Castilla J, Pintado B, Sola I, Sánchez-Morgado J M, Enjuanes L
Department of Molecular and Cell Biology, Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Nat Biotechnol. 1998 Apr;16(4):349-54. doi: 10.1038/nbt0498-349.
Protection against enteric infections can be provided by the oral administration of pathogen-neutralizing antibodies. To provide passive immunity, 18 lines of transgenic mice secreting a recombinant monoclonal antibody (Mab) neutralizing transmissible gastroenteritis coronavirus (TGEV) into the milk were generated. The genes encoding a chimeric Mab with the variable modules of the murine TGEV-specific Mab 6A.C3 and the constant modules of a human IgG, isotype Mab were expressed under the control of regulatory sequences derived from the whey acidic protein, which is an abundant milk protein. The Mab 6A.C3 binds to a highly conserved epitope present in coronaviruses of several species, which does not allow the selection of neutralization escape mutants. Antibody expression titers of 10(6) were obtained in the milk of transgenic mice that reduced TGEV infectivity 10(6)-fold. The antibody was synthesized at high levels throughout lactation. Integration of matrix attachment region sequences with the antibody genes led to a 20- to 10,000-fold increase in the antibody titer in 50% of the transgenic animals. Antibody expression levels were transgene copy number independent and related to the site of integration. The generation of transgenic animals producing virus neutralizing antibodies in milk could provide an approach to protection against neonatal infections of the enteric tract.
通过口服病原体中和抗体可预防肠道感染。为提供被动免疫,构建了18株转基因小鼠品系,这些小鼠可在乳汁中分泌一种重组单克隆抗体(Mab),该抗体可中和传染性胃肠炎冠状病毒(TGEV)。编码嵌合单克隆抗体的基因,其可变区模块来自鼠源TGEV特异性单克隆抗体6A.C3,恒定区模块来自人IgG同种型单克隆抗体,在源自乳清酸性蛋白(一种丰富的乳蛋白)的调控序列控制下表达。单克隆抗体6A.C3可结合几种冠状病毒中存在的高度保守表位,这使得无法选择中和逃逸突变体。在转基因小鼠的乳汁中获得了10⁶的抗体表达滴度,该滴度可使TGEV感染性降低10⁶倍。在整个哺乳期抗体都能高水平合成。将基质附着区序列与抗体基因整合,使50%的转基因动物的抗体滴度提高了20至10000倍。抗体表达水平与转基因拷贝数无关,而与整合位点有关。产生在乳汁中具有病毒中和抗体的转基因动物,可为预防肠道的新生儿感染提供一种方法。