# MOLECULAR ANALYSIS OF HUMAN ANTI DNA MONOCLONALS

> **NIH NIH R01** · BOSTON CHILDREN'S HOSPITAL · 1987 · $99,608

## Abstract

We shall investigate the genetic basis for the production of human anti DNA
antibodies by Epstein-Barr virus (EBV) transformed B cell lines and human
human hybrids derived from both normal individuals and patients with
systemic lupus erythematosus (SLE).  Human human hybrids and EBV cell lines
which produce anti DNA antibodies have been developed.  The specificities
and affinities of some of these lines have been fully characterized and we
will complete these analyses.  We have prepared a cDNA library from one of
the anti DNA antibody producing hybrids and have isolated a fragment
containing the heavy chain variable region.  This fragment will be
sequenced and then used as a probe for further studies.  We will examine
RNA extracted from the hybrids and B cell lines and, using Northern blots,
see whether the same VH gene is used by normals and SLE patients.  We will
examine what V-J rearrangements are used.  Similarly, we will evaluate the
light chains used in the hybrid and B cell lines, first, by using probes
for the kappa and lambda constant regions and then second, by screening our
already prepared cDNA library for the kappa light chain gene and by
constructing a cDNA library from a lambda producing hybrid and screening it
for the lambda light chain gene.  We will try to identify the genes used to
make the anti DNA antibodies by comparing them to other published antibody
sequences to see if they are similar to genes used for antibodies to known
exogenous antigens.  We will examine the anti DNA gene repertoire in
normals and patients with SLE by extracting unrearranged genomic DNA from
neutrophils, size separating it, Southern blotting it and hybridizing with
the derived variable region probes.  We will correlate our molecular
analysis and the amino acid sequence with the specificity and affinity data
for the antibodies.  We should be able to ascertain what genes are used in
the construction of anti DNA antibodies by human hybrids and EBV
transformed B cell lines, how they are combined, and whether normal derived
anti DNA producing hybrids and B cell lines differ from those derived from
patients with SLE.

## Key facts

- **NIH application ID:** 3156578
- **Project number:** 5R01AR033528-05
- **Recipient organization:** BOSTON CHILDREN'S HOSPITAL
- **SAM.gov UEI:** Z1L9F1MM1RY3
- **Principal Investigator:** SUSAN HOCH
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1987
- **Award amount:** $99,608
- **Award type:** 5
- **Project period:** 1983-09-01 → 1989-08-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3156578

## Citation

> US National Institutes of Health, RePORTER application 3156578, MOLECULAR ANALYSIS OF HUMAN ANTI DNA MONOCLONALS (5R01AR033528-05). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3156578. Licensed CC0.

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