Lipidation of Haemophilus influenzae Antigens P6 and OMP26 Improves Immunogenicity and Protection against Nasopharyngeal Colonization and Ear Infection
Lipidation of Haemophilus influenzae Antigens P6 and OMP26 Improves Immunogenicity and Protection against Nasopharyngeal Colonization and Ear Infection
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DOI：
10.1128/iai.00678-21
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发表时间：
2022-04-18
影响因子：
3.1
通讯作者：
Pichichero, Michael
中科院分区：
文献类型：
--
作者：
Kaur, Ravinder;Pichichero, Michael
无法分型的流感嗜血杆菌（NTHi）引起呼吸道感染，导致全球高发病率和死亡率，鼓励开发有效的疫苗。为了通过NTHi实现对鼻咽（NP）定殖的保护性影响，可能需要增强免疫原性，超过用重组蛋白抗原可实现的免疫原性。向重组蛋白中添加脂质部分将通过抗原呈递细胞的Toll样受体2信号传导和鼻相关淋巴组织（NALT）中的Th 17细胞应答来增强免疫原性。我们在小鼠模型中研究了重组蛋白P6和OMP 26的脂化（L）与非脂化（NL）P6和OMP 26以及作为融合构建体（L-OMP 26 phi NL-P6和L-P6 phi NL-OMP 26）的效果。在腹膜内或鼻内接种后，比较抗体应答并评估对NP定殖和中耳感染的保护。L-P6和L-OMP 26诱导的IgG抗体水平比NL-P6和NL-OMP 26高约10至100倍。融合构建体显著增加了针对两种靶蛋白的IgG抗体，即使只有一种蛋白质被脂化。在用L-P6和L-OMP 26接种后，NP定殖和中耳大疱NTHi密度比用NL-P6和NL-OMP 26接种低1至4个对数。融合构建体还导致接种后NTHi密度降低1至3个对数。来自接种脂质化蛋白构建体的小鼠的NALT细胞具有更高水平的白细胞介素-17（IL-17）、IL-22和CD 4（+）T细胞记忆。将来自L-OMP 26 phi NL-P6-接种的小鼠的血清被动转移至受体幼年小鼠减少了NP定殖和耳泡NTHi密度。我们得出结论，L-P6、L-OMP 26和融合构建体在小鼠中产生增强的抗体应答和对NP定殖和NTHi中耳感染的保护。
Nontypeable Haemophilus influenzae (NTHi) causes respiratory infections that lead to high morbidity and mortality worldwide, encouraging development of effective vaccines. To achieve a protective impact on nasopharyngeal (NP) colonization by NTHi, enhanced immunogenicity beyond that achievable with recombinantprotein antigens is likely to be necessary. Adding a lipid moiety to a recombinant protein would enhance immunogenicity through Toll-like receptor 2 signaling of antigen-presenting cells and Th17 cell response in the nasal-associated lymphoid tissue (NALT). We investigated effects of lipidation (L) of recombinant proteins P6 and OMP26 compared to nonlipidated (NL) P6 and OMP26 and as fusion constructs (L-OMP26 phi NL-P6 and L-P6 phi NL-OMP26) in a mouse model. After intraperitoneal or intranasal vaccination, antibody responses were compared and protection from NP colonization and middle ear infection were assessed. L-P6 and L-OMP26 induced approximately 10- to 100-fold-higher IgG antibody levels than NL-P6 and NL-OMP26. Fusion constructs significantly increased IgG antibody to both target proteins, even though only one of the proteins was lipidated. NP colonization and middle ear bullae NTHi density was 1 to 4 logs lower following vaccination with L-P6 and L-OMP26 than with NL-P6 and NL-OMP26. Fusion constructs also resulted in a 1- to 3-loglower NTHi density following vaccination. NALT cells from mice vaccinated with lipidated protein constructs had higher levels of interleukin-17 (IL-17), IL-22, and CD4(+) T-cell memory. Passive transfer of sera from L-OMP26 phi NL-P6-vaccinated mice to recipient infant mice reduced NP colonization and ear bulla NTHi density. We conclude that L-P6, L-OMP26, and fusion constructs generate enhanced antibody responses and protection from NP colonization and middle ear infection by NTHi in mice.