Calderon-Aranda E S, Hozbor D, Possani L D
Departamento de Bioquímica, Universidad Nacional Autónoma de México, Cuernavaca.
Toxicon. 1993 Mar;31(3):327-37. doi: 10.1016/0041-0101(93)90150-h.
Approximately 700 people die in Mexico each year from scorpion stings. The only useful therapy available is antiserum obtained from horses immunized with macerates of venomous gland from scorpions of the genus Centruroides. We report the results of experiments conducted with mice and rats in order to evaluate the relevant components of the venom from Centruroides noxius in the induction of a protective response against scorpion envenomation, either in vivo or in vitro. Gland macerates of whole telsons (stinger), soluble venom extracted by electrical stimulation, toxic fractions from gel filtration on Sephadex G-50 and highly purified toxin 2 from this scorpion venom were all used to produce hyperimmune mice and rats, which were challenged in vivo with the equivalent of the lethal dose 50% (LD50) of soluble venom, or their sera were prepared for in vitro neutralization experiments using non-immunized animals. The maximum neutralizing capacity (100%) was obtained when soluble venom was used as antigen, while purified toxin 2 produces 80% survival in vivo. The neutralizing capacity of murine antisera evaluated in vitro was: sera antifraction II > antitoxin 2 > antitotal venom > anti-gland macerates of whole telsons. Two additional aspects were further investigated in the present work. One is the demonstration by immunoblotting that proteins corresponding to the electrophoretic mobility of toxins known to block sodium channels are highly immunodominant in this venom. Second, there is a strong cross-reactivity of antisera produced with Centruroides noxius when assayed against venoms from other dangerous species of Centruroides scorpions from Mexico, but not against the Israeli scorpion Leiurus quinquestriatus. Finally, the immunodominance of toxic fractions in the immune response was observed either with immunization using Freund's adjuvant or by means of adsorption to nitrocellulose membranes. This latter vehicle was shown to be an excellent detoxifying agent, without changing the immunogenicity of the toxins, as might occur with chemical treatment of these neurotoxic peptides.
在墨西哥，每年约有700人死于蝎子蜇伤。唯一有效的治疗方法是使用抗血清，该抗血清是从用墨西哥毒蝎属蝎子的毒腺浸液免疫过的马身上获取的。我们报告了用小鼠和大鼠进行实验的结果，以评估墨西哥毒蝎毒液的相关成分在体内或体外诱导针对蝎子蜇伤的保护性反应中的作用。整个毒刺的腺体浸液、通过电刺激提取的可溶性毒液、在葡聚糖凝胶G - 50上进行凝胶过滤得到的有毒组分以及从这种蝎子毒液中高度纯化的毒素2，均用于制备超免疫小鼠和大鼠，然后用相当于可溶性毒液半数致死剂量（LD50）的剂量在体内对它们进行攻击，或者制备它们的血清用于使用未免疫动物进行的体外中和实验。当使用可溶性毒液作为抗原时，可获得最大中和能力（100%），而纯化的毒素2在体内可使80%的动物存活。在体外评估的鼠抗血清的中和能力为：抗组分II血清＞抗毒素2＞抗总毒液＞抗整个毒刺的腺体浸液。在本研究中还进一步调查了另外两个方面。一是通过免疫印迹法证明，在这种毒液中，与已知能阻断钠通道的毒素电泳迁移率相对应的蛋白质具有高度免疫显性。二是当用墨西哥毒蝎产生的抗血清检测来自墨西哥其他危险的毒蝎属蝎子的毒液时，存在强烈的交叉反应，但对以色列蝎子黄肥尾蝎则无交叉反应。最后，无论是使用弗氏佐剂免疫还是通过吸附到硝酸纤维素膜上，都观察到了有毒组分在免疫反应中的免疫显性。事实证明，后一种载体是一种极好的解毒剂，不会像对这些神经毒性肽进行化学处理那样改变毒素的免疫原性。
