Common name
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B-HLA-A2.1/hMAGEA4 MC38
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Catalog number
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322404
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Aliases
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IMD43, AMYLD6, MHC1D4; HLAA; CT1.4,
MAGE4, MAGE4A, MAGE4B, MAGE-41, MAGE-X2
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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
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Origin: The MC38 cell line is derived from C57BL/6 murine colon adenocarcinoma cells. The cell line is a commonly used murine model for colorectal carcinoma.
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Background Information: HLA-A belongs to the HLA class I heavy chain paralogues. This class I molecule is a heterodimer consisting of a heavy chain and a light chain (beta-2 microglobulin). The heavy chain is anchored in the membrane. Class I molecules play a central role in the immune system by presenting peptides derived from the endoplasmic reticulum lumen so that they can be recognized by cytotoxic T cells. MAGEA4 is a member of the MAGEA gene family. The MAGEA genes are clustered at chromosomal location Xq28. They have been implicated in some hereditary disorders, such as dyskeratosis congenita.
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Gene targeting strategy: 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 MAGEA4 CDS. Human HLA-A2.1 is highly expressed on the surface of B-HLA-A2.1/hMAGEA4 MC38. Human MAGEA4 is highly expressed on the surface of B-HLA-A2.1/hMAGEA4 MC38 cells.
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Application: B-HLA-A2.1/hMAGEA4 MC38 tumor models can be used for preclinical evaluation of cancer vaccines.
Protein expression analysis
HLA-A2.1 and MAGEA4 expression analysis in B-HLA-A2.1/hMAGEA4 MC38 cells by flow cytometry, respectively. Single cell suspensions from wild-type MC38 and B-HLA-A2.1/hMAGEA4 MC38 #1-F03 cultures were detected with species-specific anti-HLA-A2.1 antibody (Biolegend, 343306) and anti-human MAGEA4 (Invitrogen, 35-6300), respectively. Human HLA-A2.1 was detected on the surface of B-HLA-A2.1/hMAGEA4 MC38 cells but not wild-type MC38 cells(A). Human MAGEA4 was detected in the B-HLA-A2.1/hMAGEA4 MC38 cells but not wild-type MC38 cells(B).
Tumor growth curve & body weight changes
Subcutaneous tumor growth of B-HLA-A2.1/hMAGEA4 MC38 cells. B-HLA-A2.1/hMAGEA4 MC38 cells (1×106) were subcutaneously implanted into B-HLA-A2.1 mice (male, 7-week-old, n=6). Tumor volume and body weight were measured twice a week. (A) Average tumor volume. (B) Body weight. Volume was expressed in mm3 using the formula: V=0.5 × long diameter × short diameter2. Results indicate that B-HLA-A2.1/hMAGEA4 MC38 cells were able to establish tumors in vivo and can be used for efficacy studies. Values are expressed as mean ± SEM.
Protein expression analysis of tumor tissue
HLA-A2.1 and MAGEA4 expression evaluated in B-HLA-A2.1/hMAGEA4 MC38 tumor cells by flow cytometry, respectively. B-HLA-A2.1/hMAGEA4 MC38 cells were subcutaneously transplanted into B-HLA-A2.1 mice (male, 7-week-old, n=6). Upon conclusion of the experiment, tumor cells were harvested and assessed with species-specific anti-HLA-A2.1 antibody (Biolegend, 343306) and anti-human MAGEA4 (Invitrogen, 35-6300), respectively. Human HLA-A2.1 was highly expressed on the surface of tumor cells(A). Human MAGEA4 was detected in the tumor cells(B). Therefore, B-HLA-A2.1/hMAGEA4 MC38 cells can be used for in vivo efficacy studies evaluating cancer vaccines.