc-LDLによるマクロファージ中コレステロールエステルの増加は著明に抑制され, 遊離コレステロールは増加した。その際, この細胞中のSCP_2含量とmRNAレベルの増加が認められた。さらに, この増加が細胞内遊離コレステロールの増加によるものかどうかを調べる目的で25-OH cholesterolを添加したところSCP_2含量, mRNAレベルは増加した。しかし, Ac-LDLによるSCP_2増加がAc-LDLがマクロファージのスカベンジャー受容体を介したシグナルによるものかどうかを明らかにするためのmaleylated bpvine serum albumin (Mal-BSA)の添加は影響を与えなかった。これらの成績からAc-LDLによるマクロファージの泡沫細胞形成過程におけるSCP_2の増加はその遺伝子発現の増強によること, その発現増強した機序としてスカベンジャー受容体からのシグナルよりも細胞内遊離コレステロールの, One of the most important steps in the atherosclerosis is the foam cell formation in macrophages by modified LDL. Sterol carrier protein_2 (SCP_2) is thought to be one of highly touted candidate for an intracellular cholesterol carrier. It has been previously reported that incubations with Ac-LDL cause an increase of SCP_2 content in rat peritoneal macrophages during foam cell foamation. The present study was designed to clarify the mechanism of the Ac-LDL-induced increase in SCP_2 content in macrophages. Western blot analysis and Northern blot analysis demonstrated that both SCP_2 protein and its mRNA are expressed in rat peritoneal macrophages. Incubations with Ac-LDL caused an increase of cellular cholesterol (free cholesterol and cholesterol esters) and SCP_2 content in rat peritoneal macrophages. Furthermore, SCP_2 mRNA level was also increased by Ac-LDL treatment. The inhibitor of acyl-CoA: cholesterol acyltransferase further enhanced Ac-LDL-induced increase of SCP_2 protein and its mRNA. Incubations with 25-OH cholesterol also caused a dose-dependent stimulation of SCP_2 gene expression in macrophages, while incubation with Mal-BSA had no effect. These results suggest that increased cellular free cholesterol content is responsible for the enhancing SCP_2 and its gene expression in macrophages.

国立情報学研究所の「学術雑誌公開支援事業」により電子化されました。

In order to clarify the expression and tissue distribution of scavenger receptors, we have performed immunohistochemical and immunoelectron microscopical studies as well as in situ hybridization. Scavenger receptors are a trimeric glycoprotein mediating endocytosis of chemically modified low density lipoproteins (LDL). The receptors were detected specifically in macrophages in various tissues by immunohistochemistry using anti-bovine scavenger receptor monoclonal antibody IgG-D2, although hepatic sinusoidal cells are known to incorporate modified LDL actively. In contrast, scavenger receptor protein was detected both in Kupffer cells and endothelial cells of the hepatic sinusoid and lymphatic sinus by immunohistochemistry using anti-bovine scavenger receptor antibody IgG-D1 and anti-murine scavenger receptor antibody 2F8. In situ hybridization demonstrated the expression of scavenger receptor mRNA in Kupffer cells and sinusoidal endothelial cells. Scavenger receptors were not expressed in endothelial cells of other tissues than the liver and lymph nodes. These findings indicate that scavenger receptors are expressed both in macrophages and specific endothelial cells and that endothelial cells in various tissues constitute heterogeneous populations with regard to lipoprotein metabolism.

サイトカインは最初免疫機構に対する制御物質として発見された. しかし その作用は免疫だけでなく種々の細胞に及んでおり, 生体内の多くの事象を制御している. 腫瘍に対してもその増殖分化, 転移あるいはその小腿など幅広く関与している. B細胞増殖因子として発見されたIL-4はその多岐にわたる機能とともに, 腫瘍免疫にも大きくかかわっていることが解ってきた. 種々の腫瘍におけるIL-4Rの存在, in vitro において一部の腫瘍系に対する直接の成長阻害作用. CTL, マクロファージ, 好酸球などのエフェクター細胞の活性化などである. ここではIL-4の抗腫瘍免疫における役割について簡単に現在の知見をまとめてみたい.

"Osteopetrotic（OP／OP） mouse is an animal model for osteopetrosis and was demonstrated to be a mutation within the coding region of M－CSF gene itself. The OP／OP mouse serves as a model for investigating the differentiation mechanism of macrophage populations in the absence of functional M－CSF. The OP／OP mice also provide evidence to show the role of M-CSF in physiological and pathological conditions. The OP／OP mice are severely monocytopenic and show marked reduction and abnormal differentiation of tissue macrophages and osteoclasts. Most of these macrophages are ultrastructurally immature. Compared with the tissues of normal littermates, those of mutants contained about 30％ of macrophages in many tissues, suggesting that the heterogeneity of macrophages is generated by their different dependency to M－CSF. In contrast, the numbers of dendritic cells in the epidermis and lymphoid apparatus of OP／OP mice were not reduced comparing to those in normal littermates, indicating that dendritic cells are an M－CSF－independent population. After daily M-CSF injection the numbers of monocytes, tissue macrophages, and osteoclasts showed remarkable increases and macrophages demonstrated morphological maturation. However, the numbers of macrophages in the ovary and uterus were not increased. After glucan injection hepatic granulomas in OP/OP mice were formed but smaller, less numerous, and more irregular in shape than those of normal littermates. Kupffer cells in the mutant mice exhibited an active proliferation\ncapacity, particularly just before the stage of granuloma foration. After administration of M-CSF, numbers of monocytes and Kupffer cells increased rapidly in OP/OP mice and granuloma formation was enhanced in these mice. These results indicate that M-CSF-independent Kupffer cells and M-CSF-dependent macrophage populations play an important role in granuloma formation. In conclusion, M-CSF is an important molecule for proliferation and differentiation of not only M-CSF-dependent macrophages but also M-CSF-independent macrophages in\nphysiological conditions. Furthermore, M-CSF is largely responsible for providing a microenvironment for generating macrophage heterogeneity in vivo. "

The present study was carried out to examine the effects of Maharishi Amrit Kalash 4 (MAK-4) and 5 (MAK-5) on non-specific immunological mechanisms in mice. Mice aged 4 weeks were divided into 3 groups : a no treatment group (control), MAK-4 treated group (MAK-4 group) and MAK-5 treated group (MAK-5 group). The MAK-4 and MAK-5 were given p.o. at a dose of 50 mg/kg per day for 8 weeks. The following results were obtained : 1. MAK-4 and MAK-5 did not affect the body weight gain and food consumption of the mice. 2. The acid phosphatase (APH) and lactate dehydrogenase (LDH) activities of peritoneal macrophages (Mφ) in the MAK-4 and MAK-5 groups increased significantly compared to the control group. 3. Concanavalin A (Con A) induced cell proliferation in the spleen was high in the MAK-4 and MAK-5 groups. These results suggested that MAK-4 and MAK-5 has a stimulatory effect on Mφ because of the increases in the APH and LDH activities in the peritoneal Mφ of mice. MAK-4 and MAK-5 also intensified the T-cell function represented by Con A-induced splenocyte proliferation.

研究科: 千葉大学大学院医学薬学府, 学位:千大院医薬博乙第薬46号, 修了年:2011年

Morphological changes in the uterine lumen and the localization of macrophages were studied during the implantation period in the rat uterus. There were dynamic changes in the morphological features at the uterine part where the embryo existed

Ultrastructural and cytochemical features of osteoclasts of mice homozygous for the osteopetrosis (op/op) mutation were studies. The mutant mice are characterized by the prominent osteosclerosis and defective differentiation of osteoclasts, monocytes, and tissue macrophages due to a lack of functional macrophage colony-stimulating factor (M-CSF/CSF-1) activity. In op/op mice, osteoclalsts were markedly reduced and showed poorly developed ruffled borders. Daily administration of M-CSF induced the development and ultrastructural differentiation of osteoclasts, followed by the resorption of bones. These results indicate that the differentiation and proliferation of osteoclasts are mediated by M-CSF.

Numerous macrophages are aggregated at the villus tips in the small intestine of guinea pigs and monkeys. The macrophages are characterized by rich existence of large phagosomes which contain cellular elements. Morphological observation shows that effete enterocytes are not simply exfoliated into the lumen but phagocytosed by the subepithelial macrophages. The epithelium of the villus tips is infiltrated with numerous large granular lymphocytes. The cytotoxic lymphocytes with an intense perforin immunoreactivity are actively interdigitated with apoptotic enterocytes, suggesting the lymphocyte-mediated cell death. In the rat and mouse, no aggregation of phagocytotic macrophages is found in the lamina propria of villus tips. The enterocytes undergoing apoptosis are exfoliated as total, nucleus-containing cells into the lumen. Ultrastructural and cytochemical studies suggest that in the rat and mouse, lamina propria macrophages rather than intraepithelial lymphocytes are involved in cell death of enterocytes. In the human, the aggregation of macrophages is observed in the large intestine, especially the rectum. The intimate relationship between the lamina propria macrophages and the ulcerative colitis is shown experimentally and clinically.

マクロファージは正常ヒト腹腔内に常在し、腹腔内における免疫機能の中心的役割を担っている。マクロファージは、その活性化状態によって、抗腫瘍活性を示すM1マクロファージと腫瘍増殖を促進するM2マクロファージに二大別されるが、腹腔マクロファージの、腹腔内環境によってM1マクロファージあるいはM2マクロファージに分化することが報告されている。多量の腹水貯留と腹腔内播種をきたす進行卵巣癌においては、腹水中でマクロファージが増加していることが既に知られている。しかし、進行卵巣癌細胞とマクロファージの相互関連については、十分に明らかとなっていない。私は、進行卵巣癌腹水中におけるマクロファージの分化と、それに関与するsignal transducer and activator of transcription 3 (Stat3) の活性化について解析を加えるとともに、マクロファージが癌細胞増殖に及ぼす影響について検討を加えた。, Ascites macrophages in advanced epithelial ovarian cancer are involved in cancer metastasis and progression by modifying the tumor microenvironment. However, the precise mechanisms of cell-to-cell interaction between macrophages and tumor cells are still unclear. This study focused on the activation of signal transducer and activator of transcription 3 (Stat3) which is a critical signal transduction molecule at a point of convergence for numerous oncogenic signaling pathways as well as controlling the M2-poralization of macrophages.

急性冠症候群は動脈硬化のプラークが破綻することにより引き起こされる、冠動脈の血流の減少や途絶により生じる疾患の総称である。プラークの不安定化が破綻の基盤になっていると考えられているが、不安定化から破綻に至るメカニズムはよくわかっていない。近年、動脈硬化病変のマクロファージに小胞体ストレスが生じているとする報告が散見される。CHOP はC/EBP（CCAAT/enhancer-binding protein）ファミリーに属する転写因子で、小胞体ストレスによりその発現は著しく増加する。CHOP はアポトーシスの促進に重要な役割を果たしていると考えられており、アポトーシスを誘導するBim、Bax を誘導、活性化し、アポトーシスを抑制するBcl-2 の生成を抑制する。CHOP は小胞体ストレスにおけるアポトーシスの中心的役割を担っていると考えられる。本研究では、動脈硬化病変のマクロファージに小胞体ストレスおよび小胞体ストレス誘導性アポトーシスが生じていることを確認し、プラークの不安定化及び破綻に小胞体ストレス、特にCHOP が関与しているか検討した。

We investigated the preparation of macrophage monolayer from carp peritoneal exudate cells (PEC) to clarify the biosynthesis and processing of cathepsins. PEC collected at 24h interval after elicitation with 6% sodium caseinate or thioglycolate medium were transferred to culture dish in RPMI 1640 (RPMI), and the cells were incubated in CO₂ incubator for 2h. To distinguish macrophage and neutrophil, the adherent cells washed with RPMI to remove non-adherent cells were analyzed histochemically by the staining of two different esterase activities. The number of PEC was maximally increased at 3 days after injection with 15 ml of sodium caseinate, whereas that of macrophages was at 4 days after injection. Most. (about 90%) of cells cultured for 24h were positive for α-Naphthyl butyrate esterase activity but they were not positive for Naphthol AS-D chloroacetate esterase activity, confirming them to be macrophage. They were able to culture for 48h in RPMI containing 5% heat inactivated calf serum. It was possible to prepare two culture dishes of macrophage monolayer (1.33 x 10⁵/cm²) from one carp (800-900g) by this method.

Phagocytic activities of intraperitoneally exuded neutrophil against live cells of Edwardsiella tarda were studied in the Japanese flounder, Paralichthys olivaceus, injected with lipopolysaccharide of the bacterium. Addition of normal serum, or normal serum and antiserum did not increase the phagocytic rate or phagocytic index of the neutrophil, while it stimulated the phagocytic activities of the macrophage coexisted with the neutrophil. The present results suggest that against Edwardsiella tarda complement or antibody does not act as an opsonin on the phagocytosis of flounder neutrophil.

Article, 信州医学雑誌 41(4): 305-315(1993)

Macrophages are heterogenous populations existing in various tissues and organs, performing multifacetted functions in physiological and pathological conditions. In ontogeny, primitive/fetal macrophages first develop in yolk sac hematopoiesis without passing through the differentiation pathway of monocytic series. Monocytic cells are a minor cell population in the early fetal period, and increase in the late stage. Primitive/fetal macrophages proliferate and survive in loco. Tissue macrophages in adult animals also proliferate and maintain their population by self renewal. In monocytopenic mice induced by administration of bone-seeking isotope, strontium-89, tissue resident macrophages do not decrease. In macrophage colony stimulating factor （M-CSF）-deficient mice （op/op）, monocytes as well as tissue macrophages are deficient. However, M-CSF-independent tissue macrophages and Langerhans/dendritic cells are present in the defective condition of monocyte differentiation into macrophages. These findings demonstrate that differentiation pathways of tissue macrophages and nonlymphoid dendritic cells are different from that of monocytes. As an example of the specific functions of macrophages, tissue distribution, intracellular localization and expression of scavenger receptors were presented with special reference to the role of macrophages in endocytosis of chemically modified low density lipoprotein and atherogenesis.

本研究では、LCH におけるマクロファージの活性型と病態形成における役割を解析するため、ヒトLCH のパラフィン包埋標本 20 症例を用いて免疫組織化学的検討を行った。

動脈硬化病巣にみられるコレステロールエステル(CE)を蓄積した泡沫細胞の出現機構を検討した。病理組織学的検討からは,泡沫細胞の起源は中膜平滑筋細胞(SMC)とマクロファージに共に生じていることが観察されているが,マクロファージは変性低比重リポ蛋白(変性LDL)を取り込み,泡沫細胞化することが示された。しかしSMC由来の泡沫細胞形成機構は不明である。従来より内皮細胞とマクロファージ,あるいは内皮細胞とSMCなどとの細胞の相互作用を介した代謝機能調節が示されてきた。本研究ではSMCとマクロファージの細胞間相互作用を介したSMCへの脂質蓄積機構を明かにすることを試みた。SMCとマクロファージのco-culture系に本来マクロファージで代謝されるβ-migrating very low density lipoprotein(β-VLDL)を添加すると,マクロファージのみならず,SMCにも脂質の蓄積を認めた。これはSMCとマクロファージの細胞間相互作用を介したSMCへの脂質蓄積機構の存在する可能性が示唆された。その機構を検討するため[^3H]cholesteryl linoleate labeled β-VLDL([^3H]β-VLDL)添加培養液でマクロファージを培養したconditioned medium(標識Mβ-CM)をSMCに添加し,培養すると,SMCへの放射活性の取り込みは増加し,SMCのCE含量は増加した。このことからマクロファージにより生じた[^3H]β-VLDLの代謝産物がSMCに脂質蓄積をもたらしている可能性が推測された。次にこの代謝産物の性質を明かにすることを試みた。Zonal超遠心法で標識Mβ-CMを分析したところ,比重1.006(peak I)の他に1.150(peak II)にも放射活性のピークを認めた。このpeak IIはagarose gel電気泳動ではLDLに一致した泳動度を示し,CE(32%),free cholesterol(8%),phospholipid(46%)からなっていた。放射活性は[^3H]CE(80%)と[^3H〯

2007年度～2008年度科学研究費補助金(基盤研究(C))研究成果報告書, 研究概要：本研究では、自然免疫担当細胞であるマクロファージ(MФ)が産生する新規アポトーシス抑制因子(AIM)を欠損するマウスを用い、マラリア原虫感染におけるNKT細胞及びγδT細胞の活性化機能制御とMФの関係を明らかにし、新しい視点での感染防御機構だけでなく病態形成の機序について検討を加えた。その結果、マラリア感染症においては、MФの機能としての貪食作用による病原体の排除やそれに伴う炎症反応の惹起にAIMが深く関与しており、その機能制御が予防・治療に有効である可能性が示唆された。

粥状動脈硬化初期病変では、血管内皮細胞下にマクロファージ由来泡沫細胞の集簇が認められ、この泡沫細胞は様々な動脈硬化惹起因子を産生しており、動脈硬化症の発症・進展に寄与している。近年、血管内皮下においてマクロファージやマクロファージ由来泡沫細胞が増殖していることが分かり、動脈硬化症の発症・進展に重要な役割を果たすことが示唆されている。著者らの教室ではこれまでに、酸化低比重リポタンパク質（酸化LDL）がマウス腹腔マクロファージを増殖させる事、その機序として酸化LDLによる granulocyte/macrophage colony-stimulating factor(GM-CSF)の発現と分泌が必要である事、さらにGM-CSFの発現には extracellular-signal regulated kinase 1/2 (ERK1/2) の活性化が関与すること、を報告した。本研究では、酸化LDLによるマクロファージ増殖に対するAMP-activated protein kinase (AMPK) 活性化の影響を検討した。

The purpose of this study was to determine the exact cell type and site(s) involved in interleukin-1 (IL-1) production during gingival inflammation. By combining immunohistochemistry and in situ hybridization, IL-1messenger RNA (mRNA) –expressing cells in human inflamed gingival were identified as macrophages. The rate of IL-1α mRNA expression in these macrophages was the same as IL-1βmRNA expression. Additionally, the rate of IL-1 mRNA expression was higher in tissue sections furthest from sulcular areas than in those closest of sulcular lesions, although more macrophages were present at the sulcular site. The IL-1 activity in gingival crevicular fluid (GCF) obtained from inframed gingival was higher than that from healthy gingival and decreased after periodontal therapy. The IL-1 activity in GCF was almost completely abolished by the addition of anti-IL-1α antibody but not anti-IL-1βantibody, indicating that IL-1α is the predominant form in GCF. However, the IL-1 activity in GCF was unrelated to the number of IL-1 mRNA-expressing macrophages in the same gingival site where the GCF was obtained at the same time. The results suggest that macrophages in the oral side contribute to the production of IL-1 but those in the sulcular side play a different role in the generation of gingival inflammation., 歯周炎罹患者の歯肉組織中におけるインターリュウキン1 (IL-1) mRNA発現細胞の局在及び歯肉溝滲出液 (GCF) 中のIL-1活性について検索した｡歯周炎罹患者28名のうち19名から歯周外科時に歯肉組織を採取し､18名からGCFを採取した｡対照群としての歯肉組織は､本学矯正科に来院した患者から歯の便宜抜去時に採取し､GCFは歯肉健常者のボランティアから採取した｡採取後の歯肉組織はパラフィン包埋切片としPAP法による免疫組織化学的にマクロファージ､T細胞､B細胞､内皮細胞を染色し､同一切片上でin situ hybridizationをIL-1α､IL-1βについて^35S標識のプローブを用いて行った｡GCFのIL-1活性はC3H/HeJマウスの胸腺細胞の増殖を^3H-thymidinの取り込み量で測定した｡また､α/βをみるために中和実験を行った｡その結果､歯周炎罹患歯肉組織中のIL-1mRNA発現マクロファージはポケット側より口腔側に多く認められ､その局在に関しては､炎症局所における機能的対応が示唆された｡GCF中のIL-1活性は健常者に比べ歯周炎罹患者で有意に高く､歯周治療により有意に減少した｡また､α タイプが圧倒的に多くその活性を占めていた｡GCF中のIL-1活性は､歯周炎の病態変化と深い関係が認められることから､新しい臨床診断法としての可能性が期待される。

Article, 信州医学雑誌 39(1): 91-104(1991)

動脈硬化性プラーク内に存在するマクロファージにはurokinasetype plasminogen activator (uPA) とuPA受容体(uPAR) が発現しているが、その発現調節については十分に解明されていない。今回私は動脈硬化原性脂質であるlysophosphatidylcholine (lysoPC) を用いて、培養ヒト単球由来マクロファージにおけるuPAおよびuPARの発現について検討した。