A Novel LNP-Based Chlamydia Subunit Vaccine Formulation That Induces Th1 Responses without Upregulating IL-17 Provides Equivalent Protection in Mice as Formulations That Induced IL-17 and Th1 Cytokines

M. A. Boddicker, R. Kaufhold, K. Cox, Bob J. Lucas, Jinfu Xie, Debbie D. Nahas, Sinoeun Touch, Amy S. Espeseth, K. Vora, Julie M. Skinner
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引用次数: 2

Abstract

Sexually transmitted Chlamydia infections can lead to the development of debilitating diseases such as chronic pelvic pain, ectopic pregnancy, pelvic inflammatory disease, and infertility. It has been proposed that immunity against Chlamydia infection and disease may be determined by a balance of IL-17 signaling. We sought to evaluate novel formulations for a candidate Chlamydia vaccine, consisting of Chlamydia major outer membrane protein (MOMP) alone or in combination with polymorphic membrane protein D (PmpD) and polymorphic membrane protein G (PmpG) as target immunogens. Native MOMP (nMOMP) isolated from C. muridarum elementary bodies (EBs) and recombinant PmpD and PmpG proteins were adjuvanted with Monophosphoryl lipid A (MPLA), in one of two formulations containing either lipid nanoparticles (LNPs) or the cationic lipid dimethyldioctadecylammonium bromide (DDA). Antibody titers to C. muridarum, nMOMP, and EBs were evaluated by ELISA, and T-cell responses by intracellular cytokine staining (ICS). Protection from challenge was determined by qPCR and gross pathology. All mice immunized with the new vaccine formulations showed significantly higher antibody titers to nMOMP (P<0.001) and C. muridarum EBs (P<0.001), when compared to the negative control group (adjuvant alone). Antibody titers in vaccine groups with Monophosphoryl lipid A (MPLA)+LNP were higher as compared to the MPLA+DDA group (P<0.001) except for groups 6 (Cm nMOMP+PmpG+PmpD p73+PmpD p82+MPLA+DDA) vs 7 (Cm nMOMP+PmpG+PmpD p73+PmpD p82+MPLA+LNP) for both C. muridarum EBs and PmpG; the groups were not statistically significant. ICS analysis showed more robust CD4+ T-cell responses (IFN-γ/IL-2/TNF-α) in the dimethyldioctadecylammonium bromide (DDA) and LNP groups compared to the adjuvant alone group. The combination of DDA and MPLA gave robust Th17 responses in comparison to MPLA and LNP group. Immunized groups also showed protection from challenge with C. muridarum , as evidenced by a reduction in bacterial shedding from the vagina for all groups (P<0.003) compared to shedding from the adjuvant control, Group 1. Both vaccine formulations generated robust immunological responses and both vaccine formulations were protective by reducing bacterial shedding after challenge. This data indicates equal protection can be achieved without the induction of Th17 responses.
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一种新的基于LNP的衣原体亚单位疫苗制剂在不上调IL-17的情况下诱导Th1应答,在小鼠中提供与诱导IL-17和Th1细胞因子的制剂同等的保护
性传播的衣原体感染会导致慢性盆腔疼痛、异位妊娠、盆腔炎和不孕等衰弱性疾病的发展。有人提出,对衣原体感染和疾病的免疫力可以通过IL-17信号的平衡来确定。我们试图评估候选衣原体疫苗的新配方,该疫苗由衣原体主要外膜蛋白(MOMP)单独或与多态性膜蛋白D(PmpD)和多态性膜蛋白质G(PmpG)组合作为靶免疫原组成。在含有脂质纳米颗粒(LNPs)或阳离子脂质二甲基二十八烷基溴化铵(DDA)的两种制剂中的一种中,用单磷酸脂质A(MPLA)对从鼠伤寒杆菌基本体(EBs)分离的天然MOMP(nMOMP)和重组PmpD和PmpG蛋白进行佐剂。通过ELISA评估对鼠伤寒杆菌、nMOMP和EB的抗体滴度,并通过细胞内细胞因子染色(ICS)评估T细胞反应。通过qPCR和大体病理学测定对攻击的保护作用。与阴性对照组(单独佐剂)相比,用新疫苗制剂免疫的所有小鼠对nMOMP(P<0.001)和鼠伤寒杆菌EB(P<0.001。与MPLA+DDA组相比,单磷酰脂质A(MPLA)+LNP疫苗组的抗体滴度更高(P<0.001),除了第6组(Cm nMOMP+PmpG+PmpDp73+PmpDp 82+PLA+DDA)与第7组(Cm-MOMP+PmpG+PmpD p73+PmPDp p82+MPLA+LNP)的鼠伤寒杆菌EBs和PmpG;两组间无统计学意义。ICS分析显示,与单独佐剂组相比,二甲基二十八烷基溴化铵(DDA)和LNP组的CD4+T细胞反应(IFN-γ/IL-2/TNF-α)更强。与MPLA和LNP组相比,DDA和MPLA的组合提供了强大的Th17应答。免疫组也显示出对鼠伤寒杆菌攻击的保护作用,与佐剂对照组(第1组)相比,所有组的阴道细菌脱落减少(P<0.003)证明了这一点。两种疫苗制剂都产生了强大的免疫反应,并且两种疫苗配方都通过减少攻击后的细菌脱落而具有保护作用。该数据表明,在不诱导Th17反应的情况下可以实现同等的保护。
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