Effect of Substrate on Indirect Immunofluorescence Test for Canine Pemphigus Foliaceus

Abstract

The effect of substrate on indirect immunofluorescence (IIF) tests for the detection of circulating autoantibodies was studied by examining sera from 14 canine pemphigus foliaceus patients, six sera with non-pemphigus dermatoses and ten normal dog sera against five different substrates from three species. These substrates included bovine esophagus, bovine nose, bovine tongue, monkey esophagus, and canine nose skin. Nine out of 14 (64.3%) sera from patients with canine pemphigus foliaceus showed intercellular space staining by indirect immunofluorescence using bovine esophagus as substrate. However, sera from nonpemphigus dermatoses and normal dog did not react with bovine esophagus. In other substrates, only bovine tongue showed 1/8 (12.5%) positive reaction at the intercellular space by sera from canine pemphigus foliaceus. Dog nose skin showed the intercellular space staining against ten of ten (100%) normal dog serum. Monkey esophagus showed the fluorescent deposit at the intercellular space in four of nine (44.4%) of pemphigus foliacues dog sera, however, four of ten (40%) of normal dog sera revealed nonspecificity intercellular staining. These results indicate that the sensitivity and the specifity of IIF test in canine pemphigus foliaceus depend on the substrate. The best substrate for detecting circulating autoantibody in canine pemphigus foliaceus patients among five different substrates was bovine esophagus because of its sensitivity and high specificity. The diagnosis of canine pemphigus foliaceus should be made on the basis of a combination of clinical signs, histopathology, direct immunofluorescence, and the detection of circulating autoantibody.

Comparison of three monoclonal and three polyclonal antibodies in the immunohistochemical diagnosis of canine autoimmune skin diseases

“…Both immunofluorescence and immunoperoxidase methods have been used 4–13 . More recently, circulating autoantibodies have also been detected 14,15 …”

A Canine Pemphigus Foliaceus Case Showing Parallel Relationship of Disease Activity and Titer of Serum Anti-keratinocyte Cell Surface Antibodies

“…2) [5,7]. Fluorescent deposition on the cell surface of keratinocytes could be observed when both substrates were used, but maximum IIF titer on bovine esophagus (1:10240) was considerably higher than that on living MCA-B1 cells (1:80).…”

“…However, the sensitivity of IIF for diagnosis of canine PF could be improved when cryosectioned bovine esophagus was used as substrate, although a few dogs with non-pemphigus dermatoses showed false-positive immunoreactivity [5,7]. In addition, despite of its low sensitivity, living keratinocyte staining using a canine keratinocyte cell line, MCA-B1, has provided a highly reliable serological diagnostic tool to detect serum autoantibodies in canine PF with its high specificity [5].…”

Neutrophils Contact to Plasma Membrane of Keratinocytes Including Desmosomal Structures in Canine Pemphigus Foliaceus

“…Circulating autoantibodies against keratinocyte cell surfaces were recognized in dogs with PF, as determined by indirect immunofluorescence (IIF) examination on cryosections of normal epithelia or cultured canine keratinocytes [6,9]. Recent paper described that injection of serum IgG from three dogs with PF to neonatal mice caused acantholytic blisters at the superficial layer of epidermis [12].…”