• CXCL16 is a member of the chemokine superfamily, which could induce a strong chemotactic response and have a role in leukocytes migration. CXCL16 could also trigger calcium mobilization.
• The exons 2-5 of mouse Cxcl16 gene that encode extracellular domain and transmembrane domain is replaced by human counterparts in B-hCXCL16 mice. The genomic region of mouse Cxcl16 gene that encodes signal peptide is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric Cxcl16 expression is driven by endogenous mouse Cxcl16 promoter, while mouse Cxcl16 gene transcription and translation will be disrupted.
• CXCR6 is a receptor for the chemokine CXCL16, which is a 39 kD G-protein coupled chemokine receptor with seven transmembrane-spanning regions.
• The exons 2 of mouse Cxcr6 gene that encode the whole molecule (ATG to STOP codon were replaced by human counterparts in B-hCXCR6 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human CXCR6 expression is driven by endogenous mouse Cxcr6 promoter, while mouse Cxcr6 gene transcription and translation will be disrupted.

Gene targeting strategy for B-hCXCL16hCXCR6 mice.

The exons 2 of mouse Cxcr6 gene that encode the whole molecule (ATG to STOP codon were replaced by human counterparts in B-hCXCR6 mice. The promoter, 5’UTR and 3’UTR region of the mouse gene are retained. The human CXCR6 expression is driven by endogenous mouse Cxcr6 promoter, while mouse Cxcr6 gene transcription and translation will be disrupted.

The exons 2-5 of mouse Cxcl16 gene that encode extracellular domain and transmembrane domain is replaced by human counterparts in B-hCXCL16 mice. The genomic region of mouse Cxcl16 gene that encodes signal peptide is retained. The promoter, 5’UTR and 3’UTR region of the mouse gene are also retained. The chimeric Cxcl16 expression is driven by endogenous mouse Cxcl16 promoter, while mouse Cxcl16 gene transcription and translation will be disrupted.

Strain specific CXCR6 expression analysis in homozygous B-hCXCL16/hCXCR6 mice by flow cytometry. Splenocytes were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hCXCL16/hCXCR6 mice(H/H;H/H) and analyzed by flow cytometry with species-specific anti-CXCR6 antibody(Biolegend, 151105; Biolegend, 356003). mCXCR6 was detectable in T cells of wild-type mice, hCXCR6 was detectable in T cells of homozygous B-hCXCL16/hCXCR6 mice.

Strain specific CXCR6 expression analysis in homozygous B-hCXCL16/hCXCR6 mice by flow cytometry. Liver cells were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hCXCL16/hCXCR6 mice(H/H;H/H) and analyzed by flow cytometry with species-specific anti-CXCR6 antibody(Biolegend, 151105; Biolegend, 356003). mCXCR6 was detectable in T cells of wild-type mice, hCXCR6 was detectable in T cells of homozygous B-hCXCL16/hCXCR6 mice.

Strain specific CXCR6 expression analysis in homozygous B-hCXCL16/hCXCR6 mice by flow cytometry. Lung cells were collected from wild-type C57BL/6N mice (+/+) and homozygous B-hCXCL16/hCXCR6 mice(H/H;H/H) and analyzed by flow cytometry with species-specific anti-CXCR6 antibody(Biolegend, 151105; Biolegend, 356003). mCXCR6 was detectable in T cell of wild-type mice, hCXCR6 was detectable in T cells of homozygous B-hCXCL16/hCXCR6 mice.

Strain specific CXCL16 expression analysis in wild-type C57BL/6N mice and homozygous humanized B-hCXCL16/hCXCR6 mice by ELISA. Serum was collected from wild-type C57BL/6N mice (+/+) (male, n=3, 6-week-old) and heterozygous B-hCXCL16/hCXCR6 mice(H/H;H/H) (male, n=3, 6-week-old). Expression level of mouse and human CXCL16 were analyzed by ELISA (Human CXCL16 ELISA Kit : Abcam, ab187397; Mouse CXCL16 ELISA Kit : Abcam, ab197744). Mouse CXCL16 was exclusively detectable in wild-type C57BL/6N mice (n=3). Human CXCL16 was exclusively detectable in homozygous B-hCXCL16/hCXCR6 mice (n=3). Values are expressed as mean ± SEM.