TGFα‑PE38 enhances cytotoxic T‑lymphocyte killing of breast cancer cells

Wright,Stephen E.; Rewers‑Felkins,Kathleen A.; Quinlin,Imelda; Chowdhury,Nazrul I.; Ahmed,Jewel; Eldridge,Paul W.; Srivastava,Sanjay K.; Pastan,Ira

doi:10.3892/ol.2014.1969

TGFα‑PE38 enhances cytotoxic T‑lymphocyte killing of breast cancer cells

Authors:
- Stephen E. Wright
- Kathleen A. Rewers‑Felkins
- Imelda Quinlin
- Nazrul I. Chowdhury
- Jewel Ahmed
- Paul W. Eldridge
- Sanjay K. Srivastava
- Ira Pastan
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Affiliations: Women's Health Research Institute, Department of Internal Medicine, Texas Tech University Health Sciences Center, Amarillo, TX 79106, USA, Harrington Cancer Center, Amarillo, TX 79106, USA, Department of Biomedical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, Amarillo, TX 79106, USA, Laboratory of Molecular Biology, Center for Cancer Research, National Cancer Institute, National Institutes of Health, Bethesda, MD 20892‑4264, USA
Published online on: March 12, 2014 https://doi.org/10.3892/ol.2014.1969
Pages: 2113-2117

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Abstract

The aim of the present study was to determine whether the combination of two modalities of immunotherapy, targeting two different tumor antigens, may be feasible and non‑toxic, yet enhance the killing of a human breast cancer cell line. The first modality was tumor growth factor α‑Pseudomonas exotoxin 38 (TGFα‑PE38), which specifically targets and kills tumor cells that express the epidermal growth factor receptor. The second modality was mucin‑1 (MUC1)‑specific cytotoxic T lymphocytes (CTLs), generated by MUC1 stimulation of peripheral blood mononuclear cells, to target the human breast cancer cell line, MCF7. TGFα‑PE38 exhibited specific lysis of the MCF7 cells in a concentration‑ and time‑dependent manner. TGFα‑PE38 did not kill the normal hematopoietic stem cells or CTLs. Furthermore, TGFα‑PE38 was not inhibitory for the growth or differentiation of the normal human hematopoietic stem cells into erythroid and myeloid colonies. In addition, TGFα‑PE38 did not inhibit the killing function of CTLs, either when preincubated or co‑incubated with CTLs. Finally, therapeutic enhancement was observed, in that TGFα‑PE38 and CTLs were additive in the specific lysis of the MCF7 cells. These two modalities of immunotherapy may be beneficial for humans with breast cancer with or without other therapies, including autologous hematopoietic stem cell transplantation, specifically for purging cancer cells from hematopoietic stem cells prior to transplantation.

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1	Siegall CB, FitzGerald DJ and Pastan I: Selective killing of tumor cells using EGF or TGF alpha-Pseudomonas exotoxin chimeric molecules. Semin Cancer Biol. 1:345–350. 1990.PubMed/NCBI
2	Siegall CB, Xu YH, Chaudhary VK, Adhya S, Fitzgerald D and Pastan I: Cytotoxic activities of a fusion protein comprised of TGF alpha and Pseudomonas exotoxin. FASEB J. 3:2647–2652. 1989.PubMed/NCBI
3	Prestrelski SJ, Arakawa T, Wu CS, O’Neal KD, Westcott KR and Narhi LO: Solution structure and dynamics of epidermal growth factor and transforming growth factor alpha. J Biol Chem. 267:319–322. 1992.PubMed/NCBI
4	Chiron MF, Fryling CM and FitzGerald D: Furin-mediated cleavage of Pseudomonas exotoxin-derived chimeric toxins. J Biol Chem. 272:31707–31711. 1997.PubMed/NCBI
5	Phillips PC, Levow C, Catterall M, Colvin OM, Pastan I and Brem H: Transforming growth factor-α-Pseudomonas exotoxin fusion protein (TGF-α-PE38) treatment of subcutaneous and intracranial human glioma and medulloblastoma xenografts in athymic mice. Cancer Res. 54:1008–1015. 1994.
6	Wright SE, Khaznadar R, Wang Z, et al: Generation of MUC1-stimulated mononuclear cells using optimized conditions. Scand J Immunol. 67:24–29. 2008.PubMed/NCBI
7	Wright SE, Rewers-Felkins KA, Quinlin IS, et al: Adoptive immunotherapy of mucin1 expressing adenocarcinomas with mucin1 stimulated human peripheral blood mononuclear cells. Int J Mol Med. 9:401–404. 2002.PubMed/NCBI
8	Quinlin IS, Burnside JS, Dombrowski KE, Phillips CA, Dolby N and Wright SE: Context of MUC1 epitope: immunogenicity. Oncol Rep. 17:453–456. 2007.PubMed/NCBI
9	Wright SE, Kilinski L, Talib S, et al: Cytotoxic T lymphocytes from humans with adenocarcinomas stimulated by native MUC1 mucin and a mucin peptide mutated at a glycosylation site. J Immunother. 23:2–10. 2000. View Article : Google Scholar : PubMed/NCBI
10	Wright SE, Rewers-Felkins KA, Quinlin IS, et al: MHC-unrestricted lysis of MUC1-expressing cells by human peripheral blood mononuclear cells. Immunol Invest. 37:215–225. 2008. View Article : Google Scholar : PubMed/NCBI
11	Alajez NM, Schmielau J, Alter MD, Cascio M and Finn OJ: Therapeutic potential of a tumor-specific, MHC-unrestricted T-cell receptor expressed on effector cells of the innate and the adaptive immune system through bone marrow transduction and immune reconstitution. Blood. 105:4583–4589. 2005. View Article : Google Scholar
12	Kanof ME and Smith PD: Preparation of human mononuclear cell populations and subpopulations. Current Protocols in Immunology. Coligan JE: John Wiley & Sons, Inc; New York, NY: pp. 72009
13	Barnd DL, Lan MS, Metzgar RS and Finn OJ: Specific, major histocompatibility complex-unrestricted recognition of tumor-associated mucins by human cytotoxic T cells. Proc Natl Acad Sci USA. 86:7159–7163. 1989. View Article : Google Scholar
14	Roehm NW, Rodgers GH, Hatfield SM and Glasebrook AL: An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J Immunol Methods. 142:257–265. 1991. View Article : Google Scholar : PubMed/NCBI
15	Jost LM, Kirkwood JM and Whiteside TL: Improved short- and long-term XTT-based colorimetric cellular cytotoxicity assay for melanoma and other tumor cells. J Immunol Methods. 147:153–165. 1992. View Article : Google Scholar : PubMed/NCBI
16	Altman SA, Randers L and Rao G: Comparison of Trypan blue dye exclusion and fluorometric assays for mammalian cell viability determinations. Biotechnol Prog. 9:671–674. 1993. View Article : Google Scholar : PubMed/NCBI
17	Bode AM, Ma W-Y, Surh Y-J and Dong Z: Inhibition of epidermal growth factor-induced cell transformation and activator protein 1 activation by [6]-gingerol. Cancer Res. 61:850–853. 2001.
18	Vennstrom L, Bysell C, Bjorkelund H, Lundqvist H and Andersson K: Real-time viability assay based on 51Cr retention in adherent cells. Biotechniques. 44:237–240. 2008. View Article : Google Scholar : PubMed/NCBI
19	Wu MH, Liebowitz DN, Smith SL, Williams SF and Dolan ME: Efficient expression of foreign genes in human CD34+ hematopoietic precursor cells using electroporation. Gene Therapy. 8:3842001.
20	Petty AP, Wright SE, Rewers-Felkins KA, Yenderrozos MA, Vorderstrasse BA and Lindsey JS: Targeting migration inducting gene-7 inhibits carcinoma cell invasion, early primary tumor growth, and stimulates monocyte oncolytic activity. Mol Cancer Ther. 8:2412–2423. 2009. View Article : Google Scholar : PubMed/NCBI
21	Ross AA, Cooper BW, Lazarus HM, et al: Detection and viability of tumor cells in peripheral blood stem cell collections from breast cancer patients using immunocytochemical and clonogenic assay techniques. Blood. 82:2605–2610. 1993.PubMed/NCBI
22	Rill DR, Santana VM, Roberts WM, et al: Direct demonstration that autologous bone marrow transplantation for solid tumors can return a multiplicity of tumorigenic cells. Blood. 84:380–383. 1994.PubMed/NCBI
23	Braun S, Vogl FD, Naume B, et al: A pooled analysis of bone marrow micrometastasis in breast cancer. N Engl J Med. 353:793–802. 2005. View Article : Google Scholar : PubMed/NCBI