Transforming growth factor beta-1 and beta-2 in human tear fluid
Transforming growth factor beta-1 and beta-2 in human tear fluid
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DOI：
10.3109/02713689609008900
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发表时间：
1996-06-01
影响因子：
2
通讯作者：
Pflugfelder, SC
中科院分区：
文献类型：
--
作者：
Gupta, A;Monroy, D;Pflugfelder, SC
目的。目的：评价人泪液中转化生长因子β亚型1和2 （tgf - β 1和tgf - β 2）的含量。为此，采用定量抗体夹心ELISA （sELISA）、水貂肺上皮细胞（MLEC）生长抑制生物测定法和western blotting检测人泪液中tgf - β s的含量。在这些实验中，使用了各种物理和化学处理来激活tgf - β。tgf - β 5在未处理或加热的泪液中无法通过sELISA检测到；然而，在酸激活泪液中，sELISA检测到tgf - β 1的平均浓度为38.5 ng/ml， tgf - β 2的平均浓度为2.32 ng/ml。此外，在溶解黏液剂乙酰半胱氨酸处理的泪液中检测到10.54 ng/ml的tgf - β 1和3.98 ng/ml的tgf - β 2。MLEC法测定的人泪液中总tgf - β生物活性发现，未经处理的泪液为13.04 ng/ml，酸激活的泪液为24.85 ng/ml。泪液标本中约有一半的tgf - β具有生物活性（平均52%，范围39-71%）。总泪液tgf - β生物活性可被重组人tgf - β 1潜伏期相关肽（rh tgf - β 1 LAP）完全中和。tgf - β 1特异性抗血清平均中和泪液tgf - β生物活性为83%，tgf - β 2特异性抗血清平均中和泪液tgf - β生物活性为13%。在减少酸化泪液的免疫印迹中观察到大约12.5和95 kD的免疫反应性tgf - β带。在未治疗的泪液中发现高分子量（MW） tgf - β带（>203 kD）；但经乙酰半胱氨酸治疗后，该条带消失。这些研究结果表明，tgf - β 1和tgf - β 2存在于人泪液中，tgf - β 1是主要亚型。人类眼泪中似乎有能够结合tgf - β的因子。
Purpose. To evaluate human tear fluid for transforming growth factor beta isoforms 1 and 2 (TGF-beta 1 and TGF-beta 2).Methods. To accomplish this, human tears were evaluated for TGF-beta s by quantitative antibody sandwich ELISA (sELISA), mink lung epithelial cell (MLEC) growth inhibition bioassay and western blotting. Various physical and chemical treatments were used to activate TGF-beta in these assays.Results. TGF-beta s could not be detected in untreated or heated tears by sELISA; however, mean TGF-beta 1 concentrations of 38.5 ng/ml and TGF-beta 2 concentrations of 2.32 ng/ml were detected in acid-activated tears by sELISA. Furthermore, 10.54 ng/ml of TGF-beta 1 and 3.98 ng/ml of TGF-beta 2 were detected in tears treated with the mucolytic agent, acetylcysteine. Total TGF-beta bioactivity in human tears measured by the MLEC assay was found to be 13.04 ng/ml in untreated tears and 24.85 ng/ml in acid-activated tears. Approximately one-half TGF-beta in tear speci mens was biologically active (mean = 52%, range 39-71%). Total tear TGF-beta bioactivity could be completely neutralized by recombinant human TGF-beta 1 latency associated peptide (rh TGF-beta 1 LAP). Mean neutralization of tear TGF-beta bioactivity was 83% by TGF-beta 1-specific antisera, and was 13% by TGF-beta 2-specific antisera. Immunoreactive TGF-beta bands at approximately 12.5 and 95 kD were observed in immunoblots of reduced acidified tears. A high molecular weight (MW) TGF-beta band (>203 kD) was noted in untreated tears; however, this band disappeared following treatment with acetylcysteine.Conclusions. The results of these studies indicate that TGF-beta 1 and TGF-beta 2 are present in human tear fluid, and TGF-beta 1 is the predominant isoform. There appears to be factors in human tears capable of binding TGF-beta.