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. 2014 Sep;30(9):913-6.

[Mouse macrophage-derived chemokine as an adjuvant enhances the protective effect of P6 protein vaccine of nontypeable Haemophilus influenzae]

[Article in Chinese]
Xiaoyan Han  1 , Chenhong Wang  1 , Haixia Qiao  1 , Yanxia Zhang  1 , Yueli Chang  1 , Yutuo Zhang  1
Affiliations
  • PMID: 25200152

[Mouse macrophage-derived chemokine as an adjuvant enhances the protective effect of P6 protein vaccine of nontypeable Haemophilus influenzae]

[Article in Chinese]
Xiaoyan Han et al. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2014 Sep.

Abstract

Objective: To express P6 protein of nontypeable Haemophilus influenzae (NTHi) in prokaryotic cells and observe the immune effect of macrophage-derived chemokine (MDC) as an adjuvant on NTHi-P6 protein vaccine.

Methods: The encoding sequence of NTHi-P6 protein was inserted into plasmid PGEX-6P2, and the recombinant plasmid PGEX-6P2/P6 was transformed into E.coli XL1-Blue. In the expression system, the P6 protein was induced and expressed. BALB/c mice were randomized into three groups that were respectively inoculated intraperitoneally with P6 protein combined with Freund's adjuvant and MDC, P6 protein combined with Freund's adjuvant and only PBS, 3 times at 0, 14 and 28 days, respectively. Sera were collected from 12 mice of each group 14 days after each injection. The titer of IgG antibodies was detected by ELISA. Splenocytes were isolated from 3 mice of each group for determining the levels of IL-4 and IFN-γ by ELISA. Furthermore, the other 15 mice of each group were challenged with 10×LD50 NTHi ATCC49247 to observe the protective effect of the vaccine.

Results: The IgG antibody could be induced by p6 protein combined with Freund's adjuvant and MDC and P6 protein combined with Freund's adjuvant, and the titers were 1:1 140.25 and 1:3 044.38, respectively. The differences in the antibody titers and the levels of IFN-γ between the two groups were significant statistically (P<0.05). There was no difference in IL-4 levels between the two groups (P>0.05). After challenged with 10×LD50 NTHi ATCC49247, the survival rate of mice in P6 protein combined with Freund's adjuvant and MDC group was 80%, and significantly higher than that in the PBS group (P<0.05), but no statistical difference was noted when compared with P6 protein combined with Freund's adjuvant group.

Conclusion: MDC as an adjuvant could enhance the immune protective effect of NTHi-P6 protein vaccine to an extent.

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