# Towards GLIMPSE: Enabling technology to map cytokines in inflamed tissue

> **NIH NIH R21** · UNIVERSITY OF VIRGINIA · 2021 · $225,135

## Abstract

Immunity and inflammation are fundamental to diseases as diverse as cancer, autoimmunity, and
neurodegeneration. In each of these systems, secreted cytokines coordinate an intricate dance between
lymphocytes and tissue-resident cells, to correctly balance proliferation, differentiation, migration, and cell death.
Despite the critical mechanistic role of cytokines and their promise as a drug target, there are currently no tools
to detect their spatial distribution in tissue without genetic modification. We speculate that knowledge of where
and when cytokines are secreted would dramatically accelerate the design of effective immunotherapies by
revealing pockets of cellular activity. However, attempts to image cytokine distribution are frustrated by the rapid
mass transport of small proteins, the hindered delivery of large antibody-based reagents, and the requirement
that the reagents not alter the state of live cells and tissue.
 The goal of this project is to create the first method to quantify local cytokine concentrations in live tissue
samples ex vivo, while preserving the state and structure of the tissue. Our approach is to gently capture a
fraction of the secreted cytokine directly onto cells near where it was secreted, label it with a fluorescently-tagged
antibody fragment, and image the distribution of the protein by live-tissue fluorescence imaging. To avoid
inadvertently activating the tissue during the assay, we will work with antibody fragments and small peptide
binders, rather than intact antibodies that could bind Fc receptors. Protein signaling in the lymph node will be
used as a case study because the key proteins (cytokines) involved in inflammation are well-studied, have
quantifiable effects, and are of interest as drug targets for inflammatory diseases. In this pilot proposal, we will
develop a proof-of-concept assay to detect T cell-specific cytokine secretion, using antibody fragments (Aim 1),
and simultaneously develop novel peptide-based reagents to detect lymphocyte surface markers and cytokines
(Aim 2). The assay will be benchmarked against immunostaining of fixed tissues, flow cytometry, and ELISA.
The assay and new reagents will be tested in two different species, mouse and human, to demonstrate proof-of-
principle of cytokine detection during inflammatory disease.
 If successful, this high-risk, high-reward project will show the feasibility of the first chemical imaging
method to generate “maps” of cytokine secretion in unfixed tissue. The proposed technology is deliberately
modular to allow ready transposition to detect other proteins in other tissues, and is easily multiplexed for multiple
cytokines simultaneously. The technology can, in principle, be adopted to visualize any protein for which an
antibody pair or peptide binder is available, and used in any soft tissue, to revolutionize the understanding of
protein-based cross-talk in the immune system. The data will enable a future R01 application to expand the
metho...

## Key facts

- **NIH application ID:** 10218665
- **Project number:** 1R21AI160547-01A1
- **Recipient organization:** UNIVERSITY OF VIRGINIA
- **Principal Investigator:** Kimberly A. Kelly
- **Activity code:** R21 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 2021
- **Award amount:** $225,135
- **Award type:** 1
- **Project period:** 2021-03-10 → 2023-02-28

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/10218665

## Citation

> US National Institutes of Health, RePORTER application 10218665, Towards GLIMPSE: Enabling technology to map cytokines in inflamed tissue (1R21AI160547-01A1). Retrieved via AI Analytics 2026-05-23 from https://api.ai-analytics.org/grant/nih/10218665. Licensed CC0.

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