羊口疮病毒B2L截短重组蛋白刺激小鼠产生特异性抗体的研究

为研究羊口疮病毒(ORFV)新型亚单位疫苗,设计了ORFV B2L优势抗原表位截短重组蛋白,命名为HBBH并经原核表达和鉴定.以不同剂量HBBH不加或加不同佐剂,经滴鼻或颈部皮下注射免疫BALB/c小鼠,其中滴鼻共4组(HBBH 10 μg、20 μg、30μg和20 μg+LTB),注射组(HBBH 20 μg+20...

Study on specific antibody production stimulated by B2L truncated recombinant protein of orf virus

In order to research the sheep aphtha virus(ORFV)new subunit vaccine.Bioinformatics software were used to analyze the immune protective antigen epitopes of ORFV B2 L protein,for designing the truncated recombinant B2 L antigen epitope protein containing histidine label and flexible connecting area,the prokaryotic expression vector and recombinant ORFV truncated recombinant B2 L were constructed and expressed.BALB/c mice were immunized three times with different doses of HBBH(10 g,20 g,30 g)without or with different adjuvants(LTB or SEPEC 201 adjuvant)through nasal drop or neck subcutaneous injection.At the same time,ORFV tissue inactivated antigen containing SEPEC 201 adjuvant were set for control.ORFV Specific serum IgG,IgA and secretary IgA(sI gA)antibody levels were de-tected by indirect ELISA based on ORFV B2 L truncated recombinant protein HBBH.The truncated recombinant protein HBBH containing ORFV B2 L dominant antigen epitopes were efficiently expressed,which could bind to ORFV specific antibodies,indicating that the protein had good reactivity.The results showed that different doses of HBBH could stimulate the production of ORFV specific IgG antibody in mice by nasal drop or neck subcutaneous injection,and 20 g HBBH with SEPEC 201 adjuvant could stimulate the highest production of serum IgG antibody in mice.Lung lavage fluid specific sI gA was detected only in the three groups immuned with HBBH≥20 g of at 14 d post the third inmmune by nasal drop,among which the 30 g HBBH group stimulated the highest level of sI gA.When the HBBH nasal drop does were 20 g,the addition of LTB adjuvant stimulated mice to produce higher levels of sI gA.The results showed that a certain dose of HBBH plus LTB adjuvant by nasal immunization could better stimulate the production of sI gA in mice,however,the serum IgA produced by the HBBH immuned mice was maintained only for a short time.The highest titer of cell neutralizing antibody of the indrect ORFV HBBH ELISA positive IgG serum could reach 1∶128 and neutralize the 4 ORFV isolates used in this experiment.BALB/c mice were stimulated to produce high levels of anti-ORFV serum neutralizing IgG antibody by 20 g HBBH with SEPEC 201 adjuvant by neck subcutaneous injection immune and that of sI gA by 20 g HBBH with LTB adjuvant by nasal drop immune.This study provided data support for ORFV B2 L dominant antigen epitope truncated recombinant protein as subunit vaccine.