New developments in the serological diagnosis of Candida infection.
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Antibodies, Fungal↗
Biomedical subjects
R C Matthews
Publications and source records attributed to R C Matthews.
Immunoblot analysis: a new method for fingerprinting hospital pathogens.
Immunoblotting has recently become popular as a way of fingerprinting those hospital pathogens where other more conventional typing systems are deficient. Extracts of microorganisms are prepared by chemical or enzymic means, run on a standard SDS-PAGE gel and transferred onto nitrocellulose membrane. They are then probed either by a hyperimmune antiserum raised in a rabbit or by serum from a patient who has been previously infected by the organism. The pattern of antigenic bands which stain forms the basis of the method. This article discusses the limitations of the system, makes recommendations for further systems and outlines a typical fingerprinting protocol.
Animals↗
Transient abrogation of immunosuppression in a patient with chronic mucocutaneous candidiasis following vaccination with Candida albicans.
A patient with long-standing chronic mucocutaneous candidiasis had reversed T helper/suppressor (TH/TS) cell ratios, hypergammaglobulinaemia E and serum inhibitors of lymphocyte transformation to mitogens and candida antigens. Following vaccination with whole heat-killed yeasts of Candida albicans, temporary clinical improvement coincided with the return of the TH/TS cell ratio to normal, reduction in concentration of IgE and reduced serum inhibitory activity to Concanavalin A and candida antigens. These changes were not permanent and 6 months after vaccination all indices had reverted to their pretreatment values. The production of antibody to a 47 kDa antigen of C. albicans has been shown to coincide with recovery from systemic disease. High concentrations of this antibody were demonstrated initially in the patient's serum and were unaffected by vaccination. If the 47 kDa antibody is protective, the results of this study suggest that separate immune responses may protect against mucocutaneous and systemic candidiasis and that defective immune responses against mucocutaneous disease need not imply lack of protection against systemic spread.
Antibodies, Fungal↗
Isolation of immunodominant antigens from sera of patients with systemic candidiasis and characterization of serological response to Candida albicans.
Candidal antigens were isolated by affinity chromatography from the sera of patients with disseminated Candida albicans infections. The immunodominant 47-kilodalton (kDa) antigen appeared to be a heat-stable breakdown product of several larger heat-labile components (84 to 92, 74 to 79, and 66 to 72 kDa). It was undetectable in normal sera and sera from four patients with systemic C. parapsilosis, C. tropicalis, and C. krusei infections. Serum samples from 92 patients with proven systemic C. albicans infections were examined by the immunoblot technique. Seventy-four patients had detectable antibody, and 92% of these produced antibody to the 47-kDa antigen. All survivors had major serological responses to this antigen, whereas patients who died had no, minor, or fading responses. Fifty-five of the patients were neutropenic following cytotoxic chemotherapy for malignancies, usually lymphoproliferative disorders (hematological patients). The remainder were surgical or medical patients (nonhematological). Hematological patients differed from nonhematological patients in the range of antigens that were commonly recognized by their immune systems, although antibodies to the 47- and 60-kDa antigens were frequently present in both groups. They also differed in that they produced mainly an immunoglobulin M (IgM) response, failing to seroconvent to IgG. This did not reduce survival rates, which were similar in both groups. It may be responsible, however, for the lower antigen titers that were observed in hematological patients when measured by reverse passive latex agglutination.
Antibodies, Fungal↗
Role of immunoblotting in the diagnosis of culture negative and enterococcal endocarditis.
Serum samples from patients with endocarditis and septicaemia due to Enterococcus faecalis, Enterococcus faecium, Streptococcus bovis, and Streptococcus sanguis were immunoblotted against antigenic extracts from all four species. In E faecalis endocarditis there was a strong IgM response to E faecalis antigenic bands of 112, 88-90, and 45-47 Kd and a strong IgG response to 88-90 and 45-47 Kd bands. In E faecium endocarditis there was a pronounced IgG response to an E faecium band of 82-90 Kd. For S bovis endocarditis, there was a strong IgG response to several components of S bovis including bands of 66, 58, 52 and 4 Kd. For S sanguis, there was a strong IgG response to bands of 80-82, 76, 60 and 45 Kd. These patterns of antibody production were absent in patients with uncomplicated septicaemia and in controls. The delineation of these patterns enabled confirmation of the final diagnosis in seven patients initially suspected of having culture negative endocarditis.
Antibodies, Bacterial↗
Analysis of the antibody response developing in an infant with Candida albicans meningitis.
A case of meningitis due to Candida albicans in a neonate is described. This is the first report of an immunoblot analysis of both serial samples of serum and cerebrospinal fluid in relation to C. albicans antigens. The western blot technique demonstrated that the mother had IgM antibody against five candidal antigens, including one of molecular weight 47 kDaltons, but only a small amount of IgG antibody against two antigens of molecular weight 104 kDaltons and 60 kDaltons respectively. The baby produced IgM antibody and then IgG antibody against an antigen of 47 kDaltons molecular weight, and this was associated with recovery.
Antibodies, Fungal↗
Immunoblot analysis of the serological response in invasive Trichosporon beigelii and Blastoschizomyces capitatus infections.
The serological response to Trichosporon sp. was examined by the immunoblot technique. Antibodies to a range of antigens (200 to 16 kilodaltons) were detectable in three patients with invasive Trichosporon beigelii or Blastoschizomyces capitatus infections and 10 uninfected controls. High levels of preexisting antibodies may contribute to the rarity of systemic infections.
Antibodies, Fungal↗
Immunoblotting to demonstrate antigenic and immunogenic differences among nine standard strains of Clostridium difficile.
The epidemiology of Clostridium difficile-associated disease is being elucidated with the development of typing schemes for the organism. We recently described a new typing scheme based on the incorporation of [35S]methionine into bacterial proteins followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and autoradiography. Nine standard strains were identified. We report here some observations on the antigenic differences among these nine strains when studied by immunoblotting. Type-specific rabbit antiserum was raised against each of the nine standard strains. Immunoblotting of the strains with these antisera demonstrated, in addition to the presence of shared, common proteins, a type-specific response with homologous antisera. When [35S]methionine-labeled C. difficile proteins were immunoblotted with homologous and heterologous antisera, both the immunoblots and the autoradiographs demonstrated the same strain-specific response. These strain-specific proteins, which have been so useful for epidemiological and typing purposes, were also immunogenic.
Animals↗
Immunoblot analysis of serological response to Pseudomonas aeruginosa septicaemia in man.
The occurrence of an outbreak of Pseudomonas aeruginosa septicaemias on an oncology ward permitted an analysis of antibody responses in patients who were all orally exposed to the same source of infection. Seven patients became septicaemic. Serial serum samples were immunoblotted against the homologous strain. Responses were compared with those of 16 other patients with septicaemias caused by other strains and 10 healthy controls. All 18 survivors produced increasing IgG or IgA antibody, or both, against a 35,000 dalton band, whereas these antibodies were usually absent or fell in titre in the five fatal cases. These antibodies were also lacking in sera taken just before a patient became septicaemic. This band had the electrophoretic characteristics of the outer membrane porin protein F.
Adolescent↗
Control of an outbreak of systemic Candida albicans.
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Antifungal Agents↗
Use of immunoblotting to identify antigenic differences between the yeast and mycelial phases of Candida albicans.
Western blotting was applied to the analysis of Candida albicans in the yeast and mycelial phases in an attempt to recognise mycelial specific antigens which might be of serodiagnostic value. The antisera were prepared in rabbits by immunising them with pressates of C albicans type A NCTC 3153 in the yeast phase or the mycelial phase. These were blotted against C albicans in the yeast and mycelial phases and the yeast phase of C parapsilosis, C krusei, C tropicalis, and Torulopsis glabrata. Cross reactivity was greatest against C parapsilosis. One yeast specific mannoprotein was identified with a molecular weight of 49 000. No mycelial specific antigens could be identified.
Antigens, Fungal↗
Immunoblot analysis of the serological response in systemic candidosis.
To investigate the heterogeneity in antibody response to the various antigenic determinants of Candida albicans in patients with disseminated candidosis 201 serial serum samples from 45 patients with proven systemic candidosis were examined by immunoblotting (Western blotting) for antibodies to C albicans type A NCTC 3153. 40 patients had detectable antibody. 26 different antigen bands, ranging from 104 kD to 23 kD, were identified. Despite wide variation in the antigens recognised by different patients, the antibody responses fell into six patterns (A to F). 13 patients infected during an outbreak of systemic candidosis gave the same response (A). Of a further 7 patients with this response, 2 had isolates indistinguishable from the outbreak strain. Group B responses were associated with leukaemias and lymphomas. Groups C-F responses were seen in 9 patients. Production of antibody to a 47 kD antigen, common to all six groups, occurred in all those who recovered from the infection and may therefore be of prognostic significance. Whether this antigen could be used as the basis of a vaccine remains to be determined.
Antibodies, Fungal↗
New techniques in external cardiac compressions. Aquatic cardiopulmonary resuscitation.
Despite the 8,000 drownings that occur annually in the United States, procedures for aquatic first aid are currently limited to mouth-to-mouth resuscitation, while cardiopulmonary resuscitation (CPR) must be delayed until the victim is transported to a solid surface for conventional closed-chest cardiac compressions. We discuss techniques of positioning a victim on the rescuer's chest and initiating CPR, on site. Respiratory support was provided with a slightly modified scuba regulator, and procedures were tested on an instrumented aquatic CPR mannequin in the water and anesthetized dogs on land. Most results were within specified criteria for successful CPR.
Drowning↗
Feasibility study of CPR in the water.
More than 8000 drownings occur each year in the United States alone. With the increased popularity of scuba diving for commercial and sport purposes, one would predict an increase in related deaths or accidents. Yet procedures to administer first aid are limited to mouth-to-mouth, and external cardiac compressions must await moving the victim to a firm surface. This study discusses the technique of placing a victim upon the rescuer's chest and initiating full cardiopulmonary resuscitation (CPR) immediately on the site. During emergency regulator resuscitation (E.R.R.), ventilations are administered by use of a slightly modified, factory-calibrated scuba regulator. The techniques were tested on an instrumented aquatic CPR mannequin and found to meet the published criteria for successful CPR.
Diving↗
Inhaled CO2 and progressive hypoxia: ventilatory response in a yearling and a newborn harbor seal.
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Aging↗
Comparative assessment of the detection of candidal antigens as a diagnostic tool.
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Antigens, Fungal↗