To evaluate the effect of nanoemulsion as an adjuvant on immunogenicity of M.RCAg-1,a multi-epitope protein vaccine against Plasmodium falciparum.M.RCAg-1 was formulated with nanoemulsions (NE) and evaluated via IM route in BALB/c mice.Mice were immunized on days 0,14 and 28 at a dosage of 20 μg.Serum samples and spleen cells were collected 10 days after the third immunization.The serum IgG level,antibody subtype and recognition of single epitopes by serum antibody were determined by ELISA.The recognition of natural P.falciparum protein with antibody was evaluated by IFA.and the secretion level of cytokines was detected by ELISPOT.The results showed that the geometric mean titers of mice immunized with M.RCAg-1 + nanoemulsion reached 1:108.The IgGs induced were mainly IgG1.All of the 11 epitopes and natural P.falciparum were recognized significantly.The number of specific lymphocyte clones secreting IFN γ induced by various epitopes and proteins of immunized mice was larger than that secreting IL-4 (P<0.05).Results suggest that M.RCAg-1 delivered with nanoemulsion adjuvant elicited strong humoral and cellular immune responses in mice.%为了评价纳米乳作为佐剂对恶性疟疾人工重组蛋白疫苗M.RCAg-1免疫原性的影响,将纳米乳佐剂与M.RCAg-1相配伍,于第0、14、28 d肌肉注射免疫小鼠,抗原量为20 μg/只.第3次免疫后10 d,眼球取血并取脾细胞.采用ELISA法检测小鼠血清特异性IgG抗体水平、抗体亚类及其对抗原单表位的识别,用间接免疫荧光反应(IFA)检测抗体对恶性疟原虫天然蛋白的识别,用ELISPOT法检测小鼠脾淋巴细胞免疫应答水平.结果显示疫苗佐剂组小鼠血清中抗M.RCAg-1的抗体水平可达1:108;疫苗佐剂组产生的特异性IgG抗体以IgG1为主;疫苗佐剂组抗体对抗原单表位和天然虫体的识别效果显著.各表位和蛋白诱发免疫小鼠分泌IFN-γ的特异性淋巴细胞克隆数高于分泌IL-4的特异性淋巴细胞克隆数.结果提示纳米乳作为恶性疟疾疫苗M.RCAg-1的佐剂,能够显著提高机体的体液免疫和细胞免疫应答水平.
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