Monoclonal antibodies which react with the T cell receptor γ/δ recognize different subsets of CD3+WT31- T lymphocytes

S. Ferrini, I. Prigione, C. Bottino, E. Ciccone, G. Tambussi, S. Mammoliti, L. Moretta, A. Moretta

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Abstract

A polyclonal CD3+4-8-WT31- cell line (termed SFG) was utilized for mice immunization in order to produce monoclonal antibodies (mAb) specific for the T cell receptor (TcR) γ/δ. Hybrid supernatants were screened for their ability to induce SFG cells (but not conventional TcR α/β+ CTL lines) to kill the murine Fc receptor-positive P815 target cell line. Three hybrids, termed G1, A13 and F11, were isolated according to this screening. By indirect immunofluorescence G1 mAb reacted with 65% of SFG cells, while A13 stained 26% and F11 75% of cells. Double-fluorescence analysis revealed that G1 and A13 mAb identify two distinct, non-overlapping subsets of cells present in the SFG cell line. The reactivity of the mAb was also analyzed on a panel of representative TcR γ/δ clones. G1 mAb reacted with 5 clones, that were also stained by the previously described BB3 mAb (recognizing the disulfide-linked form to TcR γ/δ). These clones failed to react with A13 and δ-TCS-1 mAb (tha latter of which is known to react with a non-disulfide-linked form of TcR γ/δ). Out of six clones that reacted with A13 mAb, four were also δ-TCS-1+, whereas two were δ-TCS-1- and none of them reacted with G1, (or BB3) mAb. In contrast to the mAb above, F11 brightly stained the G1+A13- clones and more weakly the G1-A13+ clones. Moreover, F11 efficiently triggered both types of clones to kill the P815 target cells while G1 and A13 were able to trigger only G1+ or A13+ clones, respectively. None of the mAb above reacted with a large number of CD3+WT31+ clones. Antibody-induced surface antigen modulation experiments indicated that molecules recognized by G1, A13 and F11 were physically associated on cell surface with CD3 determinants. In addition, immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis (performed on 125I-surface-labeled TcR γ/δ+ clones) revealed that molecules recognized by G1, A13 and F11 displayed an apparent mol. wt. corresponding to that of CD3-associated TcR molecules, immunoprecipitated by anti-CD3 mAb from the same clones.

Original languageEnglish
Pages (from-to)57-61
Number of pages5
JournalEuropean Journal of Immunology
Volume19
Issue number1
Publication statusPublished - 1989

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T-Cell Antigen Receptor
Monoclonal Antibodies
Clone Cells
T-Lymphocytes
Cell Line
Fc Receptors
Surface Antigens
Indirect Fluorescent Antibody Technique
Immunoprecipitation
Sodium Dodecyl Sulfate
Disulfides
Polyacrylamide Gel Electrophoresis
Immunization
Fluorescence

ASJC Scopus subject areas

  • Immunology

Cite this

@article{0ab44b9ea1dd4bfe9a2ae50271613b1a,
title = "Monoclonal antibodies which react with the T cell receptor γ/δ recognize different subsets of CD3+WT31- T lymphocytes",
abstract = "A polyclonal CD3+4-8-WT31- cell line (termed SFG) was utilized for mice immunization in order to produce monoclonal antibodies (mAb) specific for the T cell receptor (TcR) γ/δ. Hybrid supernatants were screened for their ability to induce SFG cells (but not conventional TcR α/β+ CTL lines) to kill the murine Fc receptor-positive P815 target cell line. Three hybrids, termed G1, A13 and F11, were isolated according to this screening. By indirect immunofluorescence G1 mAb reacted with 65{\%} of SFG cells, while A13 stained 26{\%} and F11 75{\%} of cells. Double-fluorescence analysis revealed that G1 and A13 mAb identify two distinct, non-overlapping subsets of cells present in the SFG cell line. The reactivity of the mAb was also analyzed on a panel of representative TcR γ/δ clones. G1 mAb reacted with 5 clones, that were also stained by the previously described BB3 mAb (recognizing the disulfide-linked form to TcR γ/δ). These clones failed to react with A13 and δ-TCS-1 mAb (tha latter of which is known to react with a non-disulfide-linked form of TcR γ/δ). Out of six clones that reacted with A13 mAb, four were also δ-TCS-1+, whereas two were δ-TCS-1- and none of them reacted with G1, (or BB3) mAb. In contrast to the mAb above, F11 brightly stained the G1+A13- clones and more weakly the G1-A13+ clones. Moreover, F11 efficiently triggered both types of clones to kill the P815 target cells while G1 and A13 were able to trigger only G1+ or A13+ clones, respectively. None of the mAb above reacted with a large number of CD3+WT31+ clones. Antibody-induced surface antigen modulation experiments indicated that molecules recognized by G1, A13 and F11 were physically associated on cell surface with CD3 determinants. In addition, immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis (performed on 125I-surface-labeled TcR γ/δ+ clones) revealed that molecules recognized by G1, A13 and F11 displayed an apparent mol. wt. corresponding to that of CD3-associated TcR molecules, immunoprecipitated by anti-CD3 mAb from the same clones.",
author = "S. Ferrini and I. Prigione and C. Bottino and E. Ciccone and G. Tambussi and S. Mammoliti and L. Moretta and A. Moretta",
year = "1989",
language = "English",
volume = "19",
pages = "57--61",
journal = "European Journal of Immunology",
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TY - JOUR

T1 - Monoclonal antibodies which react with the T cell receptor γ/δ recognize different subsets of CD3+WT31- T lymphocytes

AU - Ferrini, S.

AU - Prigione, I.

AU - Bottino, C.

AU - Ciccone, E.

AU - Tambussi, G.

AU - Mammoliti, S.

AU - Moretta, L.

AU - Moretta, A.

PY - 1989

Y1 - 1989

N2 - A polyclonal CD3+4-8-WT31- cell line (termed SFG) was utilized for mice immunization in order to produce monoclonal antibodies (mAb) specific for the T cell receptor (TcR) γ/δ. Hybrid supernatants were screened for their ability to induce SFG cells (but not conventional TcR α/β+ CTL lines) to kill the murine Fc receptor-positive P815 target cell line. Three hybrids, termed G1, A13 and F11, were isolated according to this screening. By indirect immunofluorescence G1 mAb reacted with 65% of SFG cells, while A13 stained 26% and F11 75% of cells. Double-fluorescence analysis revealed that G1 and A13 mAb identify two distinct, non-overlapping subsets of cells present in the SFG cell line. The reactivity of the mAb was also analyzed on a panel of representative TcR γ/δ clones. G1 mAb reacted with 5 clones, that were also stained by the previously described BB3 mAb (recognizing the disulfide-linked form to TcR γ/δ). These clones failed to react with A13 and δ-TCS-1 mAb (tha latter of which is known to react with a non-disulfide-linked form of TcR γ/δ). Out of six clones that reacted with A13 mAb, four were also δ-TCS-1+, whereas two were δ-TCS-1- and none of them reacted with G1, (or BB3) mAb. In contrast to the mAb above, F11 brightly stained the G1+A13- clones and more weakly the G1-A13+ clones. Moreover, F11 efficiently triggered both types of clones to kill the P815 target cells while G1 and A13 were able to trigger only G1+ or A13+ clones, respectively. None of the mAb above reacted with a large number of CD3+WT31+ clones. Antibody-induced surface antigen modulation experiments indicated that molecules recognized by G1, A13 and F11 were physically associated on cell surface with CD3 determinants. In addition, immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis (performed on 125I-surface-labeled TcR γ/δ+ clones) revealed that molecules recognized by G1, A13 and F11 displayed an apparent mol. wt. corresponding to that of CD3-associated TcR molecules, immunoprecipitated by anti-CD3 mAb from the same clones.

AB - A polyclonal CD3+4-8-WT31- cell line (termed SFG) was utilized for mice immunization in order to produce monoclonal antibodies (mAb) specific for the T cell receptor (TcR) γ/δ. Hybrid supernatants were screened for their ability to induce SFG cells (but not conventional TcR α/β+ CTL lines) to kill the murine Fc receptor-positive P815 target cell line. Three hybrids, termed G1, A13 and F11, were isolated according to this screening. By indirect immunofluorescence G1 mAb reacted with 65% of SFG cells, while A13 stained 26% and F11 75% of cells. Double-fluorescence analysis revealed that G1 and A13 mAb identify two distinct, non-overlapping subsets of cells present in the SFG cell line. The reactivity of the mAb was also analyzed on a panel of representative TcR γ/δ clones. G1 mAb reacted with 5 clones, that were also stained by the previously described BB3 mAb (recognizing the disulfide-linked form to TcR γ/δ). These clones failed to react with A13 and δ-TCS-1 mAb (tha latter of which is known to react with a non-disulfide-linked form of TcR γ/δ). Out of six clones that reacted with A13 mAb, four were also δ-TCS-1+, whereas two were δ-TCS-1- and none of them reacted with G1, (or BB3) mAb. In contrast to the mAb above, F11 brightly stained the G1+A13- clones and more weakly the G1-A13+ clones. Moreover, F11 efficiently triggered both types of clones to kill the P815 target cells while G1 and A13 were able to trigger only G1+ or A13+ clones, respectively. None of the mAb above reacted with a large number of CD3+WT31+ clones. Antibody-induced surface antigen modulation experiments indicated that molecules recognized by G1, A13 and F11 were physically associated on cell surface with CD3 determinants. In addition, immunoprecipitation followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis (performed on 125I-surface-labeled TcR γ/δ+ clones) revealed that molecules recognized by G1, A13 and F11 displayed an apparent mol. wt. corresponding to that of CD3-associated TcR molecules, immunoprecipitated by anti-CD3 mAb from the same clones.

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