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L Pitzurra

Publications and source records attributed to L Pitzurra.

At least 37 records · Page 2Linked to original sources
PubMed1993

Protective effect of picolinic acid on mice intracerebrally infected with lethal doses of Candida albicans.

We have studied the effects of picolinic acid (PLA), a product of tryptophan degradation, on mouse susceptibility to intracerebral infection with Candida albicans. We show that intraperitoneal administration of PLA significantly enhances the median survival time of mice inoculated with the lethal challenge. Furthermore, intracerebral administration of this agent induces a protective state against the local lethal infection, the phenomenon depending upon the administration schedule and doses of PLA employed. According to survival data, yeast growth in the brain as well as yeast colonization of the kidneys are drastically reduced in PLA-treated mice compared with those for untreated controls. Northern (RNA) blot analysis of brain tissues demonstrates that mRNA levels specific for tumor necrosis factor and interleukin 1 are augmented and induced, respectively, after inoculation of PLA. These results indicate that PLA has a protective effect likely involving elicitation of a cytokine response in vivo against fungal infections.

Animals↗
PubMed1993

Toxic effects of tetanus toxin on GG2EE macrophages: prevention of gamma interferon-mediated upregulation of lysozyme-specific mRNA levels.

By using a nonneuronal cell system, evidence has previously been provided that tetanus toxin (TT) intoxication occurs in macrophages, impairing their secretory activity as well as their antitumoral activity. In particular, both secreted and total lysozyme (LZM) activities are reduced by TT treatment, provided that GG2EE macrophages have been preexposed to gamma interferon (IFN-gamma). In an attempt to provide insight into the molecular mechanisms underlying this phenomenon, we focused our attention on the levels of LZM-specific transcripts. GG2EE macrophages preexposed to IFN-gamma exhibited augmented levels of LZM-specific mRNA. Such an effect was detected 1 h after removal of IFN-gamma, peaked at 3 h, and gradually decreased with time in culture. Exposure of IFN-gamma-pretreated GG2EE macrophages to TT resulted in the prevention of the IFN-gamma-mediated upregulation of LZM mRNA levels. The phenomenon was mediated by the holotoxin (> or = 1 micrograms/ml) and abrogated by preexposure of the macrophages to the C fragment of TT. Protein kinase C (PKC) and Ca(2+)-calmodulin-dependent PK were likely involved in the IFN-gamma-mediated upregulation of LZM mRNA levels and biological activity, as assessed by PK inhibitors. Furthermore, PK inhibitors mimicked TT in impairing LZM activity of GG2EE macrophages, thus suggesting that impairment of PKC and/or the Ca(2+)-calmodulin-dependent PK pathway(s) may be one of the events involved in TT intoxication of macrophages.

Animals↗
PubMed1992

Candida albicans hyphal form enhances tumor necrosis factor mRNA levels and protein secretion in murine ANA-1 macrophages.

We have demonstrated that Candida albicans in its hyphal form (H-Candida) acts as a stimulating agent in the cloned macrophage population ANA-1. Both tumor necrosis factor (TNF) mRNA levels and secreted biological activity augment in ANA-1 macrophages exposed to H-Candida. Such effects are observed at an effector-to-target cell ratio of 1:1 and occur after 1 and 3 hr of coincubation, respectively. The phenomenon is independent of the metabolic status of the fungus, since viable as well as heat-killed H-Candida are comparable in inducing TNF mRNA levels. The extent and kinetics of H-Candida-mediated effects are similar to those observed following exposure of ANA-1 macrophages to lipopolysaccharide (LPS). This implies that C. albicans in its hyphal form is a potent macrophage modulator; whether it acts through the same mechanism(s) as LPS remains to be elucidated.

Animals↗
PubMed1992

Early differential molecular response of a macrophage cell line to yeast and hyphal forms of Candida albicans.

The dimorphic transition of Candida albicans from the yeast (Y-Candida) to the hyphal (H-Candida) form is a complex event; the relevance of this transition in fungal pathogenicity is still poorly understood. By using a cloned macrophage cell line (ANA-1), we questioned whether the interaction between macrophages and Y-Candida or H-Candida could affect specific cell functions, i.e., tumor necrosis factor and lysozyme production. We found that ANA-1 macrophages selectively responded to H-Candida with increased tumor necrosis factor and downregulated lysozyme, as assessed by measurement of relative mRNA levels and secreted biological activities. The H-Candida-mediated effects were (i) dependent upon the ratio between ANA-1 macrophages and H-Candida, (ii) detectable after 1 h of coincubation, and (iii) accomplished without fungal ingestion. Conversely, Y-Candida, which was found inside the ANA-1 macrophages, did not affect tumor necrosis factor and lysozyme production, nor did it prevent the macrophage response to other stimuli. Overall, these results indicate that a macrophage can distinguish between Y-Candida and H-Candida and that only the latter is able to modulate specific functions. H-Candida is recognized and probably processed as an extracellular target. The possible implication of macrophages as autocrine and paracrine regulatory cells during Candida infections is discussed.

Animals↗
PubMed1990

A rapid objective immunofluorescence microassay. Application for detection of surface and intracellular antigens.

An indirect immunofluorescence microassay, which permits automated reading, has been employed for simple, rapid and objective detection of surface and intracellular antigens. Initially, the cells, spun in microplates, are fixed with glutaraldehyde (0.25% v/v in PBS). Following fixation, the cells can be stored at 4 degrees C for up to 2 weeks before being used in the immunofluorescence microassay. The fixed cells are then stained according to standard procedures using appropriate first and fluorescein-conjugated second antibodies. An automated and quantitative evaluation of the fluorescence intensity of the cell samples was achieved using the Titertek Fluoroskan II automatic reader. This microassay was shown to be suitable for the detection of the surface MAC1 antigen and intracellular v-myc protein in the GG2EE macrophage cell line.

Animals↗
PubMed1990

Gamma interferon-induced specific binding of tetanus toxin on the GG2EE macrophage cell line.

We have recently demonstrated that tetanus toxin (TT) selectively inhibited gamma interferon-(IFN-gamma)-induced, but not basal, lysozyme activity in the GG2EE macrophage cell line. By an indirect immunofluorescence technique, we show that tetanus toxin, as well as the BIIb tetanus toxin-derived fragment, selectively binds to IFN-gamma-treated cells but fails to bind to control cells. Moreover, BIIb fragment and TT share the same binding sites, as demonstrated by competition/displacement experiments. These data indicate that IFN-gamma pretreatment results in the acquisition of tetanus toxin binding sites on GG2EE cells by a mechanism(s) yet to be identified.

Animals↗
PubMed1989

Selective inhibition of cytokine-induced lysozyme activity by tetanus toxin in the GG2EE macrophage cell line.

This study was designed to evaluate the effects of tetanus toxin (TT) on lysozyme (LZM) activity by the GG2EE macrophage cell line. GG2EE cells spontaneously produced low amounts of LZM, which were mostly secreted into the culture medium. Upon treatment with various cytokines, GG2EE cells exhibited altered LZM activity. In particular, exposure of GG2EE cells to alpha/beta interferon (IFN-alpha/beta) reduced LZM activity, as opposed to treatment with gamma interferon (IFN-gamma) or colony-stimulating factor 1, which potentiated LZM activity. Spontaneous LZM activity of GG2EE cells was not susceptible to TT action; in contrast, when IFN-gamma- or colony-stimulating factor 1-susceptible cells were treated with TT, a significant reduction on LZM activity was observed. The TT inhibitory effect was dose dependent and manifested only after a 6-h incubation of GG2EE cells with TT. Treatment of GG2EE cells with heat-inactivated TT as well as Ibc- and B-IIb-TT-derived fragments was found to be ineffective, while pretreatment with B-IIb- but not with Ibc-TT-derived fragment abrogated the TT effect. Overall, these data indicate the existence of a specific TT-GG2EE cell interaction, leading to selective inhibition of cytokine-induced LZM activity.

Animals↗
PubMed1983

Comparison of passive hemagglutination with Turkey erythrocyte assay, enzyme-linked immunosorbent assay, and counterimmunoelectrophoresis assay for serological evaluation of tetanus immunity.

Antibody titers to tetanus toxin in human sera were assayed by passive hemagglutination with turkey erythrocytes, enzyme-linked immunosorbent assay, and counterimmunoelectrophoresis. The first two of these tests were shown to be the most sensitive for antibody detection, having the same range of sensitivity and reproducibility. The antibody levels determined by these assays were up to 400-fold higher than those determined by counterimmunoelectrophoresis. The turkey erythrocyte hemagglutination assay requires only 40 min, whereas the immunosorbent assay method requires 24 h. These results suggest that the hemagglutination assay is the more appropriate method for rapid and sensitive determination of tetanus antibody levels.

Journal Article↗
PubMed1983

Use of turkey red blood cells in the passive haemagglutination test for studying tetanus immunity.

Passive haemagglutination (HA) assays were performed using turkey red blood cells (TRBC-HA) on sera from normal healthy people, from normal people previously immunized against tetanus, and from tetanus patients receiving human antitetanus immunoglobulins. The TRBC-HA test was compared with haemagglutination assays using sheep red blood cells (SRBC-HA) and with the neutralization (NT) test, and was found to be more sensitive than the SRBC-HA test and showed good correlation with the NT test.While the SRBC-HA assay calls for adsorption of sheep red blood cell agglutinins from the sera to be tested, the use of turkey red blood cells does not require any such adsorption. In addition, the TRBC-HA assay can be performed in 40 minutes compared with 6 hours for the SRBC-HA assay. All these advantages make the TRBC-HA assay a more useful test for screening large numbers of sera in the evaluation of tetanus immunity of normal people and of patients with wounds when seen in the emergency room of hospitals.

Animals↗
PubMed1981

[Immunogenic activity of hemocyanin from the hemolymph of Helix pomatia and Octopus vulgaris].

The immunogenic activity of Helix Pomatia Haemocyanin (HPH) and Octopus vulgaris Haemocyanin (OVH) has been tested in rabbits. Primary and secondary antibody response has been evaluated in sera by agar double immuno-diffusion test (Ouchterlony), counter immunoelectrophoresis and complement fixation. Bath haemocyanins proved highly antigenic: antibody titers rising up to 1/3.200 after the second antigenic challenge. The Ouchterlony test showed a partial identity between H.P.H. and O.V.H. No adverse effect was evident in injected animals.

Animals↗
PubMed1980

[Serological studies of the immunological condition versus rebulla virus in maternal and neonatal serum samples].

In 116 serum pairs sampled from the mother and the newborn at delivery, the antibody level against Rubeola virus has been assessed haemagglutination inhibition test. Thirty one per cent of the mothers resulted not protected, as the antibody titer was below 1/16. A situation akin to that described in the same percentage of not protected women was 29,45%. In 19% of the newborns the antibody level against the virus proved higher than the mother's one by at least two dilution factors. Research work is in progress to try the reason of this result, as a possible consequence of different Ig classes or aspecific inhibitors involved.

Adult↗
PubMed1979

[Preservation at -195 degrees C of trypsinized organs for primary cultures].

Trypsinized cells from rabbit kidney can be kept at -195, 8 degrees C indefinitely. When necessary, propagation in monolayer cultures is readily obtained as primary cultures, provided that some conditions be respected: 1) only glass culture flask must be used. Falcon plastic vessels do not allow cell attachment in this type of primary culture 2) the lag period is particularly long: it taken about 20 days before the beginning of cell multiplication 3) fetal calf serum is toxic for the cells in the 2nd and 3rd change of the culture medium, during the lag period.

Animals↗
PubMed1979

[Intestinal secretory IgA and bacterial antigens in oral administration].

The proposed technique for purification and concentration of secretory IgA has been applied to intestinal wash fluids obtained by clisma from 15 patients under bacterial extracts treated (COLOPTENE-DEBAT). The collected data show that the technique is suitable for routinary use and the bacterial extracts do increase the amount of IgA in the intestinal contents.

Administration, Oral↗