• Vaccine Name: H. influenzae Hap protein vaccine
• Target Pathogen: Haemophilus influenzae
• Target Disease: Meningitis
• Vaccine Ontology ID: VO_0011499
• Type: Subunit vaccine
• Status: Research
• Antigen: H. influenzae Hap
• hap gene engineering:
  • Type: Recombinant protein preparation
  • Description: Bacteria were grown in 10 liters of BHI broth for 18 h at 35°C with aeration. All the following steps were performed at 4°C. Bacterial cells were removed by centrifugation at 10,000 × g. The culture supernatant was concentrated 20-fold by using an Amicon stir cell and was fractionated overnight with ammonium sulfate at 60% saturation. After centrifugation at 17,000 × g for 1 h, the precipitate was dissolved in 20 mM Tris buffer at pH 7.4 containing 50 mM NaCl and 1 mM EDTA, dialyzed against the same buffer, and then centrifuged at 100,000 × g for 1 h to remove insoluble material. The resulting supernatant was loaded at a flow rate of 2 ml/min onto a 20-ml SP Sepharose column (Amersham Pharmacia Biotech) equilibrated with the same buffer. The column was washed until the OD280 reached the baseline, and nHapS-P860295 was eluted at a flow rate of 3 ml/min with a linear gradient of NaCl (from 55 to 500 mM) in 20 mM Tris at pH 7.5 with 1 mM EDTA. Based on sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) gels, fractions containing nHapS-P860295 were pooled (Liu et al., 2004).
  • Detailed Gene Information: Click Here.
• Adjuvant:
  • Adjuvant name:
  • VO adjuvant ID: VO_0000143
  • Description: Mutant cholera toxin CT-E29H
• Immunization Route: Subcutaneous injection

Mouse Response

• Host Strain: BALB/c
• Vaccination Protocol: Groups of 10 female, 6- to 8-week-old Swiss Webster or BALB/c mice (Taconic Farms, Germantown, N.Y.) were immunized subcutaneously at weeks 0 and 4 with protein antigens. To prepare the protein antigens for vaccination, 10 μg of rCBD or 2 μg of an induced E. coli BL21(DE23)/pLysS/pGEX-6P-1 cell lysate was absorbed onto 100 μg of aluminum phosphate at 37° for 1 h, and then 50 μg of 3-O-deacylated monophosphoryl lipid A (MPL; Corixa Corp., Hamilton, Mont.) was added. Sera collected at weeks 0, 4, and 6 were pooled for analysis (Liu et al., 2004).
• Challenge Protocol: Three weeks after the final immunization, mice were challenged intranasally with approximately ×10^6 CFU of strain TN106.P2 (Liu et al., 2004).
• Efficacy: When mice immunized intranasally with recombinant protein corresponding to the C-terminal region of Hap(S) from H. influenzae strains N187, P860295, and TN106 plus mutant cholera toxin CT-E29H were challenged with strain TN106, they were protected against nasopharyngeal colonization (Liu et al., 2004).