B CELL REPERTOIRE OF NORMAL AND AUTOIMMUNE MODEL

Project: Research project

Description

This is a proposal to investigate the orderly diversification of the murine
B cell specificity repertoire during ontogeny in both conventional and
autoimmune strains. Initially, this will be accomplished by analyzing the
specificities and idiotypes of antibodies elicited from neonatal B cells
from conventional mice responsive to the antigenic determinant
(4-hydroxy-3,5-dinitrophenyle)acetyl (NNP). Previously, distinct B cell
clonotypes specific for NNP have been found to be expressed in a temporally
regulated manner during neonatal development in Ighb bearing mice. The in
vitro splenic focus assay will be employed in order to antigen stimulate
individual B cells or their precursors from the spleen and liver of
neonatal mice of various ages. The monospecific antibodies thus elicited
will be analyzed with respect to their fine specificity for antigen and
idiotype and the frequency of B cells within the developing neonatal
repertoire which possess these specificities will be assessed. Hybridomas
which secrete antibodies representative of the neonatal B cell repertoire
will also be constructed; these will provide a complimentary means to
examine the repertoire of antigen responsive B cells during ontogeny. The
heavy and light chain variable region genes expressed by these neonatal
hybridomas will be analyzed by restriction enzyme analysis on Southern
blots, by assignment to particular variable region gene families, and,
ultimately, by sequence analysis. These experiments will provide insight
into the mechanisms responsible for the temporal acquisition of the B cell
repertoire during ontogeny and the relationships among the variable region
genes which encode antibody specificities acquired at different ages.
These studies will be extended to include inbred strains, such as the NZB
and BXSB mice, which develop spontaneous autoimmune disease. Such strains
exhibit unusual patterns of B cell development which include an early,
precocious development of B lineage cells during ontogeny followed by a
diminution in the proportion of B cell progenitors in the bone marrow of
adults. The influence of the abnormal B cell development apparent in these
strains upon the diversification and expression of the B cell repertoire
both during ontogeny and in the mature and generative (pre-receptor) B cell
pools of adults will be assessed. These experiments will constitute the
first comprehensive study of the development of the B cell repertoire in
the autoimmune strains and will contribute to an understanding of the
immune defects present in autoimmunity.
StatusFinished
Effective start/end date9/30/8612/31/93

Funding

  • National Institutes of Health
  • National Institutes of Health: $120,861.00
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health
  • National Institutes of Health: $112,319.00

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B-Lymphocytes
B-Lymphoid Precursor Cells
Bone Marrow
Antigens
Antibodies
Lymphopoiesis
T-Lymphoid Precursor Cells
Bone and Bones
Interleukin-7
SCID Mice
Restriction Mapping
Vincristine
Metaphase
Stromal Cells
Antibody Specificity
Growth and Development
Interleukin-4
Biological Assay
Systemic Lupus Erythematosus
Interleukin-2

ASJC

  • Medicine(all)
  • Immunology and Microbiology(all)