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Phase I Clinical Trial of 4-1BB-based Adoptive T-Cell Therapy for Epstein-Barr Virus (EBV)-positive Tumors.

Eom HS, Choi BK, Lee Y, Lee H, Yun T, Kim YH, Lee JJ, Kwon BS - J. Immunother. (2016)

Bottom Line: Although adoptive cell therapy using Ag-specific T cells has been tested successfully in the clinic, the production of these T cells has been challenging.By applying our simple and practical 4-1BB-based method for the generation of Ag-specific CD8 T cells, here we determined the maximum tolerated dose, toxicity profile, immunologic responses, and clinical efficacy of autologous Epstein-Barr virus (EBV)/LMP2A-specific CD8 T cells (EBV-induced Natural T cell; EBViNT) in patients with relapsed/refractory EBV-positive tumors.The strength of the second wave was related to the efficacy of the treatment.

View Article: PubMed Central - PubMed

Affiliation: *Hematologic Oncology Clinic, Center for Specific Organs Cancer, National Cancer Center, Korea †Center for Clinical Trial, National Cancer Center, Korea ‡Cancer Immunology Branch, Division of Cancer Biology §Immune Cell Production Unit, Program for Immunotherapeutic Research, Research Institute and Hospital, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi ∥Department of Hematology-Oncology, Chonnam National University Medical School, Gwangju, Korea ¶Department of Medicine, Tulane University Health Sciences Center, New Orleans, LA.

ABSTRACT
Although adoptive cell therapy using Ag-specific T cells has been tested successfully in the clinic, the production of these T cells has been challenging. By applying our simple and practical 4-1BB-based method for the generation of Ag-specific CD8 T cells, here we determined the maximum tolerated dose, toxicity profile, immunologic responses, and clinical efficacy of autologous Epstein-Barr virus (EBV)/LMP2A-specific CD8 T cells (EBV-induced Natural T cell; EBViNT) in patients with relapsed/refractory EBV-positive tumors. This was a single-center, phase I, dose-escalation trial study evaluating 4 escalating dosing schedules of single injected EBViNT. CD8 T-cell responses against different LMP2A peptides in each patient were determined, and the most effective peptides were used to produce EBViNT. The produced autologous EBViNTs were single infused to patients with EBV-associated malignancy who had failed to standard treatments and were of HLA-A02 or A24 type. Of 11 patients enrolled, 8 patients received a single infusion of EBViNT: 4 with nasopharyngeal carcinomas, 1 with Hodgkin lymphoma, 2 with extranodal NK/T lymphomas, and 1 with diffuse large B-cell lymphoma. Single infusion of EBViNT was well tolerated by all the patients and generated objective antitumor responses in 3 of them. EBViNT infusion induced 2 waves of interferon-γ response: 1 approximately 1 week and the other 4-8 weeks after the treatment. The strength of the second wave was related to the efficacy of the treatment. The current trial shows that EBViNT therapy is safe and may provide a new option for treating EBV-positive recurrent cancer patients resistant to conventional therapy.

No MeSH data available.


Related in: MedlinePlus

Changes in cytokine levels of peripheral blood mononuclear cells (PBMCs) following infusion of Epstein Barr virus (EBV)-specific CD8+ T cells. Heparinized blood samples were collected from each patient at the indicated days before and after EBV-induced Natural T cell (EBViNT) infusion. The isolated PBMCs were plated on 48-well culture plate (5×105 cells/well) and simultaneously treated with DMSO (vehicle), PMA, and ionomycine (P/I) or 5 μg/mL of EBV/LMP2A peptides (peptides) for 48 hours. Cytokines in the culture supernatant were assessed using a CBA Kit (BD Bioscience) (A–F). Gray arrows represent infusion days.
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Figure 4: Changes in cytokine levels of peripheral blood mononuclear cells (PBMCs) following infusion of Epstein Barr virus (EBV)-specific CD8+ T cells. Heparinized blood samples were collected from each patient at the indicated days before and after EBV-induced Natural T cell (EBViNT) infusion. The isolated PBMCs were plated on 48-well culture plate (5×105 cells/well) and simultaneously treated with DMSO (vehicle), PMA, and ionomycine (P/I) or 5 μg/mL of EBV/LMP2A peptides (peptides) for 48 hours. Cytokines in the culture supernatant were assessed using a CBA Kit (BD Bioscience) (A–F). Gray arrows represent infusion days.

Mentions: We measured cytokine production in PBMCs that were stimulated with EBV/LMP2A peptides or PMA/ionomycin for 48 hours. Stimulation of PBMCs with the LMP2A peptides that were used to produce EBViNT did not increase the levels of any of the cytokines tested, namely IL-1β, IL-6, IL-10, IL-4, TNF, and IFN-γ, whereas pan-T-cell activation with PMA/ionomycin did induce cytokine production (Figs. 4A–F). It was notable that the pan-T-cell activation resulted in 2 waves of IFN-γ production: the first at 1 week postinfusion, and the second at around 4 weeks postinfusion. As IFN-γ production was induced by pan-T cell activation but not by the EBV/LMP2A peptides, the secondary IFN-γ expression was probably due to T cells that were reactive with various other tumor Ags, not the LMP2A peptides. It is interesting to note that as shown in patients NCC-04 and NCC-18, prognoses and clinical outcomes were improved when the secondary IFN-γ expression was strong and extended (Fig. 4F) and there was less production of inflammatory cytokines such as IL-1β, IL-6, IL-10, and IL-4 (Figs. 4A–D). Two waves of IFN-γ production were also seen in patients NCC-03 and NCC-13, but the second IFN-γ peak was not strong. These patients appeared not to have benefited from the EBViNT therapy (Fig. 4 and Table 2).


Phase I Clinical Trial of 4-1BB-based Adoptive T-Cell Therapy for Epstein-Barr Virus (EBV)-positive Tumors.

Eom HS, Choi BK, Lee Y, Lee H, Yun T, Kim YH, Lee JJ, Kwon BS - J. Immunother. (2016)

Changes in cytokine levels of peripheral blood mononuclear cells (PBMCs) following infusion of Epstein Barr virus (EBV)-specific CD8+ T cells. Heparinized blood samples were collected from each patient at the indicated days before and after EBV-induced Natural T cell (EBViNT) infusion. The isolated PBMCs were plated on 48-well culture plate (5×105 cells/well) and simultaneously treated with DMSO (vehicle), PMA, and ionomycine (P/I) or 5 μg/mL of EBV/LMP2A peptides (peptides) for 48 hours. Cytokines in the culture supernatant were assessed using a CBA Kit (BD Bioscience) (A–F). Gray arrows represent infusion days.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4834812&req=5

Figure 4: Changes in cytokine levels of peripheral blood mononuclear cells (PBMCs) following infusion of Epstein Barr virus (EBV)-specific CD8+ T cells. Heparinized blood samples were collected from each patient at the indicated days before and after EBV-induced Natural T cell (EBViNT) infusion. The isolated PBMCs were plated on 48-well culture plate (5×105 cells/well) and simultaneously treated with DMSO (vehicle), PMA, and ionomycine (P/I) or 5 μg/mL of EBV/LMP2A peptides (peptides) for 48 hours. Cytokines in the culture supernatant were assessed using a CBA Kit (BD Bioscience) (A–F). Gray arrows represent infusion days.
Mentions: We measured cytokine production in PBMCs that were stimulated with EBV/LMP2A peptides or PMA/ionomycin for 48 hours. Stimulation of PBMCs with the LMP2A peptides that were used to produce EBViNT did not increase the levels of any of the cytokines tested, namely IL-1β, IL-6, IL-10, IL-4, TNF, and IFN-γ, whereas pan-T-cell activation with PMA/ionomycin did induce cytokine production (Figs. 4A–F). It was notable that the pan-T-cell activation resulted in 2 waves of IFN-γ production: the first at 1 week postinfusion, and the second at around 4 weeks postinfusion. As IFN-γ production was induced by pan-T cell activation but not by the EBV/LMP2A peptides, the secondary IFN-γ expression was probably due to T cells that were reactive with various other tumor Ags, not the LMP2A peptides. It is interesting to note that as shown in patients NCC-04 and NCC-18, prognoses and clinical outcomes were improved when the secondary IFN-γ expression was strong and extended (Fig. 4F) and there was less production of inflammatory cytokines such as IL-1β, IL-6, IL-10, and IL-4 (Figs. 4A–D). Two waves of IFN-γ production were also seen in patients NCC-03 and NCC-13, but the second IFN-γ peak was not strong. These patients appeared not to have benefited from the EBViNT therapy (Fig. 4 and Table 2).

Bottom Line: Although adoptive cell therapy using Ag-specific T cells has been tested successfully in the clinic, the production of these T cells has been challenging.By applying our simple and practical 4-1BB-based method for the generation of Ag-specific CD8 T cells, here we determined the maximum tolerated dose, toxicity profile, immunologic responses, and clinical efficacy of autologous Epstein-Barr virus (EBV)/LMP2A-specific CD8 T cells (EBV-induced Natural T cell; EBViNT) in patients with relapsed/refractory EBV-positive tumors.The strength of the second wave was related to the efficacy of the treatment.

View Article: PubMed Central - PubMed

Affiliation: *Hematologic Oncology Clinic, Center for Specific Organs Cancer, National Cancer Center, Korea †Center for Clinical Trial, National Cancer Center, Korea ‡Cancer Immunology Branch, Division of Cancer Biology §Immune Cell Production Unit, Program for Immunotherapeutic Research, Research Institute and Hospital, National Cancer Center, Ilsandong-gu, Goyang, Gyeonggi ∥Department of Hematology-Oncology, Chonnam National University Medical School, Gwangju, Korea ¶Department of Medicine, Tulane University Health Sciences Center, New Orleans, LA.

ABSTRACT
Although adoptive cell therapy using Ag-specific T cells has been tested successfully in the clinic, the production of these T cells has been challenging. By applying our simple and practical 4-1BB-based method for the generation of Ag-specific CD8 T cells, here we determined the maximum tolerated dose, toxicity profile, immunologic responses, and clinical efficacy of autologous Epstein-Barr virus (EBV)/LMP2A-specific CD8 T cells (EBV-induced Natural T cell; EBViNT) in patients with relapsed/refractory EBV-positive tumors. This was a single-center, phase I, dose-escalation trial study evaluating 4 escalating dosing schedules of single injected EBViNT. CD8 T-cell responses against different LMP2A peptides in each patient were determined, and the most effective peptides were used to produce EBViNT. The produced autologous EBViNTs were single infused to patients with EBV-associated malignancy who had failed to standard treatments and were of HLA-A02 or A24 type. Of 11 patients enrolled, 8 patients received a single infusion of EBViNT: 4 with nasopharyngeal carcinomas, 1 with Hodgkin lymphoma, 2 with extranodal NK/T lymphomas, and 1 with diffuse large B-cell lymphoma. Single infusion of EBViNT was well tolerated by all the patients and generated objective antitumor responses in 3 of them. EBViNT infusion induced 2 waves of interferon-γ response: 1 approximately 1 week and the other 4-8 weeks after the treatment. The strength of the second wave was related to the efficacy of the treatment. The current trial shows that EBViNT therapy is safe and may provide a new option for treating EBV-positive recurrent cancer patients resistant to conventional therapy.

No MeSH data available.


Related in: MedlinePlus