# IMMUNOGENTIC ANALYSIS OF B CELL DIFFERENTIATION

> **NIH NIH R01** · BRANDEIS UNIVERSITY · 1991 · $182,984

## Abstract

The long term goals are to determine the mechanisms that regulate shifts in
the expressed antibody repertoire during the generation of immunological
memory.  The proposed research focuses on two aspects of the repertoire
shift observed during the maturation of the immune response to (T,G)-A--L:
(1) the dominance in the primary response of B cells that recognize side-
chain specific epitopes (GT+) and use the H10/VK1 germline gene combination
and (2) the high frequency in the memory response B cells that recognize
epitopes on the A-L backbone.  To determine whether the H10/VK1 germline
combination has an intrinsically high frequency in the preimmune
repertoire, relative to other V-genes, its frequency will be measured in a
cell lysate hybridization assay that uses probes specific for these V
genes.  A mutation/selection model for repertoire shift will be examined by
measuring the affinities of primary and memory antibodies using H10/VK1; if
selection has resulted in higher affinity memory antibodies, the heavy
chains from some of these will be sequenced to determine whether/where
mutations have occurred.  Mutation/selection may also explain the higher
frequency of A-L+ clones in the memory response.  This will be determined
by analyzing for mutations the heavy chains of A-L+ antibodies derived from
clonal progeny (sibs) of a single precursor, which we have shown are in the
pauciclonal A-L+ response of mice immunized as neonates.  Since one of the
clonal bursts uses a VGam3.8 gene, which is from a VH gene family of low
complexity, its germline gene equivalent will be identified.  A second
model for repertoire shift, that primary and memory responses originate
from two separate precursor lineages, will be tested.  The frequency of GT+
vs A-L+ precursors in J11D-hi (primary lineage) and J11D-lo (memory
lineage) B cells will be measured in the splenic focus assay.  Adoptive
transfer experiments will show whether the repertoire shift is seen with
J11D-lo B cells in vivo, and the clonal frequencies of GT+ and AL-memory B
cells in the transfer recipients with time after stimulation will be
measured.  Other adoptive transfer experiments, will (a) measure the need
for continuous antigenic stimulation in the repertoire shift, and (b) show
whether removal of the dominant primary B cell clonotype (H10/VK1) from the
preimmune repertoire modulates the expressed primary and memory repertoire.
These studies have relevance for health issues concerning modes of
vaccination to establish a memory pool directed to different epitopes on
the same immunogen.

## Key facts

- **NIH application ID:** 3125545
- **Project number:** 2R01AI013725-13A2
- **Recipient organization:** BRANDEIS UNIVERSITY
- **SAM.gov UEI:** MXLZGAMFEKN5
- **Principal Investigator:** JOAN L PRESS
- **Activity code:** R01 (R01, R21, SBIR, etc.)
- **Funding institute:** NIH
- **Fiscal year:** 1991
- **Award amount:** $182,984
- **Award type:** 2
- **Project period:** 1977-09-01 → 1993-12-31

## Primary source

NIH RePORTER: https://reporter.nih.gov/project-details/3125545

## Citation

> US National Institutes of Health, RePORTER application 3125545, IMMUNOGENTIC ANALYSIS OF B CELL DIFFERENTIATION (2R01AI013725-13A2). Retrieved via MORVS 2026-10-25 from https://api.morvs.ai/grant/nih/3125545. Licensed CC0.

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