Laboratory of Infectious Disease, School of Veterinary Medicine, Azabu University, Fuchinobe, Chuo-ku, Sagamihara-shi, Kanagawa, 252-5201, Japan,
Parasitol Res. 2014 May;113(5):1681-6. doi: 10.1007/s00436-014-3812-1. Epub 2014 Feb 18.
The aim of this study is to determine the efficacy of exoantigens derived from Babesia gibsoni cultures to induce protective immunity against challenge exposure of virulent organisms. An attenuated B. gibsoni Oita strain was maintained in vitro by the microaerophilus stationary phase (MASP) method, and exoantigens-containing supernatant fluids were collected for preparation of the immunization. Two dogs received three subcutaneous immunizations with a 20-day interval of B. gibsoni exoantigens plus 0.5 mg saponin (Quil A). On day 68 after the prime immunization, the immunized dogs and control dogs were challenged intravenously with 2 × 10(8) virulent parasites of a homologous B. gibsoni strain. The results showed that exoantigens could induce a high degree of protection against virulent homologous challenge exposure. Two dogs immunized with exoantigens showed a lower parasitemia, accompanied by a slight decrease in the PCV that returned to normal values. Control dogs developed typical acute clinical signs, including severe anemia and hyperthermia. The immunization elicited humoral immune responses. In dogs immunized with exoantigens, the maximum antibody titer was 2,560 and 5,120 by indirect fluorescent antibody test (IFAT), respectively. Preliminary Western blot analysis of the immunogen revealed five dominant proteins of molecular weights of 18, 37, 43, 50, and 57 kDa. These results suggested that the culture-derived exoantigens were candidates for non-viable vaccine.
本研究旨在确定源自巴贝西原虫培养物的外抗原诱导针对强毒生物体攻击暴露的保护性免疫的功效。通过微需氧定殖相(MASP)方法在体外维持减毒巴贝西原虫大分株,收集含外抗原的上清液用于制备免疫接种。两只狗接受了三次皮下免疫接种,间隔 20 天,用巴贝西原虫外抗原加 0.5mg 皂素(Quil A)。在初次免疫接种后第 68 天,免疫狗和对照狗经静脉内用同源巴贝西原虫强毒寄生虫 2×10(8)进行攻击。结果表明,外抗原可以诱导对强毒同源攻击暴露的高度保护。用外抗原免疫的两只狗表现出较低的寄生虫血症,伴随 PCV 轻微下降,但恢复到正常水平。对照狗出现典型的急性临床症状,包括严重贫血和发热。免疫接种引起体液免疫反应。在用外抗原免疫的狗中,间接荧光抗体试验(IFAT)的最大抗体滴度分别为 2,560 和 5,120。对免疫原的初步 Western blot 分析显示,有五种主要的蛋白质分子量为 18、37、43、50 和 57 kDa。这些结果表明,培养物衍生的外抗原是非存活疫苗的候选物。