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 language | English |
|---|---|
| Pages (from-to) | 535-544 |
| Number of pages | 10 |
| Journal | Scandinavian Journal of Immunology |
| Volume | 23 |
| Issue number | 5 |
| Publication status | Published - 1986 |
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- Immunology
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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 journal › Article
}
TY - JOUR
T1 - 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
AU - Leca, G.
AU - Boumsell, L.
AU - Fabbi, M.
AU - Reinherz, E. L.
AU - Kanellopoulos, J. M.
PY - 1986
Y1 - 1986
N2 - 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.
AB - 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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M3 - Article
C2 - 3085210
AN - SCOPUS:0022494569
VL - 23
SP - 535
EP - 544
JO - Scandinavian Journal of Immunology
JF - Scandinavian Journal of Immunology
SN - 0300-9475
IS - 5
ER -