Yoon J W, Selvaggio S, Onodera T, Wheeler J, Jenson A B
Diabetologia. 1981 Apr;20(4):462-7. doi: 10.1007/BF00253408.
The capacity of reovirus type 3 to infect insulin-producing B cells was studied in human pancreatic cell cultures. Antibody to reovirus was labelled with fluorescein isothiocyanate and antibody to insulin was labelled with tetramethyl rhodamine isothiocyanate. By using a double-labelled immunofluorescent antibody technique, it was shown that only about 6% of the insulin-containing human B cells in culture became infected when inoculated with unpassaged reovirus type 3. However, by repeated passage of the virus in human pancreatic B cell cultures, the percentage of infected B cells increased to 27%, and the virus titre in cultures rose from 8.0 X 19(4) pfu/ml in the first passage, to 4.9 X 10(6) pfu/ml in the 5th passage. As measured by radioimmunoassay, the intracellular immunoreactive insulin began to decrease at 24 h after infection. This decrease roughly paralleled the increase in virus titre. In contrast, there was relatively little change in immunoreactive insulin in cultures inoculated with unpassaged reovirus type 3. These studies show that the ability of reovirus type 3 to infect human B cells is enhanced by serial passage in human pancreatic cell cultures and that the infection resulted in the destruction of B cells and release of insulin.
在人胰腺细胞培养物中研究了3型呼肠孤病毒感染产生胰岛素的B细胞的能力。将呼肠孤病毒抗体用异硫氰酸荧光素标记,胰岛素抗体用异硫氰酸四甲基罗丹明标记。通过使用双标记免疫荧光抗体技术,结果显示,接种未传代的3型呼肠孤病毒时,培养物中仅约6%含胰岛素的人B细胞被感染。然而,通过在人胰腺B细胞培养物中反复传代该病毒,被感染B细胞的百分比增加到27%,并且培养物中的病毒滴度从第一代的8.0×10⁴pfu/ml上升到第5代的4.9×10⁶pfu/ml。通过放射免疫测定法检测,细胞内免疫反应性胰岛素在感染后24小时开始减少。这种减少大致与病毒滴度的增加平行。相比之下,接种未传代的3型呼肠孤病毒的培养物中免疫反应性胰岛素变化相对较小。这些研究表明,3型呼肠孤病毒感染人B细胞的能力通过在人胰腺细胞培养物中连续传代而增强,并且这种感染导致B细胞的破坏和胰岛素的释放。