Single-Cell Proteomics Identifies Upregulated Polyfunctionality in Expanded Tumor-Infiltrating Lymphocytes, Indicating Increased Anti-Tumor Activity

  • At the September 2020 ESMO Virtual Congress, data was presented in which IsoPlexis’ single-cell proteomics was used to assess the functionality of tumor-infiltrating lymphocytes (TILs) expanded using Iovance’s Gen2 protocol.
  • IsoPlexis’ platform found that TILs expanded for 22 days using the Gen2 protocol had upregulated polyfunctionality compared to endogenous TILs isolated from tumors, indicating increased anti-tumor behavior.
  • IsoPlexis’ single-cell secreted proteomics validated the efficacy of Iovance’s expansion protocol for reversing TIL dysfunction to treat cancers.

Researchers Presented Data Utilizing Functional Single-Cell Proteomics to Assess Anti-Tumor Function of Expanded TILs

IsoPlexis’ single-cell proteomics was highlighted in a presentation at the ESMO Virtual Congress in September 2020, titled “Iovance Generation-2 Tumor-infiltrating Lymphocyte (TIL) Product is Reinvigorated During the Manufacturing Process.”1 IsoPlexis’ functional proteomics assessed the functionality of TILs generated with Iovance’s protocol and compared them to endogenous TILs.

Adoptive cell therapy (ACT), a powerful method for treating cancers, may reverse dysfunction in endogenous TILs to improve their anti-tumor function. By isolating the TILs from the immunosuppressive tumor microenvironment, scientists can promote ex vivo expansion and later reinfuse these invigorated TILs into the patient. These reinvigorated tumor-specific T cells can drive a robust anti-tumor response after reinfusion. Researchers sought to determine the efficacy of one method of promoting TIL expansion, Iovance’s second generation (Gen2) protocol. IsoPlexis’ technology was used to compare the anti-tumor function of the T cells resulting from Iovance’s TIL expansion protocol to those isolated directly from the tumor.1

Highly Functional Subsets of TILs Correlate to Potent Anti-Tumor Response

Researchers isolated endogenous CD3+ TILs (D0) from digested tumor biopsies. These TILs were compared to TILs expanded for 22 days using Iovance’s Gen2 protocol (D22). The researchers looked at 7 pairs of D0 and D22 TILs from melanoma, cervical cancer, and head and neck squamous cell carcinoma. Ex vivo expansion with the Iovance Gen2 protocol restored the ability of TILs to secrete multiple cytokines in response to TCR engagement, as determined by IsoPlexis’ functional single-cell proteomics and polyfunctional strength index. D0 T cells and D22 TILs were also assessed for their secretion of IFNg in response to stimulation with αCD3/αCD28/αCD137-coated beads. D22 TILs secreted much higher levels of IFNg in response to stimulation.1

The results suggest that Iovance’s Gen2 process for ex vivo expansion of TILs “reverses the dysfunctional state of the T cells from the tumor microenvironment, by transitioning their phenotypic, functional and tumor-reactive profile, thereby rendering the cells reinvigorated and capable of producing a potent and effective anti-tumor response in vivo.”1 IsoPlexis’ proteomics platform identified high functionality in these cells as well as increased anti-tumor activity, indicating their potential for in vivo activation and allowing these scientists to accelerate their research.

To learn how IsoPlexis technology was used in another study looking at the impact of combination PEG-IL2 with ACT therapy on T cell persistence in solid tumor, click here.


  1. Simpson-Abelson M, et al. Iovance generation-2 tumour-infiltrating lymphocyte (TIL) product is reinvigorated during the manufacturing process. Presented at ESMO Virtual Congress September 19-21, 2020.
Share This Article
Subscribe to Follow the Data

Weekly email to keep you at the forefront of single-cell & multiplexed proteomics.