Fukushima J, Tani K, Atsumi Y, Hamajima K, Kawamoto S, Okuda K
Department of Bacteriology, Yokohama City University School of Medicine, Japan.
J Immunol. 1988 Jul 1;141(1):302-7.
We have engineered a bovine papilloma virus to carry the class II MHC (A beta d) gene. The recombinant plasmid was introduced into various mouse B cell hybridomas by co-transfecting with the neomycin-resistance gene. Several transfectants which received only the beta-chain gene of I-Ad, expressed I-Ad proteins on the cell surface. Their presence was determined by direct immunofluorescence and by Northern blot hybridization as well as RNA dot blot hybridization. These I-Ad molecules could induce IL-2 production in I-Ad restricted T cell hybridomas. When I-Ad molecules are expressed on the cell surface of the transfectants, they become fully active in inducing the I-Ad restricted T cell response. The original B cell hybridomas used for transfection possessed the alpha and beta chain genes of the I-Ad molecule but there are no transcripts of these genes in the cell. We suppose that many copies of the transfected beta-chain gene or its product may also induce production of cellular alpha-chain gene products and thereby induce expression of the I-Ad molecule on the cell surface.
我们构建了一种携带II类主要组织相容性复合体(Aβd)基因的牛乳头瘤病毒。通过与新霉素抗性基因共转染,将重组质粒导入各种小鼠B细胞杂交瘤。几个仅接受I-Adβ链基因的转染子在细胞表面表达了I-Ad蛋白。通过直接免疫荧光、Northern印迹杂交以及RNA斑点印迹杂交确定了它们的存在。这些I-Ad分子可在I-Ad限制性T细胞杂交瘤中诱导白细胞介素-2的产生。当I-Ad分子在转染子的细胞表面表达时,它们在诱导I-Ad限制性T细胞反应中变得完全活跃。用于转染的原始B细胞杂交瘤拥有I-Ad分子的α链和β链基因,但细胞中没有这些基因的转录本。我们推测,转染的β链基因或其产物的多个拷贝也可能诱导细胞α链基因产物的产生,从而诱导I-Ad分子在细胞表面的表达。