Paracoccidioidomycosis in silica-treated rats

An attempt is made to evaluate the importance of macrophages in the early local reaction to infection caused by the fungus Paracoccidioides brasiliensis. With this purpose, an attempt was made to impair macrophagic activity in the site of the fungal inoculum, by means of previous injections of silica. Twenty male rats were intraperitoneally infected with yeast cells of the fungus. Ten of the rats were injected intraperitoneally with silica particles twice, namely 24 and 4 h before the infection. All the animals were killed 4 wk after fungal inoculum. The results showed that the disease was aggravated in rats pre-treated with silica, clearly demonstrated by the 100% metastatic pulmonary involvement, in contrast with only 40% observed in the non-treated controls.     

Gpnmb在小鼠M1和M2型骨髓源性巨噬细胞中表达的差异

目的了解小鼠M1和M2型骨髓源性巨噬细胞（ BMMφs）中非转移性黑色素瘤糖蛋白b（ Gpnmb）表达的差异。方法原代培养小鼠BMMφs,免疫荧光染色F4/80和流式细胞仪检测CD11b鉴定巨噬细胞;用IFN-γ和LPS诱导BMMφs向M1型分化,用IL-4诱导向M2型分化。实时荧光定量PCR检测M1型巨噬细胞标志物（ TNF-α、iNOS）、M2型巨噬细胞标志物（ MMR、Arg-1）和Gpn-mb的mRNA表达;免疫荧光双染色、Western印迹、流式细胞仪检测Gpnmb与MMR的蛋白表达。结果（1）免疫荧光染色结果示BMMφs中F4/80高表达;流式细胞仪检测结果示BMMφs中有（92．7±6．1）％细胞表达CD11b,提示BMMφs培养成功;（2）相对于未分化的M0型BMMφs,TNF-α、iNOS mRNA在M1型BMMφs中高表达（P均＜0．01）,而MMR、Arg-1 mRNA在M2型BMMφs中高表达（P均＜0．01）,提示原代M1、M2型BMMφs分化成功;（3）M2型BMMφs 的Gpnmb mRNA和蛋白表达均较M0型和M1型BMMφs显著增高（P均＜0．01）;免疫荧光双染色及流式细胞仪结果显示,BMMφs中Gpnmb与MMR共表达,在M2型BMMφs中MMR阳性BMMφs有（83．2±9．7）％表达Gpnmb。结论 M2型BMMφs的Gpnmb表达较M1型BMMφs显著增高,提示Gpnmb可能作为鉴别M1、M2型巨噬细胞的标志物,在巨噬细胞的表型分化中起作用。     

M1和M2型巨噬细胞表型的比较分析

通过对M1和M2型巨噬细胞表型相关指标的比较分析,评价各鉴定巨噬细胞类型的表型指标及其意义。按常规方法以IFN-γ及LPS将骨髓来源巨噬细胞诱导成M1型巨噬细胞,以IL-4诱导出M2型巨噬细胞。分别以RT-PCR和酶活性定量方法检测精氨酸代谢相关酶的表达和活性;以ELISA检测IL-12和IL-10的分泌;以FACS检测巨噬细胞膜分子的表达。结果显示:M1型巨噬细胞诱导性一氧化氮合酶(inducible nitric oxide synthase,iNOS)表达和活性水平较未刺激组明显升高,IL-12产生显著增加,CD16/32表达上调;而M2型巨噬细胞I型精氨酸酶(arginase 1,Arg-1)的表达水平和酶活性较未刺激巨噬细胞显著提高,IL-10分泌轻度增加,并且表达高水平的CD206和DECTIN-1。表型比较分析结果表明,iN-OS表达和活性、IL-12的分泌和膜蛋白CD16/32可用于鉴定M1型巨噬细胞,而Arg-1、CD206和DECTIN-1是鉴定M2型巨噬细胞较为理想的表型指标。     

Macrophage heterogeneity in human fetal tissue. Fetal macrophages.

The immunophenotype of the macrophage population in human fetal tissue was studied, using a panel of monoclonal antibodies against cells of the macrophage/monocyte lineage. Using a double-labelling technique two main populations were observed in tissue from 14 weeks of estimated gestational age (EGA); EBM11+ DR+ and EBM11+ DR- cells of which a small proportion were also RFD7+. Most macrophages were negative with 3.9, an antibody specific for the adhesion molecule P150.95 and LP9 which is specific for a lysosomal enzyme. The exception to this was a small population of positive cells in the thymus. Small numbers of 3.9+ cells were also infrequently observed in tissue at and above 17 weeks of EGA, while occasional RFD9+ cells were only observed in most tissues, before this time. The higher percentage of macrophages were DR+ DQ- DP-, with a few DQ+ cells appearing at 15 weeks of EGA. In the thymus, DQ+ cells outnumbered DP+ cells especially in the medulla. These results indicate the heterogeneous and immature nature of the fetal macrophage population and point to the importance of age, tissue-specific factors and probable immune mediators in macrophage differentiation.     

Factors of the extrinsic pathway of blood coagulation in fetal placental macrophages

Term human placentae were examined to identify and localize the factors contributing to extravascular fibrin formation. In addition to blood vessels, components of the extrinsic coagulation cascade were also demonstrated in intracellular localization. These cells in double and treble labeling systems expressed macrophage marker antigens, recognized by DAKO antimacrophage, RFD7, Amersham antimacrophage, and KiM7 monoclonal antibodies and showed positivity for alpha-naphthyl-acetate-esterase (ANAE). The fact that factors of the extrinsic coagulation system can be demonstrated in fetal macrophages of the chorionic stroma suggests that their role is not restricted to cellular defense and phagocytosis, but they may be involved in extravascular intraplacental fibrin formation as well.     

A new monoclonal antibody, PM-2K, specifically recognizes tissue macrophages but not blood monocytes

A new monoclonal antibody, PM-2K, was raised against 24-h cultured human peritoneal macrophages. Immunohistochemically, PM-2K recognized most tissue macrophages in lymphoreticular organs such as the thymus, spleen, lymph node, and tonsil. Kupffer cells of the liver, alveolar macrophages, and macrophages in the interstitial tissue of the kidney, pancreas, and many other organs were also positively labelled. On the other hand, PM-2K failed to recognize blood monocytes, freshly isolated peritoneal macrophages, microglial cells, osteoclasts, and dendritic cells such as Langerhans cells, interdigitating cells, and follicular dendritic cells. In various pathological conditions, PM-2K labelled a wide variety of proliferating macrophages. Reaction products of PM-2K were observed by immunoelectron microscopy on the cytoplasmic membrane of cultured peritoneal macrophages. The molecular weight of the antigen recognized by PM-2K was determined to be 150 kD by Western blotting. As no cells other than macrophages were reactive with PM-2K, this antibody is considered to be very useful not only in the investigation of macrophage differentiation and maturation, but also in the diagnosis of various proliferative disorders of macrophages.     

Effect of degree of malignancy on sensitivity of cells to cytostatic action of resident syrian hamster peritoneal macrophages

Laboratory of Antitumor Immunity, Institute of Carcinogenesis, All-Union Oncologic Scientific Center, Academy of Medical Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR N. N. Trapenzikov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 110, No. 7, pp. 83–85, July, 1990.     

Sensitive Detection of Two Igg Fc Receptors of Mouse Macrophages by Chemiluminescence Analysis

Abstract

Luminol-enhanced chemiluminescence assay was used to detect the surface expression and the consequent activation of receptors (FcRI and FcRII) of murine macrophages (Møs). When murine IgG_2a was used for the specific detection of FcRI and IgG_2b for FcRII, a newly established procedure enabled us to detect the activation of each receptor with as few as 3×10⁵ Møs. Briefly, TNP-SRBC coated with monoclonal IgG_2a or IgG_2b antibodies directed to TNP (sensitized SRBC) were used as reagent, in the presence of 1× 10^?5M luminol, and the emission was measured with a liquid scintillation counter.

When results obtained by chemiluminescence counting were compared to the results obtained by the rosette formation by adding the same SRBC reagent to peritoneal Møs obtained after ip injection of Listeria, fortified chemiluminescence counting allowed us to obtain a more definite answer about the activation of each receptor.

Under the conditions established, the specific activation of FcRI was obtained by the addition of rIFNαrA/D to the resident Møs in vitro and the specific activation of spleen Mø FcRII by iv injection of IAP (Immunosuppresive acidic protein) into mice. These two results supported the independence of the two receptors detected by the assay.     

β2-microglobulin triggers NLRP3 inflammasome activation in tumor-associated macrophages to promote multiple myeloma progression

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M2-polarized tumor-associated macrophages are associated with poor prognoses resulting from accelerated lymphangiogenesis in lung adenocarcinoma

OBJECTIVES:

Tumor-associated macrophages have been implicated in promoting tumor growth, progression and metastasis. However, the activated phenotype (M1 or M2) of tumor-associated macrophages remains unknown in solid tumors. Therefore, this study examined the density and prognostic significance of M2-polarized tumor-associated macrophages in lung adenocarcinoma.

METHODS:

Tumor specimens from 65 lung adenocarcinoma patients were assessed by ELISA for Th1/Th2 cytokine concentrations. The activated phenotype (M1 or M2) of tumor-associated macrophages was determined utilizing immunofluorescence staining. Additionally, to evaluate lymphangiogenesis, peritumoral lymphatic microvessel density was measured using D2-40. The correlation between tumor-associated macrophage subtype and overall patient survival was analyzed using the Kaplan-Meier method and compared using the log-rank test.

RESULTS:

A shift toward Th2 cytokine expression was detected within lung adenocarcinoma microenvironments. Approximately 79.71±16.27% of tumor-associated macrophages were M2 polarized; the remaining 20.35±5.31% were M1 polarized. The infiltration of M2-polarized macrophages was significantly associated with P-TNM staging and lymph node metastasis. The peritumoral lymphatic microvessel density was significantly higher in the high M2-polarized tumor-associated macrophage group than in the low M2-polarized tumor-associated macrophage group. A significant difference in overall patient survival was detected not only between patients with tumors with high and low macrophage counts but also between patients with tumors with high and low counts of M2-polarized macrophages.

CONCLUSION:

Tumor-associated macrophages in lung adenocarcinoma have an M2-polarized subtype and are associated with poor prognoses, perhaps resulting from accelerated lymphangiogenesis and lymph node metastasis.     

Divergent roles of IFNs in the sensitization to endotoxin shock by lactate dehydrogenase-elevating virus

The effect of mouse infection with lactate dehydrogenase-elevating virus (LDV), a usually non-pathogenic virus, on concomitant bacterial endotoxin shock was analyzed, in terms of lethality and cytokine production. A strong enhancement of susceptibility to the shock was observed in mice acutely infected with this virus. It correlated with a sharp increase of tumor necrosis factor and leukemia inhibitory factor production and was controlled by the mouse genetic background. The viral infection led to an imbalance in the cytokine response to LPS, with an enhancement of pro-inflammatory cytokines, including IL-18 and IFN-gamma and a delayed secretion of anti-inflammatory IL-10 that could result in exacerbated macrophage activation. Enhanced IFN-gamma production was involved in the virus-induced susceptibility to shock. In sharp contrast with other viral infections, IFN-alpha/beta diminished IFN-gamma production and the resulting increased response to LPS in LDV-infected animals.     

Conditions of preparation and mechanism of action of macrophagal pyrogen

Activation of mononuclear phagocytes by staphylococciin vitro leads to the formation of an endogenous pyrogen. The macrophagal pyrogen does not possess specific pyrogenic specificity, and on intracisternal injection sensitivity to it is enhanced by more than 100 times compared with that observed after intravenous injection. An even sharper increase in sensitivity to pyrogen was observed in animals after elevation of the body level of cyclic AMP as a result of preliminary injection of theophylline.Department of General Pathology, Institute of Experimental Medicine, Academy of Medical Sciences of the USSR, Leningrad. (Presented by Academician of the Academy of Medical Sciences of the USSR P. M. Veselkin.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 89, No. 3, pp. 278–281, March, 1980.