RESUMO
PURPOSE: To investigate the proteolytic effect of mast cell tryptase on eotaxin-1/CCL11, eotaxin-2/CCL24 and eotaxin-3/CCL26 produced by conjunctival fibroblasts. STUDY DESIGN: Experimental. METHODS: The production of eotaxin-1, -2 and -3 by conjunctival fibroblasts stimulated both with and without IL-4/IL-13 or/and TGF-ß1 was assessed by ELISA. The proteolytic activity of tryptase on eotaxins derived from conjunctival fibroblasts and recombinant eotaxins was also estimated by enzyme-linked immunosorbent assay (ELISA) and polymerase chain reaction (PCR). RESULTS: Conjunctival fibroblasts produced eotaxin-1 and -3, but not eotaxin-2. Stimulation with IL-4/IL-13 and TGF-ß1 synergistically increased eotaxin-1 and -3 production. Tryptase reduced the immunoreactivity of eotaxin-1 and -3 but not of eotaxin-2, due to the proteolysis of these eotaxins but not the inhibition of their m-RNA expression. CONCLUSION: Mast cell tryptase may exercise proteolytic activity on eotaxin-1 and -3 produced by conjunctival fibroblasts, resulting in partial suppression of the ability of eotaxin-1 and -3 to accumulate eosinophils in the conjunctiva. Eotaxin-2 in the tears may be a suitable biomarker of severity of allergic conjunctival disease.
Assuntos
Quimiocina CCL11/biossíntese , Quimiocina CCL24/biossíntese , Quimiocina CCL26/biossíntese , Túnica Conjuntiva/patologia , Conjuntivite Alérgica/metabolismo , Triptases/metabolismo , Células Cultivadas , Quimiocina CCL11/genética , Quimiocina CCL24/genética , Quimiocina CCL26/genética , Túnica Conjuntiva/metabolismo , Conjuntivite Alérgica/genética , Conjuntivite Alérgica/patologia , Ensaio de Imunoadsorção Enzimática , Fibroblastos/metabolismo , Fibroblastos/patologia , Humanos , Mastócitos/metabolismo , Mastócitos/patologia , Proteólise , RNA/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
BACKGROUND: Automated nontreponemal and treponemal test reagents based on the latex agglutination method (immunoticles auto3 RPR: ITA3RPR and immunoticles auto3 TP: ITA3TP) have been developed to improve the issues of conventional manual methods such as their subjectivity, a massive amount of assays, and so on. We evaluated these reagents in regards to their performance, reactivity to antibody isotype, and their clinical significance. METHODS: ITA3RPR and ITA3TP were measured using a clinical chemistry analyzer. Reactivity to antibody isotype was examined by gel filtration analysis. RESULTS: ITA3RPR and ITA3TP showed reactivity to both IgM- and IgG-class antibodies and detected early infections. ITA3RPR was verified to show a higher reactivity to IgM-class antibodies than the conventional methods. ITA3RPR correlated with VDRL in the high titer range, and measurement values decreased with treatment. ITA3RPR showed a negative result earlier after treatment than conventional methods. ITA3TP showed high specificity and did not give any false-negative reaction. Significant differences in the measurement values of ITA3RPR between the infective and previous group were verified. CONCLUSIONS: The double test of ITA3RPR and ITA3TP enables efficient and objective judgment for syphilis diagnosis and treatments, achieving clinical availability.