Abstract
Anti-CD3 recombinant diphtheria immunotoxin, A-dmDT390-bisFv(UCHT1), consists of the catalytic and translocation domains of diphtheria toxin fused to two single chain Fv fragments of an anti-CD3ɛ monoclonal antibody (UCHT1). A-dmDT390-bisFv(UCHT1) is capable of killing CD3+ T-lymphoma cells and normal T cells specifically in the femtomolar concentration range. To study pharmacology of A-dmDT390-bisFv(UCHT1) in patients with CD3+ T-cell lymphoma in a phase I clinical trial, (1) highly sensitive bioassay using Jurkat cells for measuring drug levels, (2) ELISA for measuring anti-DT antibody titer, and (3) 5-color FACS analysis method for measuring changes of subtype T-cell population were developed. In addition to evaluating drug efficacy and pharmacokinetics in patients, it is important to correlate pre-existing anti-DT antibody levels with maximum drug concentration in serum and extent of T-cell depletion because pre-existing anti-DT antibodies due to DPT (Diphtheria, Pertussis, and Tetanus) immunization can neutralize diphtheria immunotoxin. We observed that at the lowest treatment dose (2.5 μg/kg: twice daily for 4 days) A-dmDT390-bisFv(UCHT1) depletes greater than 99.0% of normal T cells in all six patients for a short period of time (2–3 days) and that there is no association of C max and extent of T-cell depletion with the pre-existing anti-DT antibody titer.
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References
Rizvi, M. A., Evens, A. M., Tallman, M. S., Nelson, B. P., Rosen, S. T. (2006) T-cell non-Hodgkin lymphoma. Blood 107, 1255–1264.
Hymes, K. B. (2007) Choices in the treatment of cutaneous T-cell lymphoma. Oncology (Williston Park) 21, 18–23.
Savage, K. J. (2007) Peripheral T-cell lymphomas. Blood Rev 21, 201–216.
Woo, J. H., Liu, J. S., Kang, S. H., Singh, R., Park, S. K., Su, Y., Ortiz, J., Neville, D. M., Jr., Willingham, M. C., Frankel, A. E. (2008) GMP production and characterization of the bivalent anti-human T cell immunotoxin, A-dmDT390-bisFv(UCHT1) for phase I/II clinical trials. Protein Expr Purif 58, 1–11.
Frankel, A. E., Zuckero, S. L., Mankin, A. A., Grable, M., Mitchell, K., Lee, Y. J., Neville, D. M., Woo, J. H. (2009) Anti-CD3 recombinant diphtheria immunotoxin therapy of cutaneous T cell lymphoma. Curr Drug Targets 10, 104–109.
Woo, J. H., Liu, Y. Y., Mathias, A., Stavrou, S., Wang, Z., Thompson, J., Neville, D. M., Jr. (2002) Gene optimization is necessary to express a bivalent anti-human anti-T cell immunotoxin in Pichia pastoris. Protein Expr Purif 25, 270–282.
Woo, J. H., Liu, Y. Y., Stavrou, S., Neville, D. M., Jr. (2004) Increasing secretion of a bivalent anti-T-cell immunotoxin by Pichia pastoris. Appl Environ Microbiol 70, 3370–3376.
Woo, J. H., Liu, Y. Y., Neville, D. M., Jr. (2006) Minimization of aggregation of secreted bivalent anti-human T cell immunotoxin in Pichia pastoris bioreactor culture by optimizing culture conditions for protein secretion. J Biotechnol 121, 75–85.
Woo, J. H., Neville, D. M., Jr. (2003) Separation of bivalent anti-T cell immunotoxin from Pichia pastoris glycoproteins by borate anion exchange. Biotechniques 35, 392–398.
Neville, D. M., Jr., Srinivasachar, K., Stone, R., Scharff, J. (1989) Enhancement of immunotoxin efficacy by acid-cleavable cross-linking agents utilizing diphtheria toxin and toxin mutants. J Biol Chem 264, 14653–14661.
Thompson, J., Hu, H., Scharff, J., Neville, D. M., Jr. (1995) An anti-CD3 single-chain immunotoxin with a truncated diphtheria toxin avoids inhibition by pre-existing antibodies in human blood. J Biol Chem 270, 28037–28041.
Hall, P. D., Virella, G., Willoughby, T., Atchley, D. H., Kreitman, R. J., Frankel, A. E. (2001) Antibody response to DT-GM, a novel fusion toxin consisting of a truncated diphtheria toxin (DT) linked to human granulocyte-macrophage colony stimulating factor (GM), during a phase I trial of patients with relapsed or refractory acute myeloid leukemia. Clin Immunol 100, 191–197.
Kivisakk, P., Mahad, D. J., Callahan, M. K., Trebst, C., Tucky, B., Wei, T., Wu, L., Baekkevold, E. S., Lassmann, H., Staugaitis, S. M., Campbell, J. J., Ransohoff, R. M. (2003) Human cerebrospinal fluid central memory CD4+ T cells: evidence for trafficking through choroid plexus and meninges via P-selectin. Proc Natl Acad Sci U S A 100, 8389–8394.
Gollob, J. A., Schnipper, C. P., Orsini, E., Murphy, E., Daley, J. F., Lazo, S. B., Frank, D. A., Neuberg, D., Ritz, J. (1998) Characterization of a novel subset of CD8(+) T cells that expands in patients receiving interleukin-12. J Clin Invest 102, 561–575.
Soler, D., Chapman, T. R., Poisson, L. R., Wang, L., Cote-Sierra, J., Ryan, M., McDonald, A., Badola, S., Fedyk, E., Coyle, A. J., Hodge, M. R., Kolbeck, R. (2006) CCR8 expression identifies CD4 memory T cells enriched for FOXP3+ regulatory and Th2 effector lymphocytes. J Immunol 177, 6940–6951.
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Woo, J.H., Lee, YJ., Neville, D.M., Frankel, A.E. (2010). Pharmacology of Anti-CD3 Diphtheria Immunotoxin in CD3 Positive T-Cell Lymphoma Trials. In: Yotnda, P. (eds) Immunotherapy of Cancer. Methods in Molecular Biology, vol 651. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-60761-786-0_10
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DOI: https://doi.org/10.1007/978-1-60761-786-0_10
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