Fukushima A, Nishino K, Yoshida H, Takata M, Ueno H
Department of Ophthalmology, Kochi Medical School, Nankoku City, Japan.
Br J Ophthalmol. 1999 Aug;83(8):973-9. doi: 10.1136/bjo.83.8.973.
Covalent conjugates consisting of diverse antigens coupled to optimal numbers of monomethoxypolyethylene glycol (mPEG) molecules have been shown to suppress antigen specific antibody formation. In this study, the possibility was examined that the same conjugates might prevent experimental immune mediated blepharoconjunctivitis (EC, formerly EAC) which had been shown to be caused by CD4(+) T cells-that is, to cell mediated immunity.
6-8 week old male Lewis rats were used. The test groups of rats received two intravenous injections, each of 300 microg, of a conjugate of ovalbumin mPEG (OVA(mPEG)(11)) in phosphate buffered saline (PBS), 14 and 28 days before the single immunisation with OVA in complete Freund's adjuvant. The rats were challenged 3 weeks later by eye drops containing OVA; 24 hours later they were sacrificed, and their eyes, blood, and lymph nodes were harvested for histological examination and determination of anti-OVA antibody titres and levels of cellular immunity. Two control groups received PBS or OVA in PBS before immunisation. Furthermore, the possibility that OVA(mPEG)(11) may have induced OVA specific suppressor cells was tested by establishing the effects of the co-transfer of splenocytes from OVA(mPEG)(11) treated rats with OVA primed lymph node cells on the manifestations of EC.
Either PBS or OVA pretreated rats, which had not received OVA(mPEG)(11), developed high levels of antibodies and cell mediated immune responses to OVA, and application of eye drops led to blepharoconjunctivitis with massive cellular infiltration. In contrast, pretreatment with OVA(mPEG)(11) prevented cellular infiltration into the lids and conjunctivas, as well as the formation of detectable humoral and cellular immunity against OVA. Co-transfer of splenocytes from OVA(mPEG)(11) treated rats with OVA primed lymph node cells suppressed the cellular infiltration on application of OVA on the conjunctiva.
These data indicate that intravenous injection of OVA(mPEG)(11) conjugates suppressed both humoral and cellular immunity by the effects of antigen specific suppressor cells, thus leading to the inhibition of development of EC.
由多种抗原与最佳数量的单甲氧基聚乙二醇（mPEG）分子偶联而成的共价缀合物已被证明可抑制抗原特异性抗体的形成。在本研究中，检验了相同缀合物可能预防实验性免疫介导性睑结膜炎（EC，原称EAC）的可能性，该疾病已被证明由CD4(+) T细胞引起，即由细胞介导的免疫反应引起。
使用6 - 8周龄的雄性Lewis大鼠。在大鼠用完全弗氏佐剂单次免疫卵清蛋白（OVA）前14天和28天，给试验组大鼠静脉注射两次，每次300微克磷酸盐缓冲盐水（PBS）中的卵清蛋白mPEG缀合物（OVA(mPEG)(11)）。3周后，用含OVA的滴眼液对大鼠进行攻击；24小时后处死大鼠，采集其眼睛、血液和淋巴结用于组织学检查以及测定抗OVA抗体滴度和细胞免疫水平。两个对照组在免疫前分别接受PBS或PBS中的OVA。此外，通过确定将OVA(mPEG)(11)处理的大鼠的脾细胞与OVA致敏的淋巴结细胞共同转移对EC表现的影响，来测试OVA(mPEG)(11)是否可能诱导OVA特异性抑制细胞。
未接受OVA(mPEG)(11)的PBS预处理或OVA预处理大鼠对OVA产生了高水平的抗体和细胞介导的免疫反应，应用滴眼液导致睑结膜炎并伴有大量细胞浸润。相比之下，用OVA(mPEG)(11)预处理可防止细胞浸润到眼睑和结膜，以及针对OVA的可检测的体液和细胞免疫的形成。将OVA(mPEG)(11)处理的大鼠的脾细胞与OVA致敏的淋巴结细胞共同转移可抑制在结膜上应用OVA时的细胞浸润。
这些数据表明，静脉注射OVA(mPEG)(11)缀合物通过抗原特异性抑制细胞的作用抑制了体液免疫和细胞免疫，从而导致EC发展的抑制。
