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.
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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.
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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.
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