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CD34 splice variant: an attractive marker for selection of gene-modified cells

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CD34 splice variant: an attractive marker for selection of gene-modified cells. / Fehse, B; Richters, A; Putimtseva-Scharf, K; Klump, H; Li, Z; Ostertag, W; Zander, A R; Baum, Christopher.

In: MOL THER, Vol. 1, No. 5 Pt 1, 05.2000, p. 448-456.

Research output: SCORING: Contribution to journal › SCORING: Journal article › Research › peer-review

Harvard

Fehse, B, Richters, A, Putimtseva-Scharf, K, Klump, H, Li, Z, Ostertag, W, Zander, AR & Baum, C 2000, 'CD34 splice variant: an attractive marker for selection of gene-modified cells', MOL THER, vol. 1, no. 5 Pt 1, pp. 448-456. https://doi.org/10.1006/mthe.2000.0068

APA

Fehse, B., Richters, A., Putimtseva-Scharf, K., Klump, H., Li, Z., Ostertag, W., Zander, A. R., & Baum, C. (2000). CD34 splice variant: an attractive marker for selection of gene-modified cells. MOL THER, 1(5 Pt 1), 448-456. https://doi.org/10.1006/mthe.2000.0068

Vancouver

Fehse B, Richters A, Putimtseva-Scharf K, Klump H, Li Z, Ostertag W et al. CD34 splice variant: an attractive marker for selection of gene-modified cells. MOL THER. 2000 May;1(5 Pt 1):448-456. https://doi.org/10.1006/mthe.2000.0068

Bibtex

@article{636a7bc19d284542b55ed8cbce38c8fd,
title = "CD34 splice variant: an attractive marker for selection of gene-modified cells",
abstract = "This study presents a promising selection system for gene-modified cells other than human hematopoietic progenitor and endothelial cells based on transgenic expression of human CD34. Three retrovirally transduced variants of CD34 were compared, differing in the length of their cytoplasmic domains. These were the full-length transmembrane protein (flCD34), a truncated form (tCD34) that is found as a naturally occurring splice variant and has a partial deletion of the cytoplasmic domain for signal transduction, and an engineered variant which is completely deprived of its cytoplasmic tail (dCD34). All three variants allowed selection of gene-modified cells using commercially available immunoaffinity technology. However, examination by flow cytometry as well as by Southern, Northern, and Western blot revealed that dCD34, as opposed to tCD34, is not stably anchored in the membrane and thus is expressed at low levels on the surface of transduced cells. Therefore, tCD34 was chosen as the more promising candidate for a clinically applicable cell surface marker. We show that gene-modified human primary T lymphocytes expressing tCD34 can be enriched to high purity (>95%) using clinically approved immunoaffinity columns. In addition, we demonstrate the utility of tCD34 for surface marking of murine hematopoietic cells in vivo, including primary T lymphocytes detected 9 weeks after bone marrow transplantation.",
keywords = "Animals, Antigens, CD34/biosynthesis, Antigens, Surface/biosynthesis, Blotting, Western, Bone Marrow/metabolism, Cell Separation/methods, DNA/analysis, DNA Primers/chemistry, DNA, Recombinant, Flow Cytometry, Gene Expression, Genetic Vectors, Humans, Immunomagnetic Separation, Jurkat Cells, Mice, Mice, Inbred C57BL, Nucleotide Mapping, Retroviridae/genetics, T-Lymphocytes/cytology, Transfection",
author = "B Fehse and A Richters and K Putimtseva-Scharf and H Klump and Z Li and W Ostertag and Zander, {A R} and Christopher Baum",
year = "2000",
month = may,
doi = "10.1006/mthe.2000.0068",
language = "English",
volume = "1",
pages = "448--456",
journal = "MOL THER",
issn = "1525-0016",
publisher = "NATURE PUBLISHING GROUP",
number = "5 Pt 1",

}

RIS

TY - JOUR

T1 - CD34 splice variant: an attractive marker for selection of gene-modified cells

AU - Fehse, B

AU - Richters, A

AU - Putimtseva-Scharf, K

AU - Klump, H

AU - Li, Z

AU - Ostertag, W

AU - Zander, A R

AU - Baum, Christopher

PY - 2000/5

Y1 - 2000/5

N2 - This study presents a promising selection system for gene-modified cells other than human hematopoietic progenitor and endothelial cells based on transgenic expression of human CD34. Three retrovirally transduced variants of CD34 were compared, differing in the length of their cytoplasmic domains. These were the full-length transmembrane protein (flCD34), a truncated form (tCD34) that is found as a naturally occurring splice variant and has a partial deletion of the cytoplasmic domain for signal transduction, and an engineered variant which is completely deprived of its cytoplasmic tail (dCD34). All three variants allowed selection of gene-modified cells using commercially available immunoaffinity technology. However, examination by flow cytometry as well as by Southern, Northern, and Western blot revealed that dCD34, as opposed to tCD34, is not stably anchored in the membrane and thus is expressed at low levels on the surface of transduced cells. Therefore, tCD34 was chosen as the more promising candidate for a clinically applicable cell surface marker. We show that gene-modified human primary T lymphocytes expressing tCD34 can be enriched to high purity (>95%) using clinically approved immunoaffinity columns. In addition, we demonstrate the utility of tCD34 for surface marking of murine hematopoietic cells in vivo, including primary T lymphocytes detected 9 weeks after bone marrow transplantation.

AB - This study presents a promising selection system for gene-modified cells other than human hematopoietic progenitor and endothelial cells based on transgenic expression of human CD34. Three retrovirally transduced variants of CD34 were compared, differing in the length of their cytoplasmic domains. These were the full-length transmembrane protein (flCD34), a truncated form (tCD34) that is found as a naturally occurring splice variant and has a partial deletion of the cytoplasmic domain for signal transduction, and an engineered variant which is completely deprived of its cytoplasmic tail (dCD34). All three variants allowed selection of gene-modified cells using commercially available immunoaffinity technology. However, examination by flow cytometry as well as by Southern, Northern, and Western blot revealed that dCD34, as opposed to tCD34, is not stably anchored in the membrane and thus is expressed at low levels on the surface of transduced cells. Therefore, tCD34 was chosen as the more promising candidate for a clinically applicable cell surface marker. We show that gene-modified human primary T lymphocytes expressing tCD34 can be enriched to high purity (>95%) using clinically approved immunoaffinity columns. In addition, we demonstrate the utility of tCD34 for surface marking of murine hematopoietic cells in vivo, including primary T lymphocytes detected 9 weeks after bone marrow transplantation.

KW - Animals

KW - Antigens, CD34/biosynthesis

KW - Antigens, Surface/biosynthesis

KW - Blotting, Western

KW - Bone Marrow/metabolism

KW - Cell Separation/methods

KW - DNA/analysis

KW - DNA Primers/chemistry

KW - DNA, Recombinant

KW - Flow Cytometry

KW - Gene Expression

KW - Genetic Vectors

KW - Humans

KW - Immunomagnetic Separation

KW - Jurkat Cells

KW - Mice

KW - Mice, Inbred C57BL

KW - Nucleotide Mapping

KW - Retroviridae/genetics

KW - T-Lymphocytes/cytology

KW - Transfection

U2 - 10.1006/mthe.2000.0068

DO - 10.1006/mthe.2000.0068

M3 - SCORING: Journal article

C2 - 10933966

VL - 1

SP - 448

EP - 456

JO - MOL THER

JF - MOL THER

SN - 1525-0016

IS - 5 Pt 1

ER -