# CYTOLYTIC T LYMPHOCYTE HELPER  FACTOR

> **NIH NIH R01** · UNIVERSITY OF MINNESOTA · 1988 · $186,569

## Abstract

We have developed an assay for cytokines needed for the in vitro
generation of primary mouse cytolytic T lymphocytes (CTL)
against alloantigens on tumor cells.  One of the factors which
functions in this assay is called RDCHF.

We have recently purified RDCHF to a single band by sodium
dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE).
The cytokine could be eluted from the gel with retention of
biological activity.  From a preliminary sequence of the first 10
amino acids, RDCHF is different from all sequenced cytokines.
We have also demonstrated that at least one RDCHF clone exists
in a cDNA library we have made from RDM4 cells producing
RDCHF.  We have begun to subdivide the library to isolate cDNA
clones.

We propose to refine the RDCHF purification to achieve a
reproducible technique for obtaining pure material.  We will bind
radiolabeled RDCHF to cells from established lines or from
preparations of crude cells separated by fluorescent activated cell
sorter (FACS).  In this way, we will identify the immediate target
cells of the factor.  We will also bind radiolabeled RDCHF
covalently to its receptor(s) on target cells to study the nature of
the receptor(s).

We also plan to complete the isolation of cDNA clones to RDCHF.
We will generate transient expression vectors to express the
cytokine in large amounts in the COS cell line to produce large
amounts of protein for biological studies.  We will use the cDNA
clone to isolate genomic clones to study the structure of the gene
coding for the factor.  We will perform hybridization studies to
identify the cells making RDCHF and to study the expression of
the RDCHF gene.  We will also attempt to isolate a homologous
gene from the human genome.

Once we can make large amounts of purified RDCHF, we will
attempt to make monoclonal antibodies to this cytokine both for
biological studies and for simplification for factor purification.

## Key facts

- **NIH application ID:** 3172813
- **Project number:** 2R01CA035152-05
- **Recipient organization:** UNIVERSITY OF MINNESOTA
- **SAM.gov UEI:** KABJZBBJ4B54
- **Principal Investigator:** DAVID P FAN
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1988
- **Award amount:** $186,569
- **Award type:** 2
- **Project period:** 1984-04-01 → 1991-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3172813

## Citation

> US National Institutes of Health, RePORTER application 3172813, CYTOLYTIC T LYMPHOCYTE HELPER  FACTOR (2R01CA035152-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3172813. Licensed CC0.

---

*[NIH grants dataset](/datasets/nih-grants) · CC0 1.0*