# MECHANISM OF LYMPHOCYTE ACTIVATION BY THIOL COMPOUNDS

> **NIH NIH R01** · SCRIPPS CLINIC AND RESEARCH FOUNDATION · 1988 · $246,474

## Abstract

A comprehensive group of experiments has been designed to study the
interactions with cellular components of a unique, intracellular class of
lymphocyte stimulator, the C8-substituted guanine ribonucleosides.  First,
the existence of intracellular recognition structures or receptors specific
for 8bromoguanosine (8BrGuo) will be evaluated.  Using high specific
activity (5-3H)8BrGuo, the saturability of binding to cellular components
will be assessed.  Average receptor number, apparent affinity, binding
kinetics and receptor specificity will also be determined.  These
parameters will then be evaluated in B cells from nucleoside nonresponder
mice and in a variety of B cell lines.  Second, cellular receptors for
8BrGuo will be isolated and characterized.  Cells will be biosynthetically
labeled, disrupted by nitrogen cavitation, and passed over affinity
columns.  The eluate will be analyzed on HPLC gel filtration and reverse
phase columns.  Receptors in nucleoside nonresponder mice will be studied
similarly.  Alternatively, cellular constituents will be separated by
electrophoresis, blotted onto nitrocellulose, overlaid with (U-14C)8BrGuo,
and assessed by autoradiography.  Receptors will be characterized by
relative molecular mass, isoelectric point, pH sensitivity, apparent
affinity, and rate of turnover.  Third, direct interactions between 8BrGuo
and cellular nucleic acid will be investigated.  Binding studies with
purified DNA and RNA will be performed, using nucleic acid from nucleoside
nonresponder strains as controls.  If binding is observed, it will be
characterized in terms of the parameters listed below.  Fourth, the
interactions of C8-substituted guanine ribonucleosides with enzymes of
purine metabolism will be studied.  The role of phosphorylation of 8BrGuo,
both by salvage pathway enzymes and by direct phosphorylation will be
determined.  The biologic and metabolic activity of a substituted synthetic
nucleoside which cannot be directly phosphorylated will be evaluated.  The
effects of 8BrGuo on purine nucleotide generation and degradation, on the
activity of salvage pathway enzymes, and on de novo purine biosynthesis
will be evaluated.  Finally, the biological interactions of C8-substituted
guanine ribonucleosides with cytokines will be investigated both in terms
of their ability to synergize with T cell-derived lymphokines and to induce
secretion of monokines and lymphokines.  Initial physical characterization
of these activities will be undertaken.

## Key facts

- **NIH application ID:** 3126131
- **Project number:** 5R01AI015284-11
- **Recipient organization:** SCRIPPS CLINIC AND RESEARCH FOUNDATION
- **Principal Investigator:** MICHAEL G GOODMAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $246,474
- **Award type:** 5
- **Project period:** 1978-08-01 → 1989-07-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3126131

## Citation

> US National Institutes of Health, RePORTER application 3126131, MECHANISM OF LYMPHOCYTE ACTIVATION BY THIOL COMPOUNDS (5R01AI015284-11). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3126131. Licensed CC0.

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