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Updated: Jun 25, 2025
In Vivo Immunogenicity Screening of Tumor-Derived Extracellular Vesicles by Flow Cytometry of Splenic T Cells
Published on: September 23, 2021
Immune cell populations in the tumour environment following calcium electropora-tion for cutaneous metastasis: a
Mille Vissing1, Sandra Sinius Pouplier2, Lars Munch Larsen3
1Centre for Experimental Drug and Gene Electrotransfer (C*EDGE), Department of Clinical Oncology and Palliative Care, Zealand University Hospital, Roskilde and Næstved, Næstved, Denmark; Department of Clinical Medicine, Faculty of Health and Medical Sciences, University of Copenhagen, Copenhagen, Denmark.
Background And Purpose:
Calcium electroporation (CaEP) involves injecting calcium into tumour tissues and using electrical pulses to create membrane pores that induce cell death. This study assesses resultant immune responses and histopathological changes in patients with cutaneous metastases.
Patients/Materials And Methods:
The aimed cohort comprised 24 patients with metastases exceeding 5 mm. Tumours were treated once with CaEP (day 0) or twice (day 28). Biopsies were performed on days 0 and 2, with additional samples on days 7, 28, 30, 35, 60, and 90 if multiple tumours were treated. The primary endpoint was the change in tumour-infiltrating lymphocytes (TILs) two days post-treatment, with secondary endpoints evaluating local and systemic immune responses via histopathological analysis of immune markers, necrosis, and inflammation.
Results:
Seventeen patients, with metastases primarily from breast cancer (14 patients), but also lung cancer (1), melanoma (1), and urothelial cancer (1), completed the study. Of the 49 lesions treated, no significant changes in TIL count or PD-L1 expression were observed. However, there was substantial necrosis and a decrease in FOXP3-expression (p = 0.0025) noted, with a slight increase in CD4+ cells but no changes in CD3, CD8, or CD20 expressions. Notably, four patients showed reduced tumour invasiveness, including one case of an abscopal response.
Interpretation:
This exploratory study indicates that CaEP can be an effective anti-tumour therapy potentially enhancing immunity. Significant necrosis and decreased regulatory lymphocytes were observed, although TIL count remained unchanged. Several patients exhibited clinical signs of immune response following treatment.
Insights
Calcium electroporation (CaEP) shows potential as an anti-tumour therapy by causing significant necrosis and decreasing regulatory lymphocytes, though tumour-infiltrating lymphocytes (TILs) did not change. Some patients displayed clinical signs of immune response.
Area of Science:
- Oncology
- Immunology
- Dermatology
Background:
- Calcium electroporation (CaEP) utilizes calcium injection and electrical pulses to induce cancer cell death.
- Investigating CaEP's impact on immune responses and histopathological changes in cutaneous metastases is crucial.
Purpose of the Study:
- To assess the immune responses and histopathological alterations in patients with cutaneous metastases treated with CaEP.
- To evaluate the efficacy of CaEP as an anti-tumour therapy.
Main Methods:
- A cohort of 24 patients with metastases >5 mm received one or two CaEP treatments.
- Tumour biopsies were collected at multiple time points post-treatment for analysis.
- Primary endpoint: change in tumour-infiltrating lymphocytes (TILs); Secondary endpoints: immune markers, necrosis, and inflammation.
Main Results:
- Seventeen patients completed the study, primarily with breast cancer metastases.
- CaEP induced substantial tumour necrosis and decreased FOXP3+ regulatory lymphocytes (p=0.0025).
- No significant changes in TIL count or PD-L1 expression were observed, but four patients showed reduced tumour invasiveness.
Conclusions:
- CaEP demonstrates potential as an effective anti-tumour therapy with immunomodulatory effects.
- The treatment led to significant necrosis and a reduction in regulatory lymphocytes.
- Clinical signs of immune response were observed in several patients, suggesting therapeutic promise.
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