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Cancer Immunology Research
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Intratumoral Plasmid IL12 Electroporation Therapy in Patients with Advanced Melanoma Induces Systemic and Intratumoral T-cell Responses

Samantha K. Greaney, Alain P. Algazi, Katy K. Tsai, Kathryn T. Takamura, Lawrence Chen, Christopher G. Twitty, Li Zhang, Alan Paciorek, Robert H. Pierce, Mai H. Le, Adil I. Daud and Lawrence Fong
Samantha K. Greaney
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
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  • ORCID record for Samantha K. Greaney
Alain P. Algazi
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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Katy K. Tsai
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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Kathryn T. Takamura
3OncoSec Medical Incorporated, San Diego, California.
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Lawrence Chen
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
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Christopher G. Twitty
3OncoSec Medical Incorporated, San Diego, California.
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Li Zhang
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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Alan Paciorek
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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Robert H. Pierce
3OncoSec Medical Incorporated, San Diego, California.
4University of Washington, Seattle, Washington.
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Mai H. Le
3OncoSec Medical Incorporated, San Diego, California.
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Adil I. Daud
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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  • For correspondence: Lawrence.Fong@ucsf.edu adil.daud@ucsf.edu
Lawrence Fong
1Division of Hematology and Oncology, University of California, San Francisco, San Francisco, California.
2Helen Diller Family Comprehensive Cancer Center, University of California, San Francisco, San Francisco, California.
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  • For correspondence: Lawrence.Fong@ucsf.edu adil.daud@ucsf.edu
DOI: 10.1158/2326-6066.CIR-19-0359 Published February 2020
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    Figure 1.

    Modulation of circulating T cells with tavo treatment. Frequencies of circulating immune cell subsets were assessed on serial PBMCs by flow cytometry. CD4 Teff (A), Tregs (B), PD-1+ CD4 Teff (C), PD-1+Ki67+ CD4 Teff (D), CD8+ T cells (E), PD-1+ CD8 T cells (F), PD-1+Ki67+ CD8+ T cells (G) were gated and expressed as a percentage of total lymphocytes (n = 20; Wilcoxon matched pairs test.). Pre-tx, pretreatment.

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    Figure 2.

    Antigen-specific T-cell responses following tavo therapy in patients with melanoma. IFNγ response of circulating PBMCs was measured by ELISpot. Specificity to gp100 (A), NY-ESO–1 (B), MAGE-A3 (C), and Melan-A/MART-1 (D) was tested pre- and posttreatment in response to the respective peptide pools. Each line represents a single subject (n = 24; Wilcoxon matched pairs test.) Antigen-specific T-cell responses following tavo therapy between clinical responders (R) and nonresponders (NR), with specificity to gp100 (E), NY-ESO–1 (F), MAGE-A3 (G), and Melan-A/MART-1 (H; n = 24). Lines indicate median ± interquartile range. Significance measured by Mann–Whitney U test. Pre-tx, pretreatment.

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    Figure 3.

    T-cell infiltration posttreatment. Comparison of CD3+ (A) and CD3+CD8+ (B) tumor-infiltrating lymphocytes (TIL) between clinical responders (R) and nonresponders (NR) pretreatment, 11 days after treatment, and 39 days after treatment. Each dot represents a single subject (n = 19). Treated lesions are denoted with filled circles, whereas untreated lesions are denoted with open circles. Lines indicate median ± interquartile range. Significance measured by Mann–Whitney U test.

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    Figure 4.

    Association between intratumoral T-cell infiltration and circulating immune responses. Correlation of CD3+CD8+ tumor-infiltrating lymphocytes (TIL) with antigen-specific IFNγ T-cell responses before treatment with plasmid IL12 electroporation (A, C, E, and G), and 39 days after treatment (B, D, F, and H) for all patients. Responders are denoted by filled circles, whereas nonresponders are denoted by open circles (n = 12). Best-fit lines are overlaid on the graphs. P and r values were determined by Spearman rank correlation.

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    Figure 5.

    Diversity and dynamics of TCR repertoire in tumors. TCR sequencing was performed on tumor biopsies prior to or following treatment (n = 9). A, Clonality of biopsy samples obtained pre- and posttreatment samples was assessed (P = 0.088). B, Convergent frequency of pre- and posttreatment samples was also assessed (P = 0.019). C, Morisita's distance (MD) was calculated between baseline and posttreatment time points. Boxplots indicate median ± interquartile range, lines represent SE. Significance measured by Wilcoxon matched pairs test. D, The correlation between convergent frequency and clonality was assessed (r = 0.80, P < 0.001; line fitted by a linear regression model).

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Cancer Immunology Research: 8 (2)
February 2020
Volume 8, Issue 2
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Intratumoral Plasmid IL12 Electroporation Therapy in Patients with Advanced Melanoma Induces Systemic and Intratumoral T-cell Responses
Samantha K. Greaney, Alain P. Algazi, Katy K. Tsai, Kathryn T. Takamura, Lawrence Chen, Christopher G. Twitty, Li Zhang, Alan Paciorek, Robert H. Pierce, Mai H. Le, Adil I. Daud and Lawrence Fong
Cancer Immunol Res February 1 2020 (8) (2) 246-254; DOI: 10.1158/2326-6066.CIR-19-0359

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Intratumoral Plasmid IL12 Electroporation Therapy in Patients with Advanced Melanoma Induces Systemic and Intratumoral T-cell Responses
Samantha K. Greaney, Alain P. Algazi, Katy K. Tsai, Kathryn T. Takamura, Lawrence Chen, Christopher G. Twitty, Li Zhang, Alan Paciorek, Robert H. Pierce, Mai H. Le, Adil I. Daud and Lawrence Fong
Cancer Immunol Res February 1 2020 (8) (2) 246-254; DOI: 10.1158/2326-6066.CIR-19-0359
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