Anagnostopoulos I, Hummel M, Kreschel C, Stein H
Institute of Pathology, Klinikum Benjamin Franklin, Free University Berlin, Germany.
Blood. 1995 Feb 1;85(3):744-50.
The present study was undertaken to unequivocally demonstrate the morphology, immunophenotype, and localization of Epstein Barr virus (EBV)-infected cells as well as the type of infection (latent versus productive) in tonsils of acute infectious mononucleosis. Paraffin sections from nine cases with clinical, serologic, and morphologic evidence of EBV infection were analyzed for the detection of small transcripts, designated EBER1 & 2, and BHLF1 by in situ hybridization (ISH) using nonisotopically labeled probes. ISH was combined with immunohistology, employing a broad panel of antibodies against B-, T-, epithelial-, macrophage-, and follicular dendritic cell (FDC)-antigens. All EBER-positive cells could be identified as lymphocytes, as they did not exhibit any morphologic or immunologic characteristics of epithelial cells, macrophages, or FDCs. A preferential accumulation of EBER-positive cells was noted around crypts, within surface squamous epithelium, and in the surroundings of necrosis. The majority of these lymphocytes could be shown to be B cells, which morphologically included Reed-Sternberg (RS)-like cells, immunoblasts, medium-sized lymphoid cells, as well as cells with plasmacytoid differentiation. In all cases, a varying number of EBER-positive T cells could be identified. ISH for BHLF1-RNA detection showed that almost all cases contained single positive small lymphoid cells, indicating a transition from latent to productive infection cycle. Such cells could also be detected within the crypt epithelium reaching up to its surface. Additional screening of 123 oropharyngeal mucosa samples from patients without evidence of acute EBV-infection, using the polymerase chain reaction for EBV-DNA detection combined with EBER- and BHLF1-ISH showed single latently infected lymphocytes in only one case. Our data imply that infected lymphocytes and not epithelial cells are, in fact, the reservoir for EBV infection, and that these are the cells that participate in the interindividual virus transfer.
本研究旨在明确证实急性传染性单核细胞增多症扁桃体中爱泼斯坦-巴尔病毒（EBV）感染细胞的形态、免疫表型和定位，以及感染类型（潜伏性与增殖性）。对9例具有EBV感染临床、血清学和形态学证据的病例的石蜡切片进行分析，使用非同位素标记探针通过原位杂交（ISH）检测小转录本，即EBER1和EBER2以及BHLF1。ISH与免疫组织学相结合，使用了针对B细胞、T细胞、上皮细胞、巨噬细胞和滤泡树突状细胞（FDC）抗原的广泛抗体组合。所有EBER阳性细胞均可鉴定为淋巴细胞，因为它们不表现出上皮细胞、巨噬细胞或FDC的任何形态学或免疫学特征。注意到EBER阳性细胞在隐窝周围、表面鳞状上皮内以及坏死灶周围有优先聚集。这些淋巴细胞大多数可显示为B细胞，其形态学上包括里德-施特恩贝格（RS）样细胞、免疫母细胞、中等大小的淋巴细胞以及具有浆细胞样分化的细胞。在所有病例中，均可鉴定出数量不等的EBER阳性T细胞。用于检测BHLF1-RNA的ISH显示，几乎所有病例都含有单个阳性小淋巴细胞，表明从潜伏感染周期向增殖性感染周期的转变。此类细胞也可在隐窝上皮内直至其表面检测到。对123例无急性EBV感染证据患者的口咽黏膜样本进行额外筛查，使用聚合酶链反应检测EBV-DNA并结合EBER和BHLF1-ISH，仅在1例中发现单个潜伏感染的淋巴细胞。我们的数据表明，事实上，受感染的淋巴细胞而非上皮细胞是EBV感染的储存库，并且这些细胞参与个体间的病毒传播。
