Kleine-Tebbe J, Kagey-Sobotka A, MacGlashan D W, Lichtenstein L M, MacDonald S M
Department of Medicine, Division of Allergy and Clinical Immunology, Johns Hopkins University School of Medicine, Baltimore, MD 21224-6801, USA.
J Allergy Clin Immunol. 1996 Jul;98(1):181-8. doi: 10.1016/s0091-6749(96)70241-5.
It has been suggested that differential histamine-releasing activity of an IgE-dependent histamine-releasing factor (HRF), which has recently been cloned, is related to carbohydrate difference in the IgE molecule. Lectins are able to recognize specific glycoforms and might therefore be useful in characterizing the proposed heterogeneity of IgE molecules. As one test of this hypothesis, we examined the histamine release potency of several well-characterized lectins on basophils passively sensitized with serum containing IgE molecules that support HRF-induced histamine release (IgE+) or serum that does not support release by this stimulus (IgE-). Histamine release was induced by challenging basophils with different concentrations of concanavalin A, Lens culinaris (LcH), and Pisum sativum (PSA). Dose-response curves revealed that LcH caused 30% histamine release at 2 micrograms/ml with IgE+ sensitized cells, whereas the same release with IgE- cells required sixfold higher concentrations. Similar values for PSA showed a sevenfold difference. With concanavalin A, the selectivity was reversed in that it was fourfold more active on IgE- -sensitized cells. Another pair of sera (IgE+ vs IgE-) revealed the same result for concanavalin A, but no difference occurred in LcH and PSA induced release between the IgE+ - and IgE- -sensitized cells. These contrasting findings with different pairs of IgE+ -and IgE- -containing sera indicated that the lectins LcH and PSA are not able to discriminate between IgE+ -and IgE- -sensitized cells as does HRF and therefore cannot be used to further define the proposed heterogeneity of IgE. this conclusion was supported by experiments in which basophil preparations from donors possessing natural sensitivity or insensitivity to HRF (having IgE+ or IgE- on their surface) were examined fro their response to these lectins. No difference was found in the sensitivity of the cells to challenge with LcH or PSA, and the response to concanavalin A was the opposite of that found when passively sensitized cells were used (30% histamine release at 0.9 vs 3.5 micrograms/ml for IgE+ vs IgE- donors, respectively). We conclude that oligosaccharide-specific lectins do not differentiate between HRF-reactive and HRF-nonreactive IgE molecules on basophils.
最近克隆出的一种IgE依赖性组胺释放因子(HRF)具有不同的组胺释放活性,有人认为这与IgE分子中的碳水化合物差异有关。凝集素能够识别特定的糖型,因此可能有助于表征所提出的IgE分子异质性。作为对这一假设的一项检验,我们检测了几种特性明确的凝集素对用含有支持HRF诱导组胺释放的IgE分子的血清(IgE+)或不支持这种刺激释放的血清(IgE-)被动致敏的嗜碱性粒细胞的组胺释放能力。用不同浓度的伴刀豆球蛋白A、扁豆凝集素(LcH)和豌豆凝集素(PSA)刺激嗜碱性粒细胞诱导组胺释放。剂量反应曲线显示,LcH在2微克/毫升时能使IgE+致敏细胞释放30%的组胺,而对于IgE-细胞,相同释放量所需浓度高六倍。PSA的类似值显示出七倍的差异。对于伴刀豆球蛋白A,选择性则相反,它对IgE-致敏细胞的活性高四倍。另一对血清(IgE+与IgE-)对伴刀豆球蛋白A显示出相同结果,但在LcH和PSA诱导的IgE+与IgE-致敏细胞释放之间没有差异。这些用不同对含IgE+和IgE-血清得出的对比结果表明,凝集素LcH和PSA不能像HRF那样区分IgE+和IgE-致敏细胞,因此不能用于进一步界定所提出的IgE异质性。这一结论得到了实验的支持,在这些实验中,检测了来自对HRF具有天然敏感性或不敏感性(其表面有IgE+或IgE-)的供体的嗜碱性粒细胞制剂对这些凝集素的反应。未发现细胞对LcH或PSA刺激的敏感性有差异,并且对伴刀豆球蛋白A的反应与使用被动致敏细胞时相反(对于IgE+与IgE-供体,组胺释放量分别在0.9微克/毫升与3.5微克/毫升时为30%)。我们得出结论,寡糖特异性凝集素不能区分嗜碱性粒细胞上对HRF有反应和无反应的IgE分子。