Common name
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B-HLA-A2.1/hNY-ESO-1 MC38
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Catalog number
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322354
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Aliases
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HLA, NY-ESO-1
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Disease
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Colon carcinoma
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Organism
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Mouse
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Strain
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C57BL/6
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Tissue types
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Colon
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Tissue
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Colon
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Description
The B2M gene (Exon2 to Exon3) of mouse were replaced by the sequence encompassing the human B2M CDS, HLA-A*0201 gene that included leader sequence, α1 and α2 domains ligated to a fragment of the murine H-2Db gene containing the α3, transmembrane and cytoplasmic domains and NY-ESO-1 CDS. Human HLA-A2.1 is highly expressed on the surface of B-HLA-A2.1/hNY-ESO-1 MC38.
Application
B-HLA-A2.1/hNY-ESO-1 MC38 cells have the capability to establish tumors
in vivo and can be used for efficacy studies.
Targeting strategy
Gene targeting strategy for B-HLA-A2.1/hNY-ESO-1 MC38. The B2M gene (Exon2 to Exon3) of mouse were replaced by the sequence encompassing the human B2M CDS, HLA-A*0201 gene that included leader sequence, α1 and α2 domains ligated to a fragment of the murine H-2Db gene containing the α3, transmembrane and cytoplasmic domains and NY-ESO-1 CDS. Human HLA-A2.1 is highly expressed on the surface of B-HLA-A2.1/hNY-ESO-1 MC38.
Protein expression analysis
HLA-A2.1 expression analysis in B-HLA-A2.1/NY-ESO-1 MC38 by flow cytometry. Single cell suspensions from wild-type MC38 and B-HLA-A2.1/NY-ESO-1 MC38 cultures were stained with species-specific anti-HLA-A2.1 antibody. Human HLA-A2.1 was detected on the surface of B-HLA-A2.1/NY-ESO-1 MC38 cells but not wild-type MC38 cells. The 3-D04 clone of B-HLA-A2.1/NY-ESO-1 MC38 cells was used for in vivo experiments.
Tumor growth curve & Body weight changes
Subcutaneous homograft tumor growth of B-HLA-A2.1/hNY-ESO-1 MC38 cells. B-HLA-A2.1/hNY-ESO-1 MC38 cells (5x105) were subcutaneously implanted into B-HLA-A2.1 mice (male, 7-week-old, n=6). Tumor volume and body weight were measured three times a week. (A) Average tumor volume ± SEM. (B) Body weight (Mean± SEM). Volume was expressed in mm3 using the formula: V=0.5 X long diameter X short diameter2. As shown in panel AB-HLA-A2.1/hNY-ESO-1 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies.
Protein expression analysis of tumor cells
B-HLA-A2.1/hNY-ESO-1 MC38 cells were subcutaneously transplanted into B-HLA-A2.1 mice (n=6), and on 24 days post inoculation, tumor cells were harvested and assessed for human HLA-A2.1 expression by flow cytometry and NY-ESO-1 expression by western blot, respectively. As shown, human HLA-A2.1 was highly expressed on the surface of tumor cells(A). Human NY-ESO-1 was highly expressed(B). Therefore, B-HLA-A2.1/hNY-ESO-1 MC38 cells can be used for in vivo efficacy studies of novel NY-ESO-1 therapeutics.