Host-parasite interactions with peritoneal macrophages of mice and rats in vitro and in vivo.

This paper deals with the intracellular multiplication of mycobacteria in peritoneal macrophages from mice and rats immunized with tubercle bacilli or pretreated with Triton WR 1339. If unstimulated macrophages were used, almost unrestricted multiplication of mycobacteria was observed in macrophages from both vaccinated and pretreated hosts after infection of the cells in vitro. Only when the infection of the cells was perfored in the peritoneal cavity of vaccinated hosts did the macrophages display a high degree of inhibition. This striking difference in the behavior of macrophages infected in vitro and in vivo is explained by the local inflammation caused by the intraperitoneal infection, which leads to an influx of T-cell mediators. When macrophages from hosts pretreated with Triton WR 1339 were used, inhibition of the multiplication of mycobacteria within cells infected in vitro or in vivo was very slight, though this compound displayed a marked protective effect in the host. Addition of streptomycin to the culture medium caused a strong inhibition of intracellular mycobacteria even in small concentrations; there was no difference between normal and "immune" macrophages. When rats were infected with virulent tubercle bacilli, they were initially fully susceptible to the infection but showed rapid onset of a strong immune response.     

淋巴细胞缺陷对内毒素血症小鼠腹腔巨噬细胞活化的影响

目的观察淋巴细胞缺陷对内毒素血症小鼠腹腔巨噬细胞活化的影响。方法采用腹腔注射脂多糖(LPS)建立Balb/c小鼠和T、B细胞缺陷的重症联合免疫缺陷(SCID)小鼠内毒素血症模型,ELISA检测2种小鼠腹腔灌洗液TNF-α和IL-10水平,实时荧光定量PCR检测腹腔巨噬细胞(peritoneal macrophage,PMa)TNF-α、IL-10及丝裂原蛋白激酶磷酸酶-1(mito-gen-activated protein kinase phosphatase-1,MKP-1)mRNA表达;ELISA检测Balb/c及SCID小鼠PMa体外刺激后细胞因子分泌情况。结果 LPS注射后1 h,SCID小鼠腹腔灌洗液TNF-α水平高于Balb/c小鼠,注射后3 h,IL-10水平低于Balb/c小鼠;LPS注射前及注射后,SCID小鼠PMa TNF-αmRNA表达高于Balb/c小鼠PMa,IL-10 mRNA表达低于Balb/c小鼠PMa;体外实验LPS刺激下,SCID小鼠PMa较Balb/c小鼠PMa分泌更多的TNF-α,IL-10的分泌却偏少。LPS注射前及注射后,Balb/c小鼠PMa MKP-1 mRNA表达均明显高于SCID小鼠。结论淋巴细胞缺陷导致内毒素血症小鼠腹腔巨噬细胞活性的增加,淋巴细胞抑制巨噬细胞的活化并可能调控其发育及成熟;MKP-1表达的减少可能是淋巴细胞缺陷导致腹腔巨噬细胞活性增加的分子机制之一。     

The interaction of DMSO-induced murine Friend leukaemia erythroblasts with cultured syngeneic mouse peritoneal macrophages: an in vitro model for the study of ineffective erythropoiesis.

The interaction between dimethyl sulphoxide-induced Friend leukaemia erythroblasts (IFLE) and cultured syngeneic mouse peritoneal macrophages (PM phi) was assessed radiochemically as well as by light and electron microscopy. The data revealed that a small number of the IFLE either became attached to or were ingested by the PM phi and that the interacting IFLE were usually well differentiated. Interactions of a similar nature were also seen when IFLE were co-cultured with macrophages derived from mouse bone marrow. The extent of interaction between IFLE and PM phi provides a useful system in which mechanisms underlying ineffective erythropoiesis can be defined.     

Effect of zinc on peritoneal macrophages in vitro.

Rat peritoneal macrophages incubated in a medium with 10% autologous serum showed 50% inhibition of O2 consumption by 0.4 mM zinc only when activated by yeast. Zinc ions had no effect on resting macrophages. The inhibitory efficiency of zinc depended on the ratio of yeast particles to macrophages; with more activated cells, the inhibition was less. The multiple-shaped intact macrophages changed into rounded cells in the presence of 0.1 and 0.5 mM Zn2+ as shown by photomicrography and scanning electron microscopy. When the incubation medium was replaced by zinc-free medium, cells recovered and became irregularly shaped. The reversibility of zinc effect on cell morphology corresponded with uptake and release of zinc by macrophages. The presence of serum in the medium interfered with the magnitude of zinc uptake by macrophages. We conclude that besides already reported effects of zinc on mast cells, platelets, and granulocytes, zinc ions also inhibit some functions of macrophages.     

Electron microscopic study on the interaction between normal guinea pig peritoneal macrophages and Coxiella burnetii.

An electron microscopic study was conducted to explore the interaction between normal guinea pig peritoneal macrophages and phase I and II Coxeilla burnetii previously treated with either normal or immune serum. A comparison was made on the efficiency of phagocytosis and subsequent killing of rickettsiae by macrophages. Both phases of rickettsiae previously treated with normal serum multiplied within phagosomes after phagocytosis with resultant destruction of macrophages. In contrast, suspending rickettsiae in immune serum rendered them more susceptible to phagocytosis and potentiated their destruction within macrophages.     

The interaction between macrophages and suppressor cells in immunological maturation in mice

The immunological immaturity at birth characteristic of species such as the mouse has no certain explanation. Here Bertie Argyris suggests that in mice it is a consequence of high suppressor-cell activity and an absence of macrophage activity. Gradual alterations in the balance of these activities account for the gradual emergence of immunocompetence.     

Cytotoxic action of lymphocytes on peritoneal macrophages of mice with contact dermatitis caused by dinitrochlorobenzene

Contact dermatitis was produced in BALB/c mice by percutaneous application of a solution of dinitrochlorobenzene (DNCB). On the 20th day of the experiment, when the skin test with DNCB was positive, peritoneal macrophages were taken from DNCB-sensitized and intact mice and cultured in flat-bottomed tubes for 48h. A suspension of lymphocytes from peripheral lymph nodes was added to them. When incubated together with cells of DNCB-sensitized mice the lymphocytes had a cytotoxic action on the target cells (peritoneal macrophages), as shown by staining with Trypan Blue after 48 h. When cells from intact mice were incubated with lymphocytes, lysis of the macrophages was not observed.Laboratory of General Pathological Anatomy and Laboratory of Immunomorphology and Cytochemistry, Institute of Human Morphology, Academy of Medical Sciences of the USSR. Laboratory of Experimental Pathology, Central Institute of Tuberculosis, Ministry of Health of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. I. Strukov.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 83, No. 3, pp. 277–280, March, 1977.     

The production of ceroid by mouse peritoneal macrophages in vitro.

Murine resident peritoneal macrophages accumulated lipid droplets and subsequently insoluble, ceroid-like material when cultured in vitro in a medium containing 33% fetal calf serum. At least some of this insoluble lipid was membrane-bound and by light microscopy it often appeared as ''rings'' with a hollow centre. It is suggested that the production of ceroid may be the consequence of the uptake of lipids from the extracellular medium and the activity of the macrophage''s membrane-bound oxidative microbicidal mechanisms. The results indicate that macrophages are capable of rendering lipids insoluble, supporting the suggestion that this might occur in the atherosclerotic plaque.     

In vitro interaction between normal cynolmolgus monkey alveolar macrophages and Legionnaires disease bacteria.

The interaction between normal cynomolgus monkey alveolar macrophages and Legionnaires disease bacteria was studied by transmission electron microscopy. After ingestion of Legionnaires disease bacteria, the organisms replicated within macrophages and destroyed the phagocytic cell.     

Replication of porcine parvovirus in peripheral blood lymphocytes, monocytes, and peritoneal macrophages.

Porcine peripheral blood lymphocytes (PBL), peripheral blood monocytes, and peritoneal macrophages were examined for their ability to support porcine parvovirus (PPV) replication. The cell cultures were infected with the NADL-2 strain of PPV at 0.1 multiplicity of infection. PBL cultures were stimulated with the following phytomitogens: phytohemagglutinin M, concanavalin A, and pokeweed mitogen. Unstimulated PBL cultures infected with PPV and uninfected PBL stimulated with phytomitogens served as controls. All cultures were examined daily for PPV-specific immunofluorescence and hemagglutinin. PPV replicated in PBL cultures stimulated with all phytomitogens. Both viral hemagglutinin in culture fluids and nuclear immunofluorescence in cells were detected. In contrast, unstimulated PBL did not support viral replication; however, PPV antigen was detected in the cytoplasm. PPV persisted in unstimulated PBL for 21 days (duration of the experiment) without replication, but replicated each time with the addition of phytohemagglutinin M at 0, 3, 7, 14, and 21 days after infection. Uninfected PBL stimulated with phytomitogens lacked both viral hemagglutinin and immunofluorescence. Simultaneous detection of lymphocyte surface marker and viral antigens in pokeweed mitogen-stimulated PBL revealed that both T and non-T cells (B and null cells) are able to support PPV replication. Peripheral blood monocytes and peritoneal macrophages phagocytized PPV but did not support virus replication.     

Role of macrophages in tumour immunity: I. Co-operation between macrophages and lymphoid cells in syngeneic tumour immunity

Macrophages from DBA/2 mice hyperimmunized with irradiated syngeneic L5178Y or SL2 lymphoma cells inhibited the growth of these cells in vitro and killed them within 48 hours of culture. The reaction was immunologically specific. Non-immune macrophages could be rendered capable of immunologically-specific growth inhibition of target cells in vitro (arming) (a) by direct contact of hyperimmune lymphoid cells and macrophages, and (b) by incubating macrophages with a specific macrophage-arming factor (SMAF) derived by incubating spleen cells from singly immunized mice with irradiated lymphoma cells. Singly immunized spleen cells did not arm macrophages by direct cell-to-cell contact. A temporal relationship was seen between the presence of immune macrophages in hyperimmunized mice and the ability of the spleen cells to arm normal macrophages. Furthermore, macrophages could be armed in vivo by a single i.p. injection of hyper immune spleen cells. The presence of arming factors cytophilic for macrophages, but not for cells of non-macrophage origin, is briefly discussed.     

Cell interactions in the initial contact between cultured melanoma cells and syngeneic macrophages.

Thioglycolate-induces peritoneal macrophages from melanoma-bearing mice (immune macrophages) or from control mice (control macrophages) were cultured with syngeneic melanoma cells (P51) to determine the surface characteristics of the effector cells during interaction and destruction of the target cells. After a short culture period (3 hours), immune macrophages had extensive connections via filopodia and ruffled membranes to the surfaces of the melanoma cells; control macrophages did not exhibit the same behavior. A dense region in the cytoplasm immediately beneath the macrophage plasmalemma was observed at the point of contact with the target tumor cell. With longer periods of culture (24 hours), effector cells began phagocytosis of the target cells; immune macrophages, however, had more fine filopodial connections and were more cytostatic than were controls. These observations indicate that one of the initial mechanisms of tumor cell destruction was contact-induced lysis, with phagocytosis playing a minor part.     

Interaction between Coxiella burnetii and guinea pig peritoneal macrophages.

The phagocytosis and subsequent degradation of phase I and II Coxiella burnetii by macrophages obtained from immune and nonimmune guinea pigs were compared. Phase I rickettsiae were more resistant to phagocytosis than were phase II organisms. There was no significant difference in the percentage of phagocytosis of either phase of rickettsiae by macrophages from immune or nonimmune animals. After ingestion, phase I and II organisms pretreated with normal serum multiplied and destroyed normal macrophages as well as macrophages obtained from guinea pigs immunized with phase II rickettsiae. In contrast, only phase I organisms were degraded by macrophages from phase I-immunized animals in the presence of normal serum. Immune serum rendered rickettsiae more susceptible to phagocytosis and also potentiated the destruction of organisms by all types of macrophages. The specificity of macrophages from phase I animals to degrade only phase I rickettsiae was demonstrated by the ability of Rickettsia rickettsii to replicate in these macrophages.     

Antigenic differences between macrophages and lymphocytes

Antisera against guinea-pig lymphocytes and guinea-pig macrophages were prepared by injecting rabbits with purified blood lymphocytes or peritoneal macrophages. Both antisera contained antibodies of at least two different specificities, one reacting with an antigen common to lymphocytes and macrophages and one specific for either lymphocytes or macrophages.

Opsonic adherence and phagocytosis of sensitized red cells by macrophages were inhibited by the anti-macrophage serum. The anti-lymphocyte serum showed no such inhibition, when previously absorbed with macrophages.

     

A new method for separating peritoneal lymphocytes from macrophages

A simple method has been developed for separating sterile, viable mouse peritoneal lymphocytes from suspension of peritoneal cells using velocity sedimentation. Lymphocytes are obtained in 98.6–99.2 per cent purity; macrophages in 85.4–95.0 per cent purity.     

In vitro phagocytosis of Candida albicans by peritoneal mouse macrophages.

The ability of sensitized mouse peritoneal macrophages to phagocytose and inhibit Candida albicans was studied in an in vitro system. Mice were sensitized to C. albicans by intraperitoneal infection with viable organisms or by intracutaneous injection of heat-inactivated cells in Freund complete adjuvant. Development of delayed hypersensitivity to C. albicans was evaluated by footpad tests with cytoplasmic and cell wall antigens as well as by macrophage migration inhibition by these antigens and by whole heat-inactivated cells. Inhibition of macrophage migration by heat-inactivated cells was significantly greater when the mice were sensitized by viable organisms. The macrophages from these mice were also larger and showed a greaer ability to inhibit germ tube production by phagocytosed yeasts. This suggests that macrophages may play a protective role in infection by C. albicans.     

Interactions between Histoplasma capsulatum and macrophages from normal and treated mice: comparison of the mycelial and yeast phases in alveolar and peritoneal macrophages.

Interactions between macrophages (alveolar and peritoneal) from normal, vaccinated (with heat-killed yeast cells), and Mycobacterium bovis BCG-treated mice and the mycelial and yeast phases of Histoplasma capsulatum were observed. Phagocytosis of microconidia, small hyphal fragments, and yeast cells occurred 4 to 6 h after the infection of macrophage cultures. Conversion to the yeast phase began at 6 to 7 h and was complete after a 72-h incubation at 37 degrees C. Macrophages surrounded and adhered to macroconidia and large hyphal elements. More macrophages (65 to 68%) from BCG-treated mice contained fungi at 24 h than did macrophages from normal or vaccinated mice. Although there was no increase in the number of fungi in macrophages from vaccinated mice, only the macrophages from BCG-treated mice contained fewer fungi after 48 h of infection with the mycelial phase of H. capsulatum. Fungal growth was not inhibited in any of the macrophage cultures when infected with the yeast phase. The macrophages infected with yeast cells were destroyed after 48 to 72 h in the culture. Only BCG-treated macrophages survived infection with the mycelial phase, whereas macrophages from normal and vaccinated mice were destroyed by the infection.     

A possible role for antibodies against spectrin in the interaction between erythroblasts and macrophages in vitro.

The nature of serum factors which participate in the interaction in vitro between dimethylsulphoxide-induced Friend leukaemia erythroblasts (IFLE) and syngeneic mouse peritoneal macrophages was investigated. When heat-inactivated newborn calf serum (HI-NBCS) was depleted of IgG its activity to promote the association of neuraminidase-treated 59Fe-labelled IFLE (59Fe-IFLE) with macrophages was markedly reduced but could be restored by the addition of bovine IgG. Trypsin treatment of macrophages caused incomplete inhibition of their subsequent association with both untreated and neuraminidase-treated 59Fe-IFLE in the presence of HI-NBCS. When spectrin, the major red cell cytoskeleton protein, was added to HI-NBCS there was a dose-related inhibition of the association with macrophages of both untreated and neuraminidase-treated 59Fe-IFLE. Moreover a mouse monoclonal antibody against spectrin promoted the interaction of neuraminidase-treated 59Fe-IFLE with macrophages. Mouse sera which supported the association of neuraminidase-treated 59Fe-IFLE with macrophages were found to contain anti-spectrin antibodies. These results suggest that IgG antibodies mediate the interaction between erythroblasts and macrophages via trypsin-sensitive and trypsin-resistant receptors on the macrophage surface and that at least some of the antibodies show specificity for spectrin.