Quantitative analysis of differentially expressed saliva proteins in human immunodeficiency virus type 1 (HIV-1) infected individuals.
Quantitative analysis of differentially expressed saliva proteins in human immunodeficiency virus type 1 (HIV-1) infected individuals.
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DOI：
10.1016/j.aca.2013.02.038
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发表时间：
2013-04-24
影响因子：
6.2
通讯作者：
Deng H
中科院分区：
文献类型：
--
作者：
Zhang N;Zhang Z;Feng S;Wang Q;Malamud D;Deng H
在本研究中，我们建立了一种新的方法来分析唾液蛋白从HIV-1血清阳性患者的高效抗逆转录病毒治疗（HAART）和血清阴性对照。在5名HIV-1受试者和5名对照者的合并唾液样本中，分别鉴定出593种和601种蛋白质。41个蛋白质被发现差异表达。差异表达的唾液蛋白的生物信息学分析表明，增加抗菌蛋白和减少蛋白酶抑制剂后，HIV-1感染。为了验证这些差异表达的蛋白质中的一些，建立了高通量定量方法来确定HAART前20名血清阳性患者和20名血清阴性受试者的40个个体唾液样品中10种唾液蛋白质的浓度。该方法基于在1D SDS PAGE的堆积凝胶区域内有限的蛋白质分离，并使用同位素编码的合成肽作为内标。结果表明，蛋白质谱分析和靶向定量相结合的方法是一种有效的方法，以确定和验证差异表达的唾液蛋白。结果发现，在HIV-1血清学阳性唾液标本中，钙结合蛋白S100家族成员和恶性脑肿瘤缺失1蛋白（DMBT 1）的表达上调，而粘蛋白5 B的表达下调，这为HIV-1感染的监测和感染机制的研究提供了新的视角。
In the present study, we have established a new methodology to analyze saliva proteins from HIV-1-seropositive patients before highly active antiretroviral therapy (HAART) and seronegative controls. A total of 593 and 601 proteins were identified in the pooled saliva samples from 5 HIV-1 subjects and 5 controls, respectively. Forty-one proteins were found to be differentially expressed. Bioinformatic analysis of differentially expressed salivary proteins showed an increase of antimicrobial proteins and decrease of protease inhibitors upon HIV-1 infection. To validate some of these differentially expressed proteins, a high-throughput quantitation method was established to determine concentrations of 10 salivary proteins in 40 individual saliva samples from 20 seropositive patients before HAART and 20 seronegative subjects. This method was based on limited protein separation within the zone of the stacking gel of the 1D SDS PAGE and using isotope-coded synthetic peptides as internal standards. The results demonstrated that a combination of protein profiling and targeted quantitation is an efficient method to identify and validate differentially expressed salivary proteins. Expression levels of members of the calcium-binding S100 protein family and deleted in malignant brain tumors 1 protein (DMBT1) were up-regulated while that of Mucin 5B was down-regulated in HIV-1 seropositive saliva samples, which may provide new perspectives for monitoring HIV-infection and understanding the mechanism of HIV-1 infectivity.