dos Santos M C, Diniz C R, Pacheco M A, Dias da Silva W
Fundação Ezequiel Dias, Universidade Federal de Minas Gerais, Belo Horizonte (MG), Brazil.
Toxicon. 1988;26(2):207-13. doi: 10.1016/0041-0101(88)90173-0.
Phospholipase A2, purified from crotoxin obtained from C. d. terrificus venom, alone or incorporated in Freund's complete adjuvant (FCA) or in Al(OH)3 was used as an antigen to immunize mice against the lethal effects of C. d. terrificus venom. The animals were intracutaneously (i.c.) or subcutaneously (s.c.) injected with 60 micrograms of phospholipase A2, divided into three equal doses and injected every 7 days. Samples of blood were collected just before each injection and the sera used to determine the antibodies against whole venom by the ELISA method. The animals were s.c. challenged with 8 LD50 or with 16 LD50 28 or 95 days after immunization. The animals that received two s.c. doses of antigen followed by a third i.c. dose were partially resistant to 8 LD50 (58% protection). This resistance increased when the first two injections consisted of phospholipase A2, the third of whole venom, all i.c., all in Al(OH)3 (67% of protection). The maximal protection (90%) was attained when the animals were i.c. injected with phospholipase A2 in Al(OH)3 in all three immunizing doses. Antibodies against whole venom were detected 15 days after immunization, reaching a plateau on the twenty-eighth day and remaining unchanged at least until the ninety-fifth day after immunization.
从眼镜蛇属的剧毒亚种（C. d. terrificus）毒液中提取的磷脂酶A2，单独使用或与弗氏完全佐剂（FCA）或氢氧化铝混合后，用作抗原免疫小鼠，以抵抗眼镜蛇属的剧毒亚种毒液的致死作用。将动物皮内（i.c.）或皮下（s.c.）注射60微克磷脂酶A2，分为三等份，每7天注射一次。在每次注射前采集血样，并用ELISA法测定血清中针对全毒液的抗体。在免疫后28天或95天，用8倍半数致死量（LD50）或16倍LD50对动物进行皮下攻击。接受两次皮下抗原剂量，随后进行第三次皮内剂量注射的动物，对8倍LD50有部分抵抗力（58%的保护率）。当最初两次注射为磷脂酶A2，第三次为全毒液，均为皮内注射，且均在氢氧化铝中时（保护率为67%），这种抵抗力增强。当动物在所有三次免疫剂量中均皮内注射氢氧化铝中的磷脂酶A2时，可获得最大保护率（90%）。免疫后15天可检测到针对全毒液的抗体，在第28天达到平台期，并至少在免疫后第95天保持不变。
