人类副流感病毒 (HPIV) 会引起呼吸道感染，这种感染在儿童和老年人中会加剧。正确评估病毒特征对于研究对策至关重要。然而，病毒在分离或繁殖过程中对培养细胞的适应可能会选择与实验室传代相关的突变，从而改变病毒的特征。此前有报道称，在人类气道上皮细胞中避免了 HPIV3 的适应，而不是其他 HPIV。为了研究实验室传代对 HPIV1-HPIV4 基因组的影响，我们评估了原代人支气管/气管上皮细胞气液界面（HBTEC-ALI）培养物和常规培养细胞（表达 HPIV1-HPIV4 的 Vero 细胞）传代后突变的发生情况。跨膜蛋白酶、丝氨酸 2 和正常 Vero 细胞）。 HBTEC-ALI 中突变的发生率显着低于传统培养。在 HBTEC-ALI 培养中，大多数突变是沉默的或保持较低的变异频率，从而对病毒共有序列的影响较小。相比之下，传统培养中的传代通过与传代相关的独特取代以高频率诱导或选择基因突变。在所有四种 HPIV 中都普遍观察到血凝素-神经氨酸酶的高突变，甚至在单次传代中发生突变。此外，在 HPIV1 和 HPIV2 中，大蛋白的突变更为频繁。这些结果表明，HBTEC-ALI 培养物中的传代比传统培养物更适合维持所有四种 HPIV 中临床分离株的原始特征，这有助于了解病毒发病机制。病毒对培养细胞的适应会增加感染病毒的风险。对临床分离株病毒学特征的误解。在人副流感病毒（HPIV）3中，据报道，人气道上皮和肺类器官模型更适合研究无突变临床毒株的病毒特征。因此，我们分析了所有四种 HPIV 的临床分离株，在人支气管/气管上皮细胞气液界面 (HBTEC-ALI) 或常规培养物中进行五次实验室传代后发生突变。我们发现传统培养细胞中所有四种 HPIV 均存在血凝素-神经氨酸酶突变的高风险。此外，在 HPIV1 和 HPIV2 中，常规培养的细胞中大蛋白的突变也比 HBTEC-ALI 培养中更频繁。 HBTEC-ALI 培养对于维持所有四种 HPIV 中临床分离株的原始序列和特征非常有用。本研究有助于了解 HPIV 发病机制和抗病毒策略。

Human parainfluenza virus (HPIV) causes respiratory infections, which are exacerbated in children and older people. Correct evaluation of viral characteristics is essential for the study of countermeasures. However, adaptation of viruses to cultured cells during isolation or propagation might select laboratory passage-associated mutations that modify the characteristics of the virus. It was previously reported that adaptation of HPIV3, but not other HPIVs, was avoided in human airway epithelia. To examine the influence of laboratory passage on the genomes of HPIV1-HPIV4, we evaluated the occurrence of mutations after passage in primary human bronchial/tracheal epithelial cell air-liquid interface (HBTEC-ALI) culture and conventional cultured cells (Vero cells expressing the transmembrane protease, serine 2, and normal Vero cells). The occurrence of mutations was significantly lower in HBTEC-ALI than in conventional culture. In HBTEC-ALI culture, most of the mutations were silent or remained at low variant frequency, resulting in less impact on the viral consensus sequence. In contrast, passage in conventional culture induced or selected genetic mutations at high frequency with passage-associated unique substitutions. High mutagenesis of hemagglutinin-neuraminidase was commonly observed in all four HPIVs, and mutations even occurred in a single passage. In addition, in HPIV1 and HPIV2, mutations in the large protein were more frequent. These results indicate that passage in HBTEC-ALI culture is more suitable than conventional culture for maintaining the original characteristics of clinical isolates in all four HPIVs, which can help with the understanding of viral pathogenesis.Adaptation of viruses to cultured cells can increase the risk of misinterpretation in virological characterization of clinical isolates. In human parainfluenza virus (HPIV) 3, it has been reported that the human airway epithelial and lung organoid models are preferable for the study of viral characteristics of clinical strains without mutations. Therefore, we analyzed clinical isolates of all four HPIVs for the occurrence of mutations after five laboratory passages in human bronchial/tracheal epithelial cell air-liquid interface (HBTEC-ALI) or conventional culture. We found a high risk of hemagglutinin-neuraminidase mutagenesis in all four HPIVs in conventional cultured cells. In addition, in HPIV1 and HPIV2, mutations of the large protein were also more frequent in conventional cultured cells than in HBTEC-ALI culture. HBTEC-ALI culture was useful for maintaining the original sequence and characteristics of clinical isolates in all four HPIVs. The present study contributes to the understanding of HPIV pathogenesis and antiviral strategies.