• CD134, also known as OX40 and TNFRSF4, is a 50 kD type I transmembrane protein, which is a member of the TNF receptor family.
• OX40 is expressed on activated T lymphocytes including Th1, Th2, Th17, and Treg cells and the interaction of OX40 with OX40L results in B cell proliferation and antibody secretion, regulation of primary T cell expansion, and T cell survival.
• CD252 is a 35 kD member of the TNF superfamily and is also known as OX40 ligand, OX40L, and CD134L. The OX40L is expressed on activated B cells and antigen presenting cells.
• The exons 1-5 of mouse OX40 gene that  encode the extracellular domain were replaced by human OX40 exons 1-5 in B-hOX40/hOX40L mice(C).
• The exons 2-3 of mouse Ox40l gene that  encode the extracellular region were replaced by human OX40L exons 2-3 in B-hOX40/hOX40L mice(C).
• Application: This product is used for pharmacodynamics and safety evaluation of  drugs.

Gene targeting strategy for B-hOX40/hOX40L mice(C).

The exons 1-5 of mouse OX40 gene that  encode the extracellular domain were replaced by human OX40 exons 1-5 in B-hOX40/hOX40L mice(C).

The exons 2-3 of mouse Ox40l gene that  encode the extracellular region were replaced by human OX40L exons 2-3 in B-hOX40/hOX40L mice(C).

Strain specific OX40 expression analysis in homozygous B-hOX40/hOX40L mice(C) by flow cytometry. Splenocytes were collected from wild-type BALB/cCrSlcNifdc mice(+/+) and homozygous B-hOX40/hOX40L mice(C) (H/H;H/H) stimulated with anti-CD3ε in vivo, and protein expression was analyzed with anti-mouse OX40 antibody (Biolegend, 119414)  and anti-human OX40 antibody (Biolegend, 350004) by flow cytometry. mOX40 was only detectable in T cells of wild-type BALB/cCrSlcNifdc mice. hOX40 was only detectable in T cells of homozygous B-hOX40/hOX40L mice(C), but not in wild-type BALB/cCrSlcNifdc mice.

Strain specific OX40L expression analysis in homozygous B-hOX40/hOX40L mice(C) by flow cytometry. Bone marrow cells were collected from wild-type BALB/cCrSlcNifdc mice(+/+) and homozygous B-hOX40/hOX40L mice(C)(H/H;H/H). BMDCs were induced from bone marrow cells and stimulated with LPS in vitro. Then BMDCs were analyzed with anti-mouse OX40L antibody (Biolegend, 108814)  and anti-human OX40L antibody (BD Horizon™, 563766) by flow cytometry. mOX40L was only detectable in BMDCs of wild-type BALB/cCrSlcNifdc mice. hOX40L was only detectable in BMDCs of homozygous B-hOX40/hOX40L mice, but not in wild-type BALB/cCrSlcNifdc mice.

Strain specific cytokines expression analysis in wild-type BALB/cCrSlcNifdc mice and homozygous humanized B-hOX40/0X40L mice(C) by ELISA. Serum was collected from wild-type BALB/cCrSlcNifdc mice (+/+) (female, n=2, 7-week-old) and homozygous B-hOX40/0X40L mice(C) (H/H;H/H)  (female, n=3, 7-week-old) stimulated with anti-mouse CD3ε antibody (37.5 μg/mL, 200 μL/mouse, i.p.) for 2 hrs in vivo. Expression level of IL4, IL13, IL2, IL22, IL17A, and IFN-γ were analyzed by ELISA. After mCD3ε stimulation, a significant increase of mouse IL4, IL13, IL2, IL22, IL17A, and IFN-γ were detected in BALB/cCrSlcNifdc mice (n=3) and homozygous B-hOX40/0X40L mice(C)  (n=3). Values are expressed as mean ± SEM.