Binding of low-density lipoproteins from chicken egg yolk to mouse anti-phospholipid B cells which include B cells against bromelain-treated mouse erythrocytes

Low-density lipoproteins from chicken egg yolk (EyLDL), which are reactive with mouse antibodies against bromelain-treated mouse erythrocytes (BrMRBC), were conjugated with fluorescein isothiocyanate (FITC). FITC-EyLDL could bind specifically to mouse anti-phospholipid B cells, which comprised all the BrMRBC-rosette-forming cells and anti-BrMRBC lipopolysaccharide-reactive B cells, C3H/He mice at 12 weeks of age had, approximately, 7 x 10(5) EyLDL-binding cells in the peritoneal cavity, 3 x 10(5) EyLDL-binding cells in the pleural cavity, and 3 x 10(5) EyLDL-binding cells in the spleen. In ontogeny, the numbers of EyLDL-binding cells in the peritoneal cavity expanded greatly by 4 weeks. Other normal strains of mice and C3H/HeJ mice at 12 weeks of age had 4-7 x 10(5) EyLDL-binding cells in the peritoneal cavity; the numbers were large (19 x 10(5] in NZB mice, rather small (2 x 10(5] in MRL/lpr mice, and very small (0.1 x 10(5] in CBA/N mice. In some of various strains of mice at 12 months of age, more than 20% of peritoneal cells were EyLDL-binding cells; in particular, all of five older NZB mice examined had more than 10(7) EyLDL-binding cells in the peritoneal cavity.     

Formation in agar of fibroblast-like colonies by cells from the mouse pleural cavity and other sources

Colonies of elongated fibroblast-like cells (stellate colonies) developed in agar cultures of mouse pleural cavity cells mixed with whole blood. Cultures of pleural cells alone developed only abortive clusters of round cells. The frequency of colony-forming cells in the pleural cavity was highest in neonatal mice (200/10⁵ cells) and fell progressively with aging. Stellate colony-forming cells were not in cell cycle but were radiosensitive. In adult mice, only occasional colony-forming cells were detected in peritoneal cavity, thymic, spleen, lymph node or bone marrow cell populations. Stellate colony formation was not stimulated by the granulopoietic regulator, colony stimulating factor. The active component in whole blood required for stellate colony formation was present in plasma but not serum or washed red or white cells.     

Demonstration of capsule in a strain of Staphylococcus hyicus by an electron microscope

A compact type variant designated as strain ST67V was isolated by a high-temperature subculture method from strain ST67P of Staphylococcus hyicus, which was a diffuse type in serum-soft agar. The parent strain was relatively virulent in mice and resisted ingestion by mouse peritoneal cells. The variant strain, however, was avirulent in mice and no antiphagocytic activity was observed in the peritoneal cavity. In ultra-thin sections of the organisms treated with anti-ST67P rabbit anti-serum conjugated with ferritin, the outermost layer of the cell wall of the parent strain was covered with a well-defined capsule while no capsule was shown in the variant strain.     

Demonstration of capsule in a strain of Staphylococcus hyicus by an electron microscope

A compact type variant designated as strain ST67V was isolated by a high-temperature subculture method from strain ST67P of Staphylococcus hyicus , which was a diffuse type in serum-soft agar. The parent strain was relatively virulent in mice and resisted ingestion by mouse peritoneal cells. The variant strain, however, was avirulent in mice and no antiphagocytic activity was observed in the peritoneal cavity. In ultra-thin sections of the organisms treated with anti-ST67P rabbit anti-serum conjugated with ferritin, the outermost layer of the cell wall of the parent strain was covered with a well-defined capsule while no capsule was shown in the variant strain.     

Fertilization and culture of rabbit oocytes in hydrogel chambers implanted in the peritoneal cavity of mouse recipients

The purpose of this study was to determine whether in vivo fertilization and culture using a hydrogel chamber was possible. Hydrogel chambers were made from the mixture of low-acid hema, as a monomer, and tetraethylene glycol dimethacrylate (TGD) and ethylene glycol (EG), as the cross-linkers. Rabbit oocytes and spermatozoa were transferred into the lumen of the hydrogel chambers which were filled with chemically-defined culture medium containing no protein. The chambers containing the rabbit oocytes and spermatozoa were implanted within the mouse peritoneal cavity, and were examined after recovery from the chambers following 84 hours of preservation. A total of 88 oocytes preserved after 84 hours in pHema hydrogel chambers implanted in the peritoneal cavity of female mice resulted in 29 morulae and 46 blastocysts. Thus, demonstrating that fertilization and culture can occur inside of the hydrogel chamber.     

Penetration of host IgG molecules into hydatid cysts.

Host IgG was detected in 15 out of 18 peritoneal hydatid cysts obtained from experimentally infected gerbils and in 18 out of 61 mouse hydatid cysts which had been implanted into the peritoneal cavity of gerbils for periods ranging from one day to two years. A wide variation in host IgG concentration was observed among the different cysts in which these immunoglobulins were found. These findings are discussed in terms of the possible manner by which macromolecules may enter into the hydatid cysts.     

METALLIC COPPER-INDUCED GRANULOCYTE EXUDATION IN THE STUDY OF GRANULOCYTOPOIESIS

C3H/HeHa mice implanted with rods of metallic copper (CR) or glass (GR) exudate neutrophil granulocytes and mononuclear cells into the site of rod implant (peritoneal cavity). Exudation in CR mice is substantially greater than in GR animals. In CR mice there is an impressive stimulation of myelopoiesis measured in the femoral marrow subsequent to the initial accumulation in the peritoneal cavity. Serum levels of colony stimulating activity (CSA), an in vitro myeloproliferative stimulating activity, are elevated in such animals, as are femoral marrow agar-colony forming cells (CFC). The procedure is useful to the study of myelopoiesis and myelopoietic regulatory mechanisms.     

Expanding the febrigenic role of cyclooxygenase-2 to the previously overlooked responses

Previous studies on the role of cyclooxygenase (COX)-1 and -2 in fever induced by intravenous LPS have failed to investigate the role of these isoenzymes in the earliest responses: monophasic fever (response to a low, near-threshold dose of LPS) and the first phase of polyphasic fever (response to higher doses). We studied these responses in 96 mice that were COX-1 or COX-2 deficient (-/-) or sufficient (+/+). Each mouse was implanted with a temperature telemetry probe into the peritoneal cavity and a jugular catheter. The study was conducted at a tightly controlled, neutral ambient temperature (31 degrees C). To avoid stress hyperthermia (which masks the onset of fever), all injections were performed through a catheter extension. The +/+ mice responded to intravenous saline with no change in deep body temperature. To a low dose of LPS (1 microg/kg iv), they responded with a monophasic fever. To a higher dose (56 microg/kg), they responded with a polyphasic fever. Neither monophasic fever nor the first phase of polyphasic fever was attenuated in the COX-1 -/- mice, but both responses were absent in the COX-2 -/- mice. The second and third phases of polyphasic fever were also missing in the COX-2 -/- mice. The present study identifies a new, critical role for COX-2 in the mediation of the earliest responses to intravenous LPS: monophasic fever and the first phase of polyphasic fever. It also suggests that no product of the COX-1 gene, including the splice variant COX-1b (COX-3), is essential for these responses.     

食管癌细胞诱导肥大细胞迁移的小鼠模型

目的建立小鼠肥大细胞（mast cell,MC）迁移模型,探讨化疗药物三氧化二砷对人食管癌EC109细胞株生长的杀伤机理。方法利用肥大细胞的特征性蛋白酶抗体及其免疫荧光标记MC和PI标记EC109细胞内DNA;以流式细胞术分析小鼠腹腔液中肥大细胞各亚型的百分率及肿瘤细胞周期变化;使用激光扫描共聚焦显微镜显示肥大细胞内分泌颗粒的分布。并通过组织化学方法,观察各种诱导处理后小鼠肠组织肥大细胞由肠道向腹腔移动的变化。结果①根据流式细胞仪点图分布分析,将小鼠肥大细胞分为：类胰蛋白酶阳性、类糜蛋白酶阴性（MC T）;类糜蛋白酶阳性、类胰蛋白酶阴性（MC C）和类胰蛋白酶阳性、类糜蛋白酶阳性（MC TC）三种亚型,且T型MC明显多余TC型和C型（P〈0.05）;以共聚焦显微镜显示三种亚型的MC均含有丰富的分泌颗粒并分布于细胞膜内侧,为其出芽突起形成储备的状态。②经组织切片观察到诱导处理后小鼠肠组织MC由肠道向腹腔移动,且胰酶对MC的诱导作用大于食管癌细胞和As2O3。③经诱导迁移入腹腔的MC可能与癌细胞周期由S期向G2/M期跨越相关;As2O3能延迟食管癌细胞的G0/G1期,阻碍细胞向S期跨越,从而抑制食管癌细胞的生长。结论食管癌细胞移入小鼠腹腔,主要诱导T型MC参与免疫反应。在生物机体内环境（这里指MC影响）的条件下,As2O3对肿瘤细胞生长的作用主要表现为促使癌细胞周期的G0/G1期向S期跨越延迟,或G2/M期进入细胞分裂的延迟。     

In vivo cultivation of Echinococcus multilocularis protoscoleces in micropore chambers

Micropore chambers containing unevaginated protoscoleces of E. multilocularis were implanted into the peritoneal cavity of AKR mice. Transformation from protoscoleces to fertile multivesicular cysts was obtained after 210 days. Ultrastructural observations of these morphological transformations indicate that a phase of histogenesis follows a phase of dedifferentiation. This morphogenetic process raises the question of the origin of new cell populations. The results reveal the potential role of protoscoleces in secondary echinococcosis and the value of this experimental model for further studies on the larval development.     

Fluctuations of diphenylamine-DNA in yeast

A transient increase in the number of peritoneal colony-forming cells in agar (PCFC) was noted in the peritoneal cavity of mice given one intraperitoneal injection of thioglycollate medium. This increase of PCFC was not accompanied by a similar degree of increase in the number of either pluripotent hemopoietic stem cells or committed hemopoietic stem cells for granulocytes and/or macrophages that are normally present in the peritoneal cavity in small numbers. Other substances, such as glycogen, starch, Freund's adjuvant, sheep red blood cells and lectins, were also capable of increasing PCFC, but to different degrees. The ability of thioglycollate medium as well as other stimulatory agents to increase PCFC appeared to be dose-dependent.     

Trypanosoma cruzi: morphogenesis in diffusion chambers in the mouse peritoneal cavity and attempted stimulation of host immunity

Sequential morphologic changes and antigen producing capacity of Trypanosoma cruzi in peritoneally implanted diffusion chambers were studied. Diffusion chambers were equipped with two Nuclepore filters (0.20 μm pore size) sandwiched between three Lucite rings. Epimastigotes or trypomastigotes and amastigotes were placed in diffusion chambers and surgically implanted into the peritoneal cavity of mice, or placed in in vitro cell culture, or in various types of culture media and incubated at 26 or 37 C.Epimastigotes maintained in diffusion chambers in mice changed into trypomastigotes as evidenced by the presence of numerous transitional stages and the concomitant decrease in the percentage of the former and increase in the percentage of the latter in chambers removed and examined at 16, 24, 36, 48, 72, and 84 hr after implantation. The maximum of 68% trypomastigotes was noted in chambers examined at 84 hr. Amastigotes subsequently appeared, apparently arising from trypomastigotes and reached the highest percentage (49%) obtained at 132 hr. The total number of parasites in chambers decreased slightly during the first 36 hr (20%). Little change in the total number of parasites was noted during the interval of 36–108 hr. A subsequent decrease in numbers of parasites was noted until by 280 hr after implantation, chambers contained less than 2% of the original number of organisms present in the chambers. No similar transformation of epimastigotes was noted in diffusion chambers maintained in cell culture at 37 C or in a cell culture growth medium or LIT medium at 37 or 26 C.No detectable morphological change was noted when trypomastigotes and amastigotes were implanted in diffusion chambers in the peritoneal cavity of mice. The total number of these parasites decreased notably (82%) after 24 hr.Mice receiving diffusion chambers containing epimastigotes implanted at two different intervals (21 days apart), developed only marginal protective immunity when challenged with virulent T. cruzi three weeks after the second implant of chambers, and no protection was afforded those mice implanted with chambers containing trypomastigotes and amastigotes. Sera collected from mice 6 wk after the second implantation of diffusion chambers containing parasites were observed to have antibody titers to T. cruzi as demonstrated by the fluorescent antibody technique and direct agglutination procedure.     

Taenia crassiceps: Fate of cysticerci following ingestion by the mouse

Cysticerci of Taenia crassiceps were administered to mice by gavage to determine whether enteral or parenteral infections would establish consistently. Some worms survived in the small intestine up to 16 days, whereas others penetrated through the gut wall into the peritoneal cavity within 24 hr. Similar proportions of different doses of worms reached the peritoneal cavity regardless of the size of the inoculum and sex or strain of mice used. In addiiton, it was shown that mice may acquire an intraperitoneal infection with T. crassiceps by eating the carcass of an infected mouse.     

Effects of radiation on host-tumor interactions using the multicellular tumor spheroid model

Summary Use of the multicellular tumor spheroid as a tumor model allows separate host or tumor treatment with ionizing radiation and examination of the effects on host-tumor immune interactions. Spheroids of EMT6/Ro, a BALB/c mammary tumor were implanted into the peritoneal cavity of syngeneic immunized mice, recovered, and dissociated into single cells. Cytolytic activity of mature spheroid associated cells and peritoneal cells was resistant to radiation doses as high as 1000 rads when irradiated directly prior to assay. Mice irradiated (200, 400, 700 rads) 24 h prior to spheroid injection had an increased number of tumor cells and decreased number of tumor infiltrating and peritoneal host cells upon spheroid recovery. This was paralleled by an increased colony forming efficiency per spheroid. Cytolytic activity of the spheroid associated cells against radiolabeled EMT6 cells was in many cases decreased with radiation although lysis was the same on a per cell basis. Cytolytic activity by peritoneal cells from these mice increased with dose as measured on a per cell basis. This activity from irradiated animals was carried out by a Thyl⁺ cell.     

Peritoneal exudate cells. I. Growth requirement of cells capable of forming colonies in soft agar

Mouse peritoneal exudate cells induced by thioglycollate medium can form colonies in soft agar with a plating efficiency of about 5% (0.6%–10%). Cells from an unstimulated peritoneal cavity form no colonies or have a plating efficiency of less than 0.001 %. These colony-forming cells from the peritoneal exudate are similar to bone marrow colony-forming cells in vitro in that they both require a substance(s) present in conditioned medium from L-cells or mouse embryo fibroblasts or the serum from endotoxin-treated mice for the initiation and the continuation of their growth. However, peritoneal exudate colony-forming cells have a much longer initial lag period (10–14 days) and can survive longer in the absence of L-cell conditioned medium than bone marrow colony-forming cells. Only mononuclear cells, presumably macrophages, are observed in peritoneal exudate colonies, whereas bone marrow cell colonies contain both polymorphonuclear cells and macrophages.     

Peritoneal B cells respond to phorbol esters in the absence of co-mitogen

B cells obtained by irrigation of the peritoneal cavity differ from splenic B cells in signaling requirements for the initiation of DNA synthesis. Splenic B cells are stimulated to enter S phase by phorbol esters in conjunction with a second signal provided by calcium ionophore; however, splenic B cells are not stimulated by phorbol ester alone. In contrast, peritoneal B cells from NZB and BALB/c mice were stimulated to incorporate tritiated thymidine by each of the phorbol esters, PMA and phorbol dibutyrate, acting alone. Stimulation of peritoneal B cells was apparent when cells were cultured at lower than usual cell densities, and responses were unaffected by coculture with splenic B cells. Responding cells adhered to plastic petri dishes coated with anti-mouse IgM antibody, but were not completely removed by treatment with anti-Ly-1.2 antibody plus C. These results indicate that phorbol esters constitute a complete signal that stimulates some peritoneal B cells to enter S phase.     

Correlation between increase in Ia-bearing macrophages and induction of T cell-dependent antitumor activity by Lactobacillus casei in mice

Summary When Lactobacillus casei YIT 9018 (LC 9018) or Corynebacterium parvum, known to be immunomodulators possessing antitumor activity, were injected i.p. into BALB/c mice, peritoneal exudate macrophage Ia antigen detected by indirect immunofluorescence method was expressed on their cell surface, but it was not expressed following the injection of 10% proteose peptone, an inflammatory agent, or Lactobacillus fermentum YIT 0159 (LF 0159), which have no antitumor activity. The percentage and absolute number of Ia-positive peritoneal macrophages were maximum on the 7th day after the injection of LC 9018. Immunization by injection of Meth A fibrosarcoma cells treated with mitomycin C (MMC-Meth A) 7 days after LC 9018 injection suppressed the growth of Meth A implanted i.p. 14 days after MMC-Meth A injection. A shorter interval between the injections of LC 9018 and MMC-Meth A did not allow suppression of Meth A growth. These results showed that the increase in Ia-positive macrophages in the peritoneal cavity coincided with the effective interval for induction of the antitumor activity by LC 9018. The antitumor activity induced by injections of LC 9018 and MMC-Meth A did not affect the growth of RL l leukemic cells, syngeneic to BALB/c mice. Neutralization (Winn type) tests showed that peritoneal T lymphocytes possessed tumor cytotoxicity and that the antitumor capacity was reduced by in vivo treatment with anti I-A^d monoclonal antibody simultaneously with and 1 day prior to MMC-Meth A injection. These results indicate that LC 9018-induced Ia-positive macrophages, which first encounter a tumor antigen in the peritoneal cavity, play an important role in the in vivo induction of tumor specific T cell-mediated antitumor immunity.     

Chromosome-mediated resistance of Yersinia enterocolitica serotype O9 to intracellular killing by mouse peritoneal macrophages

Abstract The survival of Yersinia enterocolitica serotype O9 within mouse peritoneal macrophages was investigated. To evaluate the role of the virulence plasmid in the resistance to intracellular killing, an isogenic pair of virulent (plasmid-bearing) and avirulent (plasmid-less) O9 strains was used. The virulent strain was able to express plasmid-encoded outer membrane proteins and to colonize the Peyer's patches of orally infected mice. When mice were infected intraperitoneally, both strains were recovered at similar rates and over the same time from the peritoneal cavity. When in vitro assays were performed, both strains showed similar resistance to intracellular killing by monolayers of resident and inflammatory peritoneal macrophages. Previous opsonization of bacteria did not modify their survival within macrophage monolayers. We concluded that serotype O9 strains display a chromosome-mediated resistance to intracellular killing by mouse peritoneal macrophages. Moreover, macrophage resistance does not seem to be of importance for virulence of serotype O9 strains in mice.     

Relationship between proteolytic activity of Aspergillus fumigatus and the fungus' invasiveness of mouse brain

This experiment was carried out to determine whether proteolytic activity of Aspergillus fumigatus was enhanced in the mouse brain during advance in growth of hypae, and if any relationship might be demonstrated between the proteolytic activity of the fungus and its invasive ability for the mouse brain.The K-2 strain of A. fumigatus and male mice of ddy line weighing 22±1 g were used in this experiment. Each mouse was inoculated intravenously with 5×10⁶ conidia of the strain suspended in 0.2 ml. phosphate buffer solution supplemented with 0.01 per cent Tween 80. Mice were sacrified at 4 hours' intervals up to 40 hours after inoculation and at an hour's intervals later. Two mice were assigned for each time. Each brain obtained was fixed in 10 per cent formalin and was cut into three sections from which histopathological specimens were prepared.The brains over 40 hours after inoculation were markedly swollen and abundant hypae were observed in the specimens. The extent of the fungal growth in the specimens from the mouse brain was 1.60, 7.71 and 10.84 per cent at 32,45 and 49 hours after inoculation, respectively.For assay of the proteolytic activity in the mouse brain, three groups of fifteen mice each were tested with the K-2 strain. These groups of mice were inoculated with 5×10⁶conidia of the strain and sacrificed 0,32 and 45 hours after inoculation, respectively. Then the brains pooled in each group and four volumes of phosphate buffer solution was added. The material was homogenized and used as test samples (enzyme solution). The proteolytic activity was measured quantitatively by a modification of Anson's method.The proteolytic activity in the mouse brain sacrificed 0,32 and 45 hours after inoculation was 0.00, 0.08 and 0.13, respectively, as expressed by the optical density.In conclusion, it was possible to confirm that proteolytic activity increased in the mouse brain in proportion to the growth of hyphae in it.     

Cytophilic antibodies in mice immunized with sheep red cells

Antibodies cytophilic for homologous and heterologous (guinea pig, rat) peritoneal cells of the mononuclear phagocyte type were demonstrated in the sera of mice immunized with sheep red cells. The formation of these antibodies can be induced by immunization with red cells with and without complete Freund's adjuvant. They are also present, in varying titres, in the sera of non-immunized mice. The peritoneal cavity of normal and immunized mice contains cells which adsorb sheep red cells. This phenomenon is significantly reduced by preincubating the peritoneal cells for two hoursin vitro at 37° C. The cytophilic activity of mouse sera can be abolished by absorption by spleen homogenate, but not by pulverized guinea pig kidneys. The haemolysin and haemagglutinin titres before and after absorption are practically the same. The cytophilic component of some sera was inhibited by 2-mercaptoethanol. Tests of the sera of mice revaccinated with sheep red cells showed, after separation on Sephadex G 200, that cytophilic activity was present mainly in the 7 S fraction, but also in the 19 S fraction. Cells adsorbing cytophilic antibodies stain supravitally with neutral red; in fixed and stained preparations they are typical macrophages and monocytoid mononuclears with varying degrees of affinity for basic dyes.