PubMed HealthSearch

Biomedical subjects

B Vray

Publications and source records attributed to B Vray.

At least 19 recordsLinked to original sources

IFN-gamma receptor-deficient mice are hypersensitive to the anti-CD3-induced cytokine release syndrome and thymocyte apoptosis. Protective role of endogenous nitric oxide.

Mice with a disruption of the IFN-gamma receptor alpha-chain gene (IFN-gamma R alpha o/o mice) were found to be significantly more sensitive than their wild-type counterparts to induction of the anti-CD3-induced disease syndrome. Specifically, when given a selected dose of anti-CD3 Ab, IFN-gamma R alpha o/o mice developed severe hypothermia and hypoglycemia, leading to 100% mortality within 72 h. In contrast, wild-type mice failed to develop overt pathologic manifestations and survived. Histologic examination revealed apoptosis in thymuses and spleens, which were significantly more pronounced in the mutant than in the wild-type mice, as confirmed by flow cytometric and DNA electrophoretic analysis. Apoptosis affected mainly CD4+CD8+ and CD4+CD8- thymocytes. Other histologic alterations were steatosis in livers, and erythrocyte extravasation and infiltration of apoptotic cells in lungs, all of which were exclusively observed in IFN-gamma R alpha o/o mice. Blood levels of TNF, IL-2, IL-6, and IL-10 were slightly more elevated in IFN-gamma R alpha o/o mice, but insufficiently so to explain increased disease severity. Thus, even more elevated cytokine levels in wild-type mice receiving high doses of anti-CD3 Ab were not associated with morbidity or apoptosis. Blood levels of IFN-gamma were barely detectable in anti-CD3-challenged wild-type mice, but were relatively high in the mutant mice. Increased susceptibility of IFN-gamma R alpha o/o mice was associated with impaired nitric oxide (NO) production, as indicated by significantly lower plasma nitrite levels and by more transient expression of spleen inducible NO synthase mRNA. Moreover, treatment of wild-type mice with the NO synthase inhibitor N-nitro-L-arginine methylester resulted in increased anti-CD3-induced morbidity and mortality. The data indicate that IFN-gamma R alpha o/o mice produce less NO and are therefore more sensitive than wild-type mice to the deleterious effect of anti-CD3 Ab.

Animals

Interferon-gamma-activated immature macrophages exhibit a high Trypanosoma cruzi infection rate associated with a low production of both nitric oxide and tumor necrosis factor-alpha.

Murine peritoneal macrophages (MPM) can be subdivided into two subpopulations of mature and immature macrophages. In contrast to mature macrophages, immature ones were highly susceptible to Trypanosoma cruzi infection. This highly susceptibility was associated with a low production of alpha 2-macroglobulin. Interferon-gamma (IFN-gamma)-activated immature macrophages also exhibited a higher infection rate than did IFN-gamma-activated mature ones. This higher rate of infection was associated with a low production of both nitric oxide (N = O) and tumor necrosis factor-alpha (TNF-alpha). In contrast, mature MPM showed a lower rate of infection and produced higher levels of N = O and TFN-alpha. Taken together, these results show a clear-cut difference in the course of T. cruzi infection in relation to the macrophage maturation state.

Animals

Eicosanoids modulate CR1- and Fc-dependent bacterial phagocytosis.

It is known that macrophages produce large amounts of eicosanoids during phagocytosis and that pharmacological concentrations of prostaglandin E2 (PGE2) inhibit phagocytosis in several models. However, the physiological effect on phagocytosis of endogenous prostaglandins, produced during CR1- or FcR-mediated bacterial phagocytosis, remains unclear. In this study, we show that indomethacin inhibits the CR1- but not the FcR-dependent phagocytosis of bacteria by rat peritoneal cells in the same range of concentrations that inhibit the synthesis of PGE2, PGI2 and thromboxane A2. An exogenous supply of PGE2 and PGE1 (10(-10) to 10(-8) M) restored the CR1-mediated phagocytosis; higher concentrations were inhibitory. Our data indicate that PGE2 and/or PGI2, produced by rat peritoneal cells, are involved in CR1-dependent bacterial phagocytosis.

Alprostadil

Trypanosoma cruzi but not Trypanosoma brucei fails to induce a chemiluminescent signal in a macrophage hybridoma cell line.

Macrophage-Trypanosoma cruzi interactions were studied by using a newly generated macrophage hybridoma cell line (2C11-12) that was selected for its capacity to produce high levels of reactive oxygen intermediates. This cell line was found to be a suitable host cell for T. cruzi, and intracellular parasitic development could be inhibited by activation with gamma interferon. When exposed to opsonized Trypanosoma brucei, Micrococcus lysodeikticus, or Legionella pneumophila, the activated macrophage cell line produces a high chemiluminescent signal, indicating the release of reactive oxygen intermediates. Alternatively, when opsonized T. cruzi was added to these activated macrophages, this parasite failed to stimulate a chemiluminescent response, suggesting an impairment in the triggering of the respiratory burst.

Animals

Macrophage heterogeneity in prostaglandins and thromboxane synthesis: differential activation by Fc- and C3b-dependent bacterial phagocytosis.

Resident peritoneal macrophages, obtained from rats, were separated into subpopulations by centrifugation on a Percoll discontinuous density gradient. Nine fractions of pure macrophages were isolated. Each subpopulation was studied for Fc- and C3b-dependent bacterial phagocytosis and assayed for the related synthesis of PGE2, TxA2 and PGI2, measured by their stable metabolites TxB2 and 6-Keto-PGF1. The results show that with decreasing density, which corresponds to a greater maturity, the production of PGE2 increases and that of TxB2 and 6-Keto-PGF1 decreases. The cells of low density were mostly stimulated by IgG-opsonized bacteria, whereas those of high density responded preferentially to C3b- opsonized bacteria. This pattern is roughly similar to the one characterizing the phagocytosis via these two receptors although the correlation is not absolute. It can be concluded that enzymes involved in the metabolism of arachidonic acid, as well as receptors for C3b and IgG, are differentially expressed among resident macrophage subpopulations and thus during macrophage maturation.

Animals

Partial characterization of a Legionella pneumophila serogroup 1 immunodominant antigenic determinant recognized by a monoclonal antibody. Legionella specific antigenic determinant.

A monoclonal antibody was used to characterize a serogroup 1 specific Legionella pneumophila Philadelphia strain 1 antigenic determinant. A quantitative fluorometric assay was developed to quantitate the antibody sites (2.7 +/- 0.4 X 10(5)) on Legionella bacteria and to determine the physico-chemical parameters of the antibody-antigen interaction (at 4 degrees C: delta G = -10.9 Kcal X mol-1, delta H = 1.7 Kcal X mol-1, delta S = 45 cal X K-1 X mol-1). The same method was used to study the modification or the removal of the antigen by chemical and enzymatic means (trypsin, papain, lysozyme, acetone, chloroform-methanol and Tris-EDTA); only Tris-EDTA extraction resulted in a significant decrease in antibody binding sites. Inhibition studies of the fluorescein-labelled antibody binding were performed with different sugars of which only L-fucosylamine was inhibitory, and with other monoclonal antibodies to Legionella pneumophila serogroup 1 in order to compare their fine specificity and affinity. The results indicate that the epitope recognized was an immunodominant carbohydrate including an aminodideoxyhexose and carried by the lipopolysaccharide.

Antibodies, Bacterial

Pregnancy and humoral immune response in mice chronically infected by Trypanosoma cruzi.

The effect of pregnancy on the humoral immune response induced by Trypanosoma cruzi was studied in groups of chronically infected and pregnant mice (IP) or chronically infected and nonpregnant mice (INP) of strain BALB/c. Groups of noninfected and nonpregnant mice (NINP) or noninfected and pregnant mice (NIP) served as controls. The pregnant mice were killed on day 17 of pregnancy. Anti-T. cruzi immunoglobulin G (IgG) and IgM antibodies, detected by immunofluorescence or enzyme-linked immunosorbent assay or both, underwent a pregnancy-associated decrease of 20 to 40%, whereas complement-mediated lytic antibodies were unaffected by pregnancy. Immunoblotting analysis indicated identical specificities of the anti-T. cruzi antibodies in IP and INP groups. The levels of all the immunoglobulin isotypes (particularly IgG2a and IgG3), circulating immune complexes, rheumatoid-like factor, and anti-DNA antibodies were considerably increased during chronic infection (NINP versus INP), which could be related to the high degree of polyclonal B-cell activation occurring in T. cruzi infection. However, pregnancy significantly decreased (by 20 to 60%) such parameters. IgG levels were particularly affected (by 40 to 60%), and the decreases could be ordered as follows: IgG3 greater than IgG2a greater than IgG1 greater than IgG2b for IP versus INP. Comparisons between the noninfected groups indicated differences only in IgG levels. These results indicate the following. (i) The specific humoral anti-T. cruzi immune response is weakly affected by pregnancy, which is not sufficient to modify the course of the mother's infection. (ii) Pregnancy does not modify the expression of the anti-T. cruzi antibody repertory. (iii) Pregnancy reduces the polyclonal B-cell activation, particularly the levels of the IgG isotypes undergoing the greatest activation.

Animals

Detection of Gardnerella vaginalis in vaginal specimens by direct immunofluorescence.

The preparation of a fluorescein-labeled Gardnerella vaginalis polyclonal antibody is described, and its usefulness is assessed for the detection of this microorganism in vaginal samples obtained from 263 women attending the gynecological department of a general hospital, 66 of whom harbored an intrauterine device. The direct immunofluorescence technique was positive for G. vaginalis in 21% of the specimens, whereas only 12.5% of the total bacteriological cultures were positive. The frequency was higher in patients harboring intrauterine devices since 34.8% exhibited positive immunofluorescence and 30.3% exhibited positive cultures.

Animals

Role of vitamin E in the protection of the resident macrophage membrane against oxidative damage.

The variation in tocopherol content of resident peritoneal rat macrophages was investigated during an oxidative stress provided by superoxide anions. Fluorometric measurements showed that phagocytic cells contain 298 +/- 18 ng vit.E/mg prot. The vitamin E level remains nearly constant during 1 h of incubation: 266 +/- 46 ng vit.E/mg prot. HPLC control at 37 degrees C validates our fluorometric measurement. Superoxide anions (O2-.) synthesis was activated by phorbol myristate acetate (PMA) (0.5 microgram/ml), after 1 h of incubation a decrease of 40% of the macrophage tocopherol level was observed and assessed by HLPC control. No tocopherolquinone (TQ) was detected. To clarify this point, tocopherol oxidation was followed spectrophotometrically. Results did not show any appearance of TQ at 265 nm but appearance of a peak at 307 mm. This our results show for the first time that macrophages possess vitamin E which plays a partial role in the protection of their plasma membrane. The lack of detection of TQ is of interest and the study of this unidentified product of oxidation should help us to understand the exact metabolism of vitamin E.

Animals

Standardization for diagnostic purposes of a monoclonal antibody against Legionella pneumophila.

A monoclonal antibody against Legionella pneumophila has been produced and characterized. The antibody was of the gamma-3 isotype and recognised the lipopolysaccharides of the bacteria as was confirmed by Western blotting. Preliminary assays were performed for the detection of the antigen in clinical samples in order to set up a rapid laboratory technique for the early diagnosis of legionellosis. By using an immuno-enzymatic technique, less than 30 000 bacteria or corresponding antigens could be detected in a few hours.

Animals

Wheat germ agglutinin potentiates uptake of bacteria by murine peritoneal macrophages.

Exposure of thioglycollate-elicited murine peritoneal macrophages to wheat germ agglutinin (WGA) increased markedly the uptake of six different bacteria, which have surface receptors for the lectin. Uptake of Staphylococcus aureus H was higher by 3-5-fold, of S. aureus 52A2 by 1.8-fold, of S. aureus 52A5 by 1.7-fold, of S. albus by 2.3-fold, of Shigella flexneri by 6-fold and of Micrococcus luteus by 6.5-fold. Klebsiella pneumoniae, devoid of receptors for WGA, was not phagocytosed following pretreatment of macrophages with the lectin. Pretreatment of the bacteria with the lectin also resulted, in most cases, in an increase in phagocytosis. Interaction of WGA with the macrophages and with the bacteria, as well as the potentiation of phagocytosis, was abolished by tri-N-acetylchitotriose, a saccharide that binds specifically to WGA, but not by monosaccharides which do not interact with this lectin. With non-elicited macrophages, enhancement of phagocytosis by WGA was less pronounced, probably because of the higher number of lectin-binding sites (5-fold) on the elicited cells. Peanut agglutinin and soybean agglutinin, that bind to macrophages but not to the bacteria studied, lack the ability to potentiate phagocytosis. Macrophage surface sugars thus appear to play an important role in phagocytosis by serving as receptors for lectins that form bridges between the macrophages and the microorganisms.

Animals

[Macrophage phagocytic function].

Described a century ago by Metschnikoff , phagocytosis plays an essential role in the defence of organisms faced with bacterial infection. Phagocytosis is ensured essentially by neutrophils and macrophages. In cases of inflammatory response, chemotactic factors induce an afflux of neutrophils which predominate at the site of infection for the first few hours. Then macrophages intervene to try and eliminate the infectious agent. If the latter can be rapidly ingested and degraded, the inflammatory response will gradually dwindle . On the other hand, if the infection persists (bacteria resistant to phagocytosis, or to bactericidal agents), the inflammatory response will become chronic. Macrophage anti-bacterial activity is therefore essential and can be broken down into several stages: first chemotaxis, which initiates macrophage migration towards the inflammatory focus. Then adherence to bacteria, via opsonins and membrane receptors. This binding releases an intra-cytoplasmic signal which governs the following stages, i.e. actual ingestion (phagocytosis), then enzymatic destruction within phagolysosomes " bactericidosis ". The phase of recognition and membrane binding to bacteria thus occupies an important place in the physiological role of macrophages. Two receptors are especially involved: the Fc receptor and the C3b receptor. The ingestion phase appears to be initiated by the redistribution of these membrane receptors, which first reform in "micro-clusters" (reversible small aggregates), then in "clusters" (irreversible larger aggregates), which finally become "patches" and "caps". " Bactericidosis " then ensures in most cases the disintegration of the infectious agent, by systems that can be either dependent or independent of oxygen. The main techniques for the study of macrophage phagocytosis are briefly reviewed.

Animals

Genetic deficiency of C4 presenting with recurrent infections and a SLE-like disease. Genetic and immunologic studies.

A young girl presenting with recurrent pulmonary infections and atypical lupus erythematosus was totally deficient in C4. In one sister, also deficient in C4, the same symptoms developed. Results of family studies were consistent with an autosomal recessive mode of transmission and with linkage of the genes determining C4 deficiency to those of the major histocompatibility complex. The patient's serum and red cells were Chido- and Rodgers-negative. Humoral and cellular immunity were normal, except for a low lymphocyte response in mixed lymphocyte culture. The cellular function of the patient's polymorphonuclear leukocytes was normal, for both phagocytosis and bactericidal activity using Candida albicans. However, in the presence of C4-deficient serum, opsonin generation and bactericidal indexes were diminished. These defects were completely reversible upon addition of purified C4.

Child, Preschool

Switched allotype expression in an immunoglobulin-nonsecreting rabbit lymphoid cell line fused with rabbit gangliocytes.

The Simian virus 40-transformed rabbit spleen cell TRSC-1 synthesizes intracellular whole IgG molecules of the alb4 allotype. Two hypoxanthine-guanine phosphoryl transferase-deficient mutants were derived from this line. One of these, TRSC-1-8, was used in somatic cell fusion experiments together with gangliocytes from a rabbit immunized against beta-galactosidase. Out of nineteen hybrid clones surviving in selective medium, only one, L17, was shown to produce free gamma chains which express the a2 allotype of the donor rabbit rather than the al marker of the parents TRSC-1-8 line. The inability to restore IgG secretion in hybrids suggests that dominant regulatory controls are exerted by the TRSC-1 genome on Ig reduction. This supports the notion that the TRSC-1 line originated from a splenocyte that had not reached the final plasmocyte differentiation stage at the time of viral transformation.

Animals