Production and characterisation of a monoclonal antibody to a cell-surface, glucomannoprotein constituent of Candida albicans and other pathogenic Candida species

Cassone, Antonio; Torosantucci, Antonella; Boccanera, Maria; Pellegrini, G.; Palma, Carla; Malavasi, Fabio

doi:10.1099/00222615-27-4-233

Cited by 72 publications

(56 citation statements)

References 14 publications

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“…These molecules have been shown to elicit specific antibodies in mice (36), rats (37), rabbits (9), and humans (11,38). The construction of neoglycolipids with phosphopeptidomannan-released ␤-1,2-oligomannosides has demonstrated that they can act as epitopes for a large number of anti-C. albicans phosphopeptidomannan monoclonal antibodies (16), suggesting that C. albicans mannoglycoconjugate(s) expressing these residues are strong immunogens.…”

Section: Discussionmentioning

confidence: 99%

The Candida albicans Phospholipomannan Is a Family of Glycolipids Presenting Phosphoinositolmannosides with Long Linear Chains of β-1,2-Linked Mannose Residues

Trinel

Plancke

Gerold

et al. 1999

Journal of Biological Chemistry

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In a series of studies, we have shown that Candida albicans synthesizes a glycolipid, phospholipomannan (PLM), which reacted with antibodies specific for ␤-1,2-oligomannosides and was biosynthetically labeled by [ 3 H]mannose, [3 H]palmitic acid, and [ 32 P]phosphorus. PLM has also been shown to be released from the C. albicans cell wall and to bind to and stimulate macrophage cells. In this study, we show by thin layer chromatography scanning of metabolically radiolabeled extracts that the C. albicans PLM corresponds to a family of mannose and inositol co-labeled glycolipids. We describe the purification process of the molecule and the release of its glycan fraction through alkaline hydrolysis. Analysis of this glycan fraction by radiolabeling and methylation-methanolysis confirmed the presence of inositol and of 1,2-linked mannose units. NMR studies evidenced linear chains of ␤-1,2-oligomannose as the major PLM components. Mass spectrometry analysis revealed that these chains were present in phosphoinositolmannosides with degrees of polymerization varying from 8 to 18 sugar residues. The PLM appears as a new type of eukaryotic inositol-tagged glycolipid in relationship to both the absence of glucosamine and the organization of its glycan chains. This first structural evidence for the presence of ␤-1,2-oligomannosides in a glycoconjugate other than the C. albicans phosphopeptidomannan may have some pathophysiological relevance to the adhesive, protective epitope, and signaling properties thus far established for these residues.

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Section: Discussionmentioning

confidence: 99%

The Candida albicans Phospholipomannan Is a Family of Glycolipids Presenting Phosphoinositolmannosides with Long Linear Chains of β-1,2-Linked Mannose Residues

Trinel

Plancke

Gerold

et al. 1999

Journal of Biological Chemistry

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“…The crude mannoprotein extract and its chromatographic fractions were assayed for their reactivity with antiCandida serum and with mAb AF1, directed against an oligosaccharide epitope of the candidal cell surface Torosantucci et al, 1990). An indirect ELISA and an ELISA-inhibition assay were used, as reported previously (Ausiello et al, 1986;Scaringi et ul., 1988;Cassone et al, 1988). The polyclonal Serum was generated in rabbits by immunization with four subcutaneous administrations, at weekly intervals, of 2 x lo7 glutaraldehyde-inactivated cells of C. afbicuns strain BP (Ausiello et al, 1986).…”

Section: Sds-page Analysismentioning

confidence: 99%

“…Candida albicans is a human commensal and opportunistic pathogen which possesses several distinctive properties of a potent biological response modifier (Cassone et al, 1987;Domer et al, 1988). It is important to investigate these properties in detail, as their comprehension may offer additional insight into the nature of the Candida-host relationship, both in normal subjects and in patients with candidosis.…”

Section: Introductionmentioning

confidence: 99%

Lymphoproliferative and cytotoxic responses of human peripheral blood mononuclear cells to mannoprotein constituents of Candida albicans

Torosantucci

Palma²,

Boccanera³

et al. 1990

Journal of General Microbiology

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Two major proteoglycan constituents (designated F1 and F2) of the cell wall of Candida albicans were separated by ion-exchange chromatography from a crude carbohydrate-rich extract (GMP), and investigated for their chemical and molecular composition, antigenicity and immunomodulatory properties in cultures of human peripheral blood mononuclear cells (PBMC). Both fractions consisted predominantly of Periodic acidSchitf (PAS) and concanavalin A (Con A)-reactive material consisting of > 90 % mannose, 3-5% protein and small amounts of phosphorus; each was recognized by an anti-Candida rabbit serum as well as by a monoclonal antibody (mAb AF1) directed against an oligosaccharide epitope present on the fungal cell surface. When F1 and F2 were subjected to SDS-PAGE, transblotted and stained with enzyme-conjugated mAb AF1 or Con A, most of the antibody or lectin bound to high molecular mass (> 200 kDa) polydisperse material, some of which was present in F2 (as in the starting GMP extract) but absent in F1. This difference was also observed in PAS-stained gels of the two fractions.The F2, but not the F1, constituent was as active as the unfractionated GMP extract in inducing lymphoproliferation, production of the cytokines interleukin-2 and interferon-y, and generation of cytotoxicity against a natural-killer-sensitive target cell line (K562). These immunomodulatory properties were, like those possessed by GMP, protease-sensitive and heat-stable. Treatment of PBMC cultures with a modulatory anti-Tcell receptor antibody abolished the lymphoproliferation induced by GMP and F2 but not that induced by phytohaemagglutinin, showing that the mannoprotein materials of C. albicans acted through interaction with the antigen receptor complex.

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“…The antibodies used throughout these experiments were selected according to their ability to react with C. albicans mannan-released acid-labile oligomannosides (Trine1 e t al., 1992(Trine1 e t al., , 1993. They consisted of two mouse IgM monoclonal antibodies, mAb AF1 and mAb DF9-3, respectively furnished by Professor A. Cassone (Cassone et al, 1988) and Dr M. Borg-von-Zepelin (Borg-von-Zepelin & Grunes, 1993), and serum factor 5, a rabbit polyclonal IgG, purchased from Iatron Laboratory (Tokyo, Japan). Preparation of whole-cell extracts.…”

Section: Methodsmentioning

confidence: 99%

Mapping of -1,2-linked oligomannosidic epitopes among glycoconjugates of Candida species

et al. 1995

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The distribution of p-1,2-linked oligomannosides among glycoconjugates of various Candida species was investigated by Western blotting, using monoclonal and polyclonal antibodies which react with these epitopes. Expression of b1,2-linked oligomannosidic epitopes on a 14-18 kDa polydisperse antigen nonreactive with concanavalin A (ConA), previously identified as a C. albicans serotype A phospholipomannan (PLM), appeared to be restricted to C. albicans serotypes A and B (including var. C. stellatoidea types I and II) and C. tropicalis. In C. albicans, p1,2-linked oligomannosidic epitopes also appeared to be slightly associated with high molecular mass (> 100 kDa) polydisperse ConA-reactive mannoproteins. For all the other Candida strains investigated, belonging to the species C. parapsilosis, C. kmsei, C. glabrata and C. robusta (4 cemwisiae), &1,2=linked oligomannosidic epitopes were found to be present in association with medium molecular mass (18-1 00 kDa) and high molecular mass ConA-reactive mannoproteins, giving reproducible labelling profiles that varied between species.

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Production and characterisation of a monoclonal antibody to a cell-surface, glucomannoprotein constituent of Candida albicans and other pathogenic Candida species

Cited by 72 publications

References 14 publications

The Candida albicans Phospholipomannan Is a Family of Glycolipids Presenting Phosphoinositolmannosides with Long Linear Chains of β-1,2-Linked Mannose Residues

The Candida albicans Phospholipomannan Is a Family of Glycolipids Presenting Phosphoinositolmannosides with Long Linear Chains of β-1,2-Linked Mannose Residues

Lymphoproliferative and cytotoxic responses of human peripheral blood mononuclear cells to mannoprotein constituents of Candida albicans

Mapping of -1,2-linked oligomannosidic epitopes among glycoconjugates of Candida species

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