Quantification of human β-defensin-2 and -3 in body fluids:: Application for studies of innate immunity
Quantification of human β-defensin-2 and -3 in body fluids:: Application for studies of innate immunity
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DOI：
10.1373/clinchem.2006.081430
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发表时间：
2007-04-01
影响因子：
9.3
通讯作者：
Weinberg, Aaron
中科院分区：
文献类型：
--
作者：
Ghosh, Santosh K.;Gerken, Thomas A.;Weinberg, Aaron
背景：人β-防御素(HBDS)是上皮细胞来源的具有抗菌和免疫调节作用的阳离子多肽。我们的目的是建立一种定量检测体液中诱导型HBD-2和-3的分析工具。方法：利用商用捕获和检测抗体建立夹心ELISA，并确定最佳检测条件(含250 mmol/L CaCl2)以克服体液内源性成分的遮盖。结果：HBD-2和HBD-3的检出限分别为75 ng/L和3微克/L。健康献血员唾液样品中HBD-2和HBD-3的平均(SD范围)分别为9.5(1.2-21)mg/L和326(50-931)mg/L。我们没有在健康参与者的吸疱液(1317；n=10)或支气管肺泡灌洗(BAL；n=5)中检测到HBD-3。我们在BF和BAL中检测到低浓度的HBD-2多肽，分别为0.16(0.03-0.32)和0.04(0-0.049)mU/g总蛋白。我们没有观察到BF和唾液中HBD-2的相关性，或者BAL和同一人唾液中HBD-2的相关性。在2例健康女性阴道拭子中，HBD-2和-3的平均浓度分别为3.42和103mU g/g总蛋白。同一妇女宫颈阴道灌洗液中HBD多肽的平均浓度分别为1.46和55.5微克/克总蛋白。结论：该方法克服了阴离子分子的掩蔽作用，可用于检测体液中可诱导的HBD多肽浓度。因此，这种方法可能适用于对健康和疾病中的这些多肽进行量化。(C)2007年美国临床化学协会。
Background: Human beta-defensins (hBDs) are epithelial cell-derived antimicrobial and immunoregulatory cationic peptides. Our objective was to establish an analytical tool to quantify inducible hBD-2 and -3 in body fluids.Methods: We developed sandwich ELISAs using commercially available capture and detection antibodies and determined optimal assay conditions (with 250 mmol/L CaCl2) to overcome masking by endogenous components of body fluids. We used recombinant hBD as calibrators and for recovery testing.Results: hBD-2 and -3 detection limits were similar to 75 ng/L and similar to 3 mu g/L, respectively. Mean (SD range) values in saliva samples from healthy donors (n = 60) were 9.5 (1.2-21) mu g/L for hBD-2 and 326 (50-931) mu g/L for hBD-3. We did not detect hBD-3 in suction blister fluid (1317; n = 10) or bronchoalveolar lavage (BAL; n = 5) from healthy participants. We detected low hBD-2 peptide concentrations in BF and BAL, 0.16 (0.03-0.32) and 0.04 (0-0.049) mu g/g total protein, respectively. We observed no correlation of hBD-2 in BF and saliva or BAL and saliva from the same person. In vaginal swabs from healthy women (n = 2), mean hBD-2 and -3 concentrations were 3.42 and 103 mu g/g total protein, respectively. Cervicovaginal lavage from the same women contained mean concentrations of 1.46 and 55.5 mu g/g total protein.Conclusion: These ELISA assays can measure inducible hBD peptide concentrations in body fluids by overcoming masking effects of anionic molecules. This approach may therefore be applicable for quantifying these peptides in health and disease. (c) 2007 American Association for Clinical Chemistry.