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Expression of CD150 in tumors of the central nervous system: identification of a novel isoform.

Romanets-Korbut O, Najakshin AM, Yurchenko M, Malysheva TA, Kovalevska L, Shlapatska LM, Zozulya YA, Taranin AV, Horvat B, Sidorenko SP - PLoS ONE (2015)

Bottom Line: In addition to the full length mRNA of the mCD150 splice isoform, in glioma cells we found a highly expressed novel CD150 transcript (nCD150), containing an 83 bp insert.Both mCD150 and nCD150 cDNA variants did not contain any mutations and had the leader sequence.The nCD150 transcript was also detected in normal and malignant B lymphocytes, primary T cells, dendritic cells and macrophages; however, in glioma cells nCD150 was found to be the predominant CD150 isoform.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of signal transduction pathways, R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of NAS of Ukraine, Kyiv, Ukraine; CIRI, International Center for Infectiology Research, IbIV team, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; CNRS, UMR5308, Lyon, France; Université Lyon 1, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France.

ABSTRACT
CD150 (IPO3/SLAM) belongs to the SLAM family of receptors and serves as a major entry receptor for measles virus. CD150 is expressed on normal and malignant cells of the immune system. However, little is known about its expression outside the hematopoietic system, especially tumors of the central nervous system (CNS). Although CD150 was not found in different regions of normal brain tissues, our immunohistochemical study revealed its expression in 77.6% of human CNS tumors, including glioblastoma, anaplastic astrocytoma, diffuse astrocytoma, ependymoma, and others. CD150 was detected in the cytoplasm, but not on the cell surface of glioma cell lines, and it was colocalized with the endoplasmic reticulum and Golgi complex markers. In addition to the full length mRNA of the mCD150 splice isoform, in glioma cells we found a highly expressed novel CD150 transcript (nCD150), containing an 83 bp insert. The insert is derived from a previously unrecognized exon designated Cyt-new, which is located 510 bp downstream of the transmembrane region exon, and is a specific feature of primate SLAMF1. Both mCD150 and nCD150 cDNA variants did not contain any mutations and had the leader sequence. The nCD150 transcript was also detected in normal and malignant B lymphocytes, primary T cells, dendritic cells and macrophages; however, in glioma cells nCD150 was found to be the predominant CD150 isoform. Similarly to mCD150, cell surface expression of nCD150 allows wild type measles virus entry to the cell. Our data indicate that CD150 expression in CNS tumors can be considered a new diagnostic marker and potential target for novel therapeutic approaches.

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Cell lines transfected with mCD150 and nCD150 are susceptible to wild type measles virus infection.(A) Surface expression of mCD150 and nCD150 isoforms in HEK293T and U87 cell lines after transfection with respective plasmids measured by flow cytometry, after staining with anti-CD150 mAb IPO3. (B) Wild type MV (G954 strain) production in HEK293T cells transfected with mCD150 and nCD150, determined daily by plaque assay on Vero-SLAM cells, 24 h (1), 48 h (2), 72 h (3), 96h (4) and 120 h (5) post-infection. Vero-SLAM and non-transfected HEK293T cells were used as positive and negative control respectively. One of three independent experiments. The expression of both mCD150 and nCD150 isoforms on the surface of transfected cells allows the entry of wt measles virus to the cell.
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pone.0118302.g009: Cell lines transfected with mCD150 and nCD150 are susceptible to wild type measles virus infection.(A) Surface expression of mCD150 and nCD150 isoforms in HEK293T and U87 cell lines after transfection with respective plasmids measured by flow cytometry, after staining with anti-CD150 mAb IPO3. (B) Wild type MV (G954 strain) production in HEK293T cells transfected with mCD150 and nCD150, determined daily by plaque assay on Vero-SLAM cells, 24 h (1), 48 h (2), 72 h (3), 96h (4) and 120 h (5) post-infection. Vero-SLAM and non-transfected HEK293T cells were used as positive and negative control respectively. One of three independent experiments. The expression of both mCD150 and nCD150 isoforms on the surface of transfected cells allows the entry of wt measles virus to the cell.

Mentions: Our results demonstrated that CD150 protein remains in the cytoplasm of glial cells (Figs. 3A, 3B, 4), although both splice isoforms, mCD150 and nCD150, contain a leader sequence and a transmembrane domain that are essential for the receptor’s surface expression. Overexpression upon transfection of U87 and HEK293T cell lines with either nCD150 or mCD150 isoforms resulted in cell surface expression of both CD150 isoforms in these cell lines (Fig. 9A). Moreover, similarly to mCD150, the expression of nCD150 on cell surface allowed wt MV entry to the cell (Fig. 9B).


Expression of CD150 in tumors of the central nervous system: identification of a novel isoform.

Romanets-Korbut O, Najakshin AM, Yurchenko M, Malysheva TA, Kovalevska L, Shlapatska LM, Zozulya YA, Taranin AV, Horvat B, Sidorenko SP - PLoS ONE (2015)

Cell lines transfected with mCD150 and nCD150 are susceptible to wild type measles virus infection.(A) Surface expression of mCD150 and nCD150 isoforms in HEK293T and U87 cell lines after transfection with respective plasmids measured by flow cytometry, after staining with anti-CD150 mAb IPO3. (B) Wild type MV (G954 strain) production in HEK293T cells transfected with mCD150 and nCD150, determined daily by plaque assay on Vero-SLAM cells, 24 h (1), 48 h (2), 72 h (3), 96h (4) and 120 h (5) post-infection. Vero-SLAM and non-transfected HEK293T cells were used as positive and negative control respectively. One of three independent experiments. The expression of both mCD150 and nCD150 isoforms on the surface of transfected cells allows the entry of wt measles virus to the cell.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4339833&req=5

pone.0118302.g009: Cell lines transfected with mCD150 and nCD150 are susceptible to wild type measles virus infection.(A) Surface expression of mCD150 and nCD150 isoforms in HEK293T and U87 cell lines after transfection with respective plasmids measured by flow cytometry, after staining with anti-CD150 mAb IPO3. (B) Wild type MV (G954 strain) production in HEK293T cells transfected with mCD150 and nCD150, determined daily by plaque assay on Vero-SLAM cells, 24 h (1), 48 h (2), 72 h (3), 96h (4) and 120 h (5) post-infection. Vero-SLAM and non-transfected HEK293T cells were used as positive and negative control respectively. One of three independent experiments. The expression of both mCD150 and nCD150 isoforms on the surface of transfected cells allows the entry of wt measles virus to the cell.
Mentions: Our results demonstrated that CD150 protein remains in the cytoplasm of glial cells (Figs. 3A, 3B, 4), although both splice isoforms, mCD150 and nCD150, contain a leader sequence and a transmembrane domain that are essential for the receptor’s surface expression. Overexpression upon transfection of U87 and HEK293T cell lines with either nCD150 or mCD150 isoforms resulted in cell surface expression of both CD150 isoforms in these cell lines (Fig. 9A). Moreover, similarly to mCD150, the expression of nCD150 on cell surface allowed wt MV entry to the cell (Fig. 9B).

Bottom Line: In addition to the full length mRNA of the mCD150 splice isoform, in glioma cells we found a highly expressed novel CD150 transcript (nCD150), containing an 83 bp insert.Both mCD150 and nCD150 cDNA variants did not contain any mutations and had the leader sequence.The nCD150 transcript was also detected in normal and malignant B lymphocytes, primary T cells, dendritic cells and macrophages; however, in glioma cells nCD150 was found to be the predominant CD150 isoform.

View Article: PubMed Central - PubMed

Affiliation: Laboratory of signal transduction pathways, R.E. Kavetsky Institute of Experimental Pathology, Oncology and Radiobiology of NAS of Ukraine, Kyiv, Ukraine; CIRI, International Center for Infectiology Research, IbIV team, Université de Lyon, Lyon, France; Inserm, U1111, Lyon, France; CNRS, UMR5308, Lyon, France; Université Lyon 1, Lyon, France; Ecole Normale Supérieure de Lyon, Lyon, France.

ABSTRACT
CD150 (IPO3/SLAM) belongs to the SLAM family of receptors and serves as a major entry receptor for measles virus. CD150 is expressed on normal and malignant cells of the immune system. However, little is known about its expression outside the hematopoietic system, especially tumors of the central nervous system (CNS). Although CD150 was not found in different regions of normal brain tissues, our immunohistochemical study revealed its expression in 77.6% of human CNS tumors, including glioblastoma, anaplastic astrocytoma, diffuse astrocytoma, ependymoma, and others. CD150 was detected in the cytoplasm, but not on the cell surface of glioma cell lines, and it was colocalized with the endoplasmic reticulum and Golgi complex markers. In addition to the full length mRNA of the mCD150 splice isoform, in glioma cells we found a highly expressed novel CD150 transcript (nCD150), containing an 83 bp insert. The insert is derived from a previously unrecognized exon designated Cyt-new, which is located 510 bp downstream of the transmembrane region exon, and is a specific feature of primate SLAMF1. Both mCD150 and nCD150 cDNA variants did not contain any mutations and had the leader sequence. The nCD150 transcript was also detected in normal and malignant B lymphocytes, primary T cells, dendritic cells and macrophages; however, in glioma cells nCD150 was found to be the predominant CD150 isoform. Similarly to mCD150, cell surface expression of nCD150 allows wild type measles virus entry to the cell. Our data indicate that CD150 expression in CNS tumors can be considered a new diagnostic marker and potential target for novel therapeutic approaches.

Show MeSH
Related in: MedlinePlus