Entomology Department, Vector Biology Laboratory, Kansas State University, 1603 Old Claflin Pl, 123 Waters Hall, Manhattan, KS 66506, USA.
Department of Global Health, Duke University, 310 Trent Drive, Durham, NC 27710, USA.
Int J Mol Sci. 2020 Jan 21;21(3):691. doi: 10.3390/ijms21030691.
Insect saliva induces significant antibody responses associated with the intensity of exposure to bites and the risk of disease in humans. Several salivary biomarkers have been characterized to determine exposure intensity to Old World mosquito species. However, new tools are needed to quantify the intensity of human exposure to bites and understand the risk of malaria in low-transmission areas in the Americas. To address this need, we conducted proteomic and bioinformatic analyses of immunogenic candidate proteins present in the saliva of uninfected from two separate colonies-one originating from Central America (STECLA strain) and one originating from South America (Cartagena strain). A ~65 kDa band was identified by IgG antibodies in serum samples from healthy volunteers living in a malaria endemic area in Colombia, and a total of five peptides were designed from the sequences of two immunogenic candidate proteins that were shared by both strains. ELISA-based testing of human IgG antibody levels against the peptides revealed that the transferrin-derived peptides, TRANS-P1, TRANS-P2 and a salivary peroxidase peptide (PEROX-P3) were able to distinguish between malaria-infected and uninfected groups. Interestingly, IgG antibody levels against PEROX-P3 were significantly lower in people that have never experienced malaria, suggesting that it may be a good marker for mosquito bite exposure in naïve populations such as travelers and deployed military personnel. In addition, the strength of the differences in the IgG levels against the peptides varied according to location, suggesting that the peptides may able to detect differences in intensities of bite exposure according to the mosquito population density. Thus, the salivary peptides TRANS-P1, TRANS-P2, and PEROX-P3 are promising biomarkers that could be exploited in a quantitative immunoassay for determination of human-vector contact and calculation of disease risk.
昆虫唾液会引起显著的抗体反应，这种反应与被叮咬的强度和人类患病的风险有关。已经有几种唾液生物标志物被描述出来，用于确定人类对旧世界蚊子物种的暴露强度。然而，需要新的工具来量化人类被叮咬的强度，并了解美洲低传播地区疟疾的风险。为了满足这一需求，我们对来自两个不同殖民地的未感染蚊子唾液中的免疫原性候选蛋白进行了蛋白质组学和生物信息学分析——一个起源于中美洲（STECLA 株），另一个起源于南美洲（卡塔赫纳株）。在哥伦比亚疟疾流行地区居住的健康志愿者的血清样本中，IgG 抗体识别出一条约 65 kDa 的带，从两个菌株共有的两种免疫原性候选蛋白的序列中设计了总共 5 个肽段。针对肽段的人类 IgG 抗体水平的 ELISA 检测表明，转铁蛋白衍生肽 TRANS-P1、TRANS-P2 和一种唾液过氧化物酶肽（PEROX-P3）能够区分疟疾病例和未感染组。有趣的是，从未经历过疟疾的人对 PEROX-P3 的 IgG 抗体水平明显较低，这表明它可能是一种衡量初入人群（如旅行者和部署的军事人员）被蚊子叮咬的良好标志物。此外，针对这些肽段的 IgG 水平的差异强度根据地点而有所不同，这表明这些肽段可能能够根据蚊子种群密度检测叮咬暴露强度的差异。因此，唾液肽 TRANS-P1、TRANS-P2 和 PEROX-P3 是有前途的生物标志物，可用于定量免疫测定，以确定人与媒介的接触并计算疾病风险。
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