Wang M W, Heap R B, King I, Taussig M J, Whyte A
AFRC Institute of Animal Physiology and Genetics Research, Cambridge, UK.
Scand J Immunol. 1988 Sep;28(3):367-76. doi: 10.1111/j.1365-3083.1988.tb01462.x.
Affinity-purified rabbit and sheep anti-idiotypic antisera raised against mouse monoclonal anti-progesterone IgG1 antibody (DB3) or mouse myeloma IgG1 protein P3 (MOPC 21) showed high binding specificities to the respective idiotypes used for immunization as determined by RIA or ELISA. They have been used in an indirect immunofluorescent method to demonstrate the localization of parenterally administered idiotypes in pregnant or pseudopregnant BALB/c mouse frozen tissue preparations, at known stages post coitum after a single intraperitoneal or intravenous injection of DB3 or P3. DB3 was visualized on the surface of uterine luminal and glandular epithelia of pregnant mice 36 h after treatment; the localization was DB3-specific as it was not seen in mice treated with P3 (using sheep anti-P3 anti-idiotype as a probe) or saline. The fluorescent staining reaction in oviduct was weak and only appeared on the surface of the oviducal serosa (peritoneal side). Both DB3 and P3 were also localized in liver (granules of Kupffer cells), kidney (glomerular basement membrane), spleen (on the membrane surface of mononuclear cells in the white pulp), and peritoneal exudate cells (on the membrane surface). Staining could be completely blocked by the addition of the free idiotypes against which the anti-idiotypes were made but not by the unrelated idiotype. Anti-idiotypic labelling in vivo is more specific and selective than anti-whole immunoglobulin labelling.
用小鼠单克隆抗孕酮IgG1抗体(DB3)或小鼠骨髓瘤IgG1蛋白P3(MOPC 21)制备的亲和纯化兔和羊抗独特型抗血清,通过放射免疫分析(RIA)或酶联免疫吸附测定(ELISA)检测,显示出对用于免疫的相应独特型具有高结合特异性。它们已被用于间接免疫荧光法,以证明在怀孕或假孕的BALB/c小鼠冷冻组织制剂中,经腹腔或静脉单次注射DB3或P3后,在已知的交配后阶段,经肠胃外给药的独特型的定位。治疗36小时后,在怀孕小鼠子宫腔和腺上皮表面可见DB3;这种定位是DB3特异性的,因为在用P3处理的小鼠(使用羊抗P3抗独特型作为探针)或生理盐水处理的小鼠中未见到。输卵管中的荧光染色反应较弱,仅出现在输卵管浆膜(腹膜侧)表面。DB3和P3也定位于肝脏(库普弗细胞颗粒)、肾脏(肾小球基底膜)、脾脏(白髓中单核细胞膜表面)和腹腔渗出细胞(膜表面)。加入制备抗独特型所用的游离独特型可完全阻断染色,但加入无关独特型则不能。体内抗独特型标记比抗全免疫球蛋白标记更具特异性和选择性。