Ito Y, Bando H, Komada H, Tsurudome M, Nishio M, Kawano M, Matsumura H, Kusagawa S, Yuasa T, Ohta H
Department of Microbiology, Mie University School of Medicine, Japan.
J Gen Virol. 1994 Mar;75 ( Pt 3):567-72. doi: 10.1099/0022-1317-75-3-567.
Primary monkey kidney cells infected with human parainfluenza type 4A virus (HPIV-4A) were treated with various concentrations of formaldehyde. Formaldehyde (0.275%) treatment completely blocked virus production. However, when mouse spleen cells were cocultured with the fixed virus-infected cells, interferon was produced in the culture fluid. On the other hand, when mouse spleen cells were incubated with the fixed virus-infected cells in the presence of anti-HPIV-4A antiserum or a mixture of anti-HN protein monoclonal antibodies, interferon activity could scarcely be detected in the culture fluid. These findings indicated that the fixed virus-infected cells had an ability to induce interferon in mouse spleen cells and that the HN protein was related to interferon induction. Subsequently, a recombinant plasmid was constructed by inserting the cDNA of the HN gene of HPIV-4A into a pcDL-SR alpha expression vector. Mouse spleen cells produced interferon when cocultured with COS7 cells transfected with the recombinant plasmid, but did not when cocultured with COS7 cells transfected with the vector alone. Furthermore, we established HeLa cells constitutively expressing HPIV-4A HN (HeLa-4aHN cells) or F protein (HeLa-4aF cells). Type I (alpha/beta) interferon was detected in culture fluids of mouse spleen cells with HeLa-4aHN cells, but was not detected in those with HeLa-4aF cells. Therefore, it was concluded that the HN glycoproteins on the cell surface were sufficient for interferon induction to occur.
用人副流感4A型病毒(HPIV - 4A)感染的原代猴肾细胞用不同浓度的甲醛处理。甲醛(0.275%)处理完全阻断了病毒产生。然而,当小鼠脾细胞与固定化病毒感染的细胞共培养时,培养液中产生了干扰素。另一方面,当小鼠脾细胞在抗HPIV - 4A抗血清或抗HN蛋白单克隆抗体混合物存在的情况下与固定化病毒感染的细胞孵育时,培养液中几乎检测不到干扰素活性。这些发现表明,固定化病毒感染的细胞具有在小鼠脾细胞中诱导干扰素的能力,并且HN蛋白与干扰素诱导有关。随后,通过将HPIV - 4A的HN基因的cDNA插入pcDL - SRα表达载体构建了重组质粒。当与用重组质粒转染的COS7细胞共培养时,小鼠脾细胞产生干扰素,但与仅用载体转染的COS7细胞共培养时则不产生。此外,我们建立了组成性表达HPIV - 4A HN的HeLa细胞(HeLa - 4aHN细胞)或F蛋白的HeLa细胞(HeLa - 4aF细胞)。在与HeLa - 4aHN细胞共培养的小鼠脾细胞的培养液中检测到了I型(α/β)干扰素,但在与HeLa - 4aF细胞共培养的培养液中未检测到。因此,得出结论,细胞表面的HN糖蛋白足以引发干扰素诱导。