Rejection of worm load through singly and repeatedly sensitized peritoneal exudate cells during experimental ancylostomiasis

Summary Sensitized peritoneal exudate cells from Swiss albino mice donors infected with a single dose of 1000A. caninum larvae could expel a challenge dose of 500 larvae from recipients at a faster rate when compared to cells from repeatedly infected (250+250+500) donors. However, at 36 h after challenge, the larval expulsion was almost the same in both the groups. Because of the bowel sensitization by the cells, some larvae (not expelled) in the 1st group, readily migrated into muscles where they met allergic immobilization and death due to infiltration of inflammatory cells and their exudates at these sites.Acknowledgment. We thank Professor H. Swarup for providing facilities and to the Council of Scientific and Industrial research, New Delhi for financial assistance.     

Rejection of worm load through singly and repeatedly sensitized peritoneal exudate cells during experimental ancylostomiasis

Sensitized peritoneal exudate cells from Swiss albino mice donors infected with a single dose of 1000 A. caninum larvae could expel a challenge dose of 500 larvae from recipients at a faster rate when compared to cells from repeatedly infected (250 + 250 + 500) donors. However, at 36 h after challenge, the larval expulsion was almost the same in both the groups. Because of the bowel sensitization by the cells, some larvae (not expelled) in the 1st group, readily migrated into muscles where they met allergic immobilization and death due to infiltration of inflammatory cells and their exudates at these sites.     

Arginase expression in peritoneal macrophages and increase in circulating polyamine levels in mice infected with Schistosoma mansoni

We investigated the nitric oxide (NO) synthase and arginase pathways in resident peritoneal macrophages of mice infected with the tropical parasite Schistosoma mansoni. The two enzymes may have opposite effects, insofar as NO may be involved in the killing of the parasite whereas arginase may stimulate parasite growth via polyamine synthesis. We determined the effects of the infection on the expression and activity of the two enzymes in macrophages, before and after cytokine activation. Cells from infected mice expressed the hepatic type I arginase, whereas in control cells, the enzyme was expressed only after cytokine activation, as were NO synthase II and type II arginase in both groups of cells. Moreover, we found that in infected mice, arginase expression in macrophages was associated with a ten fold increase in the concentration of circulating ornithine-derived polyamines. This may be of pathological importance, since parasitic helminths are though to be dependent on their hosts for the uptake and interconversion of polyamines. Received 13 March 2001; received after revision 4 May 2001; accepted 7 June 2001     

A mouse system for demonstrating the presence of inflammatory factors from human peripheral blood lymphocytes

Summary An assay system is described that allows the presence of inflammatory factors in supernatants from stimulated tuberculin-sensitive human peripheral blood lymphocytes to be demonstrated, by the induction of an inflammatory exudate in the peritoneal cavity of normal C57 BL mice.     

Respiratory burst in peritoneal exudate cells in response to a modified tuftsin.

Intraperitoneal administration of tuftsin-M [Thr-Lys-Pro-Arg-NH-(CH2)2-NH-CO-C15H31] to Balb/C mice has been shown to induce a respiratory burst in the peritoneal exudate cells. The macrophages exhibited enhanced levels of O2-, H2O2, NADPH oxidase and myeloperoxidase, but the activities of superoxide dismutase, catalase and glutathione peroxidase remained virtually unchanged. The magnitude of the oxidative burst depended directly on the dose of tuftsin-M; higher activity was observed at higher doses of the peptide. Tuftsin-M enhanced the generation of both O2- and H2O2 under in vitro conditions, as did phorbol myristate acetate. These results suggest that tuftsin-M could enhance non-specific defence against infections by activating the macrophages.     

Inflammatory factors produced by sensitized guinea-pig peripheral blood lymphocytes

Summary The supernatants obtained from stimulated tuberculin-sensitive guinea-pig peripheral blood lymphocytes contain factors that induce a cutaneous inflammatory response in normal guinea-pigs similar to the tuberculin reaction and inhibit the migration of normal guinea-pigs peritoneal exudate cells. There appears to be a correlation between the presence of in vitro migration inhibitory activity and inflammatory activity in vivo.     

Antibody-induced formation of caps inToxoplasma gondii

Summary Trophozoites ofToxoplasma gondii from mouse peritoneal exudate move their surface membrane antigens towards one pole of the cell when incubated with antibodies. The phenomenon may be induced in up to 50% of incubated parasites. It is prvented by some metabolic inhibitors and low temperatures (0–4°C). These properties do not change in parasites subpassaged after repeated incubation with antibodies.We wish to thank Dr. J.lusarczyk of the National Institute of Hygiene for his help in preparation the photomicropgraphs.     

Antibody-dependent adherence of peritoneal cells of the rat to Trichinella spiralis larvae]

Washed peritoneal exudate cells from normal Rats firmly adhere to Trichinella spiralis larvae in the presence of serum containing anti-Trichinella antibodies. This effect is observed when muscle larvae, cells and dilutions of anti-sera are incubated for 1 hr. at 37 degrees C. No adherence takes place at 4 degrees C. Whole serum or its gammaglobulin fraction are active and effect is inhibited by the addition of Trichinella antigens. Complement is not essential since antiserum heated for 2 hrs. at 56 degrees C is active. Washed cells from infested animals do not adhere to the larvae in the absence of antiserum.     

A thiol protease of peritoneal macrophages in the guinea pig

Summary Proteolytic enzymes of the guinea pig peritoneal exudate macrophages were investigated using synthetic fluorogenic peptide substrates. Among several enzymes, t-butyloxycarbonyl-phenylalanyl-seryl-arginine 4-methylcoumaryl-7-amide cleaving enzymes had the highest activity, and the activity in exudate macrophages was about 3 times tronger than that in resident thiol-blocking reagents, suggesting a thiol protease.We thank Dr Yoshiaki Motozato, Kumamoto University, for kind donation of Cellulofine GC-700 and valuable discussions. This work is supported in part by the Grant-in-Aid for Scientific Research from the Ministry of Education, Science and Culture of Japan.     

Efficacy of tumor cell vaccine after incorporating monophosphoryl lipid A (MPL) in tumor cell membranes containing tumor-associated ganglioside

Murine B16 melanoma expresses the ganglioside. GM₃. GM₃ shed from tumor cells is immunosuppressive and promotes tumor growth¹. Reduction or elimination of the shed GM₃ could be therapeutic, and the anti-GM₃ antibodies may reduce and clear the shed ganglioside. To test this hypothesis, mice were challenged with tumor cells, with or without inducing anti-GM₃ antibody response. Since gangliosides are poor immunogens and T-cell independent antigens, an adjuvant (monophosphoryl lipid A (MPL), a non-toxic lipid A ofSalmonella), directed against B-cells, was employed. MPL was incorporated onto liposomes and into the surface membrane of B16 mouse melanoma cells; both are rich in GM₃. C57BL/6J mice immunized with MPL-liposomes or MPL-B16 cells responded with elevated levels of anti-GM₃ IgM. Non-immunized mice or mice immunized with B16 cells alone or ganglioside GM₃ alone (without MPL) elicited poor anti-GM₃ IgM response, confirming the GM₃'s immunologic crypticity and MPL's immunopotentiating effect. MPL's immunopotentiating effect was improved by coupling it to melanoma cell membranes C57BL/6J mice were immunized with irradiated B16 alone or MPL alone or MPL-conjugated irradiated B16. After three weekly immunizations, each mouse received a challenge dose of viable syngeneic B16. Neither MPL alone nor B16 alone had a significant effect on tumor growth or host survival; however, administration of MPL-conjugated B16 cells significantly prevented tumor growth and prolonged survival. Our results indicate that MPL-incorporated B16 cells augment the anti-GM₃ IgM response, which may reverse GM₃-induced immunosuppression by eliminating tumor-derived GM₃, and restore immunocompetence.     

A thiol protease of peritoneal macrophages in the guinea pig

Proteolytic enzymes of the guinea pig peritoneal exudate macrophages were investigated using synthetic fluorogenic peptide substrates. Among several enzymes, t-butyloxycarbonyl-phenylalanyl-seryl-arginine 4-methylcoumaryl-7-amide cleaving enzymes had the highest activity, and the activity in exudate macrophages was about 3 times stronger than that in resident macrophages. The molecular weight of the enzyme was around 35,000 and optimal pH around 6.5-7.0. It was inhibited by thiol-blocking reagents, suggesting a thiol protease.     

Stimulation of the growth of murine bone marrow colonies in vitro by an acute inflammatory exudate. Comparison with colony stimulating factor CSF)]

Formation of colonies from mice bone marrow progenitors of macrophages and granulocytes in methylcellulose culture was induced by an inflammatory pleural exudate obtained from mice injected with dextran. Mitogenic activity of this acute inflammatory exudate was compared with that of colony stimulating factor (CSF). It was found that colony and cluster counts, during 10 days of culture, were similar with the two types of stimulating factors. When used at the same dose, exudate was less active than CSF. It was concluded that inflammatory exudate showed an activity similar to that of CSF but contained a smaller amount of stimulating factor. Further cytochemical studies are necessary to specify whether or not the two factors induced the same type of colonies (monocytic or granulocytic).     

Gamma-glutamyl-transpeptidase in lymphatic tissues of Mastomys natalensis during an infection with Acanthocheilonema viteae

During Acanthocheilonema viteae infection, the specific activity of gamma-glutamyltranspeptidase (gamma-GT) increased in peritoneal exudate cells and bone marrow and decreased in lymph nodes of Mastomys natalensis throughout the course of infection. However, though there was an increase in specific activity of gamma-GT in thymus and spleen during the prepatent phase of A. viteae infection, the level either returned to normal or decreased during the latent phase of infection. A close correlation was observed between the host's immune status during A. viteae infection and the level of gamma-GT in lymphoid tissues.     

γ-Glutamyl-transpeptidase in lymphatic tissues ofMastomys natalensis during an infection withAcanthocheilonema viteae

Summary DuringAcanthocheilonema viteae infection, the specific activity of -glutamyltranspeptidase (-GT) increased in peritoneal exudate cells and bone marrow and decreased in lymphnodes ofMastomys natalensis throughout the course of infection. However, though there was an increase in specific activity of -GT in thymus and spleen during the prepatent phase ofA. viteae infection, the level either returned to normal or decreased during the latent phase of infection. A close correlation was observed between the host's immune status duringA. viteae infection and the level of -GT in lymphoid tissues.Communication No. 4535 from Central Drug Research Institute, Lucknow.     

Protective effect of Shigyaku-to,a traditional Chinese herbal medicine,on the infection of herpes simplex virus type 1 (HSV-1) in mice

The antiviral activity of Shigyaku-to (TJS-109), a traditional Chinese herbal medicine, was investigated in mice infected with herpes simplex virus type 1 (HSV-1). TJS-109 is a combination of the medicinal plant extracts fromZingiberis siccatum rhizoma,Aconiti tuber andGlycyrrhizae radix in a specific proportion. Mice infected with a 10 LD₅₀ dose of HSV-1 were treated with TJS-109 orally at doses of 1.25 to 20 mg/kg 2 days before, and 1 and 4 days after the infection. The treated groups had 80% (1.25 mg/kg), 40% (5 mg/kg) and 23% (20 mg/kg) mortality rates 25 days after the infection as compared with a 100% mortality rate in control mice treated with saline. When HSV-1 infected mice (recipients) received CD8⁺T cell fractions derived from spleens of mice treated with TJS-109 (donors), 70% of recipients survived, as compared with 0% survivors in the groups of mice treated with saline, B cell fractions, CD4⁺ T cell fractions or macrophage-enriched fractions prepared from the same donors. TJS-109 did not show any virucidal activities against HSV-1 or any virostatic activities on the growth of HSV-1 in Vero cells. These results suggest that TJS-109 protected mice exposed to lethal amounts of HSV-1 through the activation of CD8⁺ T cells.     

Genetics of early mammalian folliculogenesis

Early ovarian folliculogenesis begins with the breakdown of germ cell clusters and formation of primordial follicles. Primordial follicles are the smallest ovarian follicle units continuously recruited to grow into primary and more advanced ovarian follicles. Genes expressed in the germ cells such as Figla, Nobox, Kit and Ntrk2, as well as genes expressed in the surrounding somatic cells such as Foxl2, Kitl and Ngf, play critical functions during early folliculogenesis. Transgenic mice continue to provide important insights into the genetic pathways that regulate early mammalian folliculogenesis. Genes critical in early folliculogenesis are important determinants of reproductive life span and represent candidate genes for human ovarian failure. Received 25 August 2005; received after revision 18 October 2005; accepted 21 November 2005     

Colicin E3 enhances the oxidoreductive activity of guinea-pig leucocytes

Summary Colicin E3 increased the capacity of peritoneal exudate leucocytes to reduce iodo-nitro-tetrazolium-chloride to formazane. This effect was directly dependent on its concentration within the range 10³–10⁵ lethal units per cell. In control experiments, dectran C exerted no influence on the rate of this activity and colicin E3 did not convert INT to formazane.     

Respiratory burst in peritoneal exudate cells in response to a modified tuftsin

Intraperitoneal administration of tuftsin-M [Thr–Lys–Pro–Arg–NH–(CH₂)₂–NH–CO–C₁₅H₃₁] to Balb/C mice has been shown to induce a respiratory burst in the peritoneal exudate cells. The macrophages exhibited enhanced levels of O₂ ^–, H₂O₂, NADPH oxidase and myeloperoxidase, but the activities of superoxide dismutase, catalase and glutathione peroxidase remained virtually unchanged. The magnitude of the oxidative burst depended directly on the dose of tuftsin-M; higher activity was observed at higher doses of the peptide. Tuftsin-M enhanced the generation of both O ₂ ^– and H₂O₂ under in vitro conditions, as did phorbol myristate acetate. These results suggest that tuftsin-M could enhance non-specific defence against infections by activating the macrophages.