Zweygarth Erich, Josemans Antoinette I, Van Strijp M Fransie, Lopez-Rebollar Laura, Van Kleef Mirinda, Allsopp Basil A
Molecular Biology, Onderstepoort Veterinary Institute, Private Bag X5, Onderstepoort 0110, South Africa.
Vaccine. 2005 Feb 25;23(14):1695-702. doi: 10.1016/j.vaccine.2004.09.030.
Heartwater is a tick-borne disease of ruminants caused by the intracellular rickettsia Ehrlichia ruminantium. The only commercially available immunization procedure involves infecting animals with cryopreserved sheep blood containing virulent E. ruminantium organisms, followed by treatment with tetracyclines when fever develops. The virulent Welgevonden stock of E. ruminantium was attenuated by continuous propagation of the organisms in a canine macrophage-monocyte cell line (DH82), followed by re-adaptation to grow in a bovine endothelial cell line (BA 886). The material used for the present experiments consisted of the attenuated stock between passages 43 and 64 after re-adaptation. When inoculated into sheep or goats the attenuated organisms did not produce disease, and the only symptom observed was a rise in body temperature in most, but not all, animals. All sheep injected with 2 ml of culture suspension were subsequently found to be fully protected against a lethal needle challenge with the virulent homologous stock or with one of four different heterologous stocks (Ball 3, Gardel, Mara 87/7, Blaauwkrans). Titrations of elementary body suspensions showed that 2ml of a 1:10,000 dilution of culture suspension injected into sheep or goats was still sufficient to trigger an immune response which resisted a lethal needle challenge with the virulent Welgevonden stock. Adult Amblyomma hebraeum ticks, fed as nymphs on sheep immunized with DH82-derived organisms of passage 111, were able to transmit the attenuated stock to a naive sheep, which was found to be protected against a subsequent lethal homologous needle challenge.
心水病是一种由细胞内立克次氏体反刍动物埃立克体引起的反刍动物蜱传疾病。唯一可用于商业用途的免疫程序是用含有强毒反刍动物埃立克体的冷冻保存羊血感染动物,然后在动物发热时用四环素进行治疗。反刍动物埃立克体的强毒韦尔格冯登毒株通过在犬巨噬细胞 - 单核细胞系(DH82)中连续传代培养而减毒,随后重新适应在牛内皮细胞系(BA 886)中生长。本实验所用材料为由重新适应后第43至64代之间的减毒株组成。当接种到绵羊或山羊体内时,减毒后的病原体不会引发疾病,观察到的唯一症状是大多数(但不是全部)动物体温升高。随后发现,所有注射2毫升培养悬浮液的绵羊均能完全抵御强毒同源毒株或四种不同异源毒株(Ball 3、Gardel、Mara 87/7、Blaauwkrans)之一的致死性针刺攻击。对原体悬浮液进行滴定表明,向绵羊或山羊注射2毫升1:10,000稀释的培养悬浮液仍足以引发免疫反应,从而抵御强毒韦尔格冯登毒株的致死性针刺攻击。以第111代DH82衍生病原体免疫的绵羊若作为若虫被成年希伯来花蜱叮咬,这些蜱能够将减毒株传播给未感染的绵羊,而该绵羊随后被发现能够抵御随后的致死性同源针刺攻击。