Ihle J N, Joseph D R, Pazmino N H
J Exp Med. 1976 Dec 1;144(6):1406-23. doi: 10.1084/jem.144.6.1406.
The expression of endogenous ecotropic viruses in radiation-induced thymomas of C57BL/6 mice was examined. Competition radioimmunoassays for AKR MuLV gp71, p30, and p12 were used for viral antigen expression. 3 of 40 lymphomas had readily detectable ecotropic gp71 at levels of 95-689 ng/mg protein; the remainder of the tumors had no detectable gp71 (less than 1.0 ng/mg protein). 30 thymomas were characterized by the presence of MuLV p30 at levels of 1-10 ng/mg protein, levels that were comparable to those found in thymus extracts from age-matched, nonirradiated control. 10 tumors were characterized by having p30 levels of 10-30 ng/mg protein. In one tumor significant levels of AKR MuLV p12 were detectable. Since B-tropic and N-tropic viruses from C57BL/6 mice have glycoproteins (gp71) indistinguishable from AKR MuLV gp71 and the N-tropic virus had a p12 serologically identical to AKR MuLV p12, these results demonstrate that overt endogenous B-tropic virus was detectable in 2 of 40 thymomas and endogenous N-tropic virus was detectable in 1 of 40 thymomas. The lack of overt expression of gp71 or p12 was also confirmed by cytotoxicity assays using monospecific antisera to these viral proteins. Radiation-induced lymphomas were also examined for the presence of reverse transcriptase after chromatography of tissue extracts on poly G-Sepharose. One tumor, which was characterized by the lack of gp71, also had no detectable reverse transcriptase; whereas one tumor with gp71 was characterized by readily detectable levels of reverse transcriptase in cellular extracts. The presence of viral RNA was examined using AKR cDNA. Low levels of RNA capable of hybridizing with AKR cDNA were found in age-matched, nonirradiated mice; these hybrids had Tm's of 72 degrees C, while hybrids with AKR MuLV 70S RNA had Tm's of 80 degrees C. In 1 of 12 thymomas the concentration of hybridizable RNA and the Tm of the hybrids were identical to control values. In 9 of 12 thymomas the concentration of hybridizable sequences increased approximately three-to fivefold and the Tm of these hybrids varied from 73 to 75 degrees C. In 1 of 12 thymomas the concentration of hybridizable sequences increased over 100-fold, hybridized completely with AKR MuLV cDNA, and the hybrids had Tm's of 79 degrees C. This thymoma was also characterized by the presence of the AKR MuLV type of gp71 and p12. One tumor was characterized by a 10-to 100-fold increase in hybridizable sequences, which only partially hybridized with AKR MuLV cDNA, and hybrids had a Tm of 73 degrees C. This tumor was characterized by the presence of AKR MuLV gp71 but not AKR MuLV p12. The results taken together demonstrate that overt endogenous ecotropic virus expression is only rarely detectable in radiation-induced thymomas of C57BL/6 mice.
对C57BL/6小鼠辐射诱导的胸腺瘤中内源性嗜亲性病毒的表达进行了检测。采用针对AKR MuLV gp71、p30和p12的竞争放射免疫分析法检测病毒抗原表达。40个淋巴瘤中有3个的嗜亲性gp71水平易于检测,为95 - 689 ng/mg蛋白质;其余肿瘤未检测到gp71(低于1.0 ng/mg蛋白质)。30个胸腺瘤的特征是存在水平为1 - 10 ng/mg蛋白质的MuLV p30,该水平与年龄匹配的未受辐射对照小鼠胸腺提取物中的水平相当。10个肿瘤的特征是p30水平为10 - 30 ng/mg蛋白质。在一个肿瘤中可检测到显著水平的AKR MuLV p12。由于C57BL/6小鼠的嗜B性和嗜N性病毒的糖蛋白(gp71)与AKR MuLV gp71无法区分,且嗜N性病毒的p12在血清学上与AKR MuLV p12相同,这些结果表明,在40个胸腺瘤中有2个可检测到明显的内源性嗜B性病毒,40个胸腺瘤中有1个可检测到内源性嗜N性病毒。使用针对这些病毒蛋白的单特异性抗血清进行细胞毒性试验也证实了gp71或p12缺乏明显表达。在组织提取物在聚G - 琼脂糖上进行层析后,还检测了辐射诱导的淋巴瘤中逆转录酶的存在情况。一个以缺乏gp71为特征的肿瘤也未检测到逆转录酶;而一个有gp71的肿瘤的特征是细胞提取物中逆转录酶水平易于检测。使用AKR cDNA检测病毒RNA的存在情况。在年龄匹配的未受辐射小鼠中发现了能够与AKR cDNA杂交的低水平RNA;这些杂交体的熔解温度(Tm)为72℃,而与AKR MuLV 70S RNA的杂交体的Tm为80℃。在12个胸腺瘤中有1个,可杂交RNA的浓度和杂交体的Tm与对照值相同。在12个胸腺瘤中有9个,可杂交序列的浓度增加了约三到五倍,这些杂交体的Tm在73至75℃之间变化。在12个胸腺瘤中有1个,可杂交序列的浓度增加了100倍以上,与AKR MuLV cDNA完全杂交,杂交体的Tm为79℃。这个胸腺瘤的特征还包括存在AKR MuLV类型的gp71和p12。一个肿瘤的特征是可杂交序列增加了10到100倍,仅与AKR MuLV cDNA部分杂交,杂交体的Tm为73℃。这个肿瘤的特征是存在AKR MuLV gp71但不存在AKR MuLV p12。综合这些结果表明,在C57BL/6小鼠辐射诱导的胸腺瘤中,明显的内源性嗜亲性病毒表达很少能被检测到。