Arigita Carmen, Bevaart Lisette, Everse Linda A, Koning Gerben A, Hennink Wim E, Crommelin Daan J A, van de Winkel Jan G J, van Vugt Martine J, Kersten Gideon F A, Jiskoot Wim
Department of Pharmaceutics, Utrecht Institute for Pharmaceutical Sciences, Utrecht University, The Netherlands.
Infect Immun. 2003 Sep;71(9):5210-8. doi: 10.1128/IAI.71.9.5210-5218.2003.
The effect of targeting strategies for improving the interaction of liposomal PorA with dendritic cells (DC) on the immunogenicity of PorA was investigated. PorA, a major antigen of Neisseria meningitidis, was purified and reconstituted in different types of (targeted) liposomes, i.e., by using mannose or phosphatidylserine as targeting moieties, or with positively charged liposomes. We studied the efficiency of liposome uptake and its effect on the maturation of and interleukin 12 (IL-12) production by murine DC. Moreover, mice were immunized subcutaneously to study the localization and immunogenicity of PorA liposomes. Uptake of liposomes by DC was significantly increased for targeted liposomes and resulted in the maturation of DC, but to various degrees. Maturation markers (i.e., CD80, CD86, major histocompatibility complex class II, and CD40) showed enhanced expression on DC incubated with targeted PorA liposomes relative to those incubated with nontargeted PorA liposomes. Moreover, only the uptake of targeted PorA liposomes induced production of IL-12 by DC, with levels similar to those produced by lipopolysaccharide (LPS)-pulsed DC. Mannose-targeted PorA liposomes administered subcutaneously had an increased localization in draining lymph nodes compared to nontargeted PorA liposomes. Liposomes in draining lymph nodes interacted preferentially with antigen-presenting cells, an effect that was enhanced with targeted PorA liposomes. Immunization studies showed an improvement of the bactericidal antibody response (i.e., increased number of responders) generated by targeted PorA liposomes compared to that generated by nontargeted ones or LPS-containing outer membrane vesicles. In conclusion, the use of targeted PorA liposomes results in an improved uptake by and activation of DC and an increased localization in draining lymph nodes. These effects correlate with an enhanced immune response toward the vaccine.
研究了改善脂质体PorA与树突状细胞(DC)相互作用的靶向策略对PorA免疫原性的影响。PorA是脑膜炎奈瑟菌的主要抗原,经纯化后在不同类型的(靶向)脂质体中重构,即使用甘露糖或磷脂酰丝氨酸作为靶向部分,或使用带正电荷的脂质体。我们研究了脂质体摄取效率及其对小鼠DC成熟和白细胞介素12(IL-12)产生的影响。此外,对小鼠进行皮下免疫以研究PorA脂质体的定位和免疫原性。靶向脂质体使DC对脂质体的摄取显著增加,并导致DC成熟,但程度不同。成熟标志物(即CD80、CD86、主要组织相容性复合体II类和CD40)在与靶向PorA脂质体孵育的DC上的表达相对于与非靶向PorA脂质体孵育的DC有所增强。此外,只有靶向PorA脂质体的摄取诱导DC产生IL-12,其水平与脂多糖(LPS)刺激的DC产生的水平相似。与非靶向PorA脂质体相比,皮下注射甘露糖靶向的PorA脂质体在引流淋巴结中的定位增加。引流淋巴结中的脂质体优先与抗原呈递细胞相互作用,靶向PorA脂质体可增强这种作用。免疫研究表明,与非靶向PorA脂质体或含LPS的外膜囊泡相比,靶向PorA脂质体产生的杀菌抗体反应有所改善(即应答者数量增加)。总之,使用靶向PorA脂质体可提高DC的摄取和活化,并增加在引流淋巴结中的定位。这些效应与对疫苗的免疫反应增强相关。