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The autocrine production of transforming growth factor-beta1 during lymphocyte activation. A study with a monoclonal antibody-based ELISA.
Abstract
We describe the production and characterization of three mAb to transforming growth factor-beta (TGF-beta) and the use of two of them for the development of a TGF-beta 1-specific ELISA and for the study of the regulation of immune function in vitro. All three mAb bound recombinant human TGF-beta 1 (rHuTGF-beta 1) with high affinity and recognized the dimer form of this molecule in immunoblots. mAb 2G7 immunoprecipitated rHuTGF-beta 1, TGF-beta 2, and rHuTGF-beta 3 and neutralized the growth inhibitory activity of all three molecules in vitro on mink lung epithelial-like cells, Mv1Lu, indicating a shared neutralization epitope. mAb 4A11 neutralized and immunoprecipitated only rHuTGF-beta 1, and mAb 12H5 immunoprecipitated rHuTGF-beta 1 but had no effect on the bioactivity of either rHuTGF-beta 1, TGF-beta 2, or rHuTGF-beta 3. These results suggest that a second neutralization epitope may be unique to TGF-beta 1. The ELISA was developed with mAb 4A11 and 12H5, with a range of 0.63 to 40 ng/ml, i.e., a sensitivity of 0.63 ng/ml or 63 pg/sample. The assay is accurate, precise, and specific for the active but not the inactive or latent TGF-beta 1 complex and fails to react with TGF-beta 2, rHuTGF-beta 3, inhibin A, and activin A. Supernatants obtained from serum-free cultures of human PBMC from multiple donors contained significant quantities of TGF-beta 1 (3 to 15 ng/ml), which was detected in the ELISA only after pH 2 treatment to convert latent TGF-beta to the active form. Treatment of the PBMC with either recombinant human IL-2 (rHuIL-2) or PHA-P/PMA enhanced the production of latent TGF-beta 1. mAb 4A11 and 2G7, but not mAb 12H5 enhanced both the proliferative response of PBMC to rHuIL-2/rHuTNF-alpha and PHA-P and the development of the rHuIL-2/rHuTNF-alpha treated PBMC into LAK cells with cytotoxic activity against COLO target cells. These findings suggest that although PBMC secrete latent TGF-beta 1, mechanisms that convert the latent TGF-beta complex into an active form exist in vitro and that the endogenously produced TGF-beta can regulate immune functions in an autocrine fashion.
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Cited by
Bakin, A. V., Tomlinson, A. K., Bhowmick, N. A., Moses, H. L., & Arteaga, C. L. (2000) · Journal of Biological Chemistry · doi:10.1074/jbc.m005912200 · Cited by 992
TGF-β1 maintains suppressor function and Foxp3 expression in CD4+CD25+ regulatory T cells
Marie, J. C., Letterio, J. J., Gavin, M., & Rudensky, A. Y. (2005) · The Journal of Experimental Medicine · doi:10.1084/jem.20042276 · Cited by 985
Ziyadeh, F. N., Hoffman, B. B., Han, D. C., Iglesias-de la Cruz, M. C., Hong, S. W., Isono, M., Chen, S., McGowan, T. A., & Sharma, K. (2000) · Proceedings of the National Academy of Sciences · doi:10.1073/pnas.120055097 · Cited by 907
Mechanisms of Immune Evasion by Tumors
Drake, C. G., Jaffee, E., & Pardoll, D. M. (2006) · Advances in Immunology · doi:10.1016/s0065-2776(06)90002-9 · Cited by 688
Belghith, M., Bluestone, J. A., Barriot, S., Mégret, J., Bach, J.-F., & Chatenoud, L. (2003) · Nature Medicine · doi:10.1038/nm924 · Cited by 586
Metadata
- Authors
- Lucas, C., Bald, L. N., Fendly, B. M., Mora-Worms, M., Figari, I. S., Patzer, E. J., & Palladino, M. A.
- Venue
- The Journal of Immunology
- Publisher
- Oxford University Press (OUP)
- Published
- 1990-09-01
- Volume
- 145
- Issue
- 5
- Pages
- 1415-1422
- ISSN
- 0022-1767 (ISSN-L)
- 1550-6606
- OpenAlex
- W1517534438
- Cited by
- 164
- References
- 0
- FWCI
- 5.94
- Type
- journal article
- Language
- en
- Open access
- closed
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- CoChat. (2026). Citation check: The autocrine production of transforming growth factor-beta1 during lymphocyte activation. A study with a monoclonal antibody-based ELISA. https://app.cochat.ai/papers/10.4049/jimmunol.145.5.1415
- The paper
- Lucas, C., Bald, L. N., Fendly, B. M., Mora-Worms, M., Figari, I. S., Patzer, E. J., & Palladino, M. A. (1990). The autocrine production of transforming growth factor-beta1 during lymphocyte activation. A study with a monoclonal antibody-based ELISA. The Journal of Immunology. https://doi.org/10.4049/jimmunol.145.5.1415
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