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.

Ultrastructural immunodetection of a Pichia anomala killer toxin : a preliminary study / J.C. Cailliez, M. Gerloni, G. Morace, S. Conti, C. Cantelli, L. Polonelli. - In: BIOLOGY OF THE CELL. - ISSN 0248-4900. - 75:1(1992), pp. 19-23.

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

G. Morace;
1992

Abstract

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.
English
antibodies, monoclonal ; antigens, fungal ; cell wall ; cytoplasm ; epitopes ; immunohistochemistry ; killer factors, yeast ; microscopy, immunoelectron ; mycotoxins ; Pichia
Settore MED/07 - Microbiologia e Microbiologia Clinica
Articolo
Esperti anonimi
1992
Société de Biologie Cellulaire de France
75
1
19
23
5
Pubblicato
Periodico con rilevanza internazionale
info:eu-repo/semantics/article
Ultrastructural immunodetection of a Pichia anomala killer toxin : a preliminary study / J.C. Cailliez, M. Gerloni, G. Morace, S. Conti, C. Cantelli, L. Polonelli. - In: BIOLOGY OF THE CELL. - ISSN 0248-4900. - 75:1(1992), pp. 19-23.
none
Prodotti della ricerca::01 - Articolo su periodico
6
262
Article (author)
no
J.C. Cailliez, M. Gerloni, G. Morace, S. Conti, C. Cantelli, L. Polonelli
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/228162
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