Detection and characterization of natural and inducible lectins in human serum

Results in immunology Pub Date : 2012-01-01 Epub Date: 2012-06-06 DOI:10.1016/j.rinim.2012.05.004
Beulaja Manikandan , Manikandan Ramar
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引用次数: 9

Abstract

This study was performed to detect and characterise the possible occurrence of natural and inducible lectins in human serum by hemagglutination method, wherein, the serum was treated using exogenous elicitors, namely, proteases and detergents.

Natural and inducible lectins were detected and characterised in human serum. Untreated serum agglutinated buffalo and rabbit RBC, while serum treated with pronase, trypsin, α-chymotrypsin or SDS for the very first time, agglutinated hen/hen and sheep RBC within 15 min in a dosimetric manner. Cross adsorption test revealed that both trypsin and α-chymotrypsin-treated serum showed similar RBC adsorption pattern. The lectin activity in untreated, pronase-treated serum was cation independent and moderately sensitive/insensitive to calcium chelator EDTA, whereas, trypsin-treated serum was cation dependent as well as EDTA sensitive (sheep RBC), cation independent and EDTA insensitive (hen RBC). Hemagglutination of untreated serum was inhibited by certain glycosides and di-, oligo-saccharides, whereas, activity in pronase-treated serum was inhibited by hexosamines. By contrast, hemagglutination of trypsin-treated serum showed specificity for acetylated mannosamine as well as sialic acid for sheep RBC and certain glycoproteins for hen RBC.

Thus, we have detected inducible lectins with distinct ligand binding specificity, upon treatment of human serum with proteases, namely, pronase and trypsin. Nevertheless, lectin activity was found in untreated human serum too with different ligand specificity.

Abstract Image

人血清中天然凝集素和诱导凝集素的检测与鉴定
本研究通过血凝法检测和表征人血清中可能存在的天然和诱导凝集素,其中,使用外源性激发剂,即蛋白酶和洗涤剂处理血清。对人血清中天然凝集素和诱导凝集素进行了检测和鉴定。未经处理的血清可凝集水牛和兔子的红细胞,而首次用蛋白酶、胰蛋白酶、α-凝乳胰蛋白酶或SDS处理的血清在15分钟内可凝集母鸡/母鸡和绵羊的红细胞。交叉吸附实验显示,胰蛋白酶和α-凝乳胰蛋白酶处理的血清具有相似的红细胞吸附模式。未处理、pronase处理的血清凝集素活性对钙螯合剂EDTA具有阳离子依赖性,对EDTA敏感(羊红细胞)、阳离子依赖性和EDTA不敏感(鸡红细胞)。某些糖苷和寡糖抑制了未处理血清的血凝,而己糖胺则抑制了蛋白酶处理血清的血凝活性。相比之下,胰蛋白酶处理血清的血凝反应对绵羊红细胞和母鸡红细胞分别对乙酰化甘露糖胺和唾液酸和某些糖蛋白具有特异性。因此,我们在用蛋白酶(即蛋白酶和胰蛋白酶)处理人血清后,检测到具有明显配体结合特异性的可诱导凝集素。然而,在未经处理的人血清中也发现了凝集素活性,但具有不同的配体特异性。
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