Tengowski M W, Bjorling D E, Albrecht R M, Saban R
Department of Zoology, University of Wisconsin, Madison 53706, USA.
J Pharmacol Toxicol Methods. 1997 Feb;37(1):15-21. doi: 10.1016/s1056-8719(96)00143-8.
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.
本研究旨在评估一种将致敏组织内抗原位置与生理反应相关联的技术。通过腹腔注射卵清蛋白（OVA）进行主动致敏的豚鼠被安乐死，并取出膀胱。将金珠（直径18 nM）与OVA（OVA-Au）和牛血清白蛋白（BSA-Au）偶联。膀胱组织悬浮于组织浴中，分别暴露于OVA、OVA-Au、BSA和BSA-Au中，然后测定组织收缩和组胺释放情况。通过电子显微镜检查膀胱组织，以确定暴露后1分钟和5分钟时金标记抗原的分布。将致敏豚鼠的膀胱组织暴露于OVA后，会同时刺激组织收缩和组胺释放，且在3分钟内达到最大水平；来自对照未致敏豚鼠的膀胱组织对OVA无反应。BSA未能刺激OVA致敏或对照膀胱组织产生反应。暴露于OVA-Au 1分钟后，标记抗原附着在致敏膀胱组织的黏膜上。5分钟内，OVA-Au存在于致敏组织的黏膜和黏膜下层。OVA-Au未附着于对照组织，也未被其内化，且BSA-Au未附着于任何组织，也未被其内化。用金标记抗原能够确定组织内抗原的位置，且不影响致敏组织对抗原暴露的反应。
