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1.
Microsc Res Tech ; 38(3): 267-75, 1997 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-9264338

RESUMO

ABSTRACT Human thymic dendritic cells (DC) represent a member of the bone marrow-derived dendritic cell family. They have a dendritic shape and are found in small numbers mainly at the corticomedullary border and in medullary regions of the thymus. Human thymic DC were isolated by density gradient separation, followed by treatment with CD2, CD7, CD1, and CD11b mAb and immunobeads magnetic separation. The resulting population contains 60-75% brightly HLA-DR+ cells which present the morphological characteristics of DC observed in situ. Extensive phenotypic analysis confirmed that they are of mesenchymal origin and that some express CD11a and CD54 molecules. Freshly isolated DC do not stain with a wide variety of anti-T-B and -monocyte or -macrophage mAb. However, they acquire the CD1 molecule after a few days in culture. By using a cell sorter we obtained 90-95% of purified human thymic DC. Functional studies have shown that human thymic DC are potent activators in mixed lymphocyte reactions, act as accessory cells in mitogenic thymocyte proliferation, increase the thymocyte proliferative response to a toxin signal, and produce IL-1. They also formed spontaneous physical associations with thymocytes, which raises questions about the implication of DC in differentiation and/or maturation processes of thymocytes.


Assuntos
Células Dendríticas/ultraestrutura , Timo/ultraestrutura , Células Dendríticas/fisiologia , Humanos , Timo/citologia
2.
Allergy ; 52(4): 394-403, 1997 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-9188920

RESUMO

We performed immunoblotting experiments to determine specific IgE and IgG subclass responses to Candida albicans antigens in allergic bronchopulmonary aspergillosis (ABPA) patients. This is a first report describing C. albicans antigens recognized by serum IgE and IgG subclasses of ABPA patients sensitized to that yeast. Among the various antigens reacting with serum IgE, a 43-kDa component was recognized by all seven patients and can be considered a major antigen of C. albicans for this particular group of patients. By comparison, only 20% of a group of asthmatic atopics (25 patients) and 10% of a group of normal controls (10 subjects) were 43-kDa positive. Multiple banding patterns, revealing no major antigen, were observed for all four IgG subclasses except for IgG1 in one case. In particular, the 43-kDa component was not always recognized by all the patients. Furthermore, oral or inhaled steroid treatment appears to have no impact on the specific IgE immunopatterns obtained. Using immunoelectron-microscopy, we localized IgE-binding primarily in the mannoprotein-containing layers of the C. albicans cell wall. In conclusion, C. albicans-IgE and IgG subclasses may participate in the physiopathology of ABPA by exacerbating pulmonary infiltrates (IgE) and inducing eosinophil-mediated inflammatory reaction (IgG1, IgG3).


Assuntos
Anticorpos Antifúngicos/sangue , Aspergilose Broncopulmonar Alérgica/imunologia , Candida albicans/imunologia , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Corticosteroides/uso terapêutico , Adulto , Aspergilose Broncopulmonar Alérgica/tratamento farmacológico , Asma/tratamento farmacológico , Asma/imunologia , Estudos de Casos e Controles , Reações Cruzadas , Feminino , Humanos , Immunoblotting , Imunoglobulina G/classificação , Masculino
3.
Infect Immun ; 65(2): 661-7, 1997 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-9009328

RESUMO

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.


Assuntos
Candidíase Bucal/virologia , Síndrome de Imunodeficiência Adquirida Murina/microbiologia , Animais , Candida albicans/fisiologia , Candidíase Bucal/etiologia , Candidíase Bucal/imunologia , Portador Sadio/microbiologia , Portador Sadio/virologia , Vírus Defeituosos/fisiologia , Feminino , Vírus Auxiliares/fisiologia , Imunidade Inata , Interferon gama/biossíntese , Vírus da Leucemia Murina/fisiologia , Estudos Longitudinais , Linfonodos/imunologia , Linfonodos/virologia , Transtornos Linfoproliferativos/imunologia , Transtornos Linfoproliferativos/microbiologia , Transtornos Linfoproliferativos/virologia , Camundongos , Camundongos Endogâmicos C57BL , Síndrome de Imunodeficiência Adquirida Murina/imunologia , Baço/imunologia , Baço/virologia
4.
Eur Surg Res ; 29(3): 229-36, 1997.
Artigo em Inglês | MEDLINE | ID: mdl-9161840

RESUMO

In the presence of severe rejection, cardiac allograft perfusion has been shown to be impaired. Since a functionally reversible vasoconstrictor component has been identified in this condition and rejection does not reverse if ischemia does not, we hypothesized that diltiazem may be beneficial in this condition. Experiments were performed on dogs with heterotopic heart transplants and chronic instrumentation for the assessment of allograft perfusion. Two groups of cardiac allograft recipients were studied: untreated recipients and recipients treated with the calcium antagonist diltiazem (180 mg twice daily, orally). Allograft blood flow was monitored daily along with plasma diltiazem levels. The lymphoproliferative response to mitogens was studied at selected intervals until terminal rejection. Contractile function of the graft was assessed daily by palpation. Without immunosuppression, terminal rejection was observed within 7 days. Rejection was confirmed by histology; cellular infiltration and myocyte necrosis were present in all cardiac allografts but to a significantly lesser degree in diltiazem-treated recipients. The mean blood flow of heterotopically implanted hearts was in the range of 35-50 ml/min, which decreased steadily in untreated recipients. In contrast, significant improvement of allograft perfusion was observed in diltiazem-treated recipients at days 4-6 after transplantation. Diltiazem also significantly attenuated mitogen-induced lymphocyte proliferation at peak sensitivity (2 days after transplantation). Diltiazem plasma concentrations were in the therapeutic range (30-60 ng/ml) before and after cardiac transplantation. Results of the present study demonstrate beneficial effects of diltiazem in the course of severe cardiac rejection. Such findings support its use during rejection when maintenance of graft blood flow and myocyte protection may be important for myocardial function and viability.


Assuntos
Diltiazem/farmacologia , Rejeição de Enxerto/tratamento farmacológico , Transplante de Coração , Animais , Concanavalina A/farmacologia , Vasos Coronários/efeitos dos fármacos , Diltiazem/sangue , Cães , Coração/efeitos dos fármacos , Tamanho do Órgão , Fito-Hemaglutininas/farmacologia , Mitógenos de Phytolacca americana/farmacologia , Fatores de Tempo
5.
Nucleic Acids Res ; 25(2): 431-7, 1997 Jan 15.
Artigo em Inglês | MEDLINE | ID: mdl-9016575

RESUMO

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.


Assuntos
Candida albicans/química , Regulação Fúngica da Expressão Gênica/genética , Íntrons/genética , Antifúngicos/farmacologia , Northern Blotting , Western Blotting , Divisão Celular/efeitos dos fármacos , Flucitosina/farmacologia , Fluoruracila/farmacologia , Genes Reporter/genética , Óperon Lac/genética , Mutagênese Sítio-Dirigida/genética , Conformação de Ácido Nucleico , Splicing de RNA/genética , RNA Ribossômico/genética , beta-Galactosidase/genética , beta-Galactosidase/metabolismo
6.
Gene ; 180(1-2): 189-96, 1996 Nov 21.
Artigo em Inglês | MEDLINE | ID: mdl-8973366

RESUMO

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.


Assuntos
Candida albicans/genética , Candida/genética , Genes Fúngicos , Íntrons , Sequência de Bases , Southern Blotting , Candida/classificação , Candida/isolamento & purificação , Candida albicans/classificação , DNA Fúngico , DNA Ribossômico/genética , Humanos , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Splicing de RNA , RNA Fúngico/genética , RNA Ribossômico/genética , Homologia de Sequência do Ácido Nucleico
7.
J Clin Microbiol ; 34(9): 2106-12, 1996 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8862566

RESUMO

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.


Assuntos
Candida albicans/classificação , Feminino , Citometria de Fluxo/métodos , Humanos , Sorotipagem
8.
Virology ; 222(1): 214-26, 1996 Aug 01.
Artigo em Inglês | MEDLINE | ID: mdl-8806501

RESUMO

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.


Assuntos
Células Dendríticas/virologia , HIV-1/patogenicidade , Timo/virologia , Animais , Morte Celular , Linhagem Celular Transformada , Células Cultivadas , Chlorocebus aethiops , Testes Imunológicos de Citotoxicidade , Células Dendríticas/imunologia , Células Dendríticas/metabolismo , Células Dendríticas/ultraestrutura , Humanos , Hibridização In Situ , Fatores Matadores de Levedura , Proteínas/imunologia , Proteínas/metabolismo , RNA Viral/análise , Timo/citologia , Timo/imunologia , Timo/metabolismo
9.
J Immunol Methods ; 180(2): 225-36, 1995 Mar 27.
Artigo em Inglês | MEDLINE | ID: mdl-7714337

RESUMO

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.


Assuntos
Separação Celular/métodos , Células Dendríticas/citologia , Células Dendríticas/imunologia , Timo/citologia , Anticorpos Monoclonais/imunologia , Antígenos CD/imunologia , Células Cultivadas , Células Dendríticas/ultraestrutura , Citometria de Fluxo , Antígenos HLA-DQ/imunologia , Antígenos HLA-DR/imunologia , Humanos
10.
Antimicrob Agents Chemother ; 38(5): 1090-5, 1994 May.
Artigo em Inglês | MEDLINE | ID: mdl-8067743

RESUMO

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.


Assuntos
Pulmão/metabolismo , Infecções por Pseudomonas/metabolismo , Tobramicina/farmacocinética , Animais , Portadores de Fármacos , Intubação Intratraqueal , Lipossomos , Pulmão/microbiologia , Masculino , Infecções por Pseudomonas/tratamento farmacológico , Infecções por Pseudomonas/microbiologia , Ratos , Ratos Sprague-Dawley , Tobramicina/administração & dosagem , Tobramicina/uso terapêutico
11.
Nucleic Acids Res ; 21(25): 6020-7, 1993 Dec 25.
Artigo em Inglês | MEDLINE | ID: mdl-7904747

RESUMO

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.


Assuntos
Candida albicans/genética , DNA Ribossômico/genética , Flucitosina/farmacologia , Íntrons , Splicing de RNA , RNA Ribossômico/genética , Sequência de Bases , Candida albicans/efeitos dos fármacos , DNA Fúngico , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Polimorfismo de Fragmento de Restrição , RNA Fúngico , RNA Ribossômico/química , Sequências Repetitivas de Ácido Nucleico
13.
Infect Immun ; 61(9): 3791-802, 1993 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-8359900

RESUMO

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)


Assuntos
Aspergilose/imunologia , Aspergillus fumigatus/imunologia , Macrófagos/imunologia , Animais , Peróxido de Hidrogênio/metabolismo , Imunidade Ativa , Imunoterapia Adotiva , Ativação de Macrófagos , Macrófagos/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Neutrófilos/imunologia , Fagocitose , Baço/imunologia
14.
J Infect Dis ; 168(2): 502-7, 1993 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-8101553

RESUMO

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.


Assuntos
Candida albicans/classificação , Candidíase Vulvovaginal/epidemiologia , Adolescente , Adulto , Southern Blotting , Candida albicans/genética , Candidíase Vulvovaginal/microbiologia , DNA Fúngico/classificação , Feminino , Genótipo , Humanos , Masculino , Pessoa de Meia-Idade , Técnicas de Tipagem Micológica , Especificidade de Órgãos , Fenótipo , Polimorfismo de Fragmento de Restrição , Recidiva
16.
Pediatr Infect Dis J ; 12(2): 126-30, 1993 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-8426769

RESUMO

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.


Assuntos
Anticorpos Anti-Hepatite/biossíntese , Hepatite B/imunologia , Hepatite B/prevenção & controle , Imunização , Portador Sadio/transmissão , Feminino , Seguimentos , Hepatite B/transmissão , Antígenos de Superfície da Hepatite B/sangue , Vacinas contra Hepatite B , Humanos , Imunização Passiva , Lactente , Recém-Nascido , Masculino , Vacinação
17.
APMIS ; 100(11): 967-75, 1992 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-1472365

RESUMO

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.


Assuntos
Acetilmuramil-Alanil-Isoglutamina/farmacologia , Candidíase/imunologia , Nefropatias/imunologia , Animais , Candida albicans/crescimento & desenvolvimento , Candidíase/microbiologia , Candidíase/patologia , Feminino , Rim/imunologia , Rim/microbiologia , Rim/patologia , Nefropatias/microbiologia , Nefropatias/patologia , Camundongos , Camundongos Endogâmicos C57BL , Camundongos Endogâmicos DBA , Tamanho do Órgão , Fatores de Tempo
18.
Cell Immunol ; 142(2): 238-51, 1992 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-1377985

RESUMO

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.


Assuntos
Antígenos CD/análise , Células Dendríticas/imunologia , Antígenos HLA-DR/análise , Antígeno-1 Associado à Função Linfocitária/análise , Timo/imunologia , Antígenos CD1 , Moléculas de Adesão Celular/análise , Separação Celular/métodos , Células Cultivadas , Pré-Escolar , Citometria de Fluxo/métodos , Antígenos de Histocompatibilidade/análise , Humanos , Imunofenotipagem , Lactente , Molécula 1 de Adesão Intercelular , Antígenos Comuns de Leucócito
19.
J Leukoc Biol ; 50(6): 587-99, 1991 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-1940612

RESUMO

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.


Assuntos
Candida albicans/imunologia , Neutrófilos/microbiologia , Candida albicans/ultraestrutura , Membrana Celular/ultraestrutura , Parede Celular/ultraestrutura , Concanavalina A/metabolismo , Humanos , Imuno-Histoquímica , Técnicas In Vitro , Microscopia Eletrônica , Neutrófilos/ultraestrutura , Fagossomos/microbiologia
20.
Cell Immunol ; 135(2): 431-44, 1991 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-2036677

RESUMO

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.


Assuntos
Células Apresentadoras de Antígenos/fisiologia , Células Dendríticas/metabolismo , Interleucina-1/biossíntese , Timo/imunologia , Células Cultivadas , Células Dendríticas/imunologia , Antígenos HLA-DR/análise , Humanos , Indometacina/farmacologia , Lipopolissacarídeos/farmacologia , Ativação Linfocitária , Fenótipo
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