Surface Of Peritoneal Macrophages Research Articles

Experimental Trichinellosis in rats: Peritoneal macrophage activity

The influence of Trichinella spiralis infection on macrophage activity in rats during the first 28 days of infection was examined by measuring the production of NO and IL-6, as well as the expression of mannose receptor on the surface of peritoneal macrophages. During the course of a dynamic shift in the 3 life-cycle stages of the parasite, intermittent variations in NO production were observed but ended with increased values that coincided with the highest values for IL-6 release in the final, muscle phase of infection. No change in mannose receptor expression was observed during the course of infection. These results confirm that the Trichinella spiralis infection provokes changes in macrophage activity that could influence not only the course of the parasitic disease but also the overall immune status of the host.

Adjuvant effect of γ-inulin is mediated by C3 fragments deposited on antigen-presenting cells

The adjuvant effect of gamma-inulin, a strong activator of the alternative complement pathway, is well-known, but its exact mechanism is not revealed yet. Here, we show that macrophages, isolated from the peritoneal cavity of gamma-inulin-injected mice and used as antigen-presenting cells, enhance the proliferation of antigen-specific T-cells up to 2.5-fold when compared with macrophages of non-treated animals. This effect is abrogated by the presence of anti-C3 F(ab')2 fragments and by prior decomplementation of the donor animals with CVF. It is demonstrated that treatment of mice with the adjuvant results in deposition of C3-fragments onto the surface of peritoneal macrophages, as does in vitro incubation of the cells with gamma-inulin in the presence of fresh autologous serum. Prior incubation of macrophages with gamma-inulin plus serum in vitro enhances subsequent C3 production. Because it has been shown earlier that CR1/2 expressed on activated T-cells and interacting with covalently bound C3-fragments plays an important role in the augmentation of the adaptive response, our present results reveal a mechanism that contributes to the adjuvant effect of gamma-inulin and point to a further link between innate and adaptive immunity.

Carcinoembryonic antigen binding proteins on elicited peritoneal macrophages.

Carcinoembryonic antigen (CEA), a member of the immunoglobulin supergene family, is recognized by a specific 80-kd receptor on Kupffer cells and alveolar macrophages. Peritoneal macrophages were harvested from male Sprague-Dawley rats 72 h after intraperitoneal injection with thioglycolate broth and from unstimulated animals. The elicited peritoneal macrophages had a much greater capacity to endocytosed CEA than the cells isolated from unstimulated animals. They took up CEA in a concentration- and time-dependent manner. Cellular uptake was temperature and colchicine sensitive indicating internalization of CEA. The process was saturable and could be specifically inhibited by CEA and nonspecific crossreacting antigen (NCA), a member of the CEA supergene family. Two binding proteins of approximately 35 and 80 kd were isolated from the surface of the peritoneal macrophages by affinity chromatography on a CEA-Sepharose column. Using CEA coupled to sulfo-succininidyl-2-[p-azidosalicyl amide]-1,3'-dithiopropionate (SASD), a photoactivatable crosslinker, two binding proteins of 55 and 80 kd were identified on the surface of live peritoneal macrophages. The specificity of CEA endocytosis and binding proteins exhibited by the elicited peritoneal cells are similar to that previously reported for Kupffer cells and alveolar macrophages.

Activation of murine peritoneal macrophages by intraperitoneal administration of a traditional chinese herbal medicine, xiao-chai-hu-tang (Japanese name: Shosaiko-To)

Macrophage activation by a traditional Chinese herbal medicine, xiao-chai-hu-tang (Japanese name: shosaiko-to), was investigated. Intraperitoneal (i.p.) administration of shosaiko-to into (BALB/c × DBA/2)F 1 mice resulted in marked activation of macrophages with respect to phagocytic and lysosomal enzyme activities (acid phosphatase and N- acetyl-β- D- glucosaminidase ) compared with the control. The maximal responses were induced by an i.p. injection of 3 mg shosaiko-to 4 days previously. Enhanced activities induced by shosaiko-to were also seen in C3H/HeJ mice, which is a non-responder strain to bacterial lipopolysaccharide (LPS). Significant macrophage accumulation in the peritoneal cavity and increased lysosomal enzyme activities were observed in mice injected with shosaiko-to. Shosaiko-to exhibited significant sytostasis-inducing activity. In addition, the administration of shosaiko-to led to a moderate expression of Ia antigen on the surface of peritoneal macrophages. These results suggest that shosaiko-to is a potent macrophage activator.

誘導マクロファージの Ia 抗原に関する研究 : 第 2 報活性化 T 細胞接種による Ia 抗原量の変化について

It is known that quantity of Ia antigen on the surface of macrophages increases in the presence of stimulated T cells in cell culture. Murine spleen T cells and thymus T cells were stimulated with Concanavalin A. The T cells were injected in murine peritoneum, and then the quantity of Ia antigen on the surface of peritoneal macrophages was measured using the method of enzyme-linked immunosorbent assay (ELISA). The results were as follows ; 1. In the presence of stimulated thymus T cells, the quantity of Ia antigen did not increase on the surface of macrophages. 2. In the presence of stimulated spleen T cells, the quantity of Ia antigen increased on the surface of macrophages. 3. On the surface of resident macrophages, the quantity of Ia antigen did not increase after injection of stimulated T cells. 4. On the surface of Pepton-induced macrophages, the quantity of Ia antigen increased rapidly, but soon decreased, after injection of stimulated spleen T cells. 5. On the surface of LPS-induced macrophages, the quantity of Ia antigen decreased slightly, and then increased gradually, after injection of stimulated spleen T cells. These results showed that stimulated spleen T cells had the capacity for increasing the quantity of Ia antigen on the surface of macrophage in mice. The expression of Ia antigen on the surface of macrophages differed with the conditions of macrophages.

Glycoprotein receptors to wheat germ agglutinin on the surface of murine macrophages.

Peritoneal macrophages elicited in various ways exhibited marked wheat germ agglutinin (WGA)-dependent cytolytic activity against MM46 cells [nearly 100% lysis at effector to target ratio (E/T) = 40], while macrophage cell lines J774.1 and WEHI-3 cells showed less activity (-25% at E/T = 0.63-2.5) and no cytolysis (E/T = 40), respectively. Studies on the kinetics of WGA-binding showed that peritoneal macrophages had an association constant (Ka) of 0.09-0.18 X 10(6) M-1 and the number of their binding sites (N) was 2-7.7 X 10(7)/cell, while the Ka and N values of J774.1 and WEHI-3 cells were 1.2 X 10(6) M-1 and 4.1 X 10(7)/cell and 0.6 X 10(6) M-1 and 5.4 X 10(7)/cell, respectively. On the other hand, cytolytic target MM46 cells showed values of 0.27 X 10(6) M-1 and 1.8 X 10(8)/cell. Lower affinities and moderate numbers of binding sites coincided with the efficient WGA-dependent cytolytic activities of the effector cells. The surface of peritoneal macrophages and J774.1 and MM46 cells was iodinated, the cells lysed, the lysates fractionated on a WGA-Sepharose column and the WGA-binding surface-glycoproteins in GlcNAc-specific fractions of the eluate were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). Peritoneal macrophages gave three major bands of 153 kilo dalton (kDa), 140 and 90 kDa proteins, whereas J774.1 and MM46 cells both gave 6 bands of very similar molecular weights and intensities that were quite unlike those of peritoneal macrophages.(ABSTRACT TRUNCATED AT 250 WORDS)

Cytochemical localization of beta-galactosidase in resident and inflammatory peritoneal macrophages from C57BL mice.

A cytochemical method for the detection of beta-galactosidase (beta-Gase) in mouse peritoneal macrophages was used to study the ultrastructural localization of this enzyme in these cells. It was found that the reaction product for beta-Gase was localized in the perinuclear cisternae, the endoplasmic reticulum, the Golgi complex, lysosomes, vesicles and on the cell surface of peritoneal macrophages from untreated C57BL mice. When examined by X-ray microanalysis the crystalline reaction product was found to contain bromine, an element present in the indolyl substrate which was used to identify beta-Gase. Injection of Proprionibacterium acnes (P. acnes) intraperitoneally or BCG intravenously caused a visible loss in beta-Gase from all the organelles and from the cell surface of the macrophages.

Role of macrophages in the transmission of dengue virus-induced suppressor signal to a subpopulation of T lymphocytes

In dengue type 2 virus (DV)-infected mice, the virus-specific immunosuppression is mediated by a two-step mechanism: (1) induction of T suppressor cells (Ts 1) by the virus to produce a suppressor factor (SF) which (2) stimulates another subpopulation of T cells (Ts 2) to produce prostaglandin which finally mediates suppression. SF suppresses DV-specific IgM plaque-forming cells (PFC) in the spleen cells sensitized in vivo or in vitro, as detected by Jerne's haemolytic plaque technique. The present study enabled us to investigate the role of an intermediary cell in transmission of the suppressor signal from Ts 1 to Ts 2. It was observed that SF was adsorbed on the surface of peritoneal macrophages. Live macrophages adsorbed SF, retrieved it from that adsorbed on heat-killed macrophages and presented it to the target cells. Heat-killed macrophages adsorbed SF to the same extent as live ones, but could not present it to the target cells by themselves. The target cells of SF were unprimed splenic T lymphocytes. SF suppressed DV-specific PFC in syngeneic spleen cells and was adsorbed on syngeneic macrophages, but not on those from allogenic animals. The findings described here show that the presence of live macrophages is obligatory for transmission of the suppressor signal to the target Ts 2.

Method to prove investigation of particles by macrophages with light microscopy.

Research on phagocytosis is often hampered by the inability to distinguish whether (opsonized) particles have been ingested by phagocytes or are only attached to the surface of these cells. Treatment of the cells after phagocytes to remove all extracellular particles makes it possible to evaluate phagocytosis with certainty by light microscopy. Opsonized erythrocytes attached to the macrophage surface are usually removed by hypotonic lysis. The present report describes the advantages of the use of lysostaphin to lyse Staphylococcus aureus and of xylene, chloroform or dioxane to dissolve polystyrene latex beads on the surface of peritoneal macrophages and embryonic fibroblasts. This procedure facilitates differentiation between professional and facultative phagocytes.

DELAYED HYPERSENSITIVITY IN THE GUINEA PIG IMMUNIZED WITH KILLED TUBERCLE BACILLI IN ADJUVANT

Guinea pigs were immunized with 2.4 mg of killed tubercle bacilli in adjuvant and Tuberculin Purified Protein Derivative (PPD) was used as test antigen. Peritoneal exudate cell migration inhibition was more marked one year than one month after sensitization. Macrophage cytophilic antibody (detected by rosette formation with antigen coated sheep red cells) was found on the surface of peritoneal macrophages and in sera at one year, but was barely detectable at one month. One year sera as final media in migration chambers inhibited migration of normal peritoneal cells, so did one month sera. Incubation of normal cells in one year sera followed by washing led to migration inhibition. Sera of guinea pigs passively sensitized with the immune sera had haemagglutinating activity against PPD coated sheep red cells. In some of the animals injected with one year sera cytophilic antibody was demonstrable on the surface of peritoneal cells and the migration of these cells was slightly inhibited by antigen. The degree of migration inhibition did not correlate with haemagglutinating and haemolysing antibody titres.