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S Montplaisir

Publications and source records attributed to S Montplaisir.

At least 19 recordsLinked to original sources

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

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

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

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

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

Human thymic dendritic cell-thymocyte association: ultrastructural cell phenotype analysis.

In rodent thymus, associations between dendritic cells (DC) and thymocytes have been suggested to be implicated in differentiation and/or maturation processes. In this study, we report intimate associations formed between human thymic DC and thymocytes in culture and we analyze their ultrastructural cell phenotype. Observations by phase contrast microscopy showed that DC present long and thin dendrites and bind many thymocytes. Transmission (TEM) and scanning electron microscopy (SEM) revealed that both cellular populations were in close connection and tight membrane contact could be observed. The phenotype of DC and attached thymocytes was characterized with a series of monoclonal antibodies by protein A-gold TEM and SEM immunolabelings. Quantitative evaluation of immunolabeling (number of gold granules/microns of cellular membrane) suggests the presence of two subpopulations of CD1+ thymic DC (strong and weak), whereas this discrepancy is not observed in DR+ and CD4+ DC populations. On the other hand, the majority of thymocytes bound to DC strongly express the CD1, CD4, CD8 and CD2 antigens and weakly the CD3 antigen, indicating that they represent double-positive immature thymocytes. Uniform distribution of DC and thymocytes membrane antigens was confirmed with a backscattered SEM study. This morphological and immunolabeling TEM and SEM analysis demonstrates that human thymic DC may form associations with CD4+CD8+CD3weak thymocytes and raises questions about their physiological relationship.

Antigens, CD1

Circulating immune complexes, antibodies to Pseudomonas aeruginosa, and pulmonary status in cystic fibrosis.

Serum samples from 37 patients with cystic fibrosis (CF), whose lungs were colonized by Pseudomonas aeruginosa, were tested in a 1 yr prospective study to examine a possible relationship between levels of circulating immune complexes (CIC) and the following parameters: level of specific antibodies to P. aeruginosa; relative importance of P. aeruginosa mucoid and non-mucoid strains isolated from sputum; the forced expiratory volume (FEV1; percentage predicted); the chest X-Ray score (Brasfield system) and the clinical score (Shwachman system). Reactivity of CIC against P. aeruginosa, Staphylococcus aureus, Haemophilus influenzae and Escherichia coli antigens were also assayed. We found that the FEV1, the chest X-Ray and the clinical scores were significantly lower in patients with high levels of CIC than in those with normal levels of CIC (p less than 0.001 for each). We also found that the level of IgG antibodies against P. aeruginosa was significantly higher (p less than 0.001) in patients with high levels of CIC than in those with normal levels of CIC. 78% of patients with high levels of CIC had predominantly mucoid P. aeruginosa isolates whereas only 21% of patients with normal levels of CIC had also predominantly mucoid P. aeruginosa isolates. Specific antibodies to P. aeruginosa were detected in all CIC isolated by polyethylene glycol precipitations from CF patients exhibiting both high levels of CIC and inferior pulmonary status. Our findings support the hypothesis that a high level of CIC in association with an aggressive humoral response to P. aeruginosa correlates with defective pulmonary status in cystic fibrosis.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent

[Sterilization and quality control].

The dental profession has always considered the sterilization of surgical instruments as an important element in the prevention of infection. During the last decade the concern with blood and to a lesser degree with saliva in the transmission of infection, due primarily to the hepatitis and HIV viruses, has resulted in many corporate organizations releasing recommendations to reinforce the use of sterilization techniques and disinfection techniques when sterilization is not applicable or justifiable. As a result of the fact that the two terms are often intermingled, the authors review the fact that each one pursues different objectives and put the emphasis on sterilization by briefly reviewing the principal advantages and disadvantages of each method of sterilization presently available for use in dental offices. Subsequently, they stress the importance of introducing quality control in any infection prevention program, in order to assure that the sterilization process is not at the mercy of human error or mechanical failure. The pharmaceutical industry, food chains and hospital centres have for more than 40 years realized the importance of using chemical indicators and especially biological testing. Only biological testing can put one's mind at ease, for the professional in a dental office and the patient receiving care, that the instruments or objects that one comes in contact with are truly sterile. The dental profession did not wait for the discovery of the hepatitis B and HIV viruses to become preoccupied with the spread of disease and have sterilized their surgical instruments for many years.(ABSTRACT TRUNCATED AT 250 WORDS)

Communicable Disease Control

[Vaccination as a means of prevention].

While recognizing the fact that the use of strict hygienic techniques, disposable materials and quality control are essential for sterilizing instruments, vaccination still remains the most efficient way to combat infection. In this article, the authors discuss the principle problems associated with the use of vaccines and especially as they relate to the dental office. Hepatitis B is certainly of interest to all dentists and their personnel, especially since its primary mode of transmission is by contact of blood with the skin and the mucosa. The two forms of vaccines presently available, are derived from the human plasma of carriers or from yeast and cause a genetic reaction which produces hepatitis B virus antigens. These vaccines protect at least over 90% of all healthy individuals and do not generate any secondary infection or unfavorable reactions. Faced with the reality of hepatitis B, it is very wise to remember that it is BETTER TO BE VACCINATED THAN TO HAVE TO LIVE WITH THE EFFECTS OF HEPATITIS.

Hepatitis A

Lymphocyte subpopulation monitoring in cyclosporine-treated patients following heart transplantation.

Circulating lymphocyte subpopulations were studied in 18 consecutive patients treated with cyclosporine-prednisone immunosuppression during the first month following heart transplantation. Eleven patients showed no evidence of graft rejection. There were eight episodes of acute rejection demonstrated at endomyocardial biopsy in 7 patients. Three patients were treated with corticosteroids, 3 were treated with rabbit antithymocyte globulin (RATG), and 1 died before treatment (early mortality: 5.6%). Using the monoclonal antibody technique, 150 determinations of lymphocyte subpopulations were performed and were correlated with 72 endomyocardial biopsy specimens. Cyclosporine immunosuppression caused a significant (p less than 0.05) decrease in total lymphocyte count (38%) and in the number of OKT3 (52%) and OKT4 cells (55%). During acute rejection, total lymphocytes and OKT3, OKT4, and OKT8 cells all increased significantly, but the T4 to T8 ratio did not change significantly. Treatment of rejection with corticosteroids resulted in a moderate but not significant decrease in all T-cell types, whereas RATG caused a marked but not selective decrease in all T-cell groups. In conclusion, T cells decrease with cyclosporine immunosuppression and with treatment of rejection and increase at onset of rejection, but the T4 to T8 ratio has no predictive value for the diagnosis and severity of rejection, and the sensitivity of the method does not permit its use to assess the degree of immunosuppression with cyclosporine following heart transplantation.

Adolescent

Combined passive-active immunization against the hepatitis B virus of 132 newborns of chronic carrier mothers: long term results.

One hundred thirty-two newborns of carrier mothers were given combined passive-active immunization against the hepatitis B virus. Doses of hepatitis B immunoglobulin (HBIG) were 0.5 ml, and doses of vaccine (Heptavax B) were 10 micrograms. Schedules used were: HBIG, 0, 3 and 6 months and vaccine 1, 2 and 7 months (Schedule 1); HBIG, 0, 3 and 6 months, vaccine, 7, 8 and 13 months (Schedule 2); HBIG, 0 month and vaccine 1, 2 and 7 months (Schedule 3); HBIG, 0 month and vaccine 0, 1 and 6 months (Schedule 4). The study population consisted of 60 boys and 72 girls. Ethnic origins were the following: Haitian, 42; Cambodian, 33; French Canadian, 22; Vietnamese, 21; others, 14. Thirty-two children were born to hepatitis B e antigen-positive mothers. General outcome was: 121 became anti-hepatitis B surface antigen-positive; 5 became hepatitis B surface antigen-positive; and 6 remained negative for all markers (nonresponders). Sex, ethnic origin and immunization schedule had no influence on the outcome. Of the 32 children born to hepatitis B e antigen-positive mothers, only 4 became hepatitis B surface antigen-positive. In those who became anti-hepatitis B surface antigen-positive, mean serum anti-hepatitis B surface antigen antibody concentrations (log mIU/ml +/- SD) were: 3.47 +/- 1.03 at 8 to 9 months post-Vaccine 1; 2.89 +/- 0.91 at 14 to 17 months; 2.39 +/- 1.0 at 23 to 29 months; and 2.26 +/- 0.92 at 35 to 41 months.(ABSTRACT TRUNCATED AT 250 WORDS)

Carrier State

Human thymic dendritic cells. Characterization, isolation and functional assays.

The phenotypic analysis of human thymic dentritic cells (DC) in culture and in purified suspensions has been studied with light and electron microscopic (EM) immunolabelling techniques. Using a series of monoclonal antibodies (mAb) and a protein A-gold technique, we demonstrated that DR- and T6-positive cultured DC strongly bind the 9.3F10 mAb, an anti-DR-related antibody produced against human blood DC, and weakly express the T4 antigen, a membrane marker shared by Langerhan's cells (LC). On the other hand, thymic-cultured DC are negative for the other T-cell and monocyte-macrophage antigens. These results support the hypothesis that human thymic DC may be related to blood DC and epidermal LC. Moreover DC, unlike thymic macrophages, do not phagocytose latex particles, opsonized sheep red blood cells (SRBC) or Candida albicans. An efficient two-step technique of isolation, using a Percoll density gradient followed by an indirect panning technique, yields a purified (70-80%) thymic DC population, OKIa1-, 9.3F10- and OKT6-positive and esterase-negative. Immunolabelling and electron microscopy confirm that these isolated DC present similar phenotypic and ultrastructural features to human thymic DC in situ and in culture. Purified DC, used as stimulator cells in mixed leucocyte reaction (MLR), induce stronger proliferative responses than peripheral blood monocytes used as a control; blocking assays with OKIa1 mAb plus complement greatly reduced this stimulatory potency. These functional assays demonstrate that we obtained a purified typical DC population that can be used in immunological functional studies to elucidate the specific role of DC in human thymus.

Antigens, Surface

Genetics of resistance to infection with Candida albicans in mice.

To determine differences in susceptibility, 234 naive mice including xid and beige mutants were infected intravenously with Candida albicans and monitored with survival analysis and quantitative culture of the kidneys. By using survival time as the criterion, animals of seven inbred strains were separated into three groups. C3H/HeJ and Dw/+ were most susceptible; C57BR/cdJ, BRVR and CBA/N (xid) were intermediate in susceptibility; C57BL/KsJ and C57BL/6J were least susceptible. Mean survival times (MST) were markedly influenced by the number of Candida cells injected while the ranking of mouse strains by survival alone was unchanged. There was a dissimilar behaviour of the strains to produce organ weight changes in response to infection when compared with uninfected mice which were matched for age and genetic lineage. Black mice had lower colony forming units (CFU) per mg of tissue at the time of death than animals of other genetic lineage. Nevertheless, the finding that MST and CFU studies were loosely correlated in a few strains of mice indicated that the proliferation of the fungus in the kidneys was not always the major cause of death. The beige mutation was found to determine an increased susceptibility to systemic Candida infection. The differences in survival for beige and nonbeige mice were influenced by the genetic lineage of the host, being much greater in the C57BL/6 strain (36.7 days) than in the C3H/He strain (5 days). C57BL/6 beige-J had significantly higher CFU per organ and per unit of weight than C57BL/6 +/+ mice. These data evinced an important contribution of host genetic factors to resistance to systemic candidiasis. It is suggested that innate resistance genes regulate the differentiation in the bone marrow and the function of cells of granulocyte-macrophage lineage.

Animals

Mucoid and pigmentation characters can be suppressed by non-anti-Pseudomonas aeruginosa antibiotics in cystic fibrosis: a report of promising preliminary results.

In an effort to develop new strategies, the authors are exploring the hypothesis that non-anti-Pseudomonas aeruginosa antibiotics can affect some major virulence factors in cystic fibrosis P. aeruginosa. Recent samples (127) of P. aeruginosa isolated from the sputum of cystic fibrosis patients and manifesting a mucoid growth were stored at -70 degrees C until experimentation. The mucoid character was retested after thawing and one 24-h in-vitro passage at 37 degrees C onto 100-mm blood agar plates to prepare a bacterial suspension for inoculation (Steers or MIC-2000 plus replicator) and cultivation (48 h, 37 degrees C) onto different 100-mm or 150-mm Mueller-Hinton agar plates containing either no antibiotics or antibiotics alone, and antibiotic combinations without presumed anti-P. aeruginosa activity. Some isolates (49, 38.6%) of P. aeruginosa expressed again the mucoid character on antibiotic-free Mueller-Hinton agar and growth was prevented by antibiotics in a number of them: 7 with doxycycline (16.0 micrograms/ml), 2 with rifampicin (16.0 micrograms/ml), 6 with roxithromycin-sulfamethoxazole (16.0-304.0 micrograms/ml) and 23 with trimethoprim-sulfamethoxazole. In the remaining evaluable isolates, the mucoid was modified towards the rough character in 36 out of 42 with doxycyline, 36 out of 47 with rifampicin, 31 out of 43 with roxithromycin-sulfamethoxazole and 20 out of 26 with trimethoprim-sulfamethoxazole. In parallel, the pigmentation was lost in the presence of antibiotics in those P. aeruginosa isolates having manifested this character on Mueller-Hinton agar. These results suggest that non-anti-P. aeruginosa antibiotics can modify the in vitro markers of virulence in cystic fibrosis P. aeruginosa and that they merit further investigation.

Anti-Bacterial Agents

A monoclonal antibody directed against a serotype-specific, outer-membrane protein of Haemophilus influenzae type b.

Monoclonal antibodies (Mabs) were produced against outer-membrane proteins (OMPs) of Haemophilus influenzae type b. The clones were screened by ELISA with outer-membrane preparations of H. influenzae type b and untypable strains as coating antigens. Antibodies directed against the proteins of mol. wt (10(3)) 43, 37 and 13 were identified by immunoblotting of SDS-PAGE patterns of OMPs. Proteolytic enzyme treatments of the OMPs resulted in reduction of Mab reactivity as measured by ELISA. Furthermore, the absence of reactivity of Mab Hb-2 with a preparation of lipopolysaccharide confirmed the protein nature of its corresponding epitope. Binding assays with live bacteria showed that Hb-2 reacted with a cell surface-exposed antigenic determinant. Mab Hb-2 was bactericidal in vitro in the presence of complement. The characterisation of Hb-2 (IgG2a) by Western immunoblotting analysis revealed that it was directed against the 37 X 10(3)-mol. wt OMP. In a dot-enzyme immunoassay, Hb-2 reacted specifically with 326 strains of H. influenzae type b. It did not cross-react with the other serotypes or untypable strains of H. influenzae or with other bacterial species. This is the first report of a monoclonal antibody identifying a serotype-specific surface-exposed OMP of H. influenzae type b.

Antibodies, Monoclonal

Identification of Haemophilus influenzae type b by a monoclonal antibody coagglutination assay.

A coagglutination assay using monoclonal antibody is described for the identification of Haemophilus influenzae type b. An immunoglobulin G2a monoclonal antibody, Hb-2, directed against a serotype-specific outer membrane protein of H. influenzae type b was adsorbed to Staphylococcus aureus Cowan 1 cells. In a dot enzyme immunoassay, Hb-2 reacted with 453 of 455 H. influenzae type b isolates and did not react with H. influenzae of other serotypes, untypeable H. influenzae strains, or other bacterial species. The Hb-2 coagglutination assay was evaluated by testing 136 H. influenzae type b strains selected on the basis of multilocus enzyme genotypes, 5 strains of another serotype, and 94 untypeable H. influenzae strains. The specificity of the coagglutination assay was demonstrated by the inhibition of the reaction by free Hb-2 monoclonal antibodies. The coagglutination assay was as specific as the dot enzyme immunoassay and can be rapidly performed and easily interpreted.

Agglutination Tests

Protein supplements, mitogens and antibiotics in lymphoblast transformation and cytogenetic analysis.

The in vitro effects of four protein supplements, foetal calf serum (FCS), gammaglobulin-depleted FCS (FCS-A), human serum albumin (HSA) and autologous serum (AS), and of four antibiotics on lymphocyte blast transformation and on lymphocyte proliferation for cytogenetic studies, were evaluated. 3H-thymidine incorporation was measured in the lymphocytes obtained from sixteen healthy adults and stimulated with phytohaemagglutinin (PHA), concanavalin A (Con A), pokeweed mitogen (PWM), PWM + PHA and PWM + Con A. The most efficient protein supplement was AS, although great variability was observed between subjects. HSA came second, while FCS and FCS-A were the least effective. The presence of immunoglobulins did not modify mitogen-induced lymphocyte responses. In unsupplemented media, peak responses to PHA and Con A occurred at lower mitogen concentrations. Furthermore, the addition of the antibiotics tested did not impede blast transformation whether or not a protein supplement was present. This work also showed that for chromosome analysis, protein supplements are necessary for optimum whole-blood lymphocyte cultures and that HSA can replace FCS.

Adult