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Related Experiment Video

Updated: Jun 24, 2026

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

A Fluorescence-based Lymphocyte Assay Suitable for High-throughput Screening of Small Molecules

Published on: March 10, 2017

Framework for in vivo T cell screens.

Lauren E Milling^1,2, Samuel C Markson^1,2,3, Qin Tjokrosurjo^1,2

¹Department of Immunology, Blavatnik Institute, Harvard Medical School, Boston, MA, USA.

The Journal of Experimental Medicine

|February 27, 2024

Summary

This study introduces the Framework for In vivo T cell Screens (FITS), offering guidelines for designing robust in vivo T cell screens. FITS optimizes parameters like gene library size and cell transfer for accurate immune response studies.

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Area of Science:

Immunology
Systems Biology
Bioinformatics

Background:

In vivo T cell screens are crucial for understanding immune responses but lack standardized design parameters.
Key parameters like gene library size, cell transfer quantity, and mouse numbers require optimization for reliable results.

Purpose of the Study:

To establish a standardized framework, the Framework for In vivo T cell Screens (FITS), for designing and analyzing in vivo immune cell screens.
To provide experimental and analytical guidelines for optimizing diverse in vivo screening contexts.

Main Methods:

Developed the Framework for In vivo T cell Screens (FITS) incorporating experimental and analytical guidelines.
Applied FITS to optimize parameters for a CD8+ T cell screen in a B16-OVA tumor model.
Integrated unique molecular identifiers (UMIs) to enhance statistical power and track T cell clonal dynamics.

Main Results:

FITS successfully optimized parameters for a CD8+ T cell screen, demonstrating its practical application.
UMIs improved statistical power and enabled detailed tracking of T cell clonal dynamics across tissues for gene knockouts.
The study provides a validated framework for future in vivo immune cell screening studies.

Conclusions:

The Framework for In vivo T cell Screens (FITS) offers essential guidelines for robust and reproducible in vivo immune cell research.
The integration of UMIs significantly enhances the analytical capabilities of in vivo screens, particularly for tracking clonal dynamics and gene knockout effects.
This work establishes a crucial foundation for advancing the field of in vivo immunology studies.