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Biomedical subjects

L Varesio

Publications and source records attributed to L Varesio.

At least 109 records · Page 6Linked to original sources
PubMed1984

In vivo activation of macrophages but not natural killer cells by picolinic acid (PLA).

Intraperitoneal injection of PLA into C57BL/6 mice induced high levels of macrophage-mediated cytostatic activity. Maximal cytostatic activity was found 3 days after injection of 100 mg/kg of PLA. In contrast, natural killer cell activity was not affected by PLA. Thus, PLA selectively modulates macrophage activity without influencing NK activity.

Animals↗
PubMed1984

Potent activation of mouse macrophages by recombinant interferon-gamma.

The ability of recombinant interferon-gamma (IFN-gamma) to activate mouse macrophages was investigated. The use of recombinant IFN-gamma has the advantage of being devoid of contaminating lymphokines. Two preparations of IFN-gamma were utilized, one which was not glycosylated and which was highly purified from Escherichia coli another which was glycosylated and which was from transfected COS-7 monkey cells. Both preparations of recombinant IFN-gamma activated murine macrophages to kill lymphoma and melanoma tumor targets, suggesting that glycosylation of the protein or the presence of other mammalian proteins is not essential for activation. Significant levels of cytolytic activity were induced from IFN-gamma (1 to 10 units/ml). This activity was undiminished by treatment of the IFN-gamma preparations with polymixin B at doses which neutralized endotoxin (50 micrograms/ml). Similarly, IFN-gamma, at low concentrations, induced an inhibition of migration by macrophages. Based on antiviral activity, IFN-gamma was shown to be 100 to 1000 times more potent than was IFN-beta as a macrophage-activating agent. Taken together, these results demonstrate that murine IFN-gamma is a macrophage-activating factor which is effective at physiological concentrations. Of particular interest is the observation that the nonglycosylated E. coli-derived IFN-gamma is active and therefore may be of value for therapeutic studies, since it can be easily produced in large amounts.

Animals↗
PubMed1984

Requirement for protein synthesis for induction of macrophage tumoricidal activity by IFN-alpha and IFN-beta but not by IFN-gamma.

Peritoneal mouse macrophages elicited by proteose-peptone (pM phi) were treated in vitro with IFN-alpha, IFN-beta, or IFN-gamma in the presence or absence of cycloheximide (Cy), a reversible inhibitor of protein synthesis, and were assayed for cytolytic activity against tumor cells. Inhibition of protein synthesis during treatment of pM phi with IFN-alpha or IFN-beta prevented the development of cytotoxic activity. In contrast, IFN-gamma was fully capable of inducing cytotoxic pM phi in the presence of Cy. Moreover, pM phi treated with mixtures of IFN in the presence of Cy were activated for cytotoxicity only by IFN-gamma together with IFN-alpha or IFN-beta, but not by IFN-alpha plus IFN-beta. These results indicate that the activation of pM phi by IFN-gamma is independent of new protein synthesis, whereas the activation of pM phi by IFN-alpha and/or IFN-beta requires active protein synthesis, suggesting that the mechanism of induction of cytotoxic pM phi by IFN-gamma differs from that by the other types of IFN.

Animals↗
PubMed1983

Selective inhibition by monosaccharides of tumor cell cytotoxicity mediated by mouse macrophages, macrophage-like cell lines, and natural killer cells.

A series of monosaccharides were tested for their ability to inhibit the effector phase of macrophage-mediated cytolysis against two susceptible murine tumor target cells, L5178Y and RL male I. Two monosaccharides, D-mannose and N-acetyl-D-galactosamine, were found to decrease cytotoxicity consistently in a dose-dependent manner. However, D-mannose preferentially inhibited lysis of RL male I target cells with little effect on lysis of L5178Y target cells, while the reverse was found with N-acetyl-D-galactosamine. Neither monosaccharide interfered with the activation of macrophages by polyinosinic:polycytidylic acid. Natural killer cell activity was decreased by a 25 mM concentration of D-mannose but not by N-acetyl-D-galactosamine, although increasing concentrations of N-acetyl-D-galactosamine were inhibitory. Neither monosaccharide affected cytotoxicity by alloimmune T cells. Cytotoxicity of macrophage-like cell lines against tumor target cells was also decreased by monosaccharides but the pattern of inhibition was different from that seen with activated macrophage effector cells. Both D-mannose and N-acetyl-D-galactosamine inhibited glucose oxidation by activated macrophages but only D-mannose significantly decreased protein synthesis of activated macrophages. These results indicate that monosaccharides can inhibit macrophage-mediated cytotoxicity in a selective manner with the pattern dependent on the tumor target cell used in the assay.

Animals↗
PubMed1983

RNA synthesis in activated macrophages I. Poly(I) X poly(C)-induced triggering of cytolytic activity is associated with decrease in RNA synthesis.

The effects of polyinosinic, polycytidylic acid [poly(I) X poly(C)] on the activation and RNA metabolism in murine peritoneal macrophages (M phi) elicited by proteose-peptone (pM phi) was investigated. Poly(I) X poly(C) triggered the cytolytic activity of pM phi and augmented their glucose oxidation. In contrast, a profound depression of [3H]uridine incorporation into RNA was observed in poly(I) X poly(C)-activated pM phi. The degree of depression of RNA labeling paralleled the dose of poly(I) X poly(C) used to activate the pM phi and the expression of tumoricidal activity. This decrease in [3H]uridine incorporation into M phi RNA could not be accounted for by decreased permeability of the activated M phi to [3H]uridine, or by instability of the labeled RNA. Moreover, analysis of the specific activity of the intracellular uridine triphosphate (UTP) pool and studies on the labeling of M phi RNA with [32P] orthophosphate indicated that the decreased RNA labeling was not due to changes in the specific activity of UTP. We concluded that poly(I) X poly(C)-activated pM phi exhibit a depressed rate of RNA synthesis. We suggest that the rate of RNA synthesis may be investigated as a potential new indicator for M phi activation.

Animals↗
PubMed1983

Suppression of in vitro maintenance and interferon-mediated augmentation of natural killer cell activity by adherent peritoneal cells from normal mice.

The ability of adherent peritoneal cells (APC) to inhibit murine natural killer (NK) cell activity was examined. Nylon wool-nonadherent splenic effector cells were incubated overnight with or without different numbers of APC. NK activity was then measured against YAC-1 in a 4-hr 51Cr-release cytotoxicity assay. Proteose peptone-elicited or unstimulated resident APC from normal mice markedly suppressed NK activity of splenic effector cells in the presence or absence of exogenously added interferon. The suppression was dependent on the number of APC added with 10% APC, relative to the number of effector cells, resulting in a greater than 65% inhibition of cytotoxicity. The effector phase of cytotoxicity was not the target of the suppressor cells, because APC did not suppress NK activity when they were present only during the cytotoxicity assay. The addition of APC to alloimmune cytotoxic T cells under similar conditions resulted in no inhibition of cytotoxicity. Both syngeneic and allogeneic APC suppressed NK activity, but several murine macrophage-like cell lines lacked this property. In contrast to APC, incubation of effector cells with adherent spleen cells from normal mice resulted in no inhibition of NK activity. APC from mice injected with C. parvum were less inhibitory for NK activity than normal resident APC. In contrast, C. parvum APC suppressed concanavalin A-induced lymphoproliferation and were directly cytotoxic to tumor target cells in vitro, whereas normal APC lacked these properties. The results indicate that the peritoneum of untreated mice contains suppressor cells that can inhibit the in vitro maintenance and IFN-mediated augmentation of NK activity. In addition, these results indicate a broader spectrum of immune reactivities regulated by APC and suggest that, depending on their level of activation, APC can preferentially inhibit different immune functions.

Animals↗
PubMed1983

Distribution of peritoneal macrophage populations after intravenous injection in mice: differential effects of eliciting and activating agents.

Murine peritoneal macrophages (pM phi) elicited in vivo by intraperitoneal (IP) inoculation of various agents were tested for their homing/distribution patterns after intravenous (IV) adoptive transfer to syngeneic C57BL/6 recipients. Resident pM phi (RpM phi) obtained from normal mice and pM phi elicited by proteose peptone (PpM phi) or thioglycollate broth (TpM phi) exhibited similar homing patterns following IV transfer. After initial arrest in the lungs, these cells rapidly disseminated to liver and spleen, with minimal or no detectable migration to peripheral lymph nodes, intestine, peritoneum, kidney, heart, or retention in the blood. The pattern of results reflected the properties of pM phi themselves, since highly enriched pM phi populations obtained by treatment of crude peritoneal exudate cells with anti-Thy 1.2 + C, or by fractionation on Percoll density gradients, gave similar results. The distribution of pM phi elicited by Brewer's thioglycollate medium (BTpM phi) was markedly different from other pM phi tested. BTpM phi homed rapidly to the lungs and many remained localized there for at least 72 hr with very little migration to the spleen. The distribution of PpM phi could be altered by activation of these cells in vivo through the IP injection of the pyran copolymer, MVE-2, prior to adoptive IV transfer. Activated PpM phi contained a population of highly differentiated, low density pM phi, separable on density gradients, which arrested in the lungs for appreciably longer periods of time than did PpM phi. These cells exhibited reduced ability for migration to the spleen. Macrophage-like (M phi-like) cell lines did not exhibit migration capability, but rather were rapidly cleared from the circulation in a manner similar to other types of tumor cells.

Animals↗
PubMed1982

Suppressor macrophages in tumor-bearing mice. Inconsistency between in vivo and in vitro findings?

The growth of a spontaneous adenocarcinoma (ADK-1t) of BALB/c mice (H-2d, MIsb) induces progressive hyporesponsiveness of spleen cells stimulated with PHA or irradiated leukocytes from DBA/2 strain (H-2d, MIsa) in mixed leukocyte culture. This hyporesponsiveness is due to the suppressor activity of macrophage-like cells, since it is abrogated by passage over nylon-wool columns, pretreatment with carrageenans, or carbonyl iron and magnet. BALB/c mice bearing greater than 20 mm mean diameter ADK-1t tumors, and displaying a drastic impairment of T lymphocyte reactivity in vitro, were challenged with the P815 mastocytoma of the DBA/2 strain. These mice rejected increasing numbers of P815 cells to the same extent as normal BALB/c mice. The reactivity towards P815 is abrogated by 450 rad, and can be reconstituted by the adoptive transfer of 2 X 10(7) normal T lymphocytes in either normal or ADK-1t bearing BALB/c mice. These data show that a discrepancy exists between the capacity of T lymphocytes from ADK-1t bearing mice to react against DBA/2 cells tested in vitro or in vivo, and indicates that great caution should be exercised in predicting the responsiveness of the immune system on the basis of in vitro data on the suppressor activity of macrophages.

Adenocarcinoma↗
PubMed1982

Interferon activates macrophages to produce plasminogen activator.

Macrophages and monocytes become activated, by a variety of mechanisms, to exhibit cytotoxicity. Associated with this activation for cytotoxicity is the production of certain neutral proteases, especially plasminogen activator (PA), which may be involved in the mechanism of cytolysis. Supernatants from concanavalin A (Con A) or antigen-stimulated spleen cells contain factor(s) which render macrophages and monocytes activated in vitro (MAF/MIF). We have found that in addition to such spleen cell supernatants murine interferon was able to activate peptone-induced C57BL/6 peritoneal macrophages to produce PA but had no effect on human monocytes. Similarly, human recombinant interferon activated human monocytes but not murine macrophages to produce PA. In addition to displaying this species specificity, interferon was able to function as a monocyte/macrophage activator in this system just as in cytotoxicity and may be a regulator of monocyte/macrophage function in vivo by modulation of proteolytic enzymes.

Animals↗
PubMed1981

Systemic and in situ natural killer and suppressor cell activities in mice bearing progressively growing murine sarcoma-virus-induced tumors.

The activity of systemic and in situ natural killer (NK) cells was measured in 3- to 5-week-old CBA mice with progressively growing murine sarcoma virus (MSV)-induced tumors. Splenic NK activity was depressed in tumor-bearing mice. NK activity was quite low or not detectable in unfractionated cell suspensions from the tumors 10-12 days after inoculation of virus, but activity was observed upon depletion of phagocytic and/or adherent cells. To determine whether this in situ inhibition of NK activity was correlated with the presence of previously described suppressor cells, subpopulations of cells from the tumors were tested for their ability to suppress various responses of normal spleen cells: NK activity, lymphoproliferative responses to concanavalin A, and proliferation-independent production of macrophage inhibition factor. A similar pattern of suppressor activity was seen in all of the assays, indicating the presence of pleiotropic suppressor cells in progressively growing MSV-induced tumors.

Animals↗
PubMed1981

Microsystem to evaluate the incorporation of 3H-uridine in macrophage RNA.

A method is described for the evaluation of the total 3H-uridine incorporated by macrophages in vitro into trichloroacetic acid (TCA)-precipitable material. The technique is based upon solubilization of the macrophage monolayers by guanidine-HCl, followed by TCA precipitation. The recovery of RNA into the precipitate and the reproducibility of the results were strictly dependent on the use of filtered reagents and on incubation of the TCA precipitate for 2 or more hours at 4 degree C before harvesting. Treatment with quanidine-HCl did not affect the recovery of labeled RNA. Moreover, we observed that radioactive precipitate had the characteristics of RNA, since its recovery was sensitive to the addition of unlabeled uridine in the culture medium and to the treatment of the macrophages with inhibitors of RNA synthesis, but not of protein synthesis. Tritiated uridine incorporation in microcultures of macrophages can be assessed with this technique, by processing the cells directly in the wells. The main advantages of this procedure are: 1) the radioactivity can be measured by semiautomatic cell harvesters, 2) a small number of macrophages are required, and 3) many samples can be processed simultaneously. Overall, the technique is simple, rapid, and could be successfully adapted to study other metabolic pathways.

Animals↗
PubMed1981

In vitro induction of tumoricidal and suppressor macrophages by lymphokines: possible feedback regulation.

Proteose-peptone-induced macrophages (pM phi) from C57BL/6 (B6) mice were treated in vitro with lymphokine, and 2 functions were evaluated: the ability to suppress lymphokine production and the cytotoxic capacity against tumor target cells. When 10% to 20% pM phi treated with lymphokine were added to a lymphokine-producing system, strong inhibition of MIF and MAF production occurred. The suppression was not specific, since pM phi inhibited the production of lymphokine obtained by either mitogenic stimulation of normal spleen cells (NSC) or alloantigen stimulation of immune spleen cells (ISC). The suppressor cells were adherent, phagocytic, Thy 1.2 negative, with monocyte-macrophage-like morphology. Lymphokine-treated pM phi were also tumoricidal when tested in 18 hr microcytotoxicity assay against RL male 1 lymphoma target cells. However, using endotoxin-free reagents (less than or equal to 0.01 ng/ml of endotoxin by the LAL assay), we found that small amounts of lipopolysaccharide (LPS) were required, in addition to lymphokine, to induce tumoricidal activity in pM phi. In contrast, noncytotoxic pM phi stimulated with lymphokine alone were also able to inhibit lymphokine production, although not as effectively as pM phi stimulated by lymphokine plus LPS. Therefore, we concluded that lymphokine treatment itself is sufficient to induce M phi to become suppressor cells, whereas an additional signal is necessary to induce cytolytic activity. We suggest that there is a negative feedback mechanism of control on the lymphokine-producing cells through the activation of suppressor M phi by lymphokine.

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