The differentiation of monocytes into macrophages,epithelioid cells,and multinucleated giant cells in subcutaneous granulomas

The morphological changes occurring in monocytes during their differentiation into macrophages, epithelioid cells, Langhans-type giant cells, and foreign-body-type giant cells were investigated in foreign-body granulomas induced by subcutaneous implantation of pieces of Melinex plastic. Analysis based on Adams's (1974) criteria for discrimination between the several types of cell of the monocyte line, showed that each type has a characteristic type of granule. Primary and secondary granules, numerous in the Golgi area of monocytes were generally found close to the cell membrane and decreased in number in maturing macrophages. This was accompanied by an increase in the number of microtubules. Mature macrophages show numerous characteristic macrophage granules, which are round (average diameter: 280 nm) and have a halo between the limiting membrane and granular matrix. Mature epithelioid cells have characteristic epithelioid cell granules, and multinucleated giant cells a heterogenous population of granules. Fusing macrophages generally have their Golgi areas facing each other, and also show a reduced thickness of the cell coat. The morphology of the multinucleated giant cell is closely related to the number of nuclei present. In Langhans-type giant cells, which generally have two to ten nuclei, a giant centrosphere with numerous aggregated centrioles is found. In transition forms between Langhans-type and foreign-body-type giant cells, which generally contain 10--30 nuclei, the centrioles show less aggregation. In the foreign-body-type giant cells, which generally have more than 30 nuclei, centrioles are virtually absent and never aggregated. These differences between the Langhans-type giant cells, the foreign-body-type giant cells, and the transition forms, support our previous finding that Langhans-type giant cells are the precursors of foreign-body-type giant cells.     

Comparative study on peroxidatic activity in inflammatory cells on cutaneous and peritoneal implants

Summary Inflammatory reactions were evoked by simultaneous implantation of pieces of Melinex plastic in the subcutaneous tissues of the dorsum and in the peritoneal cavity of rats. The cellular composition of the Melinex-adherent cells and their peroxidatic (PO) activity were investigated in relation to the duration of implantation. Several striking differences were found between the subcutaneous and peritoneal implants. On the 7th and 14th days, multinucleated giant cells were abundantly present on the subcutaneous implants, whereas they were relatively rare on the peritoneal implants. The subcutaneous implants bore no mast cells and only a few eosinophilic granulocytes, but both types of cell were observed frequently on the peritoneal implants.Macrophages and multinucleated giant cells on the subcutaneous implants show PO activity only in the granules or are PO negative. On the peritoneal implants three types of macrophages can be distinguished: exudate macrophages which have PO activity restricted to granules or are PO-negative; macrophages with PO activity in granules and both the rough endoplasmic reticulum (RER) and nuclear envelope; and resident macrophages with PO activity only in the RER and nuclear envelope. In addition, two types of multinucleated giant cells are found, one with and the other without PO activity in the RER and nuclear envelope. Multinucleated giant cells with PO activity in the RER and nuclear envelope as well as exudate macrophages with PO activity in the RER and nuclear envelope were mainly found 32 h and 3 days after implantation of the Melinex in the peritoneal cavity. These findings are discussed in the light of current knowledge of the PO activity in macrophages and multinucleated giant cells. It is concluded that the appearance of PO activity in the RER and nuclear envelope of exudate macrophages and multinucleated giant cells is in all probability a transient phenomenon, and that there is no objective evidence to support the opinion that exudate macrophages with PO activity in the RER and nuclear envelope are transitional cells between exudate and resident macrophages.     

The differentiation of monocytes into macrophages,epithelioid cells,and multinucleated giant cells in subcutaneous granulomas

Summary The peroxidatic (PO) activity of monocytes differentiating into macrophages, epithelioid cells, and multinucleated giant cells in subcutaneous granulomas was investigated with three different media for the demonstration of PO activity. Irrespective of the stage of differentiation, these cells did not show PO activity in the rough endoplasmic reticulum (RER) or nuclear envelope. In addition, it was found that the morphologically characteristic types of granule of the various cells of the monocyte line (the primary granules and secondary granules of monocytes, the macrophage granules, and the epithelioid cell granules), all have distinct cytochemical characteristics.Monocytes lose their primary and secondary granules during differentiation into mature macrophages. Simultaneously, the granules of both types become elongated and the secondary granules lose their halo. In contrast to monocytes, mature macrophages may contain a few microperoxisomes. During the differentiation of macrophages into epithelioid cells or multinucleated giant cells there is an increase in the number of microperoxisomes.     

Differentiation of granuloma cells (epithelioid cells and multinucleated giant cells): a morphometric analysis. Investigations using the model of experimental autoimmune (anti-TBM) tubulo-interstitial nephritis

Morphometric analysis disclosed distinct differences between blood monocytes, tissue monocytes (i.e. immature macrophages), epithelioid cells and multinucleated giant cells as well as phagocytic macrophages (i.e. mature macrophages) in the granuloma model of autoimmune (anti-TBM) tubulo-interstitial nephritis. The numerical density of lysosomes decreased slightly in tissue monocytes compared with blood monocytes but showed a pronounced increase during the formation of epithelioid cells. The lysosomal compartments of epithelioid cells and multinucleated giant cells resembled each other very closely, but the giant cells obviously produced additional lysosomes of small diameter (80-120 nm). Phagocytic macrophages displayed a total numerical density of lysosomes similar to that of tissue monocytes but the mean diameter of the lysosomes was markedly greater. Thus the volume density of lysosomes was highest in phagocytic macrophages. The blood monocytes exhibited the smallest lysosomal compartment. In tissue monocytes, epithelioid cells, and multinucleated giant cells the volume densities of the lysosomes were greater than in blood monocytes and remained relatively constant because the increase in numerical density was counterbalanced by a decrease in mean granule diameter. We found only minor differences in mitochondrial volume densities among the five cell populations. The shape of the mitochondria, however, changed steadily from short rotational ellipsoids in the blood monocytes to rather elongated and slender bodies in the multinucleated giant cells. The results suggest that epithelioid cells and multinucleated giant cells are active cells which may contribute by their specific performances, to the immunologic microenvironment of the granuloma.     

Morphological features of milky spots of the rat omentum in inflammation

The omental milky sports of the rat have been examined with the light and electron microscope after intraperitoneal stimulation by particulate coal and zymosan. No positive correlation between cell level proliferation in milky spots and alteration of their size has been found. The increase in size of milky spots is due to the inflow of cells from the blood and bone marrow. The development in milky spots of a great number of macrophages "tubercles" and multinucleated giant cells in the experimental conditions may respond to inflammation. The administration of zymosan result in the influx of lymphocytes forming lymphatic follicle-like structure. Despite the absence of germinative centres, the appearance of a great number of lymphoblasts and plasma cells in the milky spots provides the evidence of the active antibody production aimed at immunological protection of abdominal cavity.     

Computerized video time-lapse (CVTL) analysis of the fate of giant cells produced by X-irradiating EJ30 human bladder carcinoma cells

This study was designed to examine the viability and proliferation of uninucleated and multinucleated giant cells formed after 6 Gy X irradiation. The pedigrees of 102 individual EJ30 giant cells present 5 days after irradiation were analyzed from time-lapse movies captured over 6.3 days from 100 fields (100x). Pedigree analysis enabled us to study the proliferation of giant cells. The average starting size (area) of the giant cells (14500 +/- 9100 microm(2)) was approximately 25 times larger than the normal-sized cells observed after irradiation (560 +/- 200 microm(2)). From a total of 76 pedigrees of uninucleated giant cells, 42 had giant cells that either died or were arrested, while 34 divided at least once and produced progeny that divided again (five three times and three four times) before the progeny died or were arrested. Twenty-four pedigrees contained progeny that were lost from observation after dividing at least once. While most progeny continued to have giant cell morphology, two uninucleated giant cells ultimately produced progeny that contained two normal-sized cells. From a total of 26 multinucleated giant cells, only three divided. Two divided only once, but one produced progeny that divided two times. In all, 37 out of 102 giant cells divided at least once; eight of these divided four or five times with two of these pedigrees ultimately producing two normal-sized daughter cells. These results suggest that a small fraction of giant cells might be potentially clonogenic.     

Inhibition of macrophage-lymphocyte interaction by cytochalasin B during antigen recognition by T lymphocytes.

The effects of cytochalasin B on functional and physical macrophage-lymphocyte interaction have been examined. Cytochalasin B, an inhibitor of a variety of membrane activities blocks antigen-dependent bindings of immune lymphocytes to macrophages and antigen-triggered lymphocytes proliferation if added at the initiation of culture. Cytochalasin B becomes progressively less inhibitory if addition is delayed by increasing intervals from the onset of culture. Under these conditions neither antigen handling by macrophages nor the proliferative response of lymphocytes to PHA is inhibited by cytochalasin B. These data are interpreted to suggest that cytochalasin B inhibits antigen-specific macrophage-lymphocyte interaction either by inhibition of an initial antigen-independent phase of macrophage-lymphocyte interaction or by interfering with a lymphocyte membrane event necessary for the interaction of the antigen-specific lymphocyte receptor with the macrophage-bound antigenic signal.     

Natural killer cell-dependent mycobacteriostatic and mycobactericidal activity in human macrophages

Host defense mechanisms against Mycobacterium avium complex (MAC) are poorly understood. Recent evidence suggests the role of NK cells in the host defense against some intracellular pathogens. We investigated whether NK cells play a role in MAC infection. IL-2-activated human NK cells were incubated with human monocyte-derived macrophages either before or after infection with MAC. Macrophages were lysed 3 and 5 days after infection for quantitation of viable intracellular organisms. Although no killing was observed by nonstimulated macrophages, exposure to IL-2-treated NK cells for 24 h before infection induced macrophage to kill 70 +/- 8% of intracellular MAC by 3 days, and 81% +/- 4% in 5 days (p less than 0.01 for both compared with control). Killing was not blocked by incubation with anti-TNF antibody (Ab) or anti-IFN-gamma Ab. Similarly, incubation of macrophages for 24 h with supernatant obtained from IL-2 activated NK cells was associated with 74 +/- 4% killing of intracellular MAC in 3 days and 81 +/- 6% in 5 days (p less than 0.01 for both compared with control). However, the supernatant-mediated activation was partially blocked by anti-TNF Ab (46 +/- 6%; p less than 0.05) but not by anti-IFN gamma Ab. When infected macrophages were incubated with NK cells 24 h after infection for 48 h, they killed 54 +/- 3% of intracellular M. avium in 3 days and 73 +/- 5% in 5 days (p less than 0.02 for both compared with control). This effect was also not blocked by either anti-TNF or anti-IFN gamma Ab. These results suggest that activated NK cells may have an important role in the intracellular killing of MAC and that the NK-mediated activation of macrophages is in part mediated by TNF.     

Regeneration of changes produced in rabbit tracheal epithelium by saline lavage of the airways

The authors studied the course of the repair of changes induced in the rabbit tracheal epithelium by saline lavage of the airways. The tracheal epithelium was examined 2, 24, 48 and 72 hours after treatment. Saline lavage stimulated the goblet cells to instantaneous discharge of their secretion. 2 hours after treatment 98 +/- 3% of the goblet cells were completely exhausted and had degenerated. Repair of the changes began 24 hours after lavage and was associated with massive differentiation of new goblet cells resulting in hyperplasia of the mucus-secreting elements with formation of endoepithelial mucous glands. The most pronounced injury to the ciliated cells was apparent 2 hours after lavage, then the degree of alteration of these cells gradually decreased. Saline lavage markedly impaired the ciliary border. The mean number of kinocilia per micron2 fell to 1.5 +/- 0.3. In subsequent phases the number of kinocilia rose gradually to 7.5 +/- 0.5/micron2. This value was still significantly lower (P less than 0.005) compared with controls. The first signs of impairment of the self-cleaning ability of the epithelium were recorded 2 hours after lavage. The most pronounced disturbances of the mucus flow were observed after 24 hours. At the end of the experimental period small clumps of condensed mucus and rather numerous bacteria were still present in the area of the ciliary border.     

Effector mechanism of host resistance in murine giardiasis: specific IgG and IgA cell-mediated toxicity

The role of specific serum and milk anti-Giardia muris antibodies in mediation of host-effector responses to this enteric pathogen is unknown. We have investigated antibody-dependent cell-parasite interactions, potentially important as mediators of protection against infection at the mucosal surface. Elicited mouse peritoneal neutrophils and macrophages were incubated with G. muris trophozoites in the presence of either serum or milk antibodies, and their adherence and phagocytosis of the parasites were assessed. The percentage of trophozoites with adherent neutrophils increased significantly in the presence of heat-inactivated immune rabbit serum (93.5% +/- 6.5) and immune mouse milk (54.4% +/- 11.3) and their purified IgG (35.2% +/- 9.7) and secretory IgA fractions (48.0% +/- 12.3) when compared with incubation in RPMI-10% FCS (21.7% +/- 13.9). Similarly, macrophage adherence to trophozoites increased from 49.7% +/- 14.3 in medium alone to respective values of 92.8% +/- 7.1 in immune rabbit serum and 77.3% +/- 11.0 in immune milk. Phagocytosis of parasites by macrophages also was enhanced after incubation in immune rabbit serum (48.0% +/- 4.0) and immune mouse milk (35.0% +/- 5.0) when compared with the percentage of trophozoites ingested when cells and parasites were incubated in RPMI-10% FCS (3.3% +/- 3.0). Transmission electron microscopy showed ingestion of parasites by neutrophils or macrophages after 15 min of incubation. Morphologic evidence of intracellular parasite injury was observed at 6 hr. A decrease in parasite infectivity also resulted when trophozoites were incubated with neutrophils or macrophages and a source of antibodies, and intragastrically fed to weanling mice. These observations show that both antitrophozoite IgG, secretory IgA, and mouse phagocytic cells interact in vitro to promote parasite clearance. Because both the humoral and cellular components of this system are found intraluminally in the small intestine and in milk, they may represent a biologically relevant protective response against giardiasis.     

Lack of interaction of circulating T cells with phytohemagglutinin in bacillary positive untreated lepromatous leprosy patients--identification of subpopulation of lymphocytes by shifts in electrophoretic mobility.

Incubation of human peripheral blood lymphocytes from normal healthy subjects with phytohamagglutinin (PHA), causes the reduction of the surface charge of a subpopulation of T cells by 1363 +/- 242 e.s.u./cm2. The affected subpopulation was predominantly the high charge-bearing cells identifiable with early (10 min) rosette-forming cells with sheep erythrocytes. Purified lymphocytes obtained from untreated bacillary-positive, lepromatous leprosy patients contained high charge-bearing T lymphocyte subpopulation. However, incubation with PHA did not result in the shift of electrophoretic mobility of these cells, suggesting the absence of interacting sites for the mitogen on the surface of these cells. The absence of mitogen-interacting sites is not an inherent trait of leprosy patients; the surface charge of lymphocytes from Dapsone-treated bacillary-negative subjects was reduced upon incubation with PHA. A close correlation was found between the number of cells whose charge alters on incubation with PHA and the transformation index obtained with this mitogen.     

The alveolar macrophage: Its capacity to act as an accessory cell in mitogen-stimulated proliferation of guinea pig lymphocytes

The capacity of the alveolar macrophage to act as an accessory cell in PHA-induced lymphocyte proliferation was investigated and compared with that of the peritoneal and peritoneal exudate macrophages in guinea pigs. When lymph node cells were co-cultured with autologous lung cells recovered by airway lavage, the proliferative response to PHA was greatly enhanced over that of lymph node cells alone. In the presence of peritoneal cells or peritoneal exudate (glycogen-induced) cells, the PHA response was intermediate between that of lymph node cells alone and lymph node cells cultured with lung cells. Experiments using purified macrophages (≥98%) as accessory cells demonstrated that the difference observed between lung and peritoneal accessory cells was due to differences in macrophage function. Furthermore, when lymph node cells were cultured in the upper chamber of a double-chambered Marbrook apparatus, PHA-induced proliferation was enhanced only when lung and not peritoneal macrophages were present in the lower chamber. Additional experiments showed that this difference (1) was not an artifact of the thymidine incorporation assay to measure proliferation; (2) was not affected by changing the macrophage-lymphocyte ratio; and (3) was not simply a trephocytic or growth promoting effect of macrophages which could be replaced by 2-mercaptoethanol.These findings show that macrophages from different sources differ in their abilities to act as accessory cells in PHA-induced lymphocyte proliferation. Alveolar macrophages appear to have an enhanced capacity compared to unstimulated and stimulated peritoneal macrophages in this function. At least part of this difference may be due to a difference in the elaboration of soluble factor (s) by macrophages.     

Identification of osteoclasts and their differentiation from mononuclear phagocytes by enzyme histochemistry

A double staining method is presented which allows the enzyme histochemical differentiation between osteoclasts (mono- and multinucleated forms) and mononuclear phagocytes (macrophages, multinucleated inflammatory giant cells). Osteoclasts are characterized by a strong acid phosphatase activity whereas macrophages and inflammatory giant cells show a variable non-specific esterase activity. The described method may be useful in studying the osteoclast origin and the extraosseous distribution of these cells.     

Human lymphocyte subpopulations: giant SRBC rosettes.

Human thymus-derived lymphocytes have the ability to form rosettes with sheep red blood cells (SRBC) in vitro. In the investigation of rosettes of peripheral blood lymphocytes of 10 normal subjects, the number of SRBC adhering to the lymphocyte in each of 100 rosettes was assessed. The percentage of rosettes with SRBC greater than or equal to 36 per rosette was only 1.2 +/- 0.5. These were defined as giant SRBC rosettes. Peripheral blood lymphocytes were stimulated in vitro by four mitogens: sodium periodate, neuraminidase plus galactose oxidase, pokeweed mitogen, and concanavalin A. The lymphocytes were then cultured at 37 degrees C. The giant rosette-forming lymphocytes became significantly increased 4 to 24 hr after stimulation, prior to the appearance of lymphoblasts or increased incorporation of tritiated thymidine. The giant rosettes were not caused by the hemagglutinating properties of pokeweed mitogen and concanavalin A that were adsorbed on the lymphocyte surfaces. This was shown by the fact that, on removal of the receptors by trypsinization, they were regenerated on culture in vitro in the absence of the mitogens. It was concluded that giant SRBC rosettes constituted a marker for some of the activated lymphocytes. Their appearance was independent of the increase in size of the cells or of DNA synthesis. These receptors were intrinsic to lymphocytes and not caused by mitogens adsorbed on their surfaces.     

Fine needle aspiration cytology of fibroadenoma with multinucleated stromal giant cells. A review of cases in a six-year period

OBJECTIVE: To describe the fine needle aspiration cytology findings of fibroadenoma with multinucleated stromal giant cells, with histologic correlation. STUDY DESIGN: The author reviewed the cytologic findings of two cases of fibroadenoma with multinucleated stromal giant cells from the file of Pamela Youde Nethersole Eastern Hospital in a six-year period from 1995 to the end of 2000. The diagnosis was confirmed by histologic examination of the lumpectomy specimens. RESULTS: The two cases had similar cytologic findings. The direct smears contained cohesive clusters of bland-looking ductal cells arranged in a "staghorn" pattern. Numerous naked nuclei were also seen in the background. Also, there were occasional multinucleated giant cells in isolation. These giant cells contained 5-10 randomly arranged, round to oval nuclei, fine chromatin and sometimes distinct nucleoli. The cytoplasm was abundant and pale staining, and the cell border was ill defined. Associated epithelioid histiocytes and foamy macrophages were not seen. Histologic examination of the lumpectomy specimens showed architectural features of fibroadenoma with pericanalicular and intracanalicular patterns. In addition, scattered multinucleated giant cells with focal degenerative change were noted in the tumor stroma. Their stromal nature was confirmed by immunohistochemical study. CONCLUSION: Multinucleated stromal giant cells are rarely identified in fine needle aspiration biopsies of fibroadenoma. Recognition of this peculiar finding may help to avoid misdiagnosis of other, more sinister conditions, such as phyllodes tumor and metaplastic carcinoma.     

Identification of osteoclasts and their differentiation from mononuclear phagocytes by enzyme histochemistry

Summary A double staining method is presented which allows the enzyme histochemical differentiation between osteoclasts (mono- and multinucleated forms) and mononuclear phagocytes (macrophages, multinucleated inflammatory giant cells). Osteoclasts are characterized by a strong acid phosphatase activity whereas macrophages and inflammatory giant cells show a variable non-specific esterase activity. The described method may be useful in studying the osteoclast origin and the extraosseus distribution of these cells.Supported by Deutsche Forschungsgemeinschaft, SFB 244,A1     

巨噬细胞、内皮细胞对高密度脂蛋白的氧化修饰

为了探讨高密度脂蛋白 (HDL)在体内发生氧化修饰的部位及机制 ,分别观察了培养人动脉平滑肌细胞 (SMC)、动脉内皮细胞 (EC)及巨噬细胞 (MΦ)与HDL共同温育过程中 ,HDL的琼脂糖电泳相对迁移率 (REM)、硫代巴比妥酸反应物质 (TBARS)以及溶血卵磷脂 卵磷脂 (LPC PC)值等氧化指标的变化 .结果发现 ,HDL与 3种细胞温育 12h时 ,HDL几乎不发生氧化修饰 ,而HDL与MΦ和EC温育 2 4h后 ,其REM、TBARS、LPC PC值均显著上升 (p <0 0 1) ;而HDL与SMC温育后 ,其REM、TBARS值无显著改变 ,LPC PC值增加 (p <0 0 1) .结果提示 ,活体内HDL可能主要在动脉壁的内皮细胞及巨噬细胞的作用下发生氧化修饰     

The involvement of omentum and its milky spots in the dynamics of peritoneal macrophages

The investigation has been carried out on stimulated and unstimulated peritoneal cavities of rats. China ink and Corynebacterium parvum were injected i.p. both as peritoneal stimuli and markers. Omenta were picked up at time intervals beginning with 10 min and up to seven days after the i.p. injection. The light and electronmicroscopic investigation showed after 10-30 minutes labeled macrophages stuck as monolayers on some peritoneal areas corresponding to the milky spots which developed in size and number. Days after the i.p. injection the labeled macrophages were found deeper in the milky spots. After the fourth day they appeared in the regional lymph nodes. The milky spots contained also large lymphocytes and plasma cells. The results suggest that milky spots are not only places of resident macrophages development and release in the peritoneal cavity but also their exit pathways. Therefore the omentum leads the traffic of peritoneal macrophages. The developed milky spots play also the role of lymphoid structures providing grounds for macrophage-lymphocyte contacts.