Tumor-Infiltrating Lymphocytes (TILs)
Precise and reproducible cell counting for tumor-fighting cell therapy
Tumor-infiltrierende Lymphozyten (TIL) sind eine heterogene Gruppe von Zellen, bestehend aus B-Zellen, T-Zellen und natürlichen Killerzellen (NK). Sie werden als TILs charakterisiert, wenn sie aus dem Blutkreislauf in einen Tumor wandern, wo sie sowohl im Tumor selbst als auch im Stroma, das den Tumor umgibt, zu finden sind.
Die TIL-Therapie ist eine Form der adaptiven Immuntherapie, bei der die Zellen nicht gentechnisch verändert werden, sondern lediglich aus den Tumoren der Patienten isoliert, expandiert und den Patienten zur Bekämpfung ihrer Tumore wieder zugeführt werden1. In Kultur aktiviert IL-2 die TILs, die dann bei Wiedereintritt in den Patienten den Tumor aktiv zerstören können.
Obwohl TILs nicht ohne Herausforderungen sind, sind sie derzeit die effizienteste Art der tumorbekämpfenden Zelltherapie-Methode für die Tumore, gegen die sie gerichtet sind: Die Anti-Tumor-Antwort kann im Gegensatz zu gentechnisch veränderten Zelltherapien, einschließlich chimärer Antigenrezeptoren (CAR-T) und CAR-NK-Therapien, vielfältig sein.
Darüber hinaus können TILs als prognostische Marker bei Erkrankungen wie Brustkrebs und Melanomen eingesetzt werden2,3. In Verbindung mit traditionellen Therapien wie Chemotherapie oder neueren Behandlungen wie Immun-Checkpoint-Inhibitoren (ICIs) können TILs als Biomarker dienen, um die Ergebnisse von Immuntherapien im klinischen Umfeld vorherzusagen und so den Behandlungspfad für jeden Patienten genauer zu steuern und anzupassen.
Challenges from lack of standardized protocols
One of the biggest challenges in working with TILs is that they are available in low numbers and difficult to extract. They are only found in small numbers within the tumor tissue they have infiltrated and must be extracted from the tumor itself, so it is difficult to get a good quality sample of high viability.
When surgically removing the solid tumor, the duration of time from sample extraction through TIL cell isolation is critical to the quality of the therapeutic cells. The sample is extracted from the patient, assessed under the microscope for pathogenesis, submerged in buffered medium and cooled, then transported to the lab for TIL cell isolation.
Because this therapeutic approach is still under development, there are still no standards for patient preparation, tissue handling, cell cooling and TIL isolation. Furthermore, regulations around extraction and manufacturing vary between countries. These factors make it difficult, especially for smaller biotech companies, to set up an efficient and compliant production pipeline, leaving them reliant on best assumptions and previous successes when locking in process parameters.
Alternatively, engineered TILs may prove to be a more beneficial approach to using TILs for cancer therapies, but developments within this field are still to be determined for their use and validity4.
Tackling the process challenges with a robust cell counting instrument
Through several stages of TIL therapy manufacturing, cell counting and viability assessment are crucial. The NucleoCounter® NC-202™ is the automated cell counter of choice for cell therapeutic manufacturers because it is robust and minimalizes human error. Furthermore, labs and operators can collaborate along the processing pipeline with ease as the instrument is validated against a standard, meaning all instruments count the same, no matter where in the world they, or the operator, are located.
The NC-202™ uses the Via2-Cassette™ for sample handling. This device ensures precision and reproducibility of results because the cassette’s measurement chamber has a pre-calibrated volume, detected individually for each sample analysis run. No user or pipetting errors interfere with data generation. The Via2-Cassette™ is very safe since there is no manual handling of or exposure to dyes.
Don’t waste time on tech transfer
The NucleoCounter® is GMP/21 CFR Part 11-compliant and can be used from the earliest R&D tests, through Process Development, Manufacturing and Quality Control. This means you will not waste time on laborious technical SOP and protocol transfers but can focus on cultivating the most effective cells for an optimal patient outcome.
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References
- NP Restifo, ME Dudley, S A. Rosenberg: Adoptive immunotherapy for cancer: harnessing the T cell response. Nat Rev Immunol. 2012; 12(4): 269–281.
- LG Radvanyi: Tumor-Infiltrating Lymphocyte Therapy: Addressing Prevailing Questions. Cancer J. 2015; 21(6): 450-64.
- MV Dieci, F Miglietta, V Guarneri: Immune Infiltrates in Breast Cancer: Recent Updates and Clinical Implications. Cells. 2021;10(2):223.
- A Jiménez-Reinoso, D Nehme-Álvarez, C Domínguez-Alonso et al.: Synthetic TILs: Engineered Tumor-Infiltrating Lymphocytes With Improved Therapeutic Potential. Front Oncol. 2021; 10: 593848.