# FUNCTIONAL AND METABOLIC PROPERTIES OF TOXIC NEUTROPHILS

> **NIH NIH R01** · WAKE FOREST UNIVERSITY · 1993 · $220,224

## Abstract

The interleukin-1 beta (IL-1beta) gene, which encodes for a protein that
plays a critical role in modulating inflammation, is down-regulated in
lipopolysaccharide (LPS)-tolerant neutrophils (PMN) and macrophages (MAC).
This suppression of the IL-1beta gene in LPS-tolerant PMN and MAC can be
overcome by treating the cells with cycloheximide (CHX), suggesting that
a labile repressor protein controls LPS tolerance. This project will
investigate the regulation of LPS-tolerance at the level of the IL-1beta
gene, and at earlier points in signal transduction. Models of in vivo-
induced tolerance of blood PMN and MAC obtained from patients with the
systemic inflammatory response syndrome (SIRS), and a model of in vitro-
induced LPS tolerance in THP-1 MAC, will be used. Aim 1. Studies of LPS
tolerance at the level of the IL-1beta gene will: (i) determine whether
reduced transcription or enhanced degradation of IL-1beta mRNA occurs in
LPS-tolerant leukocytes, using nuclear run-on and mRNA degradation assays,
(ii) define the specificity of LPS tolerance by investigating TNF alpha
and prostaglandin G/H synthase-2 genes, (iii) identify the cis-acting
elements of the IL-1beta gene that are responsible for LPS tolerance,
using THP-1 cells transfected with plasmids containing regulatory elements
of the IL-1beta gene and a reporter, (iv) investigate the proteins that
interact with the cis-elements responsible for LPS tolerance, using
electrophoretic mobility-shift, DNA or RNA footprinting, and immunologic
assays. Aim 2. Experiments of signal transduction events that lie proximal
to the IL-1beta gene, and that might indirectly be responsible for LPS
tolerance will assess protein kinases C (PKC), protein tyrosine kinases
(PTK), and extracellular signal related kinases (ERK). Enzymatic and
immunoassays will be used. Aim 3. Extracellularly added anti-inflammatory
agents, including dexamethasone and IL-4, will be used in an attempt to
produce the phenotypic changes found in LPS-tolerant PMN and MAC obtained
from humans with SIRS. Studies will focus on the IL-1 and Type 2 IL-1
receptor genes.  This research may provide insight into the molecular
basis of tolerance of leukocytes to LPS-induction of cytokine genes.

## Key facts

- **NIH application ID:** 3124516
- **Project number:** 2R01AI009169-21
- **Recipient organization:** WAKE FOREST UNIVERSITY
- **SAM.gov UEI:** MBU6HCLNZ431
- **Principal Investigator:** Charles Emory McCall
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1993
- **Award amount:** $220,224
- **Award type:** 2
- **Project period:** 1974-04-01 → 1998-06-30

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3124516

## Citation

> US National Institutes of Health, RePORTER application 3124516, FUNCTIONAL AND METABOLIC PROPERTIES OF TOXIC NEUTROPHILS (2R01AI009169-21). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3124516. Licensed CC0.

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