The anti-tumoral activity of human glycoprotein HGP.92: a study with the mouse Lewis-lung-tumor cell.

The ability of a purified human glycoprotein (HGP.92) to exert anti-tumor activity was investigated in a mouse model using long-term readout assays. In vitro, in the presence of inflammatory mouse macrophages incubated with HGP.

Purification of a 92 kDa human immunostimulating glycoprotein obtained from the Tamm-Horsfall glycoprotein.

A purification method for a human urinary glycoprotein (HGP92) dissociated from Tamm Horsfall protein (THP) is described. Tamm-Horsfall protein, obtained by salt precipitations, was again precipitated in presence of monovalent ions. In these conditions, Tamm-Horsfall protein displayed a tendency to form a gel.

Human glycoprotein HGP92 induces cytokine synthesis in mouse mononuclear phagocytes.

HGP92 has been shown to enhance in vitro and in vivo the bactericidal and tumoricidal activity of mouse macrophages. In this study we investigated the effect of HGP92 on the accumulation of cytokine mRNA in mouse inflammatory, peritoneal macrophages and the monocytic cell line J774. HGP92 significantly enhanced the level of cytokine mRNA for IL-1 alpha, IL-1 beta, IL-6, IL-10, IL-12, TNF-alpha and GM-CSF during the first 24 h of the incubation.

A 92-kDa human immunostimulatory protein.

We purified to apparent homogeneity a human urinary glycoprotein of 92 kDa (HGP.92) that, administered intravenously at 250 micrograms/kg, fully protected mice against a lethal inoculum of Listeria monocytogenes. Since HGP.

Differential heat-shock protein synthesis and response to stress in three avirulent and virulent Listeria species.

Two strains of Listeria monocytogenes, a virulent (V) and an avirulent (A) strain obtained by repeated in vitro cultivation at 37 degrees C, exhibited differing constitutive syntheses of heat-shock proteins (HSP) at 37 degrees C, the temperature of the infected host, and a differential response to heat treatment. These two strains also reacted differently to addition of a superoxide ion inducer and acid to treatments. Our observations were not limited to these two strains of L.

Absence of an early detectable increase in heat-shock protein synthesis by Listeria monocytogenes within mouse mononuclear phagocytes.

We investigated the behaviour of Listeria monocytogenes during the early phase of its in vitro phagocytosis by mouse resident peritoneal macrophages, and compared behaviour and modifications in protein synthesis occurring in a virulent and a non-virulent strain of L. monocytogenes. As previously shown, these two strains have differential responses to stress and heat shock in vitro.

Macrophage-induced cytotoxicity and anti-metastatic activity of a 43-kDa human urinary protein against the Lewis tumor.

Resident, inflammatory or bone-marrow macrophages from C57Bl/6 mice incubated in vitro with a pure human urinary protein (HGP.43) decreased the growth rate of Lewis tumor cells (3LL). This inhibition of 3LL growth was the result of a cytotoxic activity of these macrophages which was independent of oxygen metabolites and nitrous oxide.

Immunostimulatory human urinary protein.

We have previously extracted a protein from inflammatory mouse granuloma that fully protects normal mice against lethal Listeria monocytogenes infection. We now report that polyclonal antibodies directed against this protein react with a human urinary fraction that also provides full protection of normal mice against L. monocytogenes.

Inflammation and anti-tumor resistance. V. Production of a cytostatic factor following cooperation of elicited polymorphonuclear leukocytes and macrophages.

When Lewis tumor cells (3LL) included in a gel of polyacrylamide microbeads are injected into mice and recovered 1 day later, incorporation of 125I-UdR is strongly reduced. In contrast, no reduction is observed in irradiated mice. The cytostatic effect is non-existent in 6 hr-old granuloma (granulocytes 90%, macrophages 10%), but is maximal in 48 hr-old granuloma (granulocytes 50%, macrophages 50%).

Inflammation and anti-tumor resistance. IV. Induction of cytostatic activity of murine peritoneal cells by a mouse granuloma protein.

Peritoneal cells from C57B1/6 mice, incubated in vitro with a pure mouse granuloma protein (MGP), decreased growth of Lewis tumor cells (3LL). This cytostatic activity was due to the release of stable cytostatic factor(s) as determined by reduction of 125iododeoxyuridine (125I-UdR) incorporation in target cell cultures. Resident, inflammatory or bone-marrow-derived macrophages incubated with MGP had no cytostatic effect on tumor cells.

Remote effects of inflammation on non-specific immunity.

Following inflammation induced in mice with non-biodegradable, non-diffusible, and non-antigenic substances, host resistance is increased against bacteria, parasites and malignant cells injected at a distance from the inflammatory focus. This resistance is also increased in germ-free and nude mice. The increased resistance is correlated with (1) an increased leukopoiesis induced, at least in part, by a protein (MW = 40 kDa, pI = 5.

Radioprotective effect of an acute non-specific inflammation in mice.

Protection against 8.7 Gy whole-body gamma-irradiation (lethal in 100 per cent of mice by 30 days) was observed in 90 per cent of mice bearing a one-day-old granuloma induced by polyacrylamide beads. When the inflammatory reaction was induced sooner or later a lower or null protection was obtained.

[Endogenous counterinflammation and immunostimulation].

It is known since a century that inflammation is the cornerstone of the resistance against pathogens at the site of penetration. Recently, it has been shown that this beneficial effect can be found in a remote site. Furthermore, it is known that, at a distance from the site of inflammation, a decreased inflammatory reaction is observed: a counterinflammation.

Increased phagocytic activity in mice treated by a mouse granuloma protein.

Intravenous injection of a protein extracted from a talc-induced granuloma (MGP) enhanced the blood clearance of a highly virulent strain of Salmonella typhimurium. This protein was able to enhance mouse resistance to systemic infection with Listeria monocytogenes when injected one or two days prior to infection. Furthermore, since MGP-treated athymic mice were also protected against Listeria infection, mature T cells were most likely not involved in this enhanced resistance.

[Radioprotective effect of acute non-specific inflammation in mice].

A full protection against a 8,5 Gy gamma irradiation is observed in Mice bearing a one day old granuloma induced by polyacrylamide microbeads, when these Mice are injected, 1 hr before irradiation, with the eluate obtained from a 1 day old granuloma. In these Mice, a striking increase of the uptake of 125IUdR in bone marrow is observed.

Maintenance of granuloma macrophages in serum-free medium.

Granulomas were induced by injecting polyacrylamide beads into subcutaneous pouches created by divulsion of the dorsal skin of mice. More than 10(7) phagocytic cells (60% macrophages, 40% polymorphonuclear cells) could be recovered from this granuloma. The separation of the phagocytic cells can be achieved either following sedimentation in Percoll or following the incubation of cells on plastic petri dishes.

Immunostimulatory mouse granuloma protein.

Earlier studies have shown that from subcutaneous talc-induced granuloma in mice, a fraction could be extracted that fully protected mice against Listeria monocytogenes. Using standard biochemical procedures--i.e.

[Polyacrylamide-induced subcutaneous inflammation in mice].

The subcutaneous injection of polyacrylamide beads into mice induced an inflammation which allowed counting and characterization of inflammatory cells and substances contained within the granuloma. The cellular and protein content of the inflamed site was followed up for seven days. One day after injection of the beads, a large concentration of macrophages was found with a low content of polymorphonuclear leukocytes.

[Inflammation and antibacterial resistance. III. Influence of an inflammatory reaction induced by the injection of polyacrylamide gels on the resistance of mice to Listeria monocytogenes infection].

Inflammation was induced in the dorsal area of pathogen-free mice following subcutaneous injection of polyacrylamide microbeads (Biogel) of varying particle size (200-400 mesh) and pore size (exclusion limit ranging from a molecular weight of 2,000 to 300,000). Six days after injection of the microbeads, mice were infected with 10(5) Listeria monocytogenes. Only animals injected with "Biogel P2, P4, P6" (exclusion limit 2,000, 4,000 and 6,000 respectively) survived this lethal inoculum of Listeria.

[Inflammation and host resistance against bacteria. II.--Isolation of an immunostimulating fraction extracted from mouse inflammatory granuloma, using a new technic of isoelectrofocusing (author's transl)].

Five days following the subcutaneous injection, in the dorsal area of mice, of talc embedded in a calcium phosphate gel the induced granuloma were extracted, pooled and homogenized. After centrifugation, the supernatant was precipitated by 33% SO4(NH4)2. The resulting precipitate (G33) was dialysed, lyophilized and fractionated following preparative isoelectrofocusing either with "Ultrodex" or "Pevikon".