Production and characterization of monoclonal antibodies against secretory proteinase of candida albicans CBS 2730

Borg, Margarete; Watters, Dianne Josephine; Reich, Bettina; Rüchel, R.

doi:10.1016/s0176-6724(88)80116-0

Cited by 11 publications

(10 citation statements)

References 26 publications

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“…The mice were immunised with purified C. albicans proteinase. This confirmed the lack of immunodominance of the catalytic sites and the existence of shared epitopes between the enzymes from C. albicans and C. tropicalis [26]. IgM murine monoclonals produced against C. tropicalis proteinase also showed cross‐reactivity with the proteinase of C. albicans and C. parapsilosis and with porcine pepsin [27].…”

Section: Discussionsupporting

confidence: 56%

Epitope mapping Candida albicans proteinase (SAP 2)

Ghadjari

Matthews

Burnie

2006

FEMS Immunology &Amp; Medical Microbiology

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The continuous epitopes of Candida albicans proteinase SAP 2 were derived by epitope mapping with sera from patients with oral candidiasis (n=3), necropsy‐proven disseminated candidiasis (n=5), paired sera from patients who had recovered from blood culture‐proven disseminated candidiasis (n=3) and infection due to Candida parapsilosis (n=2) and Candida tropicalis (n=2). In C. albicans infection, IgM identified epitopes in amino acid positions 57–61 (QAVPV), 146–151 (SQGTLY) and 346–351 (PYDKCQ) and IgG at position 386–390 (VKYTS). For C. tropicalis IgM and IgG were positive for the same epitopes whilst IgG also detected epitopes at 78–83 (SNNQKL) and 159–164 (GVSIKN). For C. parapsilosis, IgM was positive for SNNQKL and IgG detected no epitopes. Reactivity of two of the epitopes as peptides KTSKRQAVPVTL and SLAQVKYTSASSI was confirmed in an indirect ELISA. At a cut‐off optical density of 0.4, IgM against either peptide was associated with survival but present in only about half of the sera (n=60) from patients who recovered from disseminated candidiasis whilst IgG levels were disappointing. Human recombinant antibodies from a patient who had recovered from disseminated candidiasis against either of these peptides had no activity in a lethal mouse model of candidal infection.

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Section: Discussionsupporting

confidence: 56%

Epitope mapping Candida albicans proteinase (SAP 2)

Ghadjari

Matthews

Burnie

2006

FEMS Immunology &Amp; Medical Microbiology

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“…Candida albicans [8,9,24,31,32], Cparapsilosis [6], Cryptococcus albidus [27] and Saccharomyces cerevisiae [10, 13,15] have been proven to secrete enzymes characterizable by their corresponding substrates. They were also found to release antigens detectable at the yeast surface and in culture media using polyclonal or monoclonai antibodies [1]. Studies have been particularly directed towards pathogenic yeast species to characterize molecules which could be associated with virulence factors secreted along the cellular cycle [5,20].…”

Section: Introductionmentioning

confidence: 99%

Ultrastructural immunodetection of a Pichia anomala killer toxin: a preliminary study

Cailliez

Gerloni

Morace

et al. 1992

Biology of the Cell

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Summary— A monoclonal antibody (mAb KT4), produced against a Pichia anomala killer toxin, was used to study the secretion process of toxin producing cells. The indirect immunofluorescence assay, performed with large concentrations of mAb KT4, showed a homogeneous distribution of the epitope at the cell surface of the P anomala cells. When increasing dilutions of mAb KT4 were employed, a ‘punctuated’ labeling appeared on the yeast's cell wall which suggested a heterogeneous secretion of the killer toxin. Similar labeling was also observed by immunodetection on live yeast cells held in buffered suspension. These results confirmed that 'punctuated' labeling was not an artefact due to a distortion of the cell's shape by having been dried on glass slides. Indirect immunodetection was performed in electron microscopy on ultra‐thin sections of cells embedded in Araldite resin. The labeling thus obtained showed both the presence of the epitope in the cytoplasm and its sensitivity to strong glutaraldehyde fixation. Indirect immunodetection, performed on ultra‐thin frozen sections, showed a cytoplasmic and cell wall labelling. However, the amount of gold particles observed in the cell wall was too low to confirm the heterogeneous killer toxin secretion observed in immunofluorescence. In this case, killer cells were fixed with a low concentration of glutaraldehyde which preserved the structure of the epitope complementary with mAb KT4.

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“…The use of urine as a test fluid instead of serum apparently overcame any problems related to the formation of complexes between Sap and alpha-2-macroglobulin, previously reported to occur in the circulation (64), and no antigen-antibody complexes needed to be removed before testing. It is possible that the pH of urine contributed to the dissociation of any complexes that may have formed in the circulation (3,64) or that complexes were removed by the dialysis methods used in the present study. Nonetheless, neither we, using urine as the test fluid, nor Na and Song (50), using serum as the test fluid, found removal of these complexes to be necessary.…”

Section: Discussionmentioning

confidence: 99%

“…The use of urine specimens, unlike that of serum specimens, would allow frequent, noninvasive collection of large sample volumes which could be readily concentrated to potentially increase test sensitivity. In addition, other researchers may have used unpurified Sap antigens as immunogens, leading to the production of antibodies that may cross-react with contaminating cell wall mannoprotein or extraneous proteins, reducing test specificity (3,4,39,65,74). We therefore used anti-Sap antibodies, raised against highly purified Sap and shown previously not to react with contaminating mannoproteins (47), to develop and evaluate an enzyme-linked immunosorbent assay to detect Sap in urine and serum specimens from rabbits experimentally infected with C. albicans.…”

mentioning

confidence: 99%

Competitive Binding Inhibition Enzyme-Linked Immunosorbent Assay That Uses the Secreted Aspartyl Proteinase ofCandida albicansas an Antigenic Marker for Diagnosis of Disseminated Candidiasis

Morrison

Hurst

Reiss

2003

Clin Vaccine Immunol

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The secreted aspartyl proteinases (Saps) of Candida albicans have been implicated as virulence factors associated with adherence and tissue invasion. The potential use of proteinases as markers of invasive candidiasis led us to develop a competitive binding inhibition enzyme-linked immunosorbent assay (ELISA) to detect Sap in clinical specimens. Daily serum and urine specimens were collected from rabbits that had been immunosuppressed with cyclophosphamide and cortisone acetate and infected intravenously with 10 7 C. albicans blastoconidia. Disseminated infection was confirmed by organ culture and histopathology. Although ELISA inhibition was observed when serum specimens from these rabbits were used, more significant inhibition, which correlated with disease progression, occurred when urine specimens were used. Urine collected as early as 1 day after infection resulted in significant ELISA inhibition (mean inhibition ؎ standard error [SE] compared with preinfection control urine, 15.7% ؎ 2.7% [P < 0.01]), and inhibition increased on days 2 through 5 (29.4% ؎ 4.8% to 44.5% ؎ 3.5% [P < 0.001]). Urine specimens from immunosuppressed rabbits infected intravenously with Candida tropicalis, Candida parapsilosis, Candida krusei, Cryptococcus neoformans, Aspergillus fumigatus, or Staphylococcus aureus were negative in the assay despite culture-proven dissemination. Nonimmunosuppressed rabbits receiving oral tetracycline and gentamicin treatment were given 2 ؋ 10 8 C. albicans blastoconidia orally or intraurethrally to establish colonization of the gastrointestinal tract or bladder, respectively, without systemic dissemination; urine specimens from these rabbits also gave negative ELISA results. Dissemination to the kidney and spleen occurred in one rabbit challenged by intragastric inoculation, and urine from this rabbit demonstrated significant inhibition in the ELISA (mean inhibition ؎ SE by day 3 after infection, 32.9% ؎ 2.7% [P < 0.001]). The overall test sensitivity was 83%, the specificity was 92%, the positive predictive value was 84%, the negative predictive value was 91%, and the efficiency was 89% (166 urine samples from 33 rabbits tested). The specificity, positive predictive value, and efficiency could be increased to 97, 95, and 92%, respectively, if at least two positive test results were required for a true positive designation. The ELISA was sensitive and specific for the detection of Sap in urine specimens from rabbits with disseminated C. albicans infection, discriminated between colonization and invasive disease, reflected disease progression and severity, and has the potential to be a noninvasive means to diagnose disseminated candidiasis.

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Production and characterization of monoclonal antibodies against secretory proteinase of candida albicans CBS 2730

Cited by 11 publications

References 26 publications

Epitope mapping Candida albicans proteinase (SAP 2)

Epitope mapping Candida albicans proteinase (SAP 2)

Ultrastructural immunodetection of a Pichia anomala killer toxin: a preliminary study

Competitive Binding Inhibition Enzyme-Linked Immunosorbent Assay That Uses the Secreted Aspartyl Proteinase ofCandida albicansas an Antigenic Marker for Diagnosis of Disseminated Candidiasis

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