The sheep erythrocyte receptor and both α and β chains of the human T-lymphocyte antigen receptor bind the mitogenic lectin (phytohaemagglutinin) from Phaseolus vulgaris

G. Leca, L. Boumsell, M. Fabbi, E. L. Reinherz, J. M. Kanellopoulos

Research output: Contribution to journalArticle

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Abstract

We have studied the interaction of mitogenic lectins such as phytohaemagglutinin (PHA) and concanavalin A (Con A) with both surface molecules which, by the use of monoclonal antibodies, are known to trigger T-cell mitogenesis. Monoclonal antibodies recognizing the T-lymphocyte receptor for antigen (Ti) and/or its associated structure, CD3, activate T cells. More recently, a second pathway of activation has been described which involves the sheep erythrocyte binding glycoprotein CD2, a surface molecule distinct from Ti-CD3. Lysates from surface-iodinated T-leukaemia cell lines were treated with lectin and affinity purified anti-lectin antibodies coupled to protein A-Sepharose. We have shown that eluates from Con A/anti-Con A or PHA/anti-PHA immunoprecipitates contained Ti, since a rabbit anti-Tα serum, which recognizes the native and denatured forms of the constant region of the α chain, immunoprecipitated Ti from these eluates. Furthermore, Ti immunoprecipitated by anti-Tα serum from lysates of surface iodinated E+ lymphocytes was binding to PHA after elution from the immunoprecipitate. When the purified Ti molecule was reduced and alkylated, allowing the permanent dissociation of its α and β subunits, PHA interacted with both chains, whereas anti-Tα serum immunoprecipitated the α chain only. Altogether, these results demonstrate that PHA interacts with both chains of the T cell receptor for antigen on human peripheral T lymphocytes. With the HPB-ALL tumour line, a similar approach showed that both α and β chains of Ti bind to Con A and Ulex europaeus 1 but not Helix pomatia. Affinity chomatography on immobilized lectins and immunoprecipitation with lectin/anti-lectin antibodies were employed to test whether CD2 binds to PHA and Con A. The results show that CD2 from human peripheral T lymphocytes binds both lectins but with a lower affinity for PHA than Con A.

Original languageEnglish
Pages (from-to)535-544
Number of pages10
JournalScandinavian Journal of Immunology
Volume23
Issue number5
Publication statusPublished - 1986

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Antigen Receptors
Viral Tumor Antigens
Phytohemagglutinins
Lectins
Sheep
Erythrocytes
T-Lymphocytes
Concanavalin A
Anti-Idiotypic Antibodies
Serum
Monoclonal Antibodies
Ulex
T-Cell Antigen Receptor
Immunoprecipitation
Glycoproteins
Leukemia
Lymphocytes
Rabbits
Cell Line

ASJC Scopus subject areas

  • Immunology

Cite this

The sheep erythrocyte receptor and both α and β chains of the human T-lymphocyte antigen receptor bind the mitogenic lectin (phytohaemagglutinin) from Phaseolus vulgaris. / Leca, G.; Boumsell, L.; Fabbi, M.; Reinherz, E. L.; Kanellopoulos, J. M.

In: Scandinavian Journal of Immunology, Vol. 23, No. 5, 1986, p. 535-544.

Research output: Contribution to journalArticle

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abstract = "We have studied the interaction of mitogenic lectins such as phytohaemagglutinin (PHA) and concanavalin A (Con A) with both surface molecules which, by the use of monoclonal antibodies, are known to trigger T-cell mitogenesis. Monoclonal antibodies recognizing the T-lymphocyte receptor for antigen (Ti) and/or its associated structure, CD3, activate T cells. More recently, a second pathway of activation has been described which involves the sheep erythrocyte binding glycoprotein CD2, a surface molecule distinct from Ti-CD3. Lysates from surface-iodinated T-leukaemia cell lines were treated with lectin and affinity purified anti-lectin antibodies coupled to protein A-Sepharose. We have shown that eluates from Con A/anti-Con A or PHA/anti-PHA immunoprecipitates contained Ti, since a rabbit anti-Tα serum, which recognizes the native and denatured forms of the constant region of the α chain, immunoprecipitated Ti from these eluates. Furthermore, Ti immunoprecipitated by anti-Tα serum from lysates of surface iodinated E+ lymphocytes was binding to PHA after elution from the immunoprecipitate. When the purified Ti molecule was reduced and alkylated, allowing the permanent dissociation of its α and β subunits, PHA interacted with both chains, whereas anti-Tα serum immunoprecipitated the α chain only. Altogether, these results demonstrate that PHA interacts with both chains of the T cell receptor for antigen on human peripheral T lymphocytes. With the HPB-ALL tumour line, a similar approach showed that both α and β chains of Ti bind to Con A and Ulex europaeus 1 but not Helix pomatia. Affinity chomatography on immobilized lectins and immunoprecipitation with lectin/anti-lectin antibodies were employed to test whether CD2 binds to PHA and Con A. The results show that CD2 from human peripheral T lymphocytes binds both lectins but with a lower affinity for PHA than Con A.",
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