Enhancement of humoral immunity in mice by coupling pUCpGs10 and aluminium to the HCV recombinant immunogen

Virology Journal

He Q Zhang¹  Xi Q Yang¹  Zhen H Dai¹  Cui X Zhu¹  Xiao G Song¹  Guan Z Bai¹  Kun Chen¹  Guo H Wang¹  Bing S Xiu¹  Na Zhan² 

[1]Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Taiping Road No.27, Haidian, Beijing 100850, China

[2]Hospital 307 of PLA, Academy of Military Medical Sciences, Fengtai, Dongda Road No.7, Beijing 100072, China

关键词: FCM;    ELISPOT;    adjuvant;    humoral immunity;    HCV;   

Others  :  1155664
DOI  :  10.1186/1743-422X-8-507

 received in 2011-06-29, accepted in 2011-11-04,  发布年份 2011

【 摘 要 】

Aim

To investigate the enhancement of humoral immunity when CpG ODN (cytidine phosphate guanosine oligodeoxynucleotides) and aluminium adjuvants are complexed with the HCV (Hepatitis C virus) recombinant immunogen in mice.

Methods

After immunizing Balb/c mice with the recombination HCV antigen adjuvanted with pUCpGs10 and/or aluminium(antigen+CpG+alum, antigen+CpG, antigen+alum, antigen+PBS), enzyme-linked immunosorbent assay (ELISA) was used to measure the specific serum antibody titers of IgG, to determine the neutralization response to various peptide genotypes, and to determine the concentration of IL-6 and IL-10 in supernatants of in vitro cultured splenic lymphocytes. Enzyme-linked immunospot assay (ELISPOT) was used to quantify the non-specific and specific splenic antibody-secreting cells (ASCs), and flow cytometry (FCM) determined the ratio of different splenic lymphocytes. The serum of rabbits immunized with the recombinant pBVGST/HVR1 antigen immunoprecipitated the HCV isolated from 12 patients' serum.

Results

The sera antibody titers were 1:51200, 1:9051, 1:18102, 1:6400 respectively after the final immunization and demonstrated good neutralization responses to the six gene peptide containing 1a, 1b, 2a, 3a, 4a and 6a. The aluminum adjuvant increased the population of both specific ASCs (P < 0.01) and total ASCs(P < 0.05), with a proportional rise in concentrations of CD19⁺CD27⁺ (P < 0.05), as well as levels of IL-6, IL-10 (P < 0.05) in splenic lymphocytes. The results clearly indicated a significantly higher number of CD19⁺CD38⁺ splenic lymphocytes with the aluminum and pUCpGs10 adjuvant present compared to the control group(P < 0.05). Anti-HVR1 antibody in induced mice can cross-reactively capture HCV particles (10/12).

Conclusions

1. The aluminum adjuvant induces a potent Th2-biased immune response by increasing both the populations of specific and total ASCs and the ratio of CD19⁺CD27⁺ cells. 2. The pUCpGs10 complexed with the aluminum adjuvant boosts the population of plasma cells and increase the efficiency of the immune response. 3. The two adjuvants have synergistic effects on humoral immunity. 4. The recombinant HVR1 protein has the possibility of generating broadly reactive anti-HVR1 antibody.

【 授权许可】

   
2011 Zhan et al; licensee BioMed Central Ltd.

【 图 表 】

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