# MOLECULAR ANALYSIS OF MURINE THYMOCYTE DIFFERENTIATION

> **NIH NIH R01** · CALIFORNIA INSTITUTE OF TECHNOLOGY · 1987 · $260,967

## Abstract

Differentiation of T lymphocytes takes place in the thymus in a process
that involves programmed activation of multiple genetic loci.  Some of
these genes encode constitutive products of mature T cells, such as
components of their antigen-binding receptors.  Other genes of interest are
inducible genes that mediate T-cell responses after the cells have been
triggered by antigen, such as those encoding interleukin 2 (IL2) and the
IL2 receptor.  The goal of this proposal is to study the sequence of
activation of T-cell-specific genes in the thymus, with a particular focus
on the second (inducible) class.  The experiments will be designed to
determine separately when the genes first become inducible at the chromatin
level, and when the cells develop the signal-transduction mechanism that
can turn these genes on in response to stimuli such as antigen
recognition.  The basis for the work is recent evidence that the IL2 gene
itself becomes inducible before the develpoing T cell expresses receptors
that can trigger induction.

In situ hybridization will be used to detect IL2 induction in individual
cells, and flow cytometry using the Ca2+-sensitive dye indo-1 will be used
to measure their mobilization of intracellular Ca2+ in response to
mitogenic lectins.  Expression of high-affinity IL2 receptors will be
measured by binding of radiolabeled IL2, adapted to autoradiography for
single-cell detection.  These methods will be used to dissect heterogeneity
in the immature precursor thymocyte class, and to analyze the apparent loss
of competence in cells becoming committed to the sterile, major-cortical
differentiation pathway.  Finally, preliminary results suggest that
immature precursor thymocytes can be maintained in culture without
destroying their abiltiy to differentiate.  A vital aim of the proposal is
therefore to optimize these culture conditions so that thymic lineage
relationships can be determined and differentiation pathways manipulated
directly, under controlled conditions in vitro.

## Key facts

- **NIH application ID:** 3129153
- **Project number:** 5R01AI019752-06
- **Recipient organization:** CALIFORNIA INSTITUTE OF TECHNOLOGY
- **SAM.gov UEI:** U2JMKHNS5TG4
- **Principal Investigator:** ELLEN V. ROTHENBERG
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $260,967
- **Award type:** 5
- **Project period:** 1982-06-01 → 1989-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3129153

## Citation

> US National Institutes of Health, RePORTER application 3129153, MOLECULAR ANALYSIS OF MURINE THYMOCYTE DIFFERENTIATION (5R01AI019752-06). Retrieved via MORVS 2026-10-24 from https://api.morvs.ai/grant/nih/3129153. Licensed CC0.

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