Cargando…

Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method

B-less mice express a human immunoglobulin (Ig) lambda transgene that induces a severe deficiency of both immature pre-B and mature B lymphocytes. To understand this perturbation in B lymphopoiesis, we have devised a sensitive limiting dilution polymerase chain reaction assay that quantitates specif...

Descripción completa

Detalles Bibliográficos
Formato: Texto
Lenguaje:English
Publicado: The Rockefeller University Press 1993
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2191084/
https://www.ncbi.nlm.nih.gov/pubmed/8315387
_version_ 1782146921864888320
collection PubMed
description B-less mice express a human immunoglobulin (Ig) lambda transgene that induces a severe deficiency of both immature pre-B and mature B lymphocytes. To understand this perturbation in B lymphopoiesis, we have devised a sensitive limiting dilution polymerase chain reaction assay that quantitates specific Ig rearrangements and thus quantitates B lineage cells at various stages of differentiation within unfractionated bone marrow. We find that there are significantly reduced frequencies of both VH-to-DJH and VK-to-JK rearrangements in the transgenic strain, whereas the frequency of D-to-JH rearrangements approximates that of wild type. Since Ig gene rearrangements occur in a stepwise fashion in which D-to-JH joining precedes that of VH-to-DJH and VK-to-JK, these results indicate that the major block of B lymphocyte development in the B-less strain occurs after D-to-JH rearrangement. Interestingly, sequence analysis of residual VHDJH junctions from transgenic pre-B lymphocytes reveals that an abnormally high proportion of these are out of frame and therefore nonproductive. Taken together, these data suggest that early expression of the transgenic lambda protein specifically prevents the development of a normal-sized population of precursor B lymphocytes coexpressing functional IgH. The transgene-induced immunodeficiency appears to arise by a precocious maturation process in which precursors bypass a developmental stage associated with cellular expansion.
format Text
id pubmed-2191084
institution National Center for Biotechnology Information
language English
publishDate 1993
publisher The Rockefeller University Press
record_format MEDLINE/PubMed
spelling pubmed-21910842008-04-16 Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method J Exp Med Articles B-less mice express a human immunoglobulin (Ig) lambda transgene that induces a severe deficiency of both immature pre-B and mature B lymphocytes. To understand this perturbation in B lymphopoiesis, we have devised a sensitive limiting dilution polymerase chain reaction assay that quantitates specific Ig rearrangements and thus quantitates B lineage cells at various stages of differentiation within unfractionated bone marrow. We find that there are significantly reduced frequencies of both VH-to-DJH and VK-to-JK rearrangements in the transgenic strain, whereas the frequency of D-to-JH rearrangements approximates that of wild type. Since Ig gene rearrangements occur in a stepwise fashion in which D-to-JH joining precedes that of VH-to-DJH and VK-to-JK, these results indicate that the major block of B lymphocyte development in the B-less strain occurs after D-to-JH rearrangement. Interestingly, sequence analysis of residual VHDJH junctions from transgenic pre-B lymphocytes reveals that an abnormally high proportion of these are out of frame and therefore nonproductive. Taken together, these data suggest that early expression of the transgenic lambda protein specifically prevents the development of a normal-sized population of precursor B lymphocytes coexpressing functional IgH. The transgene-induced immunodeficiency appears to arise by a precocious maturation process in which precursors bypass a developmental stage associated with cellular expansion. The Rockefeller University Press 1993-07-01 /pmc/articles/PMC2191084/ /pubmed/8315387 Text en This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).
spellingShingle Articles
Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title_full Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title_fullStr Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title_full_unstemmed Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title_short Molecular characterization of transgene-induced immunodeficiency in B- less mice using a novel quantitative limiting dilution polymerase chain reaction method
title_sort molecular characterization of transgene-induced immunodeficiency in b- less mice using a novel quantitative limiting dilution polymerase chain reaction method
topic Articles
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2191084/
https://www.ncbi.nlm.nih.gov/pubmed/8315387