Childhood Antigen-Specific B-Cell and Cytokine Profiling

From a randomly selected subset of 20 longitudinal blood samples of the children aged 9 years, PT, FHA, Prn, and Td-specific enzyme-linked immunospot assays were performed on purified B-cells to determine the numbers of antigen-specific IgG producing memory B-cells (22 (link), 28 (link)). The same PBMC samples, depleted of CD19⁺ cells, were stimulated for 5 days with PT (heat inactivated), FHA, Prn, Td, or pokeweed mitogen and culture supernatants were stored at −80°C (23 (link)). In these supernatants, the cytokines interferon-gamma (IFN-γ) (Th1), interleukin-13 (IL-13) (Th2), IL-17 (Th17), and IL-10 (Treg) were quantified using the Bio-Plex cytokine assay kits (Bio-Rad Laboratories, Hercules, CA, USA) (27 (link)).

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van der Lee S., Hendrikx L.H., Sanders E.A., Berbers G.A, & Buisman A.M. (2018). Whole-Cell or Acellular Pertussis Primary Immunizations in Infancy Determines Adolescent Cellular Immune Profiles. Frontiers in Immunology, 9, 51.

Publication 2018

Bio-Plex cytokine assay kits

Antigen Assay B cells Blood Cells Children Cytokine Enzyme linked immunospot assays Ifn γ Il 10 Il 17 Interleukin 13 Memory Pokeweed mitogen Th17

Corresponding Organization : Wilhelmina Children's Hospital

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Variable analysis

independent variables

Purified B-cells stimulated with PT (heat inactivated), FHA, Prn, Td, or pokeweed mitogen

dependent variables

Numbers of antigen-specific IgG producing memory B-cells
Cytokines: interferon-gamma (IFN-γ) (Th1), interleukin-13 (IL-13) (Th2), IL-17 (Th17), and IL-10 (Treg)

control variables

Randomly selected subset of 20 longitudinal blood samples of the children aged 9 years

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