활성화된 마우스 대식세포가 Toxoplasma gondii의 증식에 미치는 영향

엄대자,민득영,안명희 한양대학교 의과대학 1992 한양의대 학술지 Vol.12 No.1

The present study was aimed to elucidate the effect of antitoxoplasma activity and oxidative capacity of peritoneal macrophage of immunized and/or normal mouse. BALB/c mice were immunized with dozens of cyst (Beverly of Fukaya strain) intraperitoneally. Glass adherent cells were used as macrophage from immunized and normal mouse peritoneal cells. Macrophage(?? cells/ml) and Toxoplasma tachyzoite(RH strain) were incubated in 10% fetal calf serum-minimum essential medium, 37℃, 5% CO₂ incubator. The ratio of cell to Toxoplasma was 1:1 or 2:1. After 4 and 24hr incubation, cell rupture and infection rate of macrophage, number of tachyzoite in a cell were observed under microscope. Production of hydrogen peroxide from immunized or normal macrophage after 1hr incubation of Toxoplasma or Toxoplasma with phorbol myristate acetate(PMA) were measured. After 4hr incubation of macrophage and Toxoplasma tachyzoite, 10% of cell rupture were observed in normal and immunized macrophages. One to five organisms per cell were observed 32.8~38.7% of normal macrophage and 50.7~52.6% of immunized macrophages. After 24hr incubation, Toxoplasma multiply freely in normal mouse macrophage and 90% of cells were ruptured. Meanwhile immunized mouse macrophage display toxoplasmacidal effect and 50% of cells were survived. Toxoplasma invation into cell were 34.~48.7% of normal macrophage and 59.4% of immunized macrophage. Immunized macrophages released 2~7 times of H₂O₂than those of normal macrophages. With above results, it is assumed that peritoneal macrophage from immunized mouse has high producibility of H₂O₂and then might enhance the subsequential toxoplasmastatic and/or toxoplasmacidal effect.

Regulation of inflammatory gene expression in macrophages by epithelial-stromal interaction 1 (Epsti1)

Kim, Young-Hoon,Lee, Jae-Rin,Hahn, Myong-Joon Elsevier 2018 Biochemical and biophysical research communication Vol.496 No.2

<P><B>Abstract</B></P> <P>Epithelial-stromal interaction 1 (<I>EPSTI1</I>) was first discovered as a gene induced in breast cancer epithelial cells by co-cultured stromal fibroblasts. There are many reports on the role of Epsti1 in cancer malignancy. Epsti1 is now well known in regulating cancer. Recently, the role of Epsti1 in the immune response has been reported; these reports suggest the role of Epsti1 in immune function, immune privilege, and autoimmune diseases. Furthermore, they show that Epsti1 is expressed in various types of immune cells. In this study, we observed that Epsti1 is highly expressed in macrophages exposed to IFNγ and lipopolysaccharide (LPS), which classically activates macrophages. Polarization of macrophage to classically activated (M1) or alternatively activated (M2) is important for mounting responses against various infections. The M1 and M2 types of macrophage have a distinct role in the immune system. However, the molecular mechanism of modulation of the macrophage type is not well defined. Our results showed that the M2 type macrophage phenotype is enhanced in Epsti1-deficient bone marrow-derived macrophages (BMDM). In addition, Epsti1 deficiency suppresses induction of pro-inflammatory genes in BMDMs via inhibition of Stat1 and p65 nuclear localization and phosphorylation. Surprisingly, <I>Epsti1</I>−/− mice show decreased numbers of M1 macrophages in the peritoneal cavity. These findings identify Epsti1 as a modulator of macrophage activation and polarization via the Stat1 and p65 pathways, and suggest a potentially important role of Epsti1 in immunotherapies against inflammatory diseases.</P> <P><B>Highlights</B></P> <P> <UL> <LI> Epsti1 is highly expressed in macrophages exposed to IFNγ and lipopolysaccharide (LPS). </LI> <LI> The M2 type macrophage phenotype is enhanced in Epsti1-deficient bone marrow-derived macrophages. </LI> <LI> <I>Epsti1</I>−/− mice show decreased the number of M1 macrophage in the peritoneal cavity. </LI> </UL> </P>

해당화의 과육 및 종자추출물의 대식세포 면역조절작용

강남성,손은화 한국자원식물학회 2010 한국자원식물학회지 Vol.23 No.5

Rosa rugosa has been used as a folk medicine with various pharmacological properties for a long time in Asia. Recently, it has been reported that the extract of fractions from different parts of Rosa rugosa have various pharmacological effects on diverse diseases including diabetes, inflammatory diseases and tumor. We investigated effects of fructus extracts of Rosa rugosa(RRF) and semen extracts of this herb(RRS) on macrophage to evaluate the possibilities as a biological response modifier. We showed increased effects on tumoricidal activity, phagocytic activity, TNF-α and NO production in RRF-treated groups without direct tumor cell cytotoxicity. RRS-treated groups increased direct tumor cell cytotoxicity at high dose without tumoricial activity except increasing of TNF-α release. These results provide further possibilities for the beneficial immunomodulating effects of RRF on immune system with relatively larger safety margin rather than RRS. 현재 임상적으로 적용되고 있는 항암요법들은 많은 부작용을 가지고 있으며, 부작용에 의하여 또다른 질환을 야기하는 것이 항암치료에서 문제점으로 지적되고 있다. 따라서 부작용을 줄이고 항암요법을 유지시키는 방법으로, 인체 안전하다고 보고된 천연물을 이용하여 면역력을 증가시킴으로써 인체내 항암 효과를 나타내게 하는 BRM들을 개발하는 연구가 큰 의미를 지닌다. 이에 본 연구에서는 민간요법으로 이미 사용되고 있는 해당화의 macrophage의 활성화에 대한 BRM 효과를 확인하였으며, 특히 과육(RRF)과 종자(RRS)의 부위별 추출물로 그 효과를 비교 분석하였다. RRF는 암세포 자체에 대한 세포독성은 나타내지 않았으나, macrophage의 활성화에 의한 항암 효과를 나타내었다. 이러한 항암 효과는 macrophage의 활성화에 의한 증가되는 NO 및 TNF-α와 같은 암세포 독성물질에 의한 효과를 기대할 수 있으나, RRF 처리에 의하여 활성화된 macrophage는 NO 분비에 효과를 나타내지 않았다. RRF의 처리는 TNF-α 분비를 증가시켰으나, macrophage의 활성화에 의해 암세포 독성을 나타내지 않았던 RRS에서도 TNF-α 분비가 증가한 것으로 보아 TNF-α 분비만으로는 macrophage의 항암효과에 직접적인 영향을 나타내지는 않는 것으로 보인다. 본 연구 결과들은 종합해 볼 때, RRF는 RRS와는 달리 macrophage를 활성화하여 항암효과를 나타내었으며, phagocytosis 능력도 증가시켰다. RRF는 TNF-α 등의 분비조절과 같이 RRS와는 다르게 macrophage를 활성화시키는 것으로 보이며, 임상적으로 적용시 RRF가 RRS보다 세포독성 측면에서도 안전하고, macrophage의 활성화 효과 측면에서도 유의성이 높을 것으로 평가 된다.

Agmatine Modulates the Phenotype of Macrophage Acute Phase after Spinal Cord Injury in Rats

김재환,김재영,문진희,서민아,이종은 한국뇌신경과학회 2017 Experimental Neurobiology Vol.26 No.5

Agmatine is a decarboxylated arginine by arginine decarboxylase. Agmatine is known to be a neuroprotective agent. It has been re- ported that agmatine works as a NMDA receptor blocker or a competitive nitric oxide synthase inhibitor in CNS injuries. In spinal cord injury, agmatine showed reduction of neuropathic pain, improvement of locomotor function, and neuroprotection. Macrophage is a key cellular component in neuroinflammation, a major cause of impairment after spinal cord injury. Macrophage has subtypes, M1 and M2 macrophages. M1 macrophage induces a pro-inflammatory response, but M2 inspires an anti-inflammatory response. In this study, it was clarified whether the neuroprotective effect of agmatine is related with the modulation of macrophage subdivision after spinal cord injury. Spinal cord injury was induced in rats with contusion using MASCIS. Animals received agmatine (100 mg/ kg, IP) daily for 6 days beginning the day after spinal cord injury. The proportion of M1 and M2 macrophages are confirmed with immunohistochemistry and FACS. CD206+ & ED1+ cells were counted as M2 macrophages. The systemic treatment of agmatine increased M2 macrophages caudal side to epicenter 1 week after spinal cord injury in immunohistochemistry. M2 macrophage re- lated markers, Arginase-1 and CD206 mRNA, were increased in the agmatine treatment group and M2 macrophage expressing and stimulated cytokine, IL-10 mRNA, also was significantly overexpressed by agmatine injection. Among BMPs, BMP2/4/7, agmatine significantly increased only the expression of BMP2 known to reduce M1 macrophage under inflammatory status. These results sug- gest that agmatine reduces impairment after spinal cord injury through modulating the macrophage phenotype.

Metabolic influence on macrophage polarization and pathogenesis

( Bikash Thapa ),( Keunwook Lee ) 생화학분자생물학회(구 한국생화학분자생물학회) 2019 BMB Reports Vol.52 No.6

Macrophages play an essential role not only in mediating the first line of defense but also in maintaining tissue homeostasis. In response to extrinsic factors derived from a given tissue, macrophages activate different functional programs to produce polarized macrophage populations responsible for inducing inflammation against microbes, removing cellular debris, and tissue repair. However, accumulating evidence has revealed that macrophage polarization is pivotal in the pathophysiology of metabolic syndromes and cancer, as well as in infectious and autoimmune diseases. Recent advances in transcriptomic and metabolomic studies have highlighted the link between metabolic rewiring of macrophages and their functional plasticity. These findings imply that metabolic adaption to their surrounding microenvironment instructs activation of macrophages with functionally distinct phenotypes, which in turn probably leads to the pathogenesis of a wide spectrum of diseases. In this review, we have introduced emerging concepts in immunometabolism with focus on the impact on functional activation of macrophages. Furthermore, we have discussed the implication of macrophage plasticity on the pathogenesis of metabolic syndromes and cancer, and how the disease microenvironment manipulates macrophage metabolism with regard to the pathophysiology. [BMB Reports 2019; 52(6): 360-372]

백서의 폐포 및 복강 대식세포의 세포독성에 대한 연구

이홍렬(Hong Lyeol Lee),김세규(Se Kyu Kim),장준(Joon Chang),김성규(Sung Kyu Kim),이원영(Won Young Lee),조철호(Chul Ho Cho) 대한내과학회 1992 대한내과학회지 Vol.43 No.4

N/A Background: Mechanisms involved in host resistance against malignant tumors have been found to be mediated mainly by cellular effectors. These effector mechanisms include activated macrophages, killer T-cells, natural killer cells and antibody-dependent cellmediated cytotoxicity. Activated macrophages can sup- press DNA synthesis of tumor cells and kill tumor cells in a selective but nonspecific fashiori in vitro. Lipopolysaccharides (LPS) can enhance the cytotoxicity of macrophages and the lipid A which is produced by mild acid hydrolysis of LPS, is responsible for the LPS effect on macrophages. The mechanism by which LPS modifies macrophage physiology is not known, but it is suggested that it acts at the level of the macrophage plasma membrane. LPS may make macrophages tumoricidal by altering the membrane composition or by transmitting a necessary signal from the membrane to the vacuolar system. Methods: We isolated alveolar and peritoneal macrophages by bronchoalveolar and peritoneal lavage in rats. Rat sarcoma cell line (XC) was used as the target cell. As recommended commonly we controlled the effector cell: target cell ratio at 10:1, Three groups were divided as folows; no LPS added group, LPS 5μg/ml added group and LPS 10μg/ml added group, We focused the assay of cytotoxicity on the cytolysis rather than cytostasis by measuring the [3H] thymidine released and calculated the percentage specific cytalysis, By this experiment, we examined the stimulation effect of LPS an the macrophage cytotoxicity and compared the cytotoxicity between alveolar and peritoneal macrophages. Results: The cytotoxicity of alveolar and peritoneal macrophages was significantly enhanced when stimulated both with 5μg/ml and 10μg/ml of LPS. There was no significant difference in macrophage cytotoxicity between two groups each stimulated with 5μg/ml and 10μg/ml of LPS. We could not observe the significant difference in cytotoxicity between the alveolar and peritoneal macrophages, Conclusion: Cytotoxicity was significantly enhanced by stimulation of LPS, in hoth alveolar and peritoneal macrophages but there was no significant difference in cytotoxicity enhancement between 5μg/ml and 10μg/ ml of LPS. Also there was no significant difference in cytotoxicity between alveolar and peritoneal macrophages.

Innate immune crosstalk in asthmatic airways: Innate lymphoid cells coordinate polarization of lung macrophages

Kim, Jihyun,Chang, Yuna,Bae, Boram,Sohn, Kyoung-Hee,Cho, Sang-Heon,Chung, Doo Hyun,Kang, Hye Ryun,Kim, Hye Young Elsevier 2019 The journal of allergy and clinical immunology Vol.143 No.5

<P><B>Background</B></P> <P>Recent studies have emphasized the role of innate lymphoid cells (ILCs) in the development of asthma. The involvement of group 2 innate lymphoid cells (ILC2s) in asthma is well studied: however, the participation of other types of ILCs in the development of asthma remains unclear.</P> <P><B>Objective</B></P> <P>This study aims to understand the role of various ILCs in patients with asthma, especially their effect on macrophage polarization.</P> <P><B>Methods</B></P> <P>Each subset of ILCs and macrophages in induced sputum from 51 steroid-naive patients with asthma and 18 healthy donors was analyzed by using flow cytometry. Alveolar macrophages (AM) were sorted and cocultured with each subset of ILCs to determine whether the polarization of macrophages could be regulated by ILCs.</P> <P><B>Results</B></P> <P>In addition to ILC2s, numbers of group 1 innate lymphoid cells (ILC1s) and group 3 innate lymphoid cells (ILC3s) were increased in induced sputum from asthmatic patients when compared with those in healthy control subjects. The dominance of macrophages in induced sputum was more prominent in asthmatic patients than in healthy control subjects. A positive correlation between numbers of ILC2s and numbers of M2 macrophages and those of ILC1s/ILC3s and M1 macrophages was observed. Coculture of ILC2s with AMs induced expression of M2 macrophage–related genes, whereas coculture of ILC1s and ILC3s with AMs induced expression of M1 macrophage–related genes through cytokine secretion, as well as cell-cell contact. According to the inflammatory signature, patients with eosinophilic asthma have more ILC2s and M2 macrophages, and those with noneosinophilic asthma have an M1 macrophage–dominant profile.</P> <P><B>Conclusion</B></P> <P>A different subset of ILCs regulates macrophage polarization, contributing to developing the distinct phenotype of asthma.</P> <P><B>Graphical abstract</B></P> <P>[DISPLAY OMISSION]</P>

Induction of heme oxygenase-1 with dietary quercetin reduces obesity-induced hepatic inflammation through macrophage phenotype switching

Kim, Chu-Sook,Choi, Hye-Seon,Joe, Yeonsoo,Chung, Hun Taeg,Yu, Rina The Korean Nutrition Society 2016 Nutrition Research and Practice Vol.10 No.6

BACKGROUND/OBJECTIVES: Obesity-induced steatohepatitis accompanied by activated hepatic macrophages/Kupffer cells facilitates the progression of hepatic fibrinogenesis and exacerbates metabolic derangements such as insulin resistance. Heme oxyganase-1 (HO-1) modulates tissue macrophage phenotypes and thus is implicated in protection against inflammatory diseases. Here, we show that the flavonoid quercetin reduces obesity-induced hepatic inflammation by inducing HO-1, which promotes hepatic macrophage polarization in favor of the M2 phenotype. MATERIALS/METHODS: Male C57BL/6 mice were fed a regular diet (RD), high-fat diet (HFD), or HFD supplemented with quercetin (HF+Que, 0.5g/kg diet) for nine weeks. Inflammatory cytokines and macrophage markers were measured by ELISA and RT-PCR, respectively. HO-1 protein was measured by Western blotting. RESULTS: Quercetin supplementation decreased levels of inflammatory cytokines ($TNF{\alpha}$, IL-6) and increased that of the anti-inflammatory cytokine (IL-10) in the livers of HFD-fed mice. This was accompanied by upregulation of M2 macrophage marker genes (Arg-1, Mrc1) and downregulation of M1 macrophage marker genes ($TNF{\alpha}$, NOS2). In co-cultures of lipid-laden hepatocytes and macrophages, treatment with quercetin induced HO-1 in the macrophages, markedly suppressed expression of M1 macrophage marker genes, and reduced release of MCP-1. Moreover, these effects of quercetin were blunted by an HO-1 inhibitor and deficiency of nuclear factor E2-related factor 2 (Nrf2) in macrophages. CONCLUSIONS: Quercetin reduces obesity-induced hepatic inflammation by promoting macrophage phenotype switching. The beneficial effect of quercetin is associated with Nrf2-mediated HO-1 induction. Quercetin may be a useful dietary factor for protecting against obesity-induced steatohepatitis.

Induction of heme oxygenase-1 with dietary quercetin reduces obesity-induced hepatic inflammation through macrophage phenotype switching

김추숙,최혜선,조연수,정헌택,유리나 한국영양학회 2016 Nutrition Research and Practice Vol.10 No.6

BACKGROUND/OBJECTIVES: Obesity-induced steatohepatitis accompanied by activated hepatic macrophages/Kupffer cells facilitates the progression of hepatic fibrinogenesis and exacerbates metabolic derangements such as insulin resistance. Heme oxyganase-1 (HO-1) modulates tissue macrophage phenotypes and thus is implicated in protection against inflammatory diseases. Here, we show that the flavonoid quercetin reduces obesity-induced hepatic inflammation by inducing HO-1, which promotes hepatic macrophage polarization in favor of the M2 phenotype. MATERIALS/METHODS: Male C57BL/6 mice were fed a regular diet (RD), high-fat diet (HFD), or HFD supplemented with quercetin (HF+Que, 0.5g/kg diet) for nine weeks. Inflammatory cytokines and macrophage markers were measured by ELISA and RT-PCR, respectively. HO-1 protein was measured by Western blotting. RESULTS: Quercetin supplementation decreased levels of inflammatory cytokines (TNFα, IL-6) and increased that of the anti-inflammatory cytokine (IL-10) in the livers of HFD-fed mice. This was accompanied by upregulation of M2 macrophage marker genes (Arg-1, Mrc1) and downregulation of M1 macrophage marker genes (TNFα, NOS2). In co-cultures of lipid-laden hepatocytes and macrophages, treatment with quercetin induced HO-1 in the macrophages, markedly suppressed expression of M1 macrophage marker genes, and reduced release of MCP-1. Moreover, these effects of quercetin were blunted by an HO-1 inhibitor and deficiency of nuclear factor E2-related factor 2 (Nrf2) in macrophages. CONCLUSIONS: Quercetin reduces obesity-induced hepatic inflammation by promoting macrophage phenotype switching. The beneficial effect of quercetin is associated with Nrf2-mediated HO-1 induction. Quercetin may be a useful dietary factor for protecting against obesity-induced steatohepatitis.

CCL2 Mediates Neuron–Macrophage Interactions to Drive Proregenerative Macrophage Activation Following Preconditioning Injury

Kwon, Min Jung,Shin, Hae Young,Cui, Yuexian,Kim, Hyosil,Thi, Anh Hong Le,Choi, Jun Young,Kim, Eun Young,Hwang, Dong Hoon,Kim, Byung Gon Society for Neuroscience 2015 The Journal of neuroscience Vol.35 No.48

<P>CNS neurons in adult mammals do not spontaneously regenerate axons after spinal cord injury. Preconditioning peripheral nerve injury allows the dorsal root ganglia (DRG) sensory axons to regenerate beyond the injury site by promoting expression of regeneration-associated genes. We have previously shown that peripheral nerve injury increases the number of macrophages in the DRGs and that the activated macrophages are critical to the enhancement of intrinsic regeneration capacity. The present study identifies a novel chemokine signal mediated by CCL2 that links regenerating neurons with proregenerative macrophage activation. Neutralization of CCL2 abolished the neurite outgrowth activity of conditioned medium obtained from neuron-macrophage cocultures treated with cAMP. The neuron-macrophage interactions that produced outgrowth-promoting conditioned medium required CCL2 in neurons and CCR2/CCR4 in macrophages. The conditioning effects were abolished in CCL2-deficient mice at 3 and 7 d after sciatic nerve injury, but CCL2 was dispensable for the initial growth response and upregulation of GAP-43 at the 1 d time point. Intraganglionic injection of CCL2 mimicked conditioning injury by mobilizing M2-like macrophages. Finally, overexpression of CCL2 in DRGs promoted sensory axon regeneration in a rat spinal cord injury model without harmful side effects. Our data suggest that CCL2-mediated neuron-macrophage interaction plays a critical role for amplification and maintenance of enhanced regenerative capacity by preconditioning peripheral nerve injury. Manipulation of chemokine signaling mediating neuron-macrophage interactions may represent a novel therapeutic approach to promote axon regeneration after CNS injury.</P>