Hawai'i Institute of Marine Biology, School of Ocean & Earth Science & Technology, University of Hawai'i, PO Box 1346, Kāne'ohe, Hawai'i 96744, USA.
Fish Shellfish Immunol. 2011 Apr-May;30(4-5):1152-8. doi: 10.1016/j.fsi.2011.03.004. Epub 2011 Mar 6.
The objective of this study was to determine whether immunization of Mozambique tilapia with different Cryptocaryon irritans i-antigen serotypes elicited cross-protection against challenge infection by both serotypes. Fish were directly exposed to live theronts of isolate W1 or isolate K1, that express different surface i-antigens. There was no significant difference in the number of trophonts infecting the fish between the two isolates, W1 and K1, following primary exposure. Serum from immunized fish exposed to live theronts showed higher immobilization titres and ELISA values against homologous isolates than to heterologous isolates after the primary exposure. However, mucus antibody did not immobilize theronts although the ELISA results clearly indicated that mucus antibodies recognizing C. irritans were generated. In a study with Western blot analyses, serum antibodies recognized only an antigen of the corresponding serotype and no proteins common to both serotypes were identified. Sequence analyses of 754 bases of rDNA nucleotide sequence including complete nuclear ribosomal ITS-1-5.8S rDNA-ITS-2 region were conducted and found to be identical for W1- and K1-isolates. These findings confirmed that both isolates were members of the species, C. irritans, and that rDNA analysis would not distinguish the two isolates. In conclusion, despite the fact that the immobilization assays and ELISA detected two serotypes in vitro, challenge assays provided evidence for only one type of C. irritans.
本研究的目的是确定用不同的 Cryptocaryon irritans i-抗原血清型对莫桑比克罗非鱼进行免疫是否能产生针对两种血清型的交叉保护作用。鱼直接暴露于表达不同表面 i-抗原的分离株 W1 或 K1 的活体游仆虫中。在初次暴露后,两种分离株 W1 和 K1 感染鱼的滋养体数量没有显著差异。初次暴露后,用活体游仆虫免疫的鱼血清对同源分离株的固定化效价和 ELISA 值均高于异源分离株。然而,粘液抗体不能固定游仆虫,尽管 ELISA 结果清楚地表明产生了识别 C.irritans 的粘液抗体。在 Western blot 分析研究中,血清抗体仅识别相应血清型的抗原,而未鉴定出两种血清型共有的蛋白质。对包括完整核核糖体 ITS-1-5.8S rDNA-ITS-2 区域在内的 754 个碱基的 rDNA 核苷酸序列进行了序列分析,发现 W1 和 K1 分离株的序列完全相同。这些发现证实了两种分离株均为 C.irritans 的成员,并且 rDNA 分析无法区分这两种分离株。总之,尽管固定化测定和 ELISA 在体外检测到两种血清型,但挑战测定仅提供了一种 C.irritans 的证据。