An in vitro model of differentiation of memory B cells into plasmablasts and plasma cells including detailed phenotypic and molecular characterization.
Human plasma cells (PCs) and their precursors play an essential role in humoral immune response but are rare and difficult to harvest. We report the generation of human syndecan-1(+) and immunoglobulin secreting PCs starting from memory B cells in a 3-step and 10-day (D) culture, including a 6-fold cell amplification. We report the detailed phenotypic and Affymetrix gene expression profiles of these in vitro PCs as well as of intermediate cells (activated B cells and plasmablasts) compared with memory B cells and bone marrow PCs, which is accessible through an open web ATLAS (http://amazonia.transcriptome.eu/). We show this B cell-to-PC differentiation to involve IRF4 and AICDA expressions in D4 activated B cells, decrease of PAX5 and BCL6 expressions, and increase in PRDM1 and XBP1 expressions in D7 plasmablasts and D10 PCs. It involves down-regulation of genes controlled by Pax5 and induction of genes controlled by Blimp-1 and XBP1 (unfold protein response). The detailed phenotype of D10 PCs resembles that of peripheral blood PCs detected after immunization of healthy donors. This in vitro model will facilitate further studies in PC biology. It will likewise be helpful to study PC dyscrasias, including multiple myeloma.
Jourdan
Michel
M
Inserm, U847, Montpellier, France.
Caraux
Anouk
A
De Vos
John
J
Fiol
Geneviève
G
Larroque
Marion
M
Cognot
Chantal
C
Bret
Caroline
C
Duperray
Christophe
C
Hose
Dirk
D
Klein
Bernard
B
D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D001854:Bone Marrow Cells / Q000166:cytology / Q000276:immunology / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D002478:Cells, Cultured; D003564:Cytidine Deaminase / Q000235:genetics; D004268:DNA-Binding Proteins / Q000235:genetics; D005434:Flow Cytometry; D020869:Gene Expression Profiling; D006801:Humans; D007136:Immunoglobulins / Q000378:metabolism; D007156:Immunologic Memory; D016130:Immunophenotyping; D050835:Interferon Regulatory Factors / Q000235:genetics; D008954:Models, Biological; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D051757:PAX5 Transcription Factor / Q000235:genetics; D010950:Plasma Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D051560:Proto-Oncogene Proteins c-bcl-6; D000071701:Regulatory Factor X Transcription Factors; D020133:Reverse Transcriptase Polymerase Chain Reaction; D013234:Stem Cells / Q000166:cytology* / Q000276:immunology / Q000378:metabolism; D053668:Syndecan-1 / Q000378:metabolism; D013997:Time Factors; D014157:Transcription Factors / Q000235:genetics; D000071717:X-Box Binding Protein 1; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D000099218:Interferon Regulatory Factor-4
D003564:Cytidine Deaminase; D004268:DNA-Binding Proteins; D007136:Immunoglobulins; D050835:Interferon Regulatory Factors; D051757:PAX5 Transcription Factor; D051560:Proto-Oncogene Proteins c-bcl-6; D000071701:Regulatory Factor X Transcription Factors; D053668:Syndecan-1; D014157:Transcription Factors; D000071717:X-Box Binding Protein 1; D000098830:AICDA (Activation-Induced Cytidine Deaminase); D000099218:Interferon Regulatory Factor-4; C084103:BCL6 protein, human; C497281:PAX5 protein, human; C000605543:XBP1 protein, human