Lapham C K, Bacík I, Yewdell J W, Kane K P, Bennink J R
Laboratory of Viral Diseases, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, Maryland 20892.
J Exp Med. 1993 Jun 1;177(6):1633-41. doi: 10.1084/jem.177.6.1633.
We isolated major histocompatibility complex (MHC)-specific viral peptides from cells infected with influenza virus in the continuous presence of the drug brefeldin A, which blocks exocytosis of newly synthesized MHC class I molecules. MHC-specific peptides were also isolated from cells expressing mouse Kd class I MHC molecules whose cytoplasmic domain was substituted by that of the adenovirus E3/19K glycoprotein. This molecule was retained in an intracellular pre-Golgi complex compartment as demonstrated by immunocytochemical and biochemical means. Since we show that intracellular association of antigenic peptides with such retained class I molecules is necessary for their isolation from cellular extracts, this provides direct evidence that naturally processed peptides associate with class I MHC molecules in an early intracellular exocytic compartment.
在布雷菲德菌素A持续存在的情况下，我们从感染流感病毒的细胞中分离出主要组织相容性复合体（MHC）特异性病毒肽，布雷菲德菌素A可阻断新合成的MHC I类分子的胞吐作用。我们还从表达小鼠Kd I类MHC分子的细胞中分离出MHC特异性肽，该分子的胞质结构域被腺病毒E3/19K糖蛋白的胞质结构域所取代。通过免疫细胞化学和生化方法证明，该分子保留在细胞内高尔基复合体前区室中。由于我们发现抗原肽与这种保留的I类分子在细胞内的结合对于从细胞提取物中分离它们是必要的，这提供了直接证据，表明天然加工的肽在细胞内早期胞吐区室中与I类MHC分子结合。
