Ulrich R, Sommerfeld K, Schröder A, Prokoph H, Arnold W, Krüger D H, Scherneck S
Institut für Medizinische Virologie, Universitätsklinikum Charité der Humboldt-Universität, Berlin, Germany.
Virus Genes. 1996;12(3):265-74. doi: 10.1007/BF00284647.
The hamster polyomavirus (HaPV) is associated with spontaneously appearing skin epithelioma of the Syrian hamster Z3 strain. Virus particles prepared from the skin epithelioma cause lymphoma and leukemia when injected into newborn hamsters from a distinct Syrian hamster colony (HaP); in contrast to the skin epithelioma the hemopoietic tumors are virus free but accumulate viral DNA. To study the humoral immune response of HaPV-infected Z3 hamsters we produced recombinant HaPV proteins in Escherichia coli as beta-galactosidase-, TrpE- and dihydrofolate reductase-fusion proteins or as non-fused proteins. Recombinant plasmids carried segments of all putative early and late HaPV proteins. The recombinant proteins were detected in stained SDS polyacrylamide gels and in Western blots using monoclonal anti-TrpE and anti-beta-galactosidase antibodies and sera of HaPV-infected hamsters. Sera from HaPV-infected Z3 hamsters and crude lysates of all clones were applied to Western blots to characterize the humoral immune response in the animals. HaPV-specific antibodies were found to be directed against early protein segments translated from the first common exon and from the second unique exon of LT and MT, resp., as well as against the late proteins VP1 and VP2/3. The almost complete VP2 was recognized by all sera whereas VP1 was detected only by a half of the sera. Our data suggest the presence of at least 2 immunodominant regions in VP2, one in the C-terminal VP1 and at least 4 in early proteins.
仓鼠多瘤病毒（HaPV）与叙利亚仓鼠Z3品系自发出现的皮肤上皮瘤有关。从皮肤上皮瘤制备的病毒颗粒注入来自不同叙利亚仓鼠群体（HaP）的新生仓鼠时会引发淋巴瘤和白血病；与皮肤上皮瘤不同，造血肿瘤不含病毒，但会积累病毒DNA。为了研究HaPV感染的Z3仓鼠的体液免疫反应，我们在大肠杆菌中制备了重组HaPV蛋白，分别作为β-半乳糖苷酶、色氨酸E和二氢叶酸还原酶融合蛋白或非融合蛋白。重组质粒携带了所有假定的HaPV早期和晚期蛋白的片段。使用单克隆抗色氨酸E和抗β-半乳糖苷酶抗体以及HaPV感染仓鼠的血清，在染色的SDS聚丙烯酰胺凝胶和蛋白质印迹中检测到了重组蛋白。将HaPV感染的Z3仓鼠的血清和所有克隆的粗裂解物应用于蛋白质印迹，以表征动物体内的体液免疫反应。发现HaPV特异性抗体针对分别从LT和MT的第一个共同外显子和第二个独特外显子翻译的早期蛋白片段，以及晚期蛋白VP1和VP2/3。几乎完整的VP2被所有血清识别，而VP1仅被一半的血清检测到。我们的数据表明VP2中至少存在2个免疫显性区域，一个在C端VP1中，早期蛋白中至少存在4个。
