Reitz M S, Miller N R, Wong-Staal F, Gallagher R E, Gallo R C, Gillespie D H
Proc Natl Acad Sci U S A. 1976 Jun;73(6):2113-7. doi: 10.1073/pnas.73.6.2113.
Cultured peripheral blood leukocytes from a woman (patient HL23) with acute myelogenous leukemia produced type-C RNA tumor viruses (HL23V). The viruses were analyzed by molecular hybridization experiments after transmission to five secondary cell culture lines. Using the criteria of molecular hybridization, we concluded that all of the transmitted virus isolates have nucleotide sequences related to the genome of simian sarcoma virus (SiSV). In addition, in agreement with data reported elsewhere, some of the transmitted viruses also have nucleotide sequences related to those of the baboon endogenous virus (BaEV). We also used molecular hybridization to ascertain whether both viruses could have originated from the patient HL23. Utilizing [3H] cDNA complementary to RNA from the separated BaEV-related component of HL23V and hybridizing this cDNA to DNA from tissues of the patient, we detected sequences related to BaEV in DNA obtained from the patient's spleen. These BaEV DNA sequences were also detectable when 125I-labeled RNA from BaEV was used as a probe. In agreement with earlier results, however, no SiSV-related sequences were detectable in the DNA of her tissues. Cytoplasmic viral-like particles, which had a buoyant density of 1.15-1.2 g/ml and were capable of synthesizing cDNA in association with a 35S RNA in vitro, were also found in the patient's fresh uncultured leukemic blood cells. cDNA synthesized by the cytoplasmic particles contained some sequences that hybridized to RNA from SiSV and, in addition, some that hybridized to RNA from BaEV. The cDNA also hybridized significantly to DNA isolated from the spleen of patient HL23 and to cytoplasmic RNA from the patient's leukocytes. These molecular hybridization results with nucleic acids obtained from the fresh blood cells of the patient, combined with the repeated isolation of similar viruses from different blood and bone marrow samples from the same patient, suggest that the virus come directly from the leukemic cell samples. The finding of BaEV-related DNA proviral sequences in the spleen of the patient strongly supports this interpretation. The failure so far to find a complete SiSV-related provirus is perplexing, but could be attributable to the existence of such a provirus in DNA of only a small population of cells in most leukemic patient.
从一名患有急性髓性白血病的女性(患者HL23)培养的外周血白细胞产生了C型RNA肿瘤病毒(HL23V)。将病毒接种到五个继代细胞培养系后,通过分子杂交实验对其进行了分析。根据分子杂交标准,我们得出结论,所有接种的病毒分离株都具有与猿猴肉瘤病毒(SiSV)基因组相关的核苷酸序列。此外,与其他地方报道的数据一致,一些接种的病毒还具有与狒狒内源性病毒(BaEV)相关的核苷酸序列。我们还使用分子杂交来确定这两种病毒是否都源自患者HL23。利用与HL23V中分离出的与BaEV相关成分的RNA互补的[3H]cDNA,并将该cDNA与患者组织的DNA杂交,我们在从患者脾脏获得的DNA中检测到了与BaEV相关的序列。当使用来自BaEV的125I标记RNA作为探针时,也可检测到这些BaEV DNA序列。然而,与早期结果一致,在她的组织DNA中未检测到与SiSV相关的序列。在患者新鲜的未培养白血病血细胞中也发现了细胞质病毒样颗粒,其浮力密度为1.15 - 1.2 g/ml,并且能够在体外与35S RNA一起合成cDNA。由细胞质颗粒合成的cDNA包含一些与SiSV RNA杂交的序列,此外,还有一些与BaEV RNA杂交的序列。该cDNA还与从患者HL23脾脏分离的DNA以及患者白细胞的细胞质RNA有明显的杂交。这些来自患者新鲜血细胞的核酸分子杂交结果,加上从同一患者不同血液和骨髓样本中反复分离出类似病毒,表明病毒直接来自白血病细胞样本。在患者脾脏中发现与BaEV相关的DNA前病毒序列有力地支持了这一解释。迄今为止未能找到完整的与SiSV相关的前病毒令人困惑,但这可能是由于大多数白血病患者中只有一小部分细胞的DNA中存在这样的前病毒。