Preclinical Double-Humanized CD3ε/CD28 Mouse Model

Humanized Immune Checkpoint Mouse Models

Design of the hCD3ε/hCD28 mouse

The double-humanized CD3ε/CD28 model was generated from intercrossing hCD3ε and hCD28 mice.

The CD3ε humanized model was developed to express the human epitope of the CD3ε chain, which is recognized by T-cell engagers (clone SP34). The rest of the extracellular domain is murine to preserve the amino acids involved in the interaction with CD3γ and CD3δ. Similarly, the transmembrane domains and the intracellular domains are murine to enable salt bridge interaction, interaction with the CD3ζ, and the signaling into mouse cells.

The humanized CD28 model, developed by Knockin at the mouse CD28 locus, expresses a chimeric CD28 with a human extracellular and murine transmembrane and intracellular domains. hCD28 expression displays a physiological regulation and expression pattern. This double-humanized model enables assessment of both compounds targeting human CD3ε and/or human CD28 in immunocompetent mice.

Applications in immuno-oncology

The hCD3ε/hCD28 mouse enables the in vivo efficacy assessment and profiling of immuno-oncology agents targeting the human T-cell engager CD3ε(SP-34) and/or the immune checkpoint CD28 in fully immunocompetent mice.


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hCD3ε/hCD28 features

  • Extracellular CD3ε N-terminal epitope is humanized
  • Conserved and functional interaction of CD3γ, CD3δ and CD3ζ subunits
  • The CD28 extracellular domain is entirely humanized
  • Physiological regulation and expression pattern of CD3ε and human CD28
  • Fully functional mouse immune system

hCD3ε/hCD28 validation*

The model has been co-validated by our partners of leaders in immuno-oncology and immunotherapy.

hCD3ε/hCD28 model validation 1

Anti-CD3 and anti-CD28 synergize to stimulate proliferation of CD8+ T cells

T cells from splenocytes are incubated in the presence of CD3 BsAb +/- CD28 BsAb. T-cell proliferation is measured 24h and 5 days post-activation, respectively.

* For more validation data, please consult our hCD28 and hCD3ε webpages, or  contact us.


Further reading


Ready to be shipped to your lab

  • Cohorts available upon request
  • Studies can be carried out at your site or at your favorite CRO
  • SOPF certification and worldwide delivery by professional breeders
  • Models provided with FTO on patent-protected technologies used for model generation

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