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Vaccine Detail

H. influenzae Hap protein vaccine
Vaccine Information
  • 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: cholera toxin
  • Immunization Route: Subcutaneous injection
Host Response

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 ×106 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).
References
Liu et al., 2004: Liu DF, Mason KW, Mastri M, Pazirandeh M, Cutter D, Fink DL, St Geme JW 3rd, Zhu D, Green BA. The C-terminal fragment of the internal 110-kilodalton passenger domain of the Hap protein of nontypeable Haemophilus influenzae is a potential vaccine candidate. Infection and immunity. 2004 Dec; 72(12); 6961-8. [PubMed: 15557618].