【摘要】：目的:初步探討和評(píng)價(jià)NMB0315核酸疫苗、重組蛋白疫苗及核酸疫苗+重組蛋白疫苗聯(lián)合免疫誘導(dǎo)小鼠產(chǎn)生的特異性體液/細(xì)胞免疫應(yīng)答水平及其免疫保護(hù)效果,為進(jìn)一步探索NMB0315疫苗有效的免疫方法和途徑提供實(shí)驗(yàn)依據(jù)。方法:大量制備核酸疫苗[pc DNA3.1(+)/NMB0315]和重組蛋白疫苗(p ET-30a/NMB0315),采用核酸初免-蛋白加強(qiáng)的方法聯(lián)合免疫或分別免疫雌性BALB/c小鼠,測(cè)定特異性體液/細(xì)胞免疫應(yīng)答水平、免疫血清體外殺菌效價(jià),觀察疫苗對(duì)感染B群腦膜炎球菌小鼠的免疫保護(hù)效果。結(jié)果:NMB0315核酸疫苗組(p NMB0315-Cp G)、蛋白疫苗組(r NMB0315-FA)及聯(lián)合免疫疫苗組(p NMB0315-Cp G+r NMB0315-FA)誘導(dǎo)的血清特異性Ig G、Ig G1、Ig G2a及生殖道灌洗液中特異性s Ig A水平在第八周達(dá)到峰值,A450值分別為(0.505±0.042、0.513±0.022、0.342±0.017、0.250±0.015)、(0.823±0.061、0.807±0.045、0.596±0.027、0.450±0.028)和(0.694±0.053、0.711±0.032、0.455±0.021、0.386±0.024),明顯高于PBS對(duì)照組(P0.01);其中,蛋白疫苗組的抗體水平明顯高于聯(lián)合免疫疫苗組和核酸疫苗組(P0.05)。小鼠脾淋巴細(xì)胞刺激指數(shù)及IFN-γ水平,聯(lián)合免疫疫苗組明顯高于蛋白疫苗組和核酸疫苗組(P0.05);核酸疫苗組、蛋白疫苗組和聯(lián)合免疫疫苗組免疫血清在補(bǔ)體介導(dǎo)下的體外殺菌抗體效價(jià)分別為1∶64、1∶128和1∶128,對(duì)實(shí)驗(yàn)小鼠的免疫保護(hù)率分別為70%、95%和80%。免疫2、4、6、8周時(shí),核酸疫苗組、重組蛋白疫苗組和聯(lián)合免疫疫苗組Ig G2a/Ig G1比值均小于1。結(jié)論:NMB0315疫苗誘導(dǎo)的體液免疫(包括黏膜免疫)效果從高到低為:重組蛋白疫苗組、聯(lián)合免疫疫苗組、核酸疫苗組;誘導(dǎo)細(xì)胞免疫的效果從高到低為:聯(lián)合免疫疫苗組、核酸疫苗組、重組蛋白疫苗組;NMB0315疫苗對(duì)實(shí)驗(yàn)小鼠的免疫保護(hù)效果從高到低為:重組蛋白疫苗組、聯(lián)合免疫疫苗組、核酸疫苗組。
[Abstract]:Objective: To study and evaluate the specific humoral/ cellular immune response level and the immune protective effect of NMB0315 nucleic acid vaccine, recombinant protein vaccine and nucleic acid vaccine + recombinant protein vaccine in mice. To provide an experimental basis for further exploring the effective immunization methods and methods of the NMB0315 vaccine. Methods: A large amount of nucleic acid vaccine (pcDN1 (+)/ NMB0315] and recombinant protein vaccine (pET-30a/ NMB0315) were prepared. To observe the effect of the vaccine on the immune protection of B-group meningococcal mice. Results: The serum specific Ig G, Ig G1, Ig G2a and the specific s Ig A level in the NMB0315 nucleic acid vaccine group (pNMB0315-Cp G), the protein vaccine group (r NMB0315-FA) and the combined immune vaccine group (p NMB0315-Cp G + r NMB0315-FA) reached the peak at the eighth week, A450 values were (0.505, 0.042, 0.513, 0.022, 0.342, 0.017, 0.250, 0.015), (0.823, 0.061, 0.807, 0.045, 0.596, 0.027, 0.450, 0.028), and (0.694, 0.053, 0.711, 0.032, 0.455, 0.021, 0.386, 0.024), respectively, significantly higher than in the PBS control group (P0.01); in which the level of antibody in the protein vaccine group was significantly higher than that of the combined immunovaccine group and the nucleic acid vaccine group (P0.05). The titer of the in vitro bactericidal antibody against the complement-mediated by the immune serum of the nucleic acid vaccine group, the protein vaccine group and the combined immune vaccine group was 1:64,1:128 and 1:128, respectively. The immune protection rate of the experimental mice was 70%,95% and 80%, respectively. The ratio of Ig G2a/ Ig G1 in the nucleic acid vaccine group, the recombinant protein vaccine group and the combined immune vaccine group was less than 1 at the 2,4,6 and 8 weeks of immunization. Conclusion: The humoral immunity (including mucosal immunity) induced by the NMB0315 vaccine is from high to low: the recombinant protein vaccine group, the combined immune vaccine group and the nucleic acid vaccine group; the effect of inducing the cell immunity is from high to low: the combined immune vaccine group, the nucleic acid vaccine group and the recombinant protein vaccine group; The immune protective effect of the NMB0315 vaccine against the experimental mice is from high to low: the recombinant protein vaccine group, the combined immune vaccine group and the nucleic acid vaccine group.
【作者單位】： 南華大學(xué)病原生物學(xué)研究所;長(zhǎng)沙市疾病預(yù)防控制中心;
【基金】：國(guó)家自然科學(xué)基金(81172890) 特殊病原體防控湖南省重點(diǎn)實(shí)驗(yàn)室(湘科計(jì)字2014-5號(hào)、湘教通2012-312號(hào)) 湖南省分子靶標(biāo)藥物研究協(xié)同創(chuàng)新中心資助
【分類號(hào)】：R392

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本文編號(hào)：2529420