Preparation of murine monoclonal antibodies against the yeast phase of the dimorphic fungus Sporothrix schenckii
Three murine monoclonal antibodies (Mabs) were raised against a cytoplasmic antigen of the yeast phase of the pathogenic fungus Sporothrix schenckii using a modification of standard hybridoma technology incorporating the immunosuppressive drug cyclophosphamide. When tested for species-specificity within the pathogenic dimorphic fungi one of these Mabs (S5) showed little cross-reactivity by enzyme-linked immunosorbent assay and Western blot, though there was some recognition of Paracoccidioides brasiliensis antigen. This Mab recognized a 70–75 kDa molecule on reduced Western blots of S. schenckii antigen. The other two Mabs (S12 and S15) showed cross-reactivity with all dimorphic fungal antigens tested, though they appeared to recognize a molecule of similar molecular weight. This is the first report of any attempt to raise species-specific Mabs against this important causative agent of dermatological disease.
- [Show abstract] [Hide abstract] ABSTRACT: Although the generation of protein and LPS profiles allows adequate differentiation of organisms for some purposes, a much greater degree of discrimination may be obtained by using an additional property of these and other microbial molecules - their antigenicity. The development of serotyping schemes resulted from the early realization that specific antibodies produced by immunization of laboratory animals could be used to detect even subtle antigenic differences between micro-organisms. Such differences were often not detectable by other methods, and the combination of sensitivity and specificity afforded by antibodies provided a major advance in typing methods. By using carefully characterized panels of antibodies, organisms sharing common major antigens are classified into serogroups, which are then further sub-divided into serotypes or serovars by using antibodies to antigens, or groups of antigens, whose production varies between different strains or isolates. The serotype finally allocated therefore usually reflects a combination of common and unique antigens of the organism, and can be used to differentiate and identify organisms at the genus and species level. Although all classes of microbial macromolecules are antigenic, antibodies to proteins, lipopolysaccharides and polysaccharides are used most widely. Different classes of molecule may be used to differentiate organisms at different levels.0Comments 0Citations
- [Show abstract] [Hide abstract] ABSTRACT: Monoclonal antibodies (MoAbs) have had a major impact on many areas of biomedical research and almost since their advent have been used in the characterisation and identification of diagnostically important antigens of fungal pathogens. Their main significance lies in three, often inter-related areas: a) the definition and characterisation of antigens for use in detection of antibody responses, b) their direct use in the detection of diagnostically useful antigen in body fluids c) their application in immunohistochemical diagnosis. The degree to which MoAbs have been applied varies between fungal pathogens, and they have now been used, for example, in the serodiagnosis of Aspergillus sp., Cryptococcus neoformans, Histoplasma capsulatum and Paracoccidioides brasiliensis. Their use in producing diagnostic tests for other fungi such as Sporothrix schenckii and Penicillium marneffei has been more restricted but considerable potential exists for further development.0Comments 5Citations
- [Show abstract] [Hide abstract] ABSTRACT: A peptide-polysaccharide, a peptide-rhamnomannan, was isolated from the pathogenic yeast form of the fungus Sporothrix schenckii. This substance, which may play a role in fungal virulence, was tested in an animal model of systemic disease, and depression of the immune response was observed in the animals between the 4th and 6th week of infection. Concomitantly, this compound showed mitogenic activity when challenged with normal lymphocytes and was also found to be involved in the inflammatory response. These results provide further information for the understanding of fungal implantation in tissues and of the pathogenicity of this systemic mycosis.0Comments 25Citations