The following influenza vaccine formulations were purchased from Moore Medical: FluMist Quadrivalent, 2014–2015, live-attenuated vaccine (MedImmune) and Fluzone Quadrivalent, 2014–2015, inactivated virus vaccine (Sanofi Pasteur) directed against A/California/7/2009 (H1N1)pdm09, A/Texas/ 50/2012 (H3N2), B/Massachusetts/2/2012, and B/Brisbane/60/2008; and 2015-2016 FluMist Quadrivalent and Fluzone Quadrivalent formulations directed against A/California/7/2009 (H1N1)pdm09, A/ Switzerland/9715293/2013 (H3N2), B/Phuket/3073/2013, and B/Brisbane/60/2008 strains. Fluorescently conjugated antibodies for Thy1.2 (clone 53-2.1, BioLegend), CD4 (clones RM4-5, BD Biosciences, and GK1.5, eBioscience), CD8 (clone 53-6.7, BioLegend), CD44 (clone IM7, BioLegend), CD62L (clone MEL- 14, BioLegend), CD69 (clone H1.2F3, eBioscience), and CD103 (clone 2E7, eBioscience) were purchased. The influenza NP–specific (NP366–374) H-2Db tetramer was purchased from MBL International. FTY720 was purchased from Cayman Chemical Company.
Fluzone quadrivalent
Manufactured by Sanofi
Sourced in Switzerland
Fluzone Quadrivalent is an injectable influenza vaccine that protects against four different strains of the influenza virus. It is designed to stimulate the immune system to produce antibodies against the specific strains included in the vaccine.
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Lab products found in correlation
Flumist quadrivalent, by Sanofi (2 mentions)
Fty720, by Cayman Chemical (2 mentions)
Fluarix quadrivalent, by GlaxoSmithKline (2 mentions)
Cd44 clone im7, by BioLegend (1 mentions)
Cd69 clone h1.2f3, by Thermo Fisher Scientific (1 mentions)
Cd62l clone mel 14, by BioLegend (1 mentions)
Thy1.2 clone 53 2 1, by BioLegend (1 mentions)
Cd103 clone 2e7, by Thermo Fisher Scientific (1 mentions)
Cd8 clone 53 6, by BioLegend (1 mentions)
Fluzone high dose, by Sanofi (1 mentions)
Anti cd28 clone 37, by BD (1 mentions)
Anti cd3e clone 145 2c11, by BD (1 mentions)
9 protocols using fluzone quadrivalent
1
Influenza Vaccine Formulations and Immunological Analysis
2
Influenza Vaccine Response in Healthy Children
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A study was performed during the 2016–2017 influenza season at St. Jude Children’s Research Hospital in Memphis, TN (clinical trials.gov #NCT02649192) among 22 healthy children between the ages of two and eight years. These children were participating as controls in a vitamin supplementation study. They received a placebo gummy at the time of vaccination [3 (link)]. Vaccines were Fluzone Quadrivalent (Sanofi Pasteur) for children <3 years of age and Fluarix Quadrivalent (GlaxoSmithKline) for children of ≥3 years of age, administered intramuscularly (IM). Serum samples were taken prior to vaccination and 28 days post-vaccination. The recommended composition of the 2016–2017 vaccine was A/California/7/2009 H1N1 pdm09-like virus, A/Hong Kong/4801/2014 H3N2-like virus, B/Brisbane/60/2008-like virus (B/Victoria lineage), and B/Phuket/3073/2013-like virus (B/Yamagata lineage). This study was reviewed and approved by the Institutional Review Board of St. Jude Children’s Research Hospital.
3
Influenza Vaccine Composition Changes
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In season 2015–2016, the vaccine was FluMist® (AstraZeneca, Cambridge, UK), whereas in seasons 2016–2017 and 2017–2018, the vaccines were Fluzone® Quadrivalent (Sanofi Pasteur, Lyon, France) for children < 3 years of age and Fluarix® Quadrivalent (GlaxoSmithKline, Brentford, UK) for children of ≥ 3 years of age. These changes were due to revised recommendations by the Advisory Committee on Immunization Practices (ACIP) of the Centers for Disease Control and Prevention (CDC). Vaccine components changed in each influenza virus season. In 2015–2016, components were A/CA/7/09 H1N1, A/Switzerland/9715293/13 H3N2, B/Phuket/3073/13, and B/Brisbane/60/2008. In 2016–2017 components were A/CA/7/09 H1N1, A/Hong Kong/4801/14 H3N2, B/Phuket/3073/13, and B/Brisbane/60/2008, and in 2017–2018, components were A/Michigan/45/2015 H1N1, A/Hong Kong/4801/14 H3N2, B/Phuket/3073/13, and B/Brisbane/60/2008.
4
Quantifying Antigen-Specific Antibodies
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The following antigens were used for antigen-specific IgG and IgA capturing: quadrivalent influenza vaccine 2019–20, 2020–21, 2021–22, or 2023–24 (ie for antigen-specific IgG capturing) (Fluarix Quadrivalent Influenza Vaccine 2019–20, 2020–21, 2021–22, or 2023–24 Formula, respectively; GSK Biologicals/ABO Pharmaceuticals; Afluria® Quadrivalent (Seqirus) and Fluzone® Quadrivalent (Sanofi Pasteur)), Tetanus Toxoid, Clostridium tetani (Calbiochem/Millipore Sigma or Fina Biosolutions), and SARS-CoV-2 S2P (recombinant SARS-CoV-2 soluble spike trimer protein, lot #P210721.02; Protein Expression Laboratory, Frederick National Laboratory for Cancer Research (FNLCR), Frederick, MD). For relative quantitation of Ag-specific antibody titers, standard curves were generated using monoclonal antibody (mAb) standards of anti-tetanus toxin mAb (clone TetE3; The Native Antigen Company) and SARS-CoV-2-reactive (ie spike RBD) mAb (Abeomics). For determining the concentrations of total IgG, purified human IgG (ChromePure human IgG, JacksonImmuno Research Laboratories) was used as a standard.
5
RSV Prefusion Antigen Vaccine Formulations
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RSVpreF formulations contained equal amounts of 2 stabilized prefusion F antigens, 1 from the subgroup A/Ontario strain and 1 from the subgroup B/Buenos Aires strain, totaling 60 μg, 120 μg, or 240 μg, with or without 0.2 mg Al(OH)3. Commercially available quadrivalent SIIV (Fluzone Quadrivalent, Sanofi Pasteur, Swiftwater, Pennsylvania) was administered to participants 18–49 years of age, and high-dose trivalent SIIV (Fluzone High-Dose, Sanofi Pasteur) was administered to participants 65–85 years of age [22 , 23 ]. For the 2018–2019 influenza season, high-dose trivalent SIIV contained strains A/Michigan/45/2015 (H1N1)pdm09-like, A/Singapore/INFIMH-16-0019/2016 A(H3N2)-like, and B/Colorado/06/2017-like (Victoria lineage), and the quadrivalent SIIV additionally contained a B/Phuket/3073/2013-like (Yamagata lineage) strain. Placebo was a sterile 0.9% sodium chloride solution.
6
Influenza Vaccine Characterization Assay
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FluMist Quadrivalent, 2015–2016, live-attenuated vaccine (MedImmune) and Fluzone Quadrivalent, 2015–2016, inactivated virus vaccine (Sanofi Pasteur), both containing the four strains: (1) A/California/7/2009 (H1N1) pdm09, (2) A/Switzerland/9715293/2013 (H3N2), (3) B/Phuket/3073/2013, and (4) B/Brisbane/60/2008, were purchased from Moore Medical. Anti–CD3e (clone 145-2C11) and anti–CD28 (clone 37.51) mAbs were purchased as azide-free, low-endotoxin formulations from BD for use in T cell–activation assays. Fluorescently conjugated Abs for cell-surface phenotypic staining, cell sorting, and intracellular staining were purchased from BD, BioLegend, and eBioscience and are listed, including target, clone, color, and manufacturer, in Table S6. The influenza nucleoprotein-specific (NP366–374 ASNENMDAM) MHCI H-2Db tetramer; the influenza, RNA-dependent RNA polymerase PB1 subunit-specific (PB1703-711 SSYRRPVGI) MHCI H-2Kb tetramer; and the influenza HA-specific (HA533–541 IYSTVASSL) MHCI H-2Kd tetramer, all conjugated to PE, were purchased from MBL International. FTY720 was purchased from Cayman Chemical.
7
Composition of 2016-2017 Influenza Vaccines
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In accordance with US Food and Drug Administration guidance for composition of Northern Hemisphere 2016–2017 influenza vaccines, IIV4 (Fluzone Quadrivalent; Sanofi Pasteur) contained the A/California/07/2009 X-179A (H1N1), A/Hong Kong/4801/2014 X-263B (H3N2), B/Brisbane/60/2008 (Victoria lineage) and B/Phuket/3073/2013 (Yamagata lineage) strains. Each half dose (0.25 mL) was formulated to contain 7.5 μg hemagglutinin per strain, and each full dose (0.5 mL) was formulated to contain 15 μg hemagglutinin per strain. Study vaccines were supplied, respectively, in 0.25- or 0.5-mL, prefilled single-dose syringes.
8
Double-Dose Influenza Vaccine Evaluation
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Children in stable health were recruited in the United States and Mexico during the 2014–15 influenza season (Supplementary Appendix). The double-dose IIV4 (GSK Vaccines, Quebec, Canada) contained 15 μg HA of each of the 4 strains: A/California/7/2009 (A/H1N1), A/Texas/50/2012 (A/H3N2), B/Brisbane/60/2008 (B/Victoria), and B/Massachusetts/2/2012 (B/Yamagata). The standard-dose IIV4 (Fluzone Quadrivalent; Sanofi Pasteur, Swiftwater, PA) contained 7.5 μg of HA of each of the same strains.
Children were randomized 1:1 to double-dose or standard-dose IIV4. Allocation to a study group at the investigator site was performed using an internet-based randomization system (SBIR). The randomization algorithm used a minimization procedure to balance the composition of treatment groups, accounting for age (6–17 and 18–35 months), center, and influenza vaccine priming status. The study aimed to enroll 40%–50% of children in the 6–17 months age group. Children were considered vaccine-primed if they had received 2 or more doses of influenza vaccine since July 1, 2010 or at least 1 dose of the 2013–14 influenza vaccine. Vaccine-primed children received a single dose on day 0. Vaccine-unprimed children received 1 dose on day 0 and another on day 28.
Children were randomized 1:1 to double-dose or standard-dose IIV4. Allocation to a study group at the investigator site was performed using an internet-based randomization system (SBIR). The randomization algorithm used a minimization procedure to balance the composition of treatment groups, accounting for age (6–17 and 18–35 months), center, and influenza vaccine priming status. The study aimed to enroll 40%–50% of children in the 6–17 months age group. Children were considered vaccine-primed if they had received 2 or more doses of influenza vaccine since July 1, 2010 or at least 1 dose of the 2013–14 influenza vaccine. Vaccine-primed children received a single dose on day 0. Vaccine-unprimed children received 1 dose on day 0 and another on day 28.
9
RSVpreF Vaccine Formulation and Administration
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Each RSVpreF formulation contained equal amounts of recombinant RSV prefusion F trimer from subgroups A (Ontario) and B (Buenos Aires), totaling 60, 120, or 240 µg, formulated with or without 0.2 mg Al(OH)3. Lyophilized RSVpreF was reconstituted with either sterile water or Al(OH)3 in water as diluent and administered in a 0.5-mL injection volume; sentinel and expanded cohort participants receiving the vaccine were injected into the left deltoid. In the sentinel cohort, placebo (0.9% sodium chloride) was injected into the left deltoid. In the expanded cohort, placebo or SIIV (Fluzone Quadrivalent; Sanofi Pasteur) was injected into the right deltoid at vaccination 1 and into the deltoid of the nondominant arm at vaccination 2.
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