Yeang H Y, Cheong K F, Sunderasan E, Hamzah S, Chew N P, Hamid S, Hamilton R G, Cardosa M J
Biotechnology and Strategic Research Division, Rubber Research Institute of Malaysia, Kuala Lumpur.
J Allergy Clin Immunol. 1996 Sep;98(3):628-39. doi: 10.1016/s0091-6749(96)70097-0.
Two major water-insoluble proteins are located on the surface of rubber particles in Hevea brasiliensis latex. A 14.6 kd protein (Hev b 1), found mainly on large rubber particles (> 350 mm in diameter), and a 24 kd protein (Hev b 3), found mainly on small rubber particles (average diameter, 70 nm), are recognized by IgE from patients with spina bifida and latex allergy. Although Hev b 1 (also called the rubber elongation factor [REF]) has previously been reported as a major latex allergen, this conclusion has been disputed on the basis of results from other studies. The allergenicity of Hev b 1 is verified in this study by testing the recombinant protein generated from its gene. Because allergenicity is confined to patients with spina bifida and not observed in adults sensitive to latex, it is not a major latex allergen. The identification of Hev b 3 as another allergen originating from rubber particles is confirmed by immunogold labeling and electron microscopy. Observations with the monoclonal antibody USM/RC2 developed against Hev b 3 show that the protein has a tendency to fragment into several polypeptides of lower molecular weight (from 24 kd to about 5 kd) when stored at -20 degrees C. There is also indication of protein aggregation from the appearance of proteins with molecular weights greater than 24 kd. Fragmentation of Hev b 3 is induced immediately on he addition of latex B-serum, which is normally compartmentalized in the lutoids in fresh latex. In the preparation of ammoniated latex (used for the manufacture of latex products), the lutoids are ruptured, and the released B-serum reacts with Hev b 3 on the rubber particles to give rise to an array of low molecular weight polypeptides that are allergenic to patients with spina bifida.
两种主要的水不溶性蛋白质位于巴西橡胶树胶乳的橡胶粒子表面。一种14.6千道尔顿的蛋白质(Hev b 1),主要存在于大的橡胶粒子(直径>350纳米)上,以及一种24千道尔顿的蛋白质(Hev b 3),主要存在于小的橡胶粒子(平均直径70纳米)上,能被脊柱裂和乳胶过敏患者的IgE识别。尽管Hev b 1(也称为橡胶延伸因子[REF])先前已被报道为主要的乳胶过敏原,但该结论基于其他研究结果受到了质疑。本研究通过测试由其基因产生的重组蛋白验证了Hev b 1的致敏性。由于致敏性仅限于脊柱裂患者,在对乳胶敏感的成年人中未观察到,所以它不是主要的乳胶过敏原。通过免疫金标记和电子显微镜证实了Hev b 3是另一种源自橡胶粒子的过敏原。用针对Hev b 3开发的单克隆抗体USM/RC2观察发现,该蛋白质在-20℃储存时倾向于断裂成几种较低分子量的多肽(从24千道尔顿到约5千道尔顿)。从分子量大于24千道尔顿的蛋白质出现情况也表明有蛋白质聚集。加入乳胶B血清后,Hev b 3会立即发生断裂,乳胶B血清在新鲜胶乳中通常分隔在类液泡中。在制备氨化胶乳(用于制造乳胶产品)时,类液泡破裂,释放的B血清与橡胶粒子上的Hev b 3反应,产生一系列对脊柱裂患者有致敏性的低分子量多肽。