title: Enhancement of humoral immunity in mice by coupling pUCpGs10 and aluminium to the HCV recombinant immunogen.
abstract: OBJECTIVE
  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.
authors: Institute of Basic Medical Sciences, Academy of Military Medical Sciences, Haidian, Beijing 100850, China.
authors: Na
authors: N
authors: Zhan
authors: Bing S
authors: BS
authors: Xiu
authors: Guo H
authors: GH
authors: Wang
authors: Kun
authors: K
authors: Chen
authors: Guan Z
authors: GZ
authors: Bai
authors: Xiao G
authors: XG
authors: Song
authors: Cui X
authors: CX
authors: Zhu
authors: Zhen H
authors: ZH
authors: Dai
authors: Xi Q
authors: XQ
authors: Yang
authors: He Q
authors: HQ
authors: Zhang
mesh_terms: D000276:Adjuvants, Immunologic / Q000008:administration & dosage; D000536:Aluminum Hydroxide / Q000008:administration & dosage; D000595:Amino Acid Sequence; D000818:Animals; D000921:Antibody-Producing Cells / Q000276:immunology* / Q000378:metabolism; D015703:Antigens, CD / Q000032:analysis / Q000276:immunology; D058501:Enzyme-Linked Immunospot Assay; D016174:Hepacivirus / Q000737:chemistry / Q000235:genetics / Q000276:immunology*; D019698:Hepatitis C, Chronic / Q000276:immunology / Q000517:prevention & control* / Q000821:virology; D007111:Immunity, Cellular; D056724:Immunity, Humoral; D016753:Interleukin-10 / Q000276:immunology / Q000378:metabolism; D015850:Interleukin-6 / Q000276:immunology / Q000378:metabolism; D008214:Lymphocytes / Q000276:immunology / Q000378:metabolism; D051379:Mice; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000008:administration & dosage / Q000737:chemistry; D011817:Rabbits; D013154:Spleen / Q000276:immunology / Q000378:metabolism; D014761:Viral Hepatitis Vaccines; D014764:Viral Proteins / Q000235:genetics / Q000276:immunology*
chemical_list: D000276:Adjuvants, Immunologic; D015703:Antigens, CD; C408982:CPG-oligonucleotide; C400562:HVR1 protein, Hepatitis C virus; D015850:Interleukin-6; D009838:Oligodeoxyribonucleotides; D014761:Viral Hepatitis Vaccines; D014764:Viral Proteins; D016753:Interleukin-10; D000536:Aluminum Hydroxide