Diagnostic significance of the histopathology of bone marrow macrophages in forensic autopsies

Forensic pathologists often encounter autopsies that require an assessment of antemortem general conditions (e.g., infection, metabolic disorders). To establish evaluation clues for such cases, we quantitatively examined macrophages and the general pathology of bone marrow in samples from 180 forensic autopsy cases of decedents with various conditions. Hematoxylin-eosin staining, Berlin blue staining, and immunostainings for CD163, CD138, and CD61 were performed. We determined the numbers per field (density) of total macrophages, swollen macrophages, macrophages with hemophagocytosis, and hemosiderin-laden macrophages. Each density was standardized by identifying its ratio to the total number of macrophages. The decedents' background data (cause of death, other pathological findings, postmortem interval, antemortem symptoms, and presence of resuscitation) were extracted. No correlations were found between the postmortem interval and the other decedent data, indicating that these data are not affected by postmortem changes. In the group in which inflammatory disease was the cause of death, there were significant elevations in the ratio of the swollen macrophage density to total macrophages. Significantly higher ratios of the density of swollen and hemophagocytic macrophages were observed in the group in which conditions with a prolonged agonal period were the cause of death. The group with a return of spontaneous circulation to resuscitation showed a significantly higher ratio of macrophage density with hemophagocytosis. This study provides the first statistical analysis focused on bone marrow histopathology in forensic autopsies. The results will be useful for elucidating causes of death and agonal-period conditions.     

Development and function of tissue resident macrophages in mice

Macrophages are important for tissue development, homeostasis as well as immune response upon injury or infection. For a long time they were only seen as one uniform group of phagocytes with a common origin and similar functions. However, this view has been challenged in the last decade and revealed a complex diversity of tissue resident macrophages. Here, we want to present the current view on macrophage development and tissue specification and we will discuss differences as well as common patterns between heterogeneous macrophage subpopulations.     

Inhibitory effect of phospholipase A2 isolated from Crotalus durissus terrificus venom on macrophage function

Recent work demonstrated that crotoxin, the main toxin of Crotalus durissus terrificus venom, inhibits macrophage spreading and phagocytic activities. The crotoxin molecule is composed of two subunits, an acidic non-toxic and non-enzymatic polypeptide named crotapotin and a weakly toxic basic phospholipase A₂ (PLA₂). In the present work, the active subunit responsible for the inhibitory effect of crotoxin on macrophage function was investigated. Peritoneal macrophages harvested from naive rats were used. Crotapotin (2.12, 3.75, or 8.37 nM/ml), added for 2 h to the medium of peritoneal cell incubation, did not modify the spreading and phagocytic activities of these cells. On the other hand, the PLA₂ (1.43, 2.86, or 6.43 nM/ml) subunit caused a significant reduction (30, 33, and 35%, respectively) of the spreading activity. The PLA₂ also inhibited the phagocytosis of opsonised zymosan, opsonised sheep erythrocytes, and Candida albicans, indicating that this inhibitory effect is not dependent on the type of receptor involved in the phagocytosis process. The inhibitory effect of PLA₂ was not due to loss of cell membrane integrity, since macrophage viability was higher than 95%. These findings indicate that the inhibitory effect of crotoxin on macrophage spreading and phagocytic activities is caused by the phospholipase A₂ subunit.     

Reciprocal regulation of the nitric oxide synthase/arginase balance in mouse bone marrow-derived macrophages by TH 1 and TH 2 cytokines

Activation with lipopolysaccharide induces macrophages to produce the enzymes arginase and nitric oxide (NO) synthase. Both enzymes use as a substrate the amino acid L-arginine, which can be either hydrolyzed by arginase to urea and ornithine or oxidized by NO synthase to NO and citrulline. NO is important in the bactericidal and cytotoxic activities of macrophages. An equivalent functional role of arginase and its products is not known. We tested the induction of arginase in bone marrow-derived macrophages by endogenous mediators that are known to induce NO synthase, such as interferon-γ (IFN-γ), or suppress the induction of this enzyme, such as interleukin (IL)-4, IL-10, and prostaglandin E₂ (PGE₂). We find that PGE₂ and the TH2 cytokines IL-4 and IL-10 are potent inducers of arginase. In contrast, the TH 1 cytokine IFN-γ does not induce arginase. Simultaneous application of both types of mediators leads to reduced induction of both arginase and NO synthase. Exposure of macrophage cultures to inducers of NO synthase exhausts their ability to respond subsequently to inducers of arginase. Conversely, exposure of the cells to inducers of arginase exhausts their ability to respond subsequently to inducers of NO synthase. The results are consistent with a competition of both enzymes for their substrate, L-arginine, with a reciprocal inhibition in the induction of both enzymes, or a combination of both phenomena. The enzymes NO synthase and arginase appear to define two alternate functional states of macrophages, induced by TH 1 and TH 2 cytokines, respectively.     

小檗碱对兔颈动脉粥样硬化中内膜增生和巨噬细胞趋化作用的影响

目的观察小檗碱对家兔颈动脉粥样硬化的内膜中膜比和巨噬细胞变化的影响。方法24只雄性日本大耳白兔随机分为3个试验组,正常组每天肌肉注射生理盐水,普通饲料喂养,对照组高脂喂养,1周后行颈动脉内膜空气干燥术并每日肌肉注射生理盐水,小檗碱干预组高脂喂养,1周后行颈动脉内膜空气干燥术并肌肉注射小檗碱,5周时取手术侧的颈动脉做弹力纤维染色,计算内膜中膜比;巨噬细胞免疫组化检测巨噬细胞在颈动脉粥样硬化病变中的变化,计算巨噬细胞的阳性率。结果对照组内膜厚度明显增加,中膜萎缩变薄,经计算I/M为1.20±0.007,小檗碱组的I/M为0.65±0.008。两组间有显著差异(P<0.01);巨噬细胞免疫组化染色对照组内膜下和中膜有大量巨噬细胞,小檗碱干预组内膜和中膜下也可以见有巨噬细胞沉积,通过计算巨噬细胞阳性率,小檗碱干预组的巨噬细胞阳性率明显小于对照组(P<0.01)。结论小檗碱可以降低家兔颈动脉粥样硬化中的血管内膜厚度、减少粥样斑块中的巨噬细胞数目,从而干预颈动脉粥样硬化的形成。     

328. Mechanismen der langzeitakzeptanz allogener rattenlebertransplantate: makrophagenaustausch und suppressorzellaktivierung

Summary After orthotopic rat liver transplantation in the fully allogeneic BN (RT-1n) to LEW (RT-11) combination, the phenomenon of spontaneous tolerance of donor antigen occurs. We demonstrate two different immune mechanisms that may account for this process. Using adoptive transfer assays we show the presence of donor-specific T-suppressor lymphocytes in the spleens of long-term surviving liver graft, recipients. These cells prolong - adoptively transferred into irradiated syngeneic hosts — the survival of donor-specific (BN) but not third-party (DA) renal allografts (I00 days vs 1I days in control groups). Secondly, we demonstrate the replacement of Kupffer cells in the graft by recipient macrophages using polymorphic monoclonal antibodies in an immunoperoxidase technique. This may contribute to graft adaptation and thus to long-term graft acceptance.     

球囊成形术后氯沙坦对兔血管内膜增殖及巨噬细胞表达的影响

目的　通过观察AT1受体的特异性拮抗剂氯沙坦 (Losartan)对兔腹主动脉球囊成形术后内膜增殖和巨噬细胞表达的影响 ,探讨氯沙坦治疗血管再狭窄的机制。方法　采用内皮剥脱加高脂饮食 (含 1 5 %胆固醇 )喂养 8周制作兔腹主动脉粥样硬化膜型 ,然后对狭窄部位行球囊成形术。术后氯沙坦组给予氯沙坦 10mg·kg-1·day-1口服 ,对照组喂生理盐水。 4周后取腹主动脉行组织形态学观察 ,用免疫组织化学方法分析巨噬细胞。结果　对照组内膜厚度 /中膜厚度比值为 0 65± 0 17,氯沙坦组则减少为0 44± 0 11(P <0 0 1) ,对照组内膜面积 /中膜面积比值为 0 74± 0 16,氯沙坦组则减少为 0 5 4± 0 13(P <0 0 1)。氯沙坦组新生内膜中巨噬细胞数亦较对照组显著减少 (P <0 0 1)。结论　氯沙坦通过抑制AngⅡ的作用进而抑制巨噬细胞的浸润 ,抑制炎症反应 ,从而减轻再狭窄     

银耳制剂对小鼠移植性肿瘤预防及其机理的实验研究

小鼠在移植肿瘤细胞前,注射银耳制剂,显示出银耳对荷腹水型或荷实体瘤小鼠肿瘤的生长有明显的抑制作用.正常小鼠给银耳制剂后,其腹腔巨噬细胞(M￠)数量与功能、形态有明显变化,此M￠可吞噬和杀伤肿瘤细胞.     

The role of macrophages in demyelination

Experimental allergic encephalomyelitis (EAE) is an autoimmune inflammatory disease of the central nervous system (CNS). In particular in the CsA-induced chronic relapsing form (CREAE), pronounced demyelination occurs, in temporal association with relapses. It is still a matter of discussion which cell type ultimately is responsible for the actual process of demyelination. Macrophages, cytotoxic T lymphocytes and also astrocytes are possible candidates. In this short overview, the role of macrophages in the pathogenesis of EAE is discussed. It is shown that in particular, newly recruited macrophages play a crucial role in the generation of clinical signs. Possible mechanisms by which macrophages are involved in the pathogenesis of demyelinating diseases are presented.