Cultured human thymocytes lacking CD2 and CD11a/CD18 antigens are functional and adhere to endothelial cells via CD56 or CDw49d molecules

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Abstract

The preferential growth of CD3-CD2-CD1la/CD18- thymocytes was obtained by stimulation of CD2-CD3- thymic cells with low doses of PMA (0.5 ng/ml) and subsequent culture in the presence of recombinant interleukin-2 (100 U/ml). After 2-3 weeks, CD3-CD2-CD11a/CD18- thymocytes represented 40-60% of the total proliferating cells. Highly purified CD3-CD2-CD11a/CD18- cell populations were obtained by depletion of the CD11a/CD18+ thymocytes by immunomagnetic beads. Moreover, these populations proliferated for 2-5 weeks and did not change their surface phenotype. It is of note that these cells, despite the lack of CD2 and CD11a/CD18 adhesion molecules, could bind to umbilical vein endothelial cells as efficiently as did CD3+CD2+CD11a/CD18+ thymocytes. Furthermore we demonstrate that (a) CD56 molecule is involved in the adhesion of CD3-CD2-CD11a/CD18- thymic cells, but not of peripheral CD3-CD56+ lymphocytes, to untreated or IFN-γ- and/or TNF-α-treated endothelium, (b) anti-CDw49d mAb could inhibit the adhesion of this thymus-derived population to either IFN-γ- or TNF-α-treated endothelial cells but not to untreated endothelium, and (c) CD56 antigen expressed by these cultured thymocytes has a sialic acid content different from that of peripheral lymphocytes. Indeed, isoelectrofocusing analysis showed that CD56 molecule expressed on CD3-CD2-CD11a/CD18- thymocytes displayed an isoelectric point (pI5.0) different from that of CD56 antigen expressed by peripheral NK cells (pI4.7 and 5.4). Further, we noted that CD56 antigen showed the same pI 5.8 after desialylation obtained using neuraminidase treatment. Finally, CD3-CD2-CD11a/CD18- thymocytes mobilized Ca2+ and released TNF-α and IFN-γ after treatment with lectins.

Original languageEnglish
Pages (from-to)319-330
Number of pages12
JournalCellular Immunology
Volume140
Issue number2
DOIs
Publication statusPublished - Apr 1 1992

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CD11a Antigens
CD18 Antigens
Thymocytes
Endothelial Cells
CD56 Antigens
Endothelium
Lymphocytes
Population
Umbilical Veins
Isoelectric Point
Neuraminidase
N-Acetylneuraminic Acid
CDw49d
Lectins
Natural Killer Cells
Thymus Gland
Interleukin-2

ASJC Scopus subject areas

  • Cell Biology
  • Immunology

Cite this

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title = "Cultured human thymocytes lacking CD2 and CD11a/CD18 antigens are functional and adhere to endothelial cells via CD56 or CDw49d molecules",
abstract = "The preferential growth of CD3-CD2-CD1la/CD18- thymocytes was obtained by stimulation of CD2-CD3- thymic cells with low doses of PMA (0.5 ng/ml) and subsequent culture in the presence of recombinant interleukin-2 (100 U/ml). After 2-3 weeks, CD3-CD2-CD11a/CD18- thymocytes represented 40-60{\%} of the total proliferating cells. Highly purified CD3-CD2-CD11a/CD18- cell populations were obtained by depletion of the CD11a/CD18+ thymocytes by immunomagnetic beads. Moreover, these populations proliferated for 2-5 weeks and did not change their surface phenotype. It is of note that these cells, despite the lack of CD2 and CD11a/CD18 adhesion molecules, could bind to umbilical vein endothelial cells as efficiently as did CD3+CD2+CD11a/CD18+ thymocytes. Furthermore we demonstrate that (a) CD56 molecule is involved in the adhesion of CD3-CD2-CD11a/CD18- thymic cells, but not of peripheral CD3-CD56+ lymphocytes, to untreated or IFN-γ- and/or TNF-α-treated endothelium, (b) anti-CDw49d mAb could inhibit the adhesion of this thymus-derived population to either IFN-γ- or TNF-α-treated endothelial cells but not to untreated endothelium, and (c) CD56 antigen expressed by these cultured thymocytes has a sialic acid content different from that of peripheral lymphocytes. Indeed, isoelectrofocusing analysis showed that CD56 molecule expressed on CD3-CD2-CD11a/CD18- thymocytes displayed an isoelectric point (pI5.0) different from that of CD56 antigen expressed by peripheral NK cells (pI4.7 and 5.4). Further, we noted that CD56 antigen showed the same pI 5.8 after desialylation obtained using neuraminidase treatment. Finally, CD3-CD2-CD11a/CD18- thymocytes mobilized Ca2+ and released TNF-α and IFN-γ after treatment with lectins.",
author = "Alessandro Poggi and Zocchi, {Maria Raffaella}",
year = "1992",
month = "4",
day = "1",
doi = "10.1016/0008-8749(92)90199-Y",
language = "English",
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journal = "Cellular Immunology",
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T1 - Cultured human thymocytes lacking CD2 and CD11a/CD18 antigens are functional and adhere to endothelial cells via CD56 or CDw49d molecules

AU - Poggi, Alessandro

AU - Zocchi, Maria Raffaella

PY - 1992/4/1

Y1 - 1992/4/1

N2 - The preferential growth of CD3-CD2-CD1la/CD18- thymocytes was obtained by stimulation of CD2-CD3- thymic cells with low doses of PMA (0.5 ng/ml) and subsequent culture in the presence of recombinant interleukin-2 (100 U/ml). After 2-3 weeks, CD3-CD2-CD11a/CD18- thymocytes represented 40-60% of the total proliferating cells. Highly purified CD3-CD2-CD11a/CD18- cell populations were obtained by depletion of the CD11a/CD18+ thymocytes by immunomagnetic beads. Moreover, these populations proliferated for 2-5 weeks and did not change their surface phenotype. It is of note that these cells, despite the lack of CD2 and CD11a/CD18 adhesion molecules, could bind to umbilical vein endothelial cells as efficiently as did CD3+CD2+CD11a/CD18+ thymocytes. Furthermore we demonstrate that (a) CD56 molecule is involved in the adhesion of CD3-CD2-CD11a/CD18- thymic cells, but not of peripheral CD3-CD56+ lymphocytes, to untreated or IFN-γ- and/or TNF-α-treated endothelium, (b) anti-CDw49d mAb could inhibit the adhesion of this thymus-derived population to either IFN-γ- or TNF-α-treated endothelial cells but not to untreated endothelium, and (c) CD56 antigen expressed by these cultured thymocytes has a sialic acid content different from that of peripheral lymphocytes. Indeed, isoelectrofocusing analysis showed that CD56 molecule expressed on CD3-CD2-CD11a/CD18- thymocytes displayed an isoelectric point (pI5.0) different from that of CD56 antigen expressed by peripheral NK cells (pI4.7 and 5.4). Further, we noted that CD56 antigen showed the same pI 5.8 after desialylation obtained using neuraminidase treatment. Finally, CD3-CD2-CD11a/CD18- thymocytes mobilized Ca2+ and released TNF-α and IFN-γ after treatment with lectins.

AB - The preferential growth of CD3-CD2-CD1la/CD18- thymocytes was obtained by stimulation of CD2-CD3- thymic cells with low doses of PMA (0.5 ng/ml) and subsequent culture in the presence of recombinant interleukin-2 (100 U/ml). After 2-3 weeks, CD3-CD2-CD11a/CD18- thymocytes represented 40-60% of the total proliferating cells. Highly purified CD3-CD2-CD11a/CD18- cell populations were obtained by depletion of the CD11a/CD18+ thymocytes by immunomagnetic beads. Moreover, these populations proliferated for 2-5 weeks and did not change their surface phenotype. It is of note that these cells, despite the lack of CD2 and CD11a/CD18 adhesion molecules, could bind to umbilical vein endothelial cells as efficiently as did CD3+CD2+CD11a/CD18+ thymocytes. Furthermore we demonstrate that (a) CD56 molecule is involved in the adhesion of CD3-CD2-CD11a/CD18- thymic cells, but not of peripheral CD3-CD56+ lymphocytes, to untreated or IFN-γ- and/or TNF-α-treated endothelium, (b) anti-CDw49d mAb could inhibit the adhesion of this thymus-derived population to either IFN-γ- or TNF-α-treated endothelial cells but not to untreated endothelium, and (c) CD56 antigen expressed by these cultured thymocytes has a sialic acid content different from that of peripheral lymphocytes. Indeed, isoelectrofocusing analysis showed that CD56 molecule expressed on CD3-CD2-CD11a/CD18- thymocytes displayed an isoelectric point (pI5.0) different from that of CD56 antigen expressed by peripheral NK cells (pI4.7 and 5.4). Further, we noted that CD56 antigen showed the same pI 5.8 after desialylation obtained using neuraminidase treatment. Finally, CD3-CD2-CD11a/CD18- thymocytes mobilized Ca2+ and released TNF-α and IFN-γ after treatment with lectins.

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