The rLCMV vector expressing Ag85B-TB10.4 (herein referred to as “rLCMV” throughout) was generated, and stocks were grown and titrated following established protocols [20 (link),21 (link)]. The Ag85B-TB10.4 fusion protein consisting of amino acids 42-325 of the Ag85B ORF and fused at its C-terminus to the 96 amino acid-long TB10.4 ORF was preceded by the human tissue plasminogen activator signal peptide for optimal immunogenicity [37 (link)]. The LCMV backbone was derived from the Armstrong Clone 13 variant, with asparagine at position 400 of the nucleoprotein mutated (N400S) to eliminate the immunodominant H-2Db-restricted viral epitope NP396-404 [38 (link)]. rLCMV was administered intramuscularly (i.m.) at a dose of 2 × 106 plaque forming units unless specified otherwise. BCG was administered to mice subcutaneously at a dose of 106 CFU in 100 µL PBS. For DTPa-6 immunization a total dose of 100 µL Infanrix Hexa (GlaxoSmithKline, Brentford, UK) was administered simultaneously via the i.m. route (25 µL into each leg) and intraperitoneally (50 µL).
Infanrix hexa
Manufactured by GlaxoSmithKline
Sourced in United Kingdom, Belgium
Infanrix Hexa is a combined vaccine that protects against six infectious diseases: diphtheria, tetanus, pertussis (whooping cough), hepatitis B, polio, and Haemophilus influenzae type b (Hib). It is a sterile, injectable suspension that is administered intramuscularly.
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Lab products found in correlation
Infanrix ipv hib, by GlaxoSmithKline (1 mentions)
Prevenar, by Pfizer (1 mentions)
Rotavirus, by GlaxoSmithKline (1 mentions)
Hexaxim, by Sanofi (1 mentions)
Varilrix, by GlaxoSmithKline (1 mentions)
Rotarix, by GlaxoSmithKline (1 mentions)
Engerix b, by GlaxoSmithKline (1 mentions)
Prevenar13, by Pfizer (1 mentions)
Synflorix, by GlaxoSmithKline (1 mentions)
Priorix, by GlaxoSmithKline (1 mentions)
Menitorix, by GlaxoSmithKline (1 mentions)
Rotateq, by Merck Group (1 mentions)
6 protocols using infanrix hexa
1
Recombinant LCMV Vector Vaccine Preparation
2
Childhood Vaccination Regimen and Antipyretic Use
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Participants received 3-dose primary vaccination with PHiD-CV (Synflorix™, GSK, Belgium) at 3, 4, and 5 months of age and booster dose at 12–15 months of age (intramuscular, in the right thigh, or deltoid for children >12 months); 2 doses of DTPa-HBV-IPV/Hib (Infanrix hexa™, GSK, Belgium) at 3 and 5 months of age and booster dose at 12–15 months of age (intramuscular, left thigh or deltoid); and one dose of DTPa-IPV/Hib (Infanrix-IPV/Hib™, GSK, Belgium) at 4 months of age (intramuscular, left thigh). The first dose of antipyretic (ibuprofen (Nurofen™, Reckitt Benckiser, UK) – 10 mg/kg/dose, with a maximum daily dose of 30 mg/kg, or paracetamol (Panadol Baby™, GSK, UK) – 15 mg/kg/dose with a maximum daily dose of 60 mg/kg) was administered orally either immediately after vaccination at the study site (immediate administration) or by the parents at home 4–6 hours after vaccination (delayed administration). The second and third dose of antipyretic were administered by the parents at home, 6–8 hours after the previous dose; if a child slept overnight, the dose was deferred to the following morning.
3
Infant Vaccination Protocols and Administration
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The composition and batch numbers of the vaccines administered in the primary series and booster parts are described elsewhere for Study 1 (DTaP-IPV-HB-PRP~T [Hexaxim; Sanofi Pasteur], DTwP/PRP~T [CombAct-Hib; Sanofi Pasteur], HB [Engerix B; GlaxoSmithKline], OPV [Sanofi Pasteur], MMR [ROR, Sanofi Pasteur], and V [Varilrix, GlaxoSmithKline])16 ,24 and Study 2 (DTaP-IPV-HB-PRP~T [Hexaxim; Sanofi Pasteur], DTaP-IPV-HB//PRP~T [Infanrix hexa; GlaxoSmithKline], PCV7 [Prevenar; Pfizer], and rotavirus [Rotarix; GlaxoSmithKline]).20 The DTaP-IPV-HB-PRP~T, DTaP-IPV-HB//PRP~T, PCV7, and HB vaccines were administered intramuscularly, MMR was administered either intramuscularly or subcutaneously, V was administered subcutaneously, and rotavirus vaccine was administered orally (see Figure 1 for subjects disposition).
4
Comparing PCV10 and PCV13 Vaccinations
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Children in the PCV10 group were vaccinated with Synflorix (GSK, Belgium) during regular visits to well-baby clinics. PCV10 contains 1 μg of serotypes 1, 5, 6B, 7F, 9V, 14, and 23F and 3 μg of serotypes 4, 18C, and 19F. The polysaccharides are conjugated to protein D, except for 18C (tetanus toxoid) and 19F (diphtheria toxoid). Children in the PCV13 group received Prevenar13 (Pfizer, UK) during home visits by the study team. This vaccine contains 2.2 μg of serotypes 1, 3, 4, 5, 6A, 6B, 7F, 9V, 14, 18C, 19A, 19F, and 23F, all conjugated to diphtheria toxoid CRM197. Both groups concomitantly received Infanrix-hexa (GSK, Belgium) against diphtheria, tetanus, acellular pertussis, hepatitis B, poliovirus, and Haemophilus influenzae type B (conjugated to tetanus toxoid), at 2, 3, 4, and 11 months of age.
5
Antibody Response to DTPa-6 Vaccine
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To determine antibody responses elicited by DTPa-6 (Infanrix Hexa; GlaxoSmithKline, Brentford, UK), we performed enzyme-linked immunosorbent assays, as previously described [40 (link)]. In brief, tetanus toxoid (TT), Bordetella pertussis toxoid (PT), Bordetella pertussis pertactic (PRN), and filamentous hemagglutinin (FHA) were coated to plates and overlaid with serially diluted mouse serum, and bound antibodies were detected using a mouse IgG-specific detection antibody, the binding of which was quantified by means of an electrochemiluminescent label.
6
Routine Infant Immunization Schedule
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All infants received routine immunisations according to the Australian National Immunisation Program: at birth: intramuscular HepB vaccine (H-B-Vax II Paediatric® (bioCSL)); at 6 weeks, 4 and 6 months of age: intramuscular combined diphtheria-tetanus-acellular pertussis (DTPa)-HepB-inactivated polio (IPV)-Haemophilus influenzae type b (Hib) vaccine (Infanrix® hexa (GlaxoSmithKline)), intramuscular 13-valent conjugate pneumococcal vaccine (PCV13) (conjugated to CRM197, a diphtheria toxin) (Prevenar13® (Wyeth)), and oral rotavirus vaccine (RotaTeq® (Merck)); at 12 months of age: subcutaneous measles-mumps-rubella (MMR) vaccine (Priorix® (GlaxoSmithKline)) or (M-M-R®II (Seqirus)) and intramuscular combined Hib and meningococcal C vaccine (conjugated to tetanus toxin) (Menitorix® (GlaxoSmithKline)). Approximately half of the infants were randomised to receive intradermal BCG-Denmark (Statens Serum Institute, Copenhagen) shortly after birth as part of the MIS BAIR. None of the infants received an influenza vaccine. Vaccine records were obtained from the National Australian Immunisation register and/or individual immunisation records.
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