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Efficiency of HPV 16 L1/E7 DNA immunization: Influence of cellular localization and capsid assembly

机译：HPV 16 L1 / E7 DNA免疫效率：细胞定位和衣壳装配的影响

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Infections by human papillomaviruses (HPV) are the major cause of uterine cancer in women worldwide. Aiming to develop a combined prophylactic and therapeutic vaccine we have previously demonstrated immunogenicity of chimeric virus-like particles consisting of a C-terminally truncated HPV 16 L1 capsid protein fused to an E7 portion. Here we show that genetic vaccination with a corresponding DNA was inefficient in the induction of a L1-specific prophylactic immune response. DNA immunization with C-terminally truncated HPV 16 L1 genes of different lengths revealed that only short deletions (L1(1-498)) were tolerated for eliciting a humoral immune response against viral capsids. This correlates with the observation that the C-terminal sequences are critical for nuclear localization, capsomere and capsid assembly. However, only the ability of L1 protein to form capsomeres or capsids showed a direct influence on the outcome of the immune response. C-terminal insertion of 60 amino acids of E7 was tolerated in fusion constructs, whereas insertion of full-length E7(1-98) or shuffled E7 (149 aa) completely abolished the humoral immune response. The L1(1-498)/E7(1-60) fusion construct not only induced L1-specific antibodies but also L1- and E7-specific CTL responses after DNA vaccination.
机译：人类乳头瘤病毒（HPV）感染是全世界女性子宫癌的主要原因。为了开发预防性和治疗性联合疫苗，我们先前已经证明了由C末端截短的HPV 16 L1衣壳蛋白融合到E7部分构成的嵌合病毒样颗粒的免疫原性。在这里，我们显示了用相应的DNA进行基因疫苗接种在诱导L1特异性预防性免疫应答中效率低下。用不同长度的C末端截短的HPV 16 L1基因进行的DNA免疫显示，只有短缺失（L1（1-498））可以耐受引发针对病毒衣壳的体液免疫应答。这与C端序列对于核定位，衣壳和衣壳组装至关重要的观察结果相关。然而，仅L1蛋白形成衣壳或衣壳的能力直接影响免疫应答的结果。 E7的60个氨基酸的C端插入在融合构建体中是被容许的，而全长E7（1-98）或改组的E7（149aa）的插入完全消除了体液免疫应答。 L1（1-498）/ E7（1-60）融合构建体不仅诱导L1特异性抗体，而且在DNA疫苗接种后诱导L1和E7特异性CTL反应。

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