Expression of CCL20 and granulocyte-macrophage colony-stimulating factor, but not Flt3-L, from modified vaccinia virus Ankara enhances antiviral cellular and humoral immune responses
Expression of CCL20 and granulocyte-macrophage colony-stimulating factor, but not Flt3-L, from modified vaccinia virus Ankara enhances antiviral cellular and humoral immune responses
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DOI：
10.1128/jvi.02748-05
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发表时间：
2006-08-01
影响因子：
5.4
通讯作者：
Feinberg, A. B.
中科院分区：
文献类型：
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作者：
Chavan, R.;Marfatia, K. A.;Feinberg, A. B.
虽然修饰的安卡拉牛痘病毒（MVA）目前正处于临床开发中，作为针对天花和异源传染病的安全疫苗，但由于病毒不能在哺乳动物宿主中有效复制，其免疫原性可能有限。鉴于最近的数据表明，牛痘病毒，包括MVA，优先感染抗原呈递细胞（APC），发挥关键作用，在产生抗病毒免疫，我们假设，表达特定的细胞因子和趋化因子介导的APC募集和激活重组MVA（rMVA）载体将增强这些载体的免疫原性。为了验证这一假设，我们制备了表达鼠粒细胞-巨噬细胞集落刺激因子（mGM-CSF）、人CCL 20/人巨噬细胞炎性蛋白3 α（human macrophage inflammatory protein 3 alpha（hCCL 20/hMIP-3 α）或人鳍状酪氨酸激酶3配体（hFlt 3-L），预测增加树突细胞（DC）水平、将DC募集到免疫部位或促进DC体内成熟的因子，分别这些rMVA还共表达充分表征的免疫显性淋巴细胞性脉络丛脑膜炎病毒核蛋白（NP）抗原，该抗原能够在免疫BALB/c小鼠后灵敏和定量评估抗原特异性CD 8（+）T细胞应答。我们的研究结果表明，与非佐剂rMVA引起的那些反应相比，用表达mGM-CSF或hCCL 20但不表达hFlt 3-L的rMVA免疫小鼠，导致针对载体编码抗原的细胞免疫应答增加2至4倍，MVA特异性抗体滴度增强6至17倍。值得注意的是，细胞免疫应答的细胞因子增强发生在rMVA作为初次免疫接种时，而不是当它们用作加强免疫接种时，这表明这些APC调节蛋白，当用作痘病毒编码的佐剂时，在刺激幼稚T细胞应答方面比在促进先前存在的记忆T细胞应答的回忆方面更有效。我们的研究结果表明，从rMVA载体表达特异性基因佐剂的策略可以成功地应用于增强MVA疫苗的免疫原性。
While modified vaccinia virus Ankara (MVA) is currently in clinical development as a safe vaccine against smallpox and heterologous infectious diseases, its immunogenicity is likely limited due to the inability of the virus to replicate productively in mammalian hosts. In light of recent data demonstrating that vaccinia viruses, including MVA, preferentially infect antigen-presenting cells (APCs) that play crucial roles in generating antiviral immunity, we hypothesized that expression of specific cytokines and chemokines that mediate APC recruitment and activation from recombinant MVA (rMVA) vectors would enhance the immunogenicity of these vectors. To test this hypothesis, we generated rMVAs that express murine granulocyte-macrophage colony-stimulating factor (mGM-CSF), human CCL20/human macrophage inflammatory protein 3 alpha (hCCL20/hMIP-3 alpha), or human fins-like tyrosine kinase 3 ligand (hFlt3-L), factors predicted to increase levels of dendritic cells (DCs), to recruit DCs to sites of immunization, or to promote maturation of DCs in vivo, respectively. These rMVAs also coexpress the well-characterized, immunodominant lymphocytic choriomeningitis virus nucleoprotein (NP) antigen that enabled sensitive and quantitative assessment of antigen-specific CD8(+) T-cell responses following immunization of BALB/c mice. Our results demonstrate that immunization of mice with rMVAs expressing mGM-CSF or hCCL20, but not hFlt3-L, results in two- to fourfold increases of cellular immune responses directed against vector-encoded antigens and 6- to 17-fold enhancements of MVA-specific antibody titers, compared to those responses elicited by nonadjuvanted rMVA. Of note, cytokine augmentation of cellular immune responses occurs when rMVAs are given as primary immunizations but not when they are used as booster immunizations, suggesting that these APC-modulating proteins, when used as poxvirus-encoded adjuvants, are more effective at stimulating naive T-cell responses than in promoting recall of preexisting memory T-cell responses. Our results demonstrate that a strategy to express specific genetic adjuvants from rMVA vectors can be successfully applied to enhance the immunogenicity of MVA-based vaccines.