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Differences in virulence of clinical isolates of Candida tropicalis and Candida albicans in mice.

Prior reports from this institution indicated that Candida tropicalis was more pathogenic than C. albicans in oncology patients. Pairs of clinical isolates of C. tropicalis and C. albicans recovered from similar patients at other institutions were examined to determine their relative virulence. After intravenous inoculation in normal mice, three pairs of isolates had no significant differences in the 50% lethal dose, and one C. tropicalis isolate was less virulent than its companion C. albicans isolate. In contrast, in mice treated with antibiotics and cytarabine, an antineoplastic drug which damages the gastrointestinal mucosa and produces granulocytopenia, oral inoculation of yeast cells produced striking differences in the 50% infective dose: each C. tropicalis isolate was more virulent than the companion C. albicans isolate from the same institution. The increased virulence of the C. tropicalis isolates compared with the C. albicans isolates when given orally to compromised mice parallels clinical observations in compromised patients.

Agranulocytosis↗

Enhancement of non-Candida antibody responses by Candida albicans cell wall glycoprotein.

Two cell wall glycoprotein extracts from Candida albicans (glycoprotein [GP] and peptidoglucomannan [PGM]) were tested for their influence on antibody responses to type III pneumococcal polysaccharide and sheep erythrocytes. GP was isolated from lipid-extracted cell walls with ethylenediamine, whereas PGM was extracted with dilute sodium hydroxide. Both glycoproteins increased the number of antibody-producing plaque-forming cells in the spleens of mice immunized with type III polysaccharide or sheep erythrocytes, although PGM appeared to be about 10 times more effective. PGM could be administered up to 3 days prior to immunization with sheep erythrocytes to elicit enhancement; it did not have to be administered by the same route as the immunogen to cause significant enhancement. Enhancement did not appear to be the result of a direct mitogenic effect of GP and PGM on lymphocytes, nor did these glycoproteins appear to stimulate the production of B-cell growth factors or interleukin 2.

Adjuvants, Immunologic↗

Purification of actin from Candida albicans and comparison with the Candida 48,000-Mr protein.

Actin was purified from Candida albicans cells by affinity chromatography by DNase-Sepharose and was recognized by immunoblotting with monoclonal antibody directed against chick muscle actin. The C. albicans 48-kilodalton protein recognized by sera from patients with invasive candidiasis was shown by DEAE chromatography and immunoblotting not to be identical with the purified C. albicans actin.

Actins↗

Overexpression of the Candida albicans ALA1 gene in Saccharomyces cerevisiae results in aggregation following attachment of yeast cells to extracellular matrix proteins, adherence properties similar to those of Candida albicans.

Candida albicans maintains a commensal relationship with human hosts, probably by adhering to mucosal tissue in a variety of physiological conditions. We show that adherence due to the C. albicans gene ALA1 when transformed into Saccharomyces cerevisiae, is comprised of two sequential steps. Initially, C. albicans rapidly attaches to extracellular matrix (ECM) protein-coated magnetic beads in small numbers (the attachment phase). This is followed by a relatively slower step in which cell-to-cell interactions predominate (the aggregation phase). Neither of these phases is observed in S. cerevisiae. However, expression of the C. albicans ALA1 gene from a low-copy vector causes S. cerevisiae transformants to attach to ECM-coated magnetic beads without appreciable aggregation. Expression of ALA1 from a high-copy vector results in both attachment and aggregation. Moreover, transcriptional fusion of ALA1 with the galactose-inducible promoters GALS, GALL, and GAL1, allowing for low, moderate, and high levels of inducible transcription, respectively, causes attachment and aggregation that correlates with the strength of the GAL promoter. The adherence of C. albicans and S. cerevisiae overexpressing ALA1 to a number of protein ligands occurs over a broad pH range, is resistant to shear forces generated by vortexing, and is unaffected by the presence of sugars, high salt levels, free ligands, or detergents. Adherence is, however, inhibited by agents that disrupt hydrogen bonds. The similarities in the adherence and aggregation properties of C. albicans and S. cerevisiae overexpressing ALA1 suggest a role in adherence and aggregation for ALA1 and ALA1-like genes in C. albicans.

Adhesiveness↗

Degradation of hydrocarbons by members of the genus Candida. II. Oxidation of n-alkanes and l-alkenes by Candida lipolytica.

Candida lipolytica ATCC 8661 was grown in a mineral-salts hydrocarbon medium. n-Alkanes and 1-alkenes with 14 through 18 carbon atoms were used as substrates. Ether extracts of culture fluids and cells obtained from cultures grown on the various substrates were analyzed by thin-layer and gas-liquid chromatography. Analyses of fluids from cultures grown on n-alkanes indicated a predominance of fatty acids and alcohols of the same chain length as the substrate. In addition, numerous other fatty acids and alcohols were present. Analyses of saponifiable and nonsaponifiable material obtained from the cells revealed essentially the same products. The presence of primary and secondary alcohols, as well as fatty acids, of the same chain length as the n-alkane substrate suggested that attack on both the methyl and alpha-methylene group was occurring. The significance of these two mechanisms in the degradation of n-alkanes by this organism was not evident from the data presented. Analyses of fluids from cultures grown on 1-alkenes indicated the presence of 1,2-diols, as well as omega-unsaturated fatty acids, of the same chain length as the substrate. Alcohols present were all unsaturated. Saponifiable and nonsaponifiable material obtained from cells contained essentially the same products. The presence of 1,2-diols and omega-unsaturated fatty acids of the same chain length as the substrate from cultures grown on 1-alkenes indicated that both the terminal methyl group and the terminal double bond were being attacked.

Alcohols↗

Degradation of hydrocarbons by members of the genus Candida 3. Oxidative intermediates from 1-hexadecene and 1-heptadecene by Candida lipolytica.

Candida lipolytica (Phaff) was grown in a mineral-salts medium amended with either 1-hexadecene or 1-heptadecene as substrate. Intermediates of the same chain length as the substrate were isolated and identified by various analytical procedures. The following intermediates of 16 and 17 carbon atoms were identified: omega-unsaturated acids, omega-unsaturated primary and secondary alcohols, 1,2-epoxides, 1,2-diols, and 2-hydroxy acids. Based on the chemical structure of these compounds, three oxidative mechanisms are proposed for the degradation of long-chain 1-alkenes by this yeast: (i) methyl-group oxidation, (ii) double-bond oxidation, and (iii) subterminal oxidation.

Acids↗

Candida antigenemia, as detected by passive hemagglutination inhibition, in patients with disseminated candidiasis or Candida colonization.

A passive hemagglutination inhibition assay was studied by using a hyperimmune serum from rabbits immunized with whole yeast cells (Candida albicans group A). This technique was effective at detecting small amounts of laboratory-prepared mannan or a whole-cell extract of C. albicans. Of 32 patients with documented disseminated candidiasis that were tested, 19 showed evidence of circulating antigen by passive hemagglutination inhibition. Three of these patients showed only partial, rather than complete, inhibition. Among 22 colonized patients, 4 showed partial inhibition, and none of 49 normal controls demonstrated inhibition. All of the sera were tested for antibody by agglutination, immunodiffusion, and passive hemagglutination. This last technique added increased sensitivity, but not specificity, to the standard tests already in use. Fourfold or greater titer rises by passive hemagglutination occurred in fewer than one-third of patients with invasive candidiasis and developed in more than one-half of patients who were colonized and did not require systemic anticandida therapy.

Antibodies, Fungal↗

Restriction enzyme analysis of ribosomal DNA shows that Candida inconspicua clinical isolates can be misidentified as Candida norvegensis with traditional diagnostic procedures.

We identified 29 yeast isolates from 22 patients using the API ID32C panel. Twenty-eight of these isolates were Candida norvegensis and one was C. inconspicua. Although C. norvegensis is considered a pseudohypha-producing species, only one isolate produced pseudohyphae. Restriction enzyme analysis of PCR-amplified ribosomal DNA with four different enzymes proved that all isolates were C. inconspicua.

Candida↗

Differentiation of Candida albicans and Candida dubliniensis by using recombinant human antibody single-chain variable fragments specific for hyphae.

To identify antigens specific for the filamentous form of Candida albicans, a combinatorial phage display library expressing human immunoglobulin heavy and light chain variable regions was used to select phage clones capable of binding to the surfaces of viable C. albicans filaments. Eight distinct phage clones that bound specifically to filament surface antigens not expressed on blastoconidia were identified. Single-chain antibody variable fragments (scFv) derived from two of these phage clones (scFv5 and scFv12) were characterized in detail. Filament-specific antigen expression was detected by an indirect immunofluorescence assay. ScFv5 reacted with C. dubliniensis filaments, while scFv12 did not. Neither scFv reacted with C. glabrata, C. parapsilosis, C. rugosa, C. tropicalis, or Saccharomyces cerevisiae grown under conditions that stimulated filament formation in C. albicans and C. dubliniensis. Epitope detection by the two scFv was sensitive to proteinase K treatment but not to periodate treatment, indicating that the cognate epitopes were composed of protein. The antigens reactive with scFv5 and scFv12 were extractable from the cell surface with Zymolyase, but not with sodium dodecyl sulfate (SDS) and 2-mercaptoethanol, and migrated as polydisperse, high-molecular-weight bands on SDS-polyacrylamide gel electrophoresis gels. The epitopes were detected on clinical specimens obtained from infants with thrush and urinary candidiasis without passage of the organisms on laboratory media, confirming epitope expression in human infection. The availability of a monoclonal immunologic reagent that recognizes filaments from both C. albicans and C. dubliniensis and another specific only to C. albicans adds to the repertoire of potential diagnostic reagents for differentiation between these closely related species.

Antigens, Fungal↗

Disk diffusion testing using Candida sp. colonies taken directly from CHROMagar Candida medium may decrease time required to obtain results.

We compared results of disk diffusion antifungal susceptibility testing from Candida sp. strains passaged on CHROMagar and on potato dextrose agar. The overall categorical agreements for fluconazole and voriconazole disk testing were 95% and 98% with 0% and 0.5% very major errors, respectively. Disk diffusion testing by the CLSI (formerly NCCLS) M44-A method can be performed accurately by taking inocula directly from CHROMagar.

Agar↗

[Percutaneous transhepatic intraportal administration of amphotericin B to a patient with multiple liver abscesses due to Candida albicans: with monitoring by fungal index as a parameter of Candida volume].

A 59-year-old female with acute promyelocytic leukemia in remission was admitted to our center because of an episode of incidental high fever with general fatigue. She was found to have hepatomegaly. Abdominal CT revealed multiple liver abscesses and a positive culture was obtained for candida albicans from an aspirated abscess. She was treated with percutaneous transhepatic intraportal administration of amphotericin B in addition to oral and intravenous administration. We confirmed the remission of these abscesses by means of the fungal index which is the difference between the values of the limulus test and endotoxin specific test. The fungal index appears to be useful for early diagnosis and treatment of fungal infection.

Amphotericin B↗

[Prevalence of Candida albican serotypes in blood isolates in Chile, and first report of Candida dubliniensis candidemia].

Our main goal was to determine the prevalence of C. albicans serotypes isolates from blood cultures and identify the presence of C. dubliniensis. We studied 47 strains identified as C. albicans by conventional methods, 28 were isolated from children and 19 from adult patients. The strains were re-identified by standard methods and phenotypic screening as xylose assimilation and growth at 42 degrees C. API ID 32C (bioMérieux) was employed with the C. dubliniensis suspected strains and confirmation was made by molecular fingerprinting using random amplified polymorphic DNA (RAPD). The C. albicans serotype was determined by agglutination with antiserum anti-antigen 6 from cell wall (Candida Check, Iatron Inc., Japan) and the in vitro susceptibilities were evaluated by a microdilution method. From 47 strains, 46 were confirmed as C. albicans, 31 of them (67%) were serotype A. Adult patients presented a high prevalence of serotype A (95%) and children presented a frequency of 52% of the serotype B (p<0.05). We confirmed the identification of C. dubliniensis in one strain isolated from an infant. All serotype B strains were susceptible to fluconazole, itraconazole and amphotericin B. On the other hand, 3% and 6% of serotype A strains were "susceptible dose dependent" to fluconazole and itraconazole, respectively. C. albicans serotype A was predominant in adult candidemia and its distribution was homogenous in children patients. All strains were highly susceptible to antifungals. We report here the first case of C. dubliniensis candidemia in South America.

Adult↗

[Electron microscopic presentation of immune reactions on Candida cells: asteroid bodies in Candida albicans from the urine of nephritis patients].

Candida albicans cells from the urine of two nephritis patients were concentrated and incubated with ferritin-labeled antihuman grammaglobulin (either anti-IgA, anti-IgG, or anti-IgM). Electron microscopy showed the electron-transparent yeast cell wall to be surrounded by an electron-dense capsule-like substance of remarkable volume. This must be regarded as an antigen-antibody precipitate corresponding to the "asteroid body" of previous authors. The antibodies involved in the formation of the precipitate are mainly those of the IgA and IgG classes. Considering the results of previous authors, the following definition is proposed: "Asteroid Bodies" are light microscopically visible antigen-antibody precipitates on the cell wall of fungi parasitic condition.

Antigen-Antibody Reactions↗

[Serologic specificity of various Candida membranaefaciens and Candida rugosa strains].

C. membranaefaciens (C. majoricensis) CBS-2875 and C. rugosa 829 strains were agglutinated strongly with 0 6,7 agglutinating serum of S. cholerae suis. On the other hand the same gave a negative result with C. membranaefaciens CBS-1952 and C. rugosa (C. rugosa v. elegans) CBS-1948. These results can be considered in way of identification of the mentioned Candida types.

Agglutination Tests↗

Mannoproteins from Candida albicans elicit a Th-type-1 cytokine profile in human Candida specific long-term T cell cultures.

Since T lymphocytes are active producers of regulatory cytokines, long-term T cell cultures (LTTC) specific for a major mannoproteic antigen (MP) of Candida albicans from peripheral blood mononuclear cells (PBMC) of healthy donors were generated and their cytokine profile studied. LTTC consisted of an expanded CD3CD4 T-helper (Th) populations, with a high proportion of CD45R0 T memory cells. Stimulation of LTTC by MP induced a Th-1 type cytokine profile, as indicated by the presence of IFN-gamma and the absence of IL-5 (both as mRNA and protein) in cell cultures and supernatants. These results suggest a predominant Th1 response elicited by C. albicans mannoprotein antigen in human PBMC.

Antigens, Fungal↗

Candida endocrinopathy syndrome with membranoproliferative glomerulonephritis: demonstration of glomerular candida antigen.

A boy who had the Candida endocrinopathy syndrome, consisting of mucocutaneous candidiasis, hypoparathyroidism and hypoadrenocorticism, also had hypocomplementemic membranoproliferative glomerulonephritis. A renal biopsy at age 16 showed lobular glomerulonephritis with chronic interstitial inflammation. Glomerular and focal tubular deposition of IgG and C3, but not C4, were seen by immunofluorescence microscopy. Electron microscopy revealed dense intramembranous deposits in glomerular, tubular and Bowman's capsul basement membranes. C. albicans antigenic material was detected in the glomeruli by immunofluorescence microscopy using an FITC-labelled rabbit antiserum to the patient's organism. These fidings suggest that chronic C. albicans infection led to an immune complex form of nephritis. Clinical and laboratory evidence of resolution of his nephritis and return to normal serum levels of C3 followed prolonged antifungal therapy.

Adolescent↗

Experimental and clinical studies of 5-fluorocytosine activity in candida ocular infections, II. The effect of 5-fluorocytosine in treating experimental candida infection in rabbits' eyes.

Mild and severe experimental ocular infections in rabbits' eyes were induced by inoculation of Candida albicans with or without addition of dexamycin drops. These rabbits were treated with 5-Fluorocytosine, topically, orally and in a combined topical and oral manner. The topical treatment was effective only in mild infections. Severe infections responded to oral and especially to combined oral-tropical treatment.

Administration, Oral↗