[Effect of white spot syndrome virus envelope protein Vp28 expressed in silkworm (Bombyx mori) pupae on disease resistence in Procambarus clarkii].

Shi yan sheng wu xue bao Pub Date : 2005-06-01
Ke Qiang Wei, Zi Rong Xu
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Abstract

The vaccine made of recombinant envelope protein (rVp28) of white spot syndrome virus (WSSV) expressed in silkworm (Bombyx mori) pupae using a baculovirus vector was used to investigate the efficacy of oral administration on WSSV disease resistance of Procambarus clarkii. Vaccine was mixed with diet at a ratio of 2% (w/w), and Procambarus clarkii were orally administered throughout 75 days. Vaccination with rVP28 showed the significantly higher cumulative survival compared with positive and negative control (P < 0.05) following an oral challenge on the 35th day post-vaccination (dpv), with PRP values 54.16% and 59.26%, respectively. rVP28 induced higher resistance via IM (intramuscular) injection challenge with WSSV stock, with PRP value of 46.12% and 49.99%, respectively. The survivors were subsequently re-challenged on the 55th dpv. rVP28 induced the significantly higher resistance to oral re-challenge (P < 0.05), with both PRP values 55.80% and 63.16%, respectively. rVP28 induced higher resistance to IM injection re-challenge, with both PRP values 31.25%. A DIG labeled WSSV DNA probe was used to detect WSSV by in situ hybridization. The positive cells were observed in epithelial cells of stomach, hepatopancreas and gut of the infected control crayfish, while negative reaction were observed in the tissues of survivors-vaccinated. These results indicated that vaccination of crayfish with recombinant protein had significant effect on oral infection, and had higher resistance against intramuscular injection challenge. This suggested the protection against WSSV could be induced in crayfish by recombinant protein rVp28 expressed in silkworm pupae.

家蚕蛹表达的白斑综合征病毒包膜蛋白Vp28对克氏原螯虾(Procambarus clarkii)抗病的影响
用杆状病毒载体制备白斑综合征病毒(WSSV)重组包膜蛋白(rVp28)疫苗,研究了口服克氏原螯虾(Procambarus clarkii)对WSSV的抗性效果。疫苗以2% (w/w)的比例与饲料混合,口服克氏原螯虾75 d。接种rVP28疫苗后第35天(dpv)口服攻毒,累积存活率显著高于阳性对照组和阴性对照组(P < 0.05), PRP值分别为54.16%和59.26%。rVP28肌内注射WSSV原液攻毒后产生较高的抗性,PRP值分别为46.12%和49.99%。幸存者随后在第55次dpv上再次受到质疑。rVP28对口腔再攻毒的抗性显著提高(P < 0.05), PRP值分别为55.80%和63.16%。rVP28对IM注射再攻毒的抗性较高,PRP值均为31.25%。采用DIG标记的WSSV DNA探针原位杂交检测WSSV。感染对照小龙虾的胃、肝胰脏和肠道上皮细胞呈阳性反应,而接种幸存者组织呈阴性反应。结果表明,重组蛋白免疫小龙虾对口腔感染有显著的预防作用,对肌内注射攻毒具有较高的抵抗力。这表明在家蚕蛹中表达的重组蛋白rVp28可以诱导小龙虾对WSSV的保护作用。
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