Increasing the immunogenicity of protein antigens through the genetic insertion of VQGEESNDK sequence of human IL-1 beta into their sequence.
The immunogenicity of two recombinant protein Ag containing the immunostimulatory sequence of human IL-1 beta 163-171 (VQGEESNDK) genetically engineered into their structure has been evaluated. The IL-1 beta sequence was inserted into the loop between alpha helices D and E of recombinant human ferritin H chain and into the hypervariable region of recombinant flagellin from Salmonella muenchen. The chimeric proteins were injected into mice and the level of humoral immune response developed against the native proteins was assessed by measuring the number of Ag-specific plaque forming cells/spleen or as the level of serum IgG response. The response was compared to that of mice receiving injections with wild-type protein Ag not containing the VQGEESNDK sequence or with hybrid constructs containing unrelated foreign peptide sequences of the same length. A significantly higher immune response was observed in mice immunized with chimeric constructs containing the human IL-1 beta 163-171 sequence. These data suggest that the insertion of the VQGEESNDK sequence may prove useful to increase the immune response against poorly immunogenic recombinant proteins.
Beckers
W
W
Immunobiological Research Institute, Siena, Italy.
Villa
L
L
Gonfloni
S
S
Castagnoli
L
L
Newton
S M
SM
Cesareni
G
G
Ghiara
P
P
D000276:Adjuvants, Immunologic; D000595:Amino Acid Sequence; D000818:Animals; D000917:Antibody Formation; D001483:Base Sequence; D005260:Female; D005293:Ferritins / Q000276:immunology*; D005408:Flagellin / Q000276:immunology*; D006801:Humans; D007375:Interleukin-1 / Q000737:chemistry / Q000276:immunology*; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000276:immunology*; D012475:Salmonella / Q000276:immunology
D000276:Adjuvants, Immunologic; D007375:Interleukin-1; D009838:Oligodeoxyribonucleotides; D011993:Recombinant Fusion Proteins; D005408:Flagellin; D005293:Ferritins