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'Actin-reactive' discriminated from 'non-actin-reactive' smooth muscle autoantibody by immunofluorescence reactivity
Ban-Hock Toh1, Roberta Taylor, Wendy Pollock
1Autoimmunity Laboratory, Centre for Inflammatory Diseases, Department of Medicine, Faculty of Medicine, Monash University, Victoria, Australia. Ban-Hock.Toh@med.monash.edu.au
Aims:
To compare smooth muscle antibody (SMA) patterns in tissue sections with patterns in an immunofluorescence assay (IFA) using a rat intestinal epithelial cell line and results from an F-actin IgG ELISA.
Methods:
SMA positive sera (n = 188) were classified by immunofluorescence staining of rodent kidney, stomach and liver sections as SMA-T (tubules) (n = 124) or SMA-V (vessels) (n = 64). The F-actin pattern on the rat epithelial cell line was identified by immunofluorescence staining of actin cables that was confirmed by dual immunofluorescence co-localisation with phalloidin.
Results:
Of 124 SMA-T positive sera, 123 reacted with the epithelial cell line and 120 with F-actin by ELISA, giving sensitivity for detection of anti-F-actin antibody of 99% and 97%, respectively. Of 64 SMA-V positive sera, four reacted with the epithelial cell line (6%) and 41 with F-actin by ELISA (64%). Tests of 493 normal blood donors and 100 disease controls yielded specificities of 584/593 (98.5%) and 562/593 (94.8%) for the cell line IFA and F-actin ELISA, respectively.
Conclusions:
The rat epithelial cell line IFA is a robust diagnostic assay for anti-F-actin antibody that can either replace the routine screening for actin-reactive SMA-T antibody in tissue sections or be used as a confirmatory assay for anti-F-actin antibody after screening by F-actin ELISA or on rodent tissue sections by IFA.
