ROTHBARD S, WATSON R F
J Exp Med. 1962 Sep 1;116(3):337-46. doi: 10.1084/jem.116.3.337.
Antibody to rat collagen, prepared in rabbits and injected into the circulation of normal or adjuvant-prepared rats, becomes fixed to its antigen and can then be identified in tissue sections under ultraviolet light by its fluorescence after application of fluorescein-conjugated anti-rabbit globulin. In heart, lung, liver, spleen, adrenal, kidney, jejunum, lymph node, thymus, joint synovia, peripheral nerve, aorta, skeletal muscle, eye, and brain, the antibody was found at all sites where collagen and reticulin are normally present, but except for the kidneys of the adjuvant-prepared rats, no pathological abnormalities were demonstrated. It was not found within cells. Specific fluorescence was absent from tissues of rats injected with normal rabbit serum or rabbit anti-fish collagen serum or rabbit anti-rat collagen serum after absorption with rat collagen, but was present when the anti-rat collagen serum had been absorbed with fish collagen. The reaction could be blocked by pretreatment of sections with unlabeled anti-rabbit globulin and did not occur with heterologous labeled anti-duck globulin. After serial treatment in vitro with homologous antibody to collagen and the conjugated anti-rabbit globulin, purified reconstituted collagen fibers showed the same fluorescence as fibers in the tissues; no fluorescence of the fibers occurred when heterologous antibody to collagen was applied. These findings indicate that the antibody to rat collagen is directed toward an antigen present in both collagen and reticulin.
用兔制备的抗大鼠胶原蛋白抗体,注入正常或佐剂处理过的大鼠循环系统后,会与抗原结合,然后在应用荧光素标记的抗兔球蛋白后,可通过紫外线在组织切片中识别其荧光。在心脏、肺、肝、脾、肾上腺、肾、空肠、淋巴结、胸腺、关节滑膜、外周神经、主动脉、骨骼肌、眼和脑中,在胶原蛋白和网状纤维正常存在的所有部位都发现了抗体,但除了佐剂处理过的大鼠的肾脏外,未发现病理异常。在细胞内未发现该抗体。注射正常兔血清、兔抗鱼胶原蛋白血清或用大鼠胶原蛋白吸收后的兔抗大鼠胶原蛋白血清的大鼠组织中未出现特异性荧光,但在用鱼胶原蛋白吸收抗大鼠胶原蛋白血清后则出现特异性荧光。用未标记的抗兔球蛋白预处理切片可阻断该反应,而异源标记的抗鸭球蛋白则不会引发该反应。用同源胶原蛋白抗体和共轭抗兔球蛋白进行体外连续处理后,纯化的重组胶原蛋白纤维显示出与组织中纤维相同的荧光;应用异源胶原蛋白抗体时,纤维未出现荧光。这些发现表明,抗大鼠胶原蛋白抗体针对的是胶原蛋白和网状纤维中都存在的一种抗原。