MF59和甘油-3-磷酸复合佐剂对甲型H1N1流感抗原体液免疫效果的作用

betway必威登陆网址（betway.com ）学报››2022,Vol. 43››Issue (5): 335-338.DOI:10.3969/j.issn.2097-0005.2022.05.003

MF59和甘油-3-磷酸复合佐剂对甲型H1N1流感抗原体液免疫效果的作用

王珂(), 毛双双, 马心怡, 李玉卓, 李雅林()

betway必威登陆网址（betway.com ）临床与基础医学院，山东泰安 271000

收稿日期:2021-09-22出版日期:2022-05-25发布日期:2022-06-09
通讯作者:李雅林
作者简介:王珂，betway必威登陆网址 2018级临床医学本科，E-mail：591958012@qq.com。
基金资助:
山东省重点研发计划(2019GSF108256);山东省自然科学基金(ZR2021MH163);山东省大学生创新创业训练计划(S202010439099)

Study on the effect of MF59 and glycerol-3-phosphate compound adjuvant on the humoral immune effect of influenza A H1N1 antigen

Ke WANG(), Shuangshuang MAO, Xinyi MA, Yuzhuo LI, Yalin Li()

School of Clinical and Basic Medicnie，Shandong First Medical University & Shandong Academy of Medical Sciences，Taian 271000，China

Received:2021-09-22Online:2022-05-25Published:2022-06-09
Contact:Yalin Li

摘要/Abstract

摘要： 目的

制备复合佐剂MF59和甘油-3-磷酸（glycerol-3-phosphate，G3P），探讨其用于甲型H1N1流感疫苗的效果。

方法

将G3P佐剂与MF59混合为复合佐剂，将其单独或作为复合佐剂与甲型H1N1流感抗原混匀制成疫苗。将50只小鼠分为五组：空白对照组、单纯甲型H1N1流感抗原组、甲型H1N1流感抗原 + MF59佐剂组、甲型H1N1流感抗原 + G3P佐剂组、甲型H1N1流感抗原 + MF59和G3P复合佐剂组。于第0天和第28天腹腔免疫2次。免疫后观察小鼠有无不良反应。在初次免疫后的第28及第42天取血，采用ELISA检测抗H1N1 IgG抗体水平，通过血凝抑制试验检测血凝抑制抗体滴度和假病毒中和试验评价体液免疫的保护效果。

结果

各组小鼠均无不良反应。MF59和G3P复合佐剂组通过腹腔免疫诱导抗H1N1 IgG的水平高于单纯H1N1流感抗原组、MF59佐剂流感抗原组和G3P佐剂流感抗原组，差异均具有统计学意义（P< 0.05）。复合佐剂组血凝抑制抗体和假病毒中和抗体滴度也高于传统的MF59佐剂流感抗原组和G3P佐剂流感抗原组，差异均具有统计学意义（P< 0.05）。

结论

MF59和G3P复合佐剂甲型H1N1流感疫苗保护作用最佳，主要通过诱导产生高滴度的抗H1N1 IgG，提高血凝抑制抗体滴度和中和抗体滴度发挥作用。本研究为开发用于人的新型甲型H1N1流感疫苗提供了实验基础。

关键词:MF59佐剂,甘油-3-磷酸佐剂,复合佐剂,H1N1,流感疫苗

Abstract: Objective

To prepare the combined adjuvant MF59 and glycerol-3-phosphate（G3P）， and to explore its effect on influenza A H1N1 vaccine.

Methods

The G3P adjuvant and MF59 were mixed as a composite adjuvant， and it was mixed with H1N1 influenza antigen alone or as a composite adjuvant to prepare a vaccine. Fifty mice were divide into five groups： blank control group， simple influenza A H1N1 antigen group， A H1N1 influenza antigen + MF59 adjuvant group， A H1N1 influenza antigen group + G3P adjuvant group， and A H1N1 influenza antigen group + G3P adjuvant group H1N1 influenza antigen + MF59 and G3P compound adjuvant group. Intraperitoneal immunization was performed twice on 0 and 28th days. After immunization， the mice were observed whether they had adverse reactions. Blood was collected on the 28th and 42nd days after the initial immunization， the level of anti-H1N1 IgG antibody was detected by ELISA， the titer of hemagglutination inhibitor was detected by hemagglutination inhibition test， and the protective effect of humoral immunity was evaluated by the pseudovirus neutralization test.

Results

There were no adverse reactions in each group of mice. The level of anti-H1N1 IgG induced by intraperitoneal immunization in the MF59 and G3P composite adjuvant group was significantly higher than that of the H1N1 influenza antigen group alone （P< 0.05）， the MF59 adjuvant influenza antigen group （P< 0.05） and G3P adjuvant influenza antigen group （P< 0.05）， combined adjuvant group hemagglutination inhibitory antibody and pseudovirus neutralizing antibody titers were also significantly higher than the traditional MF59 adjuvant influenza antigen group and G3P adjuvant influenza antigen group （P< 0.05）.

Conclusion

The protective effect of MF59 and G3P compound adjuvant influenza A H1N1 vaccine is the best， which mainly acts by inducing the production of high titers of IgG， increasing the titer of hemagglutination inhibitory antibodies and neutralizing antibodies. This research provides an experimental basis for the development of a new type of influenza A H1N1 vaccine for humans.

Key words:MF59 adjuvant,glycerol-3-phosphate adjuvant,composite adjuvant,H1N1,influenza vaccine

王珂, 毛双双, 马心怡, 李玉卓, 李雅林. MF59和甘油-3-磷酸复合佐剂对甲型H1N1流感抗原体液免疫效果的作用[J]. betway必威登陆网址（betway.com ）学报, 2022, 43(5): 335-338.

Ke WANG, Shuangshuang MAO, Xinyi MA, Yuzhuo LI, Yalin Li. Study on the effect of MF59 and glycerol-3-phosphate compound adjuvant on the humoral immune effect of influenza A H1N1 antigen[J]. Journal of Shandong First Medical Unversity & Shandong Academy of Medical Sciences, 2022, 43(5): 335-338.

图/表3

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	2.32 ± 0.29^a	3.32 ± 0.53^a
MF59佐剂组	2.94 ± 0.65^ab	4.67 ± 0.68^ab
G3P佐剂组	2.74 ± 0.25^ab	4.56 ± 0.64^ab
复合佐剂组	4.81 ± 0.83^abcd	7.38 ± 1.42^abcd
F	66.140	72.840
P	< 0.001	< 0.001

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	2.32 ± 0.29^a	3.32 ± 0.53^a
MF59佐剂组	2.94 ± 0.65^ab	4.67 ± 0.68^ab
G3P佐剂组	2.74 ± 0.25^ab	4.56 ± 0.64^ab
复合佐剂组	4.81 ± 0.83^abcd	7.38 ± 1.42^abcd
F	66.140	72.840
P	< 0.001	< 0.001

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	2.02 ± 0.22^a	2.82 ± 0.37^a
MF59佐剂组	2.44 ± 0.46^ab	3.46 ± 0.59^ab
G3P佐剂组	2.35 ± 0.46^ab	3.25 ± 0.45^ab
复合佐剂组	3.81 ± 0.64^abcd	4.85 ± 0.64^abcd
F	27.820	10.990
P	< 0.001	< 0.001

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	2.02 ± 0.22^a	2.82 ± 0.37^a
MF59佐剂组	2.44 ± 0.46^ab	3.46 ± 0.59^ab
G3P佐剂组	2.35 ± 0.46^ab	3.25 ± 0.45^ab
复合佐剂组	3.81 ± 0.64^abcd	4.85 ± 0.64^abcd
F	27.820	10.990
P	< 0.001	< 0.001

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	< 1∶40	< 1∶40
MF59佐剂组	< 1∶40	< 1∶320
G3P佐剂组	< 1∶40	< 1∶320
复合佐剂组	< 1∶640	< 1∶1 280

组别	初免疫后天数
空白对照组	0.00 ± 0.00	0.00 ± 0.00
甲型H1N1流感抗原组	< 1∶40	< 1∶40
MF59佐剂组	< 1∶40	< 1∶320
G3P佐剂组	< 1∶40	< 1∶320
复合佐剂组	< 1∶640	< 1∶1 280

参考文献16

1	Bella A， Gesualdo F， Orsi A， et al. Effectiveness of the trivalent MF59 adjuvated influenza vaccine in preventing hospitalization due to influenza B and A（H1N1）pdm09 viruses in the elderly in Italy， 2017-2018 season［J］. Expert Rev Vaccines， 2019， 18（6）： 671.
2	Hauser MI， Muscatello DJ， Soh ACY， et al. An indirect comparison meta-analysis of AS03 and MF59 adjuvants in pandemic influenza A（H1N1）pdm09 vaccines［J］. Vaccine， 2019， 37（31）： 4246.
3	Rashedi N， Taghizadeh M， Mohamadynejad P， et al. Evaluating the immune response of recombinant H1N1 hemagglutinin with MF59 adjuvant in animal model as a novel alternative to the influenza vaccine［J］. Iran J Allergy Asthma Immunol， 2020， 19（5）： 497.
4	Tregoning JS， Russell RF， Kinnear E. Adjuvanted influenza vaccines［J］. Hum Vaccin Immunother， 2018， 14（3）： 550.
5	Ferdman J， Palladino G， Liao HX， et al. Intra-seasonal antibody repertoire analysis of a subject immunized with an MF59®-adjuvanted pandemic 2009 H1N1 vaccine［J］. Vaccine， 2018， 36（35）： 5325.
6	Green AE， Howarth D， Chaguza C， et al. Pneumococcal colonization and virulence factors identified via experimental evolution in infection models［J］. Mol Biol Evol， 2021， 38（6）： 2209.
7	接晶. MF59联合CpG ODN1826通过促进树突状细胞成熟增强重组MUC1-MBP融合蛋白疫苗的抗肿瘤活性研究［D］. 长春：吉林大学， 2018.
8	孔祥忍，毛双双，刘欣宇，等. 新型佐剂-细菌样颗粒对百日咳抗原体液免疫的效果研究［J］. 医学信息， 2021， 34（2）： 95.
9	罗兴，刘颖，杨春亭，等. CpG-ODN和氢氧化铝复合佐剂对流感病毒裂解疫苗免疫效果的影响［J］. 中华微生物学和免疫学杂志， 2011（10）： 942.
10	Del Giudice G， Rappuoli R， Didierlaurent AM. Correlates of adjuvanticity： a review on adjuvants in licensed vaccines［J］. Semin Immunol， 2018， 39： 14.
11	Wagner R， Hildt E. ［Composition and mode of action of adjuvants in licensed viral vaccines］［J］. Bundesgesundheitsblatt Gesundheitsforschung Gesundheitsschutz， 2019， 62（4）： 462.
12	Wilkins AL， Kazmin D， Napolitani G， et al. AS03- and MF59-Adjuvanted influenza vaccines in children［J］. Front Immunol， 2017， 8： 1760.
13	李卓凡，李卫东. MF59佐剂的研究进展［J］. 国际检验医学杂志， 2016， 37（18）： 2596.
14	Zhao Y， Cao P， Cui YF， et al. Enhanced production of seed oil with improved fatty acid composition by overexpressing NAD⁺-dependent glycerol-3-phosphate dehydrogenase in soybean［J］. J Integr Plant Biol， 2021， 63（6）： 1036.
15	Zhang C， Gu HY， Ren YR， et al. GlcA-mediated glycerol-3-phosphate synthesis contributes to the oxidation resistance ofAspergillus fumigatusvia decreasing the cellular ROS［J］. Fungal Genet Biol， 2021， 149： 103531.
16	金晓，李晓红，贾森泉，等. 甘油-3-磷酸对甲肝疫苗诱导的体液免疫影响［J］. 实验动物科学， 2018， 35（2）： 21.