Lobigs M, Rothenfluh H S, Blanden R V, Müllbacher A
Division of Cell Biology, John Curtin School of Medical Research, Australian National University, P. O. Box 334, Canberra, ACT 2601, Australia.
Immunogenetics. 1995;42(5):398-407. doi: 10.1007/BF00179402.
We have already shown that in species with highly polymorphic major histocompatibility complex (MHC) class I molecules (human, mouse) no functional polymorphism of the peptide transporters TAP1 and TAP2 is detectable (Lobigs and Müllbacher 1993). Investigating the antigen-presentation machinery of the class I MHC monomorphic Syrian hamster using mouse MHC class I expression via recombinant vaccinia viruses (VV) we found that six hamster cell lines fall into two phenotypic classes. four cell lines (HaK, FF, MF-2, and HT-1) showed no defect in expressing four different H2 class I molecules (Kk, Kd, Kb, Dd) and the appropriate VV peptide recognized by mouse VV-immune cytotoxic T (Tc) cells on the cell surface. Two cell lines (BHK-21 and NIL-2) expressed Dd and Kb in association with VV peptides as recognized by VV-immune, H2-restricted Tc cells but not Kk and Kd. However, Kd was expressed on the cell surface, as shown by fluorescence-activated cell sorter (FACS) analysis and alloreactive Tc-cell recognition. Kk is only surface-expressed in these two cell lines when superinfected with two VV recombinants encoding rat TAP1 (VV-mtp1) and TAP2 (VV-mtp2). Superinfection with VV-mtp1 and VV-mtp2 rendered both cell lines, after infection with either VV-Kk and VV-Kd, susceptible to lysis by either Kk- or Kd-restricted VV-immune Tc cells. Thus Syrian hamster cell lines express functionally polymorphic peptide transporters. The TAP2 gene from FF cells was cloned and sequenced; comparison with human, mouse, and rat TAP2 sequences show 78%, 88% and 87% similarity, respectively.
我们已经表明，在具有高度多态性主要组织相容性复合体（MHC）I类分子的物种（人类、小鼠）中，未检测到肽转运体TAP1和TAP2的功能多态性（洛比格斯和米尔巴赫，1993年）。通过重组痘苗病毒（VV）利用小鼠MHC I类表达来研究I类MHC单态叙利亚仓鼠的抗原呈递机制时，我们发现六个仓鼠细胞系可分为两个表型类别。四个细胞系（HaK、FF、MF - 2和HT - 1）在表达四种不同的H2 I类分子（Kk、Kd、Kb、Dd）以及细胞表面上被小鼠VV免疫细胞毒性T（Tc）细胞识别的相应VV肽方面没有缺陷。两个细胞系（BHK - 21和NIL - 2）表达与VV肽相关的Dd和Kb，这可被VV免疫的、H2限制的Tc细胞识别，但不表达Kk和Kd。然而，如荧光激活细胞分选仪（FACS）分析和同种异体反应性Tc细胞识别所示，Kd在细胞表面表达。当用编码大鼠TAP1（VV - mtp1）和TAP2（VV - mtp2）的两种VV重组体进行超感染时，Kk仅在这两个细胞系的表面表达。用VV - mtp1和VV - mtp2进行超感染使这两个细胞系在感染VV - Kk和VV - Kd后，易被Kk或Kd限制的VV免疫Tc细胞裂解。因此，叙利亚仓鼠细胞系表达功能多态性的肽转运体。克隆并测序了来自FF细胞的TAP2基因；与人类、小鼠和大鼠TAP2序列的比较分别显示出78%、88%和87%的相似性。
