Use of gold-labeled ovalbumin to correlate antigen deposition and localization with tissue response

Abstract This study was performed to evaluate a technique for correlating the location of antigen within sensitized tissues with physiological response. Guinea pigs actively sensitized by intraperitoneal injection of ovalbumin (OVA) were euthanized, and urinary bladders were removed. Gold beads (18 nM diameter) were conjugated to OVA (OVA-Au) and bovine serum albumin (BSA-Au). Bladder tissue was suspended in tissue baths, exposed to OVA, OVA-Au, BSA, and BSA-AU, and tissue contraction and histamine release were determined. Bladder tissues were examined by electron microscopy to determine distribution of gold-labeled antigen at 1 and 5 min after exposure. Exposure of bladder tissue from sensitized guinea pigs to OVA stimulated concomitant contraction and histamine release which reached maximal levels within 3 min; bladder tissue from control, nonsensitized guinea pigs did not respond to OVA. BSA failed to stimulate response from OVA-sensitized or control bladder tissue. Labeled antigen was adhered to mucosa of sensitized bladder tissue 1 min after exposure to OVA-Au. OVA-Au was present within the mucosa and submucosa of sensitized tissues within 5 min. OVA-Au did not adhere to, or become internalized by, control tissues, and BSA-Au did not adhere to, or become internalized by, any tissues. Labeling of antigen with gold allowed the location of antigen within tissues to be determined and did not affect the response of sensitized tissues to antigen exposure.