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S Montplaisir

Publications and source records attributed to S Montplaisir.

At least 19 recordsLinked to original sources
PubMed1997

Expression of a reporter gene interrupted by the Candida albicans group I intron is inhibited by base analogs.

We previously reported the identification of an intron (CaLSU) in the 25S ribosomal RNA of some Candida albicans yeast strains. CaLSU was shown to self-splice and has the potential to adopt a secondary structure typical of group I introns. The presence of CaLSU inC. albicans strains correlates with a high degree of susceptibility to base analog antifungal agents, 5-fluorocytosine (5-FC) or 5-fluorouracil (5-FU). Cell death, resulting from addition of base analogs to growing cultures, precluded demonstration of a causal relationship between CaLSU presence and susceptibility to base analogs. In the present study, CaLSU was inserted in a non-essential lacZ reporter gene and expression was examined in Saccharomyces cerevisiae. Different mutations affecting in vitro self-splicing also had similar effects on reporter gene expression in vivo. This indicates that in vivo removal of CaLSU from the reporter gene occurs through the typical self-splicing mechanism of group I introns. Base analogs inhibited expression of the reporter gene product in a concentration-dependent manner upon their addition to the cultures. This supports a model in which disruption of intron secondary structure, consecutive to the incorporation of nucleotide analogs, is a major factor determining the susceptibility of C.albicans cells to base analogs.

Antifungal Agents↗
PubMed1997

Oral carriage of Candida albicans in murine AIDS.

Oral candidiasis is a common fungal infection in patients infected with the human immunodeficiency virus (HIV). Although rare at the time of primary HIV infection, it is frequently found throughout the asymptomatic phase and is predictive of progressive immunodeficiency. However, the precise immune defect which results in outgrowth of commensal Candida albicans in HIV infection has not been identified. Mice infected with the Du5H(G6T2) mixture of mouse leukemia viruses develop a syndrome, designated murine AIDS (MAIDS), that has many of the immune abnormalities found in HIV infection. Retrovirus-infected C57BL/6 mice were examined for their ability to resist the development of oral candidiasis from the carrier state established after a self-limiting acute infection and to clear a subsequent secondary inoculum of oral C. albicans. Most of the mice orally colonized with C. albicans and then inoculated with the retrovirus mixture maintained a low-level oral carriage of C. albicans, while 30% of coinfected mice developed recurring 2- to 3-week episodes of acute Candida proliferation, separated by transient recoveries to the carrier state. The frequencies of CD4+ and CD8+ lymphocytes were, respectively, unchanged and significantly decreased (P < 0.05) in both cervical lymph nodes and spleens of coinfected mice compared to the corresponding frequencies in C. albicans-carrying, virus-free, age-matched control animals. Secretion of gamma interferon by concanavalin A (ConA)-stimulated spleen cells from Candida-carrying, retrovirus-infected mice was significantly decreased (P < 0.05) compared to that of C. albicans-carrying, retrovirus-free mice, in accordance with known abnormalities associated with MAIDS. However, production of this cytokine by ConA-stimulated or unstimulated cervical lymph node cells from coinfected mice was enhanced compared to that of virus-free animals colonized with C. albicans. Acquired resistance to reinfection with C. albicans was maintained in retrovirus-infected mice and was associated with a mucosal recruitment of CD8+ cells not observed in control mice. These results suggest that alterations in mucosal immunity which occur in MAIDS differ substantially from defects observed at other sites and that surrogate epithelial defense mechanisms may function locally to limit Candida proliferation.

Animals↗
PubMed1996

A novel group I intron in Candida dubliniensis is homologous to a Candida albicans intron.

In the present study, we determined the sequence of group I self-splicing introns found in the large ribosomal RNA subunit of Candida albicans, Candida stellatoidea and the recently-described species Candida dubliniensis. It was found that both the intron and ribosomal RNA nucleotide sequences are almost perfectly identical between different C. albicans strains as well as between C. albicans and C. stellatoidea strains. Comparisons of ribosomal RNA sequences suggest that local isolates of atypical C. albicans from individuals infected with human immunodeficiency virus can be assigned to the C. dubliniensis species. C. dubliniensis strains also harbor a group I intron in their ribosomal RNA, as observed in about 40% of C. albicans strains and all C. stellatoidea strains. This novel C. dubliniensis group I intron is identical to the C. albicans and C. stellatoidea intron, except for two widely divergent stem-loop regions. Despite these differences, the C. dubliniensis intron possesses self-splicing ability in an in vitro assay. Taken together, these data support the idea that C. albicans and C. stellatoidea should be joined together as variants of the same species while C. dubliniensis is a distinct but closely related microorganism. To our knowledge, the C. albicans and C. dubliniensis introns are the first example of a pair of homologous group I introns differing only by the presence of apparently facultative sequences in some stem-loops suspected to be involved in stabilization of tertiary structure.

Base Sequence↗
PubMed1996

In vitro characterization of purified human thymic dendritic cells infected with human immunodeficiency virus type 1.

In the thymus, dendritic cells (DC) are functionally associated with thymocytes and are recognized to play a major role in the intrathymic differentiation of T cells. Several studies have previously investigated the role of DC during HIV-infection, but the status of thymic DC in HIV-1 pathogenesis remains unclear. In this study, we investigated the susceptibility of purified human thymic DC to HIV-1 infection in vitro. HIV-1 was not detected in cell-free supernatants collected from HIV-infected DC. However, these cultures were shown to transmit HIV-1 infection since coculture with permissive MT4 cells resulted in virus production. The exposure of DC in culture to HIV-1 was shown to promote severe DC morphological changes and killing. We also found that one or several heat labile soluble cytotoxic agents present in the HIV-1-infected DC supernatant mediated the killing of thymocytes. Our observations raise the possibility that (1) the HIV-1-induced DC killing, (2) the capacity of DC to transmit viral infection, and/or (3) the release of HIV-1-mediated cytotoxic agent(s) from DC may contribute to AIDS pathogenesis in vivo.

Animals↗
PubMed1996

Candida albicans serotype analysis by flow cytometry.

Candida albicans strains can be assigned to either of two major serogroups, A or B. Antigenic surface determinants present only in serotype A strains allow such a distinction, which has epidemiologic relevance. Reports have established that the relative distributions of the two serotypes can vary depending on the geographic origin of the isolates. A prevalence of susceptibility to an antifungal agent, flucytosine, was also observed with isolates of serotype A. More recently, it was suggested that the occurrence of serotype B isolates in various clinical forms of candidiasis is increasing. However, this latest finding remains controversial since serotyping results vary widely from one laboratory to another because of the lack of standardized methodologies. Difficulty in interpretation of results, which may lead to erroneous serotype identification, is the major setback associated with current methods. For this study, we thus devised a procedure that relies on flow cytometry and that may eliminate ambiguities in serotype determination. The validation of results was achieved with two types of serotype A-specific antisera, Iatron Factor 6 antiserum and an anti-C. albicans antiserum adsorbed on serotype B yeast cells. Agreement between results obtained with these two reagents was 100% with a wide array of Candida strains. These results confirmed the potential of the flow cytometric procedure as a reliable and reproducible method to establish the serotypes of C. albicans strains. Furthermore, some applications of this procedure to the epidemiological study of this human pathogen are presented.

Candida albicans↗
PubMed1995

An improved method for purifying human thymic dendritic cells.

Thymic dendritic cells (DC) play a prominent role in the immune response as they constitute a key element involved in the maturation of thymocytes in the thymus. Human thymic DC, like DC from other lymphoid organs, represent a minor cell population (< 2%) of the thymus. Since these cells cannot replicate in vitro, the development of efficient purification methods is an essential prerequisite for extensive functional studies. DC express high levels of HLA-DR, a cell surface marker of the MHC class II antigen which is not exclusive to DC. Since no specific human thymic DC marker has been identified so far, DC purification methods are mainly based on depletion of particular subgroups of cells. We report here an improved method for purifying human thymic dendritic cells. In contrast to prior work, CD2+ thymocytes were first depleted by rosetting with neuraminidase treated sheep red blood cells. The nonrosetted cells were separated in a Percoll gradient, and the low-density cells were subsequently depleted of nondendritic cells by using thymocyte and macrophage specific monoclonal antibodies and either magnetic bead depletion or cytofluorometry. Cell populations (18-55 x 10(6) cells) obtained following magnetic bead purification were at least 80% HLA-DR+/CD2- and exhibited ultrastructural morphological features and functional activities such as those described previously for thymic DC. This improved method was compared with different purification approaches that use various combinations of cell density-based separation techniques and cell surface specific markers antibody reactivity. The magnetic beads depletion approach provided higher yields.

Antibodies, Monoclonal↗
PubMed1994

Pulmonary retention of free and liposome-encapsulated tobramycin after intratracheal administration in uninfected rats and rats infected with Pseudomonas aeruginosa.

The pulmonary residence time of free and liposome-encapsulated tobramycin was studied with uninfected rats and rats infected with Pseudomonas aeruginosa. Chronic infection in lungs was established by intratracheal administration of 10(8) CFU of P. aeruginosa PA 508 prepared in agar beads. After 3 days, a single dose (300 micrograms) of free or liposome-encapsulated tobramycin was given intratracheally to both infected and uninfected rats. At various time intervals (0.25 to 16 h) after drug instillations, the remaining tobramycin was evaluated in blood, lungs, and kidneys by a microbiological assay. Intratracheal instillation of liposome-encapsulated tobramycin resulted in high and sustained levels of tobramycin in lungs of uninfected and infected rats over the 16-h period studied; however, the tobramycin levels were two times higher in uninfected rats. There was no tobramycin detected in the blood or kidneys from these animals. In contrast, the intratracheally instilled free tobramycin was cleared within 3 and 1 h from the lungs of uninfected and infected animals, respectively. These data suggest that the encapsulation of tobramycin in liposomes can result in a significant increase of its residence time within lungs. This study also shows that pulmonary infection was associated with a lowering of tobramycin levels in lungs.

Animals↗
PubMed1993

Correlation between the presence of a self-splicing intron in the 25S rDNA of C.albicans and strains susceptibility to 5-fluorocytosine.

Candida albicans presents a well characterized EcoRI RFLP pattern of intensely staining bands. One of these bands, the dimorphic 3.7/4.2 kbp fragment shown to originate from the ribosomal RNA-encoding regions (rDNA), has been used by several investigators to subdivide C. albicans strains in two distinct subtypes. In the present manuscript, we report that an epidemiological study of 120 C.albicans strains revealed a significant correlation between these subtypes and susceptibility to 5-fluorocytosine, an antifungal agent extensively used for biotyping C.albicans. The 4.2 kbp strains being generally more susceptible than their counterparts to this agent and one of its metabolic by-product, 5-fluorouracil. A 379 nucleotides insertion in the 25S rRNA-encoding gene of 4.2 kbp type strains was shown to be responsible for the 3.7/4.2 size difference. This intervening sequence is typical of a group I intron by its site of insertion, its predicted secondary structure, and its self-splicing capability. Assuming there is a genuine causal relationship between presence of the intron and resistance to 5-fluorocytosine, one possible mechanism suggests that inhibition of self-splicing by the insertion of 5-fluorouracil residues in the 25S rRNA precursor might be responsible for the higher susceptibility of 4.2 kbp type strains.

Base Sequence↗
PubMed1993

Application of biotyping and DNA typing of Candida albicans to the epidemiology of recurrent vulvovaginal candidiasis.

One-hundred and five Candida albicans isolates from various anatomic sites of 28 patients, obtained at the onset of two consecutive episodes of well-documented recurrent vulvovaginitis, were typed by methods relying on physiologic or genomic markers. The isolates represented a wide variety of types, and neither a single biotype nor genotype was associated with recurrent vaginitis or a particular body site. Patients generally carried similar strains at various anatomic sites that persisted over time. Genomic methods indicated an 86% rate of relapse, which suggested that most recurrent vaginal infections are of endogenous origin. A similar evaluation with biotyping methods was inconclusive because of a lack of reproducibility, resulting from clonal variation or switching, and difficulties in establishing the number of phenotypic tests necessary to distinguish between identical and different strains. Therefore, Southern hybridization was considered the ideal reference method to study the epidemiology of C. albicans infections.

Adolescent↗
PubMed1993

Combined active-passive immunization against the hepatitis B virus: five-year follow-up of children born to hepatitis B surface antigen-positive mothers.

In order to evaluate the duration of "protective" concentrations (i.e. > or = 10 IU/liter) of antibody to hepatitis B surface antigen (HBsAg) in children vaccinated against hepatitis B in infancy, we followed 146 children born to HBsAg-positive mothers from birth to age 60 months. Children were seen at yearly intervals and tested for hepatitis B virus serologic markers. Of the children included in the study, 134 were protected against infection with development of antibody to HBsAg, 5 became HBsAg-positive and 6 failed to respond to the vaccine but did not become infected. Antibody concentrations fell progressively with the passage of time. The probability of maintaining a "protective" concentration of antibody in vaccine responders at age 60 months was 86% (95% confidence interval, 80 to 93%). Gender, ethnic origin, HBeAg status of the mother and immunization schedule had no influence on the rate of antibody loss. We conclude that in developed countries, the great majority of children vaccinated in infancy remain protected against infection at least until age 60 months. The need for booster doses of vaccine in this population will be determined by long term follow-up of immunized cohorts.

Carrier State↗
PubMed1993

Acquired immunity in experimental murine aspergillosis is mediated by macrophages.

A number of studies have substantiated the pivotal role of innate defense mechanisms in protection against invasive aspergillosis. However, experiments demonstrating increased resistance to lethal intravenous (i.v.) infection with Aspergillus fumigatus conidia in cortisone-treated or untreated mice preinfected with a sublethal dose of conidia and protection of turkeys inoculated subcutaneously with a killed A. fumigatus germling vaccine against subsequent aerosol challenge led us to speculate that acquired immunity may also contribute to host defense against Aspergillus infection. Five-week-old male BALB/c mice were inoculated i.v. with 1.0 x 10(4) viable conidia or saline and challenged i.v. with 1.0 x 10(6) conidia after 7, 15, or 21 days. No protection against challenge was found after 7 days. However, significant and reproducible protection was observed after 15 and 21 days. Mortality was reduced from 90% in control mice to 53% in preinfected mice 40 days after challenge (P = 0.0002). Increased survival was correlated with decreased content of chitin in lungs, liver, and kidneys 4 and 7 days after challenge (P < 0.05). Mice were again inoculated with 1.0 x 10(4) conidia or saline, and after 21 days, 1.0 x 10(8) or 2.0 x 10(8) splenocytes were transferred to naive syngeneic recipients; 2.0 x 10(8) immune splenocytes conferred significant protection (P = 0.0001) against i.v. challenge with 1.0 x 10(6) conidia, and mortality decreased from 83 to 48% 40 days after challenge. Transfer of immune serum offered no protection despite the presence of antibody against a hyphal homogenate of A. fumigatus, which was absent in the sera of control mice. Protection by immune splenocytes was maintained after selective depletion of T cells but was abolished after removal of plastic-adherent splenocytes. Adherent cells were characterized as macrophages by using morphological criteria, nonspecific esterase, and MAC-1 monoclonal antibody. Production of hydrogen peroxide by peritoneal and splenic macrophages from preinfected mice was the same as and lower than, respectively, that from uninfected controls. However, phagocytosis of conidia by peritoneal or splenic macrophages from mice preinfected i.v. or intratracheally was significantly increased after 2 and 3 h of coculture compared with that from uninfected animals, whereas in vitro killing of conidia by splenic macrophages was unaltered. Peritoneal or splenic macrophages from control or preinfected mice failed to kill hyphae in vitro. Killing of hyphae by polymorphonuclear leukocytes was not significantly different between mice preinfected i.v. and uninfected controls. Taken together, the results indicate that acquired immunity mediated by activated macrophages can be demonstrated in experimental murine aspergillosis.(ABSTRACT TRUNCATED AT 400 WORDS)

Animals↗
PubMed1992

The human thymic dendritic cell phenotype and its modification in culture.

In order to extend our study of human thymic dendritic cells (DC) we have purified DC by density gradient separation followed by treatment with CD1 and CD2 mAb and antibody-coated immunobeads. The resulting population contains 60 to 75% brightly HLA-DR+ cells. Morphological and functional studies demonstrate that these cells share the common characteristics of dendritic cells. Extensive phenotypic analysis of the purified DC has been made using a panel of mAb. Cytofluorometric assays with mAb reactive with common leucocyte antigen confirm that the brightly HLA-DR+ cells are of mesenchymal origin. Thymic DC express HLA-DQ and HLA-class I antigens. They are also positive for the expression of CD45RA molecules and some express the ICAM-1 and the LFA-1 molecules. DC do not stain with a wide variety of anti-T, -B, and -monocyte or -M phi mAb and lack Fc gamma RIII, CR2, and CR3. Freshly isolated DC failed to stain with OKT6 mAb; however, they progressively acquire the CD1 molecule after a few days culture. The acquisition of CD1 molecule is selective since CD4, CD2, and HLA-ABC molecules are not upregulated under the same conditions. From phenotypic results, it was therefore possible to sort brightly HLA-DR+ or -DQ+ cells and so obtain greater than 90 to 95% purified human thymic DC. Such homogeneous DC populations are obviously of great interest for the study of thymic DC functions.

Antigens, CD↗
PubMed1992

Influence of muramyl dipeptide on renal candidiasis in genetically distinct mice.

Susceptible (DBA/2) and resistant (C57BL/6) mice were inoculated intravenously with Candida albicans to evaluate the effect of a four-day prophylaxis with muramyl dipeptide (MDP) on the renal burden of organisms during the first week after infection. In sham-treated DBA/2 mice injected with 8 x 10(4) candida cells, renal CFU (LOG10 +/- SEM) on days 1, 4 and 7 after infection were found to average 5.050 +/- 0.109, 4.882 +/- 0.133 and 5.482 +/- 0.245. In sham-treated C57BL/6 mice injected with 2 x 10(5) candida cells, renal CFU on days 1, 4 and 7 reached only 3.610 +/- 0.118, 3.404 +/- 0.107 and 4.176 +/- 0.580. MDP-treated DBA/2 mice achieved significant reduction in CFU of C. albicans on day 1 (1.3 log units) and day 4 (0.6 log unit), while MDP-treated C57BL/6 mice had significant reduction in CFU of C. albicans only on day 1 (0.6 log unit) after infection. Sham-treated mice of both strains had a 28.6 to 30% increase in kidney weights on day 4 only, a transient change not seen in MDP-treated mice. Histopathological examination on days 8, 15 and 21 after infection revealed a higher incidence of renal papillary necrosis in DBA/2 mice than C57BL/6 mice (approximately 70% vs 10%). The incidence of granulomas and of chronic interstitial inflammation was much higher in MDP-treated mice. We conclude that the genetic makeup of the host influences the potential effectiveness of MDP as a biological response modifier.

Acetylmuramyl-Alanyl-Isoglutamine↗
PubMed1991

Histochemical and immunochemical study of the fate of Candida albicans inside human neutrophil phagolysosomes.

To further define the ultrastructural events associated with the killing of Candida albicans by human neutrophils, four methods were used: (1) the periodate-thiocarbohydrazide-silver proteinate (PA-TCH-SP) staining of vicinal-glycol-containing complex carbohydrates; (2) the localization of thermostable immunodeterminants of the yeast cell wall, mannans or mannoproteins, using monospecific antibodies and a protein A-gold complex (monAb-gold); (3) the localization of mannose residues with concanavalin A labeled with gold particles (Con A-gold); (4) the localization of chitin oligomers using wheat germ agglutinin and ovomucoid labeled with gold particles (WGA-gold). The mannan-rich cell wall layers were progressively lost as shown by altered PA-TCH-SP reactivity and a diffuse pattern of staining with Con A-gold and monAb-gold. The de novo appearance of conspicuous amounts of glycogen-like particles near the plasmalemma and in the cell wall was interpreted as evidence of a reparative process of the yeast cell wall. Chitin was seemingly unaltered and readily demonstrated by the WGA-gold in the wall remnants of ghost cells.

Candida albicans↗
PubMed1991

IL-1 production by human thymic dendritic cells: studies on the interrelation with DC accessory function.

Thymic dendritic cells (DC) have been proposed to play a critical role in the generation of immunocompetent T lymphocytes. Since IL-1 is widely considered to be an important second signal in T cell stimulation, we have studied the ability of isolated human thymic DC to produce IL-1. Using the EL4/CTLL conversion assay standardized with recombinant IL-1 beta (rIL-1 beta), we demonstrate that upon LPS-stimulation thymic DC produce small amounts of IL-1 as compared to peripheral blood monocytes (PBM). In contrast with PBM, DC IL-1 production is not influenced by indomethacin. IL-1 activity was detected in the supernatants of DC cultures from all thymuses tested, although quantitative variability was noted among individual thymic donors. The specificity of the active factor was confirmed by neutralization assays with anti-IL-1 beta mAb. On the other hand, we demonstrate that rIL-1 beta cannot substitute for nor amplify the accessory function of thymic DC and that anti-IL-1 beta mAb fails to block the DC accessory function. Thus we conclude that IL-1 beta might not be a major factor for the efficient DC accessory function toward mature thymocytes recently demonstrated in our laboratory. Of interest, IL-1 beta was also detected in the supernatants of DC-thymocyte cocultures in the absence of mitogenic factor, suggesting that thymocyte contacts can constitute a sufficient signal to induce DC to produce IL-1. These observations indicate that human thymic DC represent an intrathymic source of IL-1 whose role in thymocyte proliferation or maturation remains to be understood.

Antigen-Presenting Cells↗
PubMed1990

Accessory function of human thymic dendritic cells in Con A-induced proliferation of autologous thymocyte subsets.

Human thymic dendritic cells (DC) have previously been shown to be intimately associated with thymocytes in situ and in culture. We report that thymic DC express LFA-3 and ICAM-1 adhesion molecules and may spontaneously associate with autologous thymocytes within mitogen-independent clusters. Moreover, the accessory activity of isolated human thymic DC was investigated in Con A-stimulation assays. By proliferation experiments, measured as [3H]TdR incorporation, we demonstrated that irradiated thymic DC strongly increase the mitogen-induced activation of autologous PBL as well as of unfractionated thymocytes. More interestingly, in coculture assays performed with purified thymocyte subsets, we have found that thymic DC greatly enhance the Con A proliferation of CD1- CD3bright thymocytes whereas the accessory activity toward the CD1+ CD3- thymocytes was very weak. Inhibition experiments demonstrated that the DC accessory activity is inhibited by anti-DR-related and anti-IL-2R mAb. However, blocking assays with anti-CD11b, anti-CD11c, anti-LFA-3, and anti-ICAM1 mAb showed that the accessory function obtained is similar to that with untreated cultures. We conclude that isolated human thymic DC may present potent DR- and IL-2-dependent accessory activity mainly directed toward the CD1- CD3bright thymocyte subpopulation, suggesting that thymic DC may be involved in the in vivo proliferation of mature thymocytes.

Antigen-Presenting Cells↗