Preclinical humanized genO‑hCD4 mouse model

Validation data

• Human CD4 replaces murine CD4. Homozygous genO‑hCD4 humanized mice display human-CD4 expression in PBMC. No murine CD4 expression is observed.

  Blood samples from wild-type and homozygous genO‑hCD4 humanized mice stained with a combination of mAbs (anti-murine CD4, anti-human CD4) and analyzed by flow cytometry.

  

• Unaltered immune cell distribution. Homozygous genO‑hCD4 humanized mice display same frequencies of B cells, NK cells and T cells in spleen.

  Splenocytes are stained with different mAb combinations to assess frequencies of lymphocyte subsets measured by flow cytometry. Black bars represent values for wild-type mice and hatched bars represent values for genO‑hCD4 humanized mice.

  

• Conserved cytokine expression profile. Homozygous genO‑hCD4 humanized mice display similar cytokine expression after ConA stimulation.

  IL-2, IFN-γ and IL-4 concentrations in culture supernatants of mouse splenocytes stimulated with Con A are measured by using a 3-Plex Luminex kit. Black bars represent values for wild-type mice and hatched bars represent data for genO‑hCD4 humanized mice.

  

• Similar Ig production. Homozygous genO‑hCD4 humanized mice display similar anti-KLH isotyping to wild-type mice.

  Anti-KLH specific IgG1, and IgG2b polyclonal antibodies are assessed by sandwich ELISA in the serum of mice at day 28 after immunization with KLH and CFA. Serum samples from wild-type (WT) or homozygous genO‑hCD4 humanized mice (HO CD4) were tested at different dilutions plotted against OD values.