Anti Glypican 3, a Novel Ancillary Maker in the Histological Assessment of Hirschsprung's Disease
Objectives: Hirschsprung's disease (HSCR) results from a malformation of the enteric nervous system. A congenital absence of intrinsic ganglion cells from the distal rectum and a variable length of the contiguous bowel is the required diagnostic feature of Hirschsprung's disease and total colonic aganglionosis (TCA) We evaluated the utility of a monoclonal antibody directed against glypican 3 (GPC 3) a membrane bound protein involved in regulation of the signaling of Wingless types (WNTs) Hedgehogs (Hh) Fibroblast Growth Factors (FGFs) and Bone Morphogenetic Proteins (BMPs) in the detection of ganglion cells in formalin fixed, paraffin embedded tissue sections. Methods: The presence/absence of ganglion cells was evaluated retrospectively by immunohistochemical staining for calretinin and GPC 3 in tissue specimens; a total of 15 patients who underwent colectomy (total or sub total) for histologically proven aganglionosis (14 HSCR, 1 TCA) and 5 rectal suction biopsies (4HSCR B, 1 TCA B) were considered. Of the 20 considered cases, a total of 60 tissue specimens (3 for each patient) were selected. A total of 30 additional normal (N) colonic mucosa biopsy samples were also included. Results: GPC 3 constantly identified ganglion cell bodies in all but 2 normal biopsies (with normal presentation of ganglion cells on hematoxylin and eosin (H&E) stain) and was negative in all 60 aganglionotic biopsies; these results were reflective of calretinin staining pattern. Conclusions: The present study indicates that monoclonal anti GPC 3 might prove to be useful immunohistochemical marker in the identification of ganglion cells in paraffin embedded rectal tissue specimens and suction biopsies. Further studies in larger series will contribute to demonstrate its utility as an ancillary marker in the histological assessment of HSCR aganglionosis.
Journal of pediatric gastroenterology and nutrition