Plant K P, LaPatra S E, Cain K D
Hagerman Fish Culture Experiment Station, University of Idaho, Hagerman, ID, USA.
J Fish Dis. 2009 Jun;32(6):521-34. doi: 10.1111/j.1365-2761.2009.01008.x. Epub 2008 Apr 29.
Flavobacterium psychrophilum heat shock proteins (Hsp) 60 and 70 are highly immunogenic and were therefore investigated as potential vaccine candidates. Recombinant Hsps were purified from Escherichia coli and rainbow trout (Oncorhynchus mykiss) were intraperitoneally injected with phosphate buffered saline/Freunds complete adjuvant (FCA), 8 microg of rHsp60/FCA, rHsp70/FCA or a combination of 4 microg each of rHsp60 and rHsp70/FCA. Antibody responses against recombinant Hsp60 and Hsp70 8 weeks post-immunization were observed, but only fish immunized with rHsp70 exhibited highly elevated antibody levels against F. psychrophilum whole cell lysate. Some cross reactivity occurred, which may have been due to the V5 tag common to both proteins. Protection against F. psychrophilum challenge was not observed in any treatments at 8 weeks post-immunization. To further investigate any protective effect of these proteins, hsps were polymerase chain reaction amplified and cloned into pVAX1. Rainbow trout were intramuscularly injected with 8 microg of pVAX1hsp60, pVAX1hsp70 or a combination of 4 microg each of pVAX1hsp60 and pVAX1hsp70. Antibody responses at 4 weeks post-immunization were low and protection was not observed following challenge at 6 or 10 weeks post-immunization. Although Hsps of F. psychrophilum have been shown to be immunodominant, these antigens do not appear to be good vaccine candidates when delivered alone or in combination.
嗜冷黄杆菌热休克蛋白(Hsp)60和70具有高度免疫原性,因此被作为潜在的疫苗候选物进行研究。从大肠杆菌中纯化重组Hsps,并将磷酸盐缓冲盐水/弗氏完全佐剂(FCA)、8微克重组Hsp60/FCA、重组Hsp70/FCA或4微克重组Hsp60和重组Hsp70/FCA的组合腹腔注射到虹鳟(Oncorhynchus mykiss)体内。观察到免疫后8周针对重组Hsp60和Hsp70的抗体反应,但只有用重组Hsp70免疫的鱼对嗜冷黄杆菌全细胞裂解物表现出高度升高的抗体水平。发生了一些交叉反应,这可能是由于两种蛋白质共有的V5标签所致。免疫后8周,在任何处理中均未观察到对嗜冷黄杆菌攻击的保护作用。为了进一步研究这些蛋白质的任何保护作用,对hsps进行聚合酶链反应扩增并克隆到pVAX1中。将8微克pVAX1hsp60、pVAX1hsp70或4微克pVAX1hsp60和pVAX1hsp70的组合肌肉注射到虹鳟体内。免疫后4周的抗体反应较低,免疫后6周或10周攻击后未观察到保护作用。尽管嗜冷黄杆菌的Hsps已被证明具有免疫优势,但单独或联合递送时,这些抗原似乎不是良好的疫苗候选物。