Chauhan D, Bharti A, Raje N, Gustafson E, Pinkus G S, Pinkus J L, Teoh G, Hideshima T, Treon S P, Fingeroth J D, Anderson K C
Department of Adult Oncology, Dana-Farber Cancer Institute and the Department of Medicine, Harvard Medical School, Brigham and Womens Hospital, Boston, MA, USA.
Blood. 1999 Mar 1;93(5):1482-6.
Whether Kaposi's sarcoma herpesvirus (KSHV) is associated with multiple myeloma (MM) remains controversial. We assayed for KSHV DNA sequences in long-term bone marrow stromal cells (BMSCs) from 26 patients with MM and 4 normal donors. Polymerase chain reaction (PCR) using primers which amplify a KSHV gene sequence to yield a 233-bp fragment (KS330233 within open reading frame 26) was negative in all cases. Aliquots of these PCR products were used as templates in subsequent nested PCR, with primers that amplify a 186-bp product internal to KS330233. BMSCs from 24 of 26 (92%) patients with MM and 1 of 4 normal donors were KSHV PCR+. DNA sequence analyses showed interpatient specific mutations (2 to 3 bp). Both Southern blot and sequence analyses confirmed the specificity of PCR results. The presence of the KSHV gene sequences was further confirmed by amplifying T 1.1 (open reading frame [ORF] K7) and viral cyclin D (ORF 72), two other domains within the KSHV genome. Immunohistochemical studies of KSHV PCR+ MM BMSCs demonstrate expression of dendritic cell (DC) lineage markers (CD68, CD83, and fascin). Serological studies for the presence of KSHV lytic or latent antibodies were performed using sera from 53 MM patients, 12 normal donors, and 5 human immunodeficiency virus (HIV)/KSHV+ patients. No lytic or latent antibodies were present in sera from either MM patients or normal donors. Taken together, these findings show that KSHV DNA sequences are detectable in BMSCs from the majority of MM patients, but that serologic responses to KSHV are not present. Ongoing studies are defining whether the lack of antibody response is caused by the absence of ongoing infection, the presence of a novel viral strain associated with MM, or underlying immunodeficiency in these patients.
卡波西肉瘤疱疹病毒（KSHV）是否与多发性骨髓瘤（MM）相关仍存在争议。我们检测了26例MM患者和4名正常供者的长期骨髓基质细胞（BMSC）中的KSHV DNA序列。使用引物进行聚合酶链反应（PCR），该引物扩增KSHV基因序列以产生233 bp片段（开放阅读框26内的KS330233），所有病例均为阴性。这些PCR产物的等分试样用作后续巢式PCR的模板，引物扩增KS330233内部的186 bp产物。26例MM患者中的24例（92%）和4名正常供者中的1例的BMSC为KSHV PCR阳性。DNA序列分析显示患者间存在特异性突变（2至3 bp）。Southern印迹和序列分析均证实了PCR结果的特异性。通过扩增KSHV基因组内的另外两个区域T 1.1（开放阅读框[ORF]K7）和病毒细胞周期蛋白D（ORF 72），进一步证实了KSHV基因序列的存在。对KSHV PCR阳性的MM BMSC进行免疫组织化学研究，显示树突状细胞（DC）谱系标志物（CD68、CD83和fascin）的表达。使用53例MM患者、12名正常供者和5名人类免疫缺陷病毒（HIV）/KSHV阳性患者的血清进行KSHV裂解或潜伏抗体存在情况的血清学研究。MM患者或正常供者的血清中均不存在裂解或潜伏抗体。综上所述，这些发现表明，大多数MM患者的BMSC中可检测到KSHV DNA序列，但对KSHV不存在血清学反应。正在进行的研究正在确定抗体反应缺失是由于持续感染的缺乏、与MM相关的新型病毒株的存在，还是这些患者潜在的免疫缺陷所致。
