Eager K B, Hackett C J, Gerhard W U, Bennink J, Eisenlohr L C, Yewdell J, Ricciardi R P
Wistar Institute of Anatomy and Biology, Philadelphia, PA 19104.
J Immunol. 1989 Oct 1;143(7):2328-35.
A retrovirus vector containing the hemagglutinin (HA) gene of influenza virus was constructed and used to infect murine cell lines of fibroblast, mastocytoma and B cell lineages which are able to present antigens to MHC-restricted T cells. Stable cell lines were selected in which the retrovirus vector integrated as a single copy in almost all of the individual cell clones examined. The HA mRNA was shown to be of the expected length by Northern blot analysis, but the levels varied among the cell clones. Although the HA transcript was difficult to detect in any of the retrovirus-infected cell clones derived from fibroblasts, HA Ag was easily detected on the cell surface by cytofluorographic analysis. Significantly, retrovirus-infected clones derived from each cell type were recognized by HA-specific class I and class II MHC-restricted T lymphocytes. HA produced in these cells was able to be acquired, processed, and presented to class II-restricted T cells by additional, non-HA-expressing APC. This indicates that HA endogenously synthesized within these cell lines is available for Ag processing by an exogenous route.
构建了一种含有流感病毒血凝素(HA)基因的逆转录病毒载体,并用于感染能够将抗原呈递给MHC限制性T细胞的成纤维细胞、肥大细胞瘤和B细胞系的小鼠细胞系。选择了稳定的细胞系,其中逆转录病毒载体在几乎所有检测的单个细胞克隆中以单拷贝形式整合。通过Northern印迹分析表明HA mRNA长度符合预期,但细胞克隆之间的水平有所不同。虽然在源自成纤维细胞的任何逆转录病毒感染的细胞克隆中都很难检测到HA转录本,但通过细胞荧光分析很容易在细胞表面检测到HA抗原。值得注意的是,源自每种细胞类型的逆转录病毒感染克隆被HA特异性的I类和II类MHC限制性T淋巴细胞识别。这些细胞中产生的HA能够被额外的非HA表达抗原呈递细胞摄取、加工并呈递给II类限制性T细胞。这表明在这些细胞系内源性合成的HA可通过外源性途径进行抗原加工。