# CYTOLYTIC T LYMPHOCYTE HELPER FACTOR

> **NIH NIH R01** · UNIVERSITY OF MINNESOTA · 1987 · $180,553

## Abstract

Recently we have developed an assay for cytokines needed for the in vitro
generation of primary murine cytolytic T lymphocytes (CTL) against
alloantigens on tumor cells.  The factors which function in the assay are
called CHF for CTL helper factor.  This assay can detect previously
uncharacterized cytokines including a factor of 30,000 molecular weight
(RDCHF) made by the cell line RDM4.

We have been able to concentrate this cytokine by adsorption to Sepralyte
and to obtain a good degree of purification using reverse phase HPLC.  We
will continue to purify this molecule to a single major protein species for
use in biological experiments and for obtaining an N terminal amino acid
sequence.

We will confirm that the protein we begin to sequence is actually RDCHF by
cloning the molecule using recombinant DNA techniques.  We have already
accomplished the key step in this procedure, the expression of the molecule
in Xenopus laeis oocytes.  Thus we are able to assay the mRNA specific for
the cytokine.  Our next steps are:  (1) Construction of cDNA libraries.
(2) Identification of RDCHF cDNA.  (3) Analysis of the nucleotide sequence
of RDCHF cDNA.  (4) Expression of recombinant RDCHF cDNA in mammalian cells.

Once we have obtained reasonably homogeneous RDCHF, we will test it for its
biochemical properties.  We will also test its biological properties
including the binding of radiolabeled RDCHF to different cells involved in
CTL generation, the addition of RDCHF to spleen cells depleted of certain
cell types, and the function of RDCHF in other help deficient systems for
the generation of CTL in vitro.

We have found novel molecules with CHF activity from not only RDM4 but also
other cell lines such as EL4 and ASFTL-1.  We propose to repeat the studies
described above with these cytokines.

## Key facts

- **NIH application ID:** 3172812
- **Project number:** 2R01CA035152-04
- **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:** 1987
- **Award amount:** $180,553
- **Award type:** 2
- **Project period:** 1984-04-01 → 1988-03-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3172812

## Citation

> US National Institutes of Health, RePORTER application 3172812, CYTOLYTIC T LYMPHOCYTE HELPER FACTOR (2R01CA035152-04). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3172812. Licensed CC0.

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