Enzyme immunoassay for the macrocyclic trichothecene roridin A: production, properties, and use of rabbit antibodies

Antisera against roridin A were prepared by using a roridin A-hemisuccinate derivative coupled to human serum albumin as the immunogen. Antibodies could be detected in the sera of the immunized rabbits as early as 4 weeks after the initial exposure. After one booster injection at week 14, high antibody titers were measured over a period of 21 weeks. The specificity and sensitivity of the antibodies were tested by using roridin A-hemisuccinate coupled to horseradish peroxidase as an enzyme-linked toxin in a competitive assay with a double-antibody solid phase. The assay was most specific for the tested macrocyclic trichothecenes, and the relative cross-reactivities with roridin A, roridin J, verrucarin A, satratoxin H, and satratoxin G were 1, 0.41, 0.15, 0.15, and 0.07, respectively. When 16 nonmacrocyclic trichothecenes were tested, only diacetylverrucarol (0.0015) and verrucarol (0.0005) showed minor cross-reactivity. The sensitivity of the enzyme immunoassay for the detection of roridin A was in the range of 5 to 50 ng/ml (0.16 to 1.6 ng per assay).     

Characteristics of heterologous antisera against stromal fibroblasts]

As shown with the use of heterologous rabbit antifibroblast sera (AFS) to stromal mechanocytes of guinea pig, the media from the cultured bone marrow, spleen, and thymus stromal fibroblasts contained specific trypsinoresistant fibroblast protein (AG-1), which was also present in the normal blood serum and on the surface of stromal fibroblasts. AG-1 was insensitive to collagenase effect and apparently differed from the chief surface protein of fibroblasts (CSP). AG-1 is referred to gamma1-globulin, and is probably a new specific surface fibroblast protein. AFS contained also antibodies to the nonspecific protein of fibroblasts (AG-2), alpha1,-globulin related to AG1.     

Immunological diagnosis of Mycoplasma pneumonias]

A complex of immunological cell tests with M. pneumoniae antigen (the lymphocyte blast-cell transformation test, the allergic neutrophil alteration test) was carried out in order to establish the correlation between the results of positive seroconversion and the sepcific immunological reactivity of lymphoid cells in pneumonia patients. Mycoplasmic cutireactive allergen, when used for the accelerated diagnosis of mycoplasmic pneumonia in humans, was shown to be specific and safe. Cuti-allergic tests with mycoplasmic allergen allowed to diagnose mycoplasmic pneumonia at early stages (beginning from days 5--7), which ensures the possibility of indicating etiotropic treatment to patients in due time.