Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells
The in vitro CD40-CD154 interaction promotes human B lymphocytes differentiation into plasma cells. Currently, CD138 is the hallmark marker enabling the detection of human plasma cells, both in vitro and in vivo; its presence can be monitored by flow cytometry using a specific antibody. We have deve...
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doaj-8fc19fa317b3483dbc5ddb7c03c69c322020-11-24T23:47:58ZengHindawi LimitedBioMed Research International2314-61332314-61412014-01-01201410.1155/2014/536482536482Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma CellsRayelle Itoua Maïga0Jennifer Lemieux1Annie Roy2Carl Simard3Sonia Néron4Héma-Québec, Ingénierie cellulaire, Recherche et Développement, 1070 Avenue des Sciences-de-la-Vie, Québec, QC, G1V 5C3, CanadaHéma-Québec, Ingénierie cellulaire, Recherche et Développement, 1070 Avenue des Sciences-de-la-Vie, Québec, QC, G1V 5C3, CanadaHéma-Québec, Ingénierie cellulaire, Recherche et Développement, 1070 Avenue des Sciences-de-la-Vie, Québec, QC, G1V 5C3, CanadaHéma-Québec, Ingénierie cellulaire, Recherche et Développement, 1070 Avenue des Sciences-de-la-Vie, Québec, QC, G1V 5C3, CanadaHéma-Québec, Ingénierie cellulaire, Recherche et Développement, 1070 Avenue des Sciences-de-la-Vie, Québec, QC, G1V 5C3, CanadaThe in vitro CD40-CD154 interaction promotes human B lymphocytes differentiation into plasma cells. Currently, CD138 is the hallmark marker enabling the detection of human plasma cells, both in vitro and in vivo; its presence can be monitored by flow cytometry using a specific antibody. We have developed a culture system allowing for the differentiation of memory B lymphocytes. In order to detect the newly formed plasma cells, we have compared their staining using five anti-CD138 monoclonal antibodies (mAbs). As a reference, we also tested human cell lines, peripheral blood mononuclear cells, and bone marrow samples. The five anti-CD138 mAbs stained RPMI-8226 cells (>98%) with variable stain index (SI). The highest SI was obtained with B-A38 mAb while the lowest SI was obtained with DL-101 and 1D4 mAbs. However, the anti-CD138 mAbs were not showing equivalent CD138+ cells frequencies within the generated plasma cells. B-A38, B-B4, and MI-15 were similar (15–25%) while DL-101 mAb stained a higher proportion of CD138-positive cells (38–42%). DL-101 and B-A38 mAbs stained similar populations in bone marrow samples but differed in their capacity to bind to CD138high and CD138lo cell lines. In conclusion, such cellular fluctuations suggest heterogeneity in human plasma cell populations and/or in CD138 molecules.http://dx.doi.org/10.1155/2014/536482 |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Rayelle Itoua Maïga Jennifer Lemieux Annie Roy Carl Simard Sonia Néron |
spellingShingle |
Rayelle Itoua Maïga Jennifer Lemieux Annie Roy Carl Simard Sonia Néron Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells BioMed Research International |
author_facet |
Rayelle Itoua Maïga Jennifer Lemieux Annie Roy Carl Simard Sonia Néron |
author_sort |
Rayelle Itoua Maïga |
title |
Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells |
title_short |
Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells |
title_full |
Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells |
title_fullStr |
Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells |
title_full_unstemmed |
Flow Cytometry Assessment of In Vitro Generated CD138+ Human Plasma Cells |
title_sort |
flow cytometry assessment of in vitro generated cd138+ human plasma cells |
publisher |
Hindawi Limited |
series |
BioMed Research International |
issn |
2314-6133 2314-6141 |
publishDate |
2014-01-01 |
description |
The in vitro CD40-CD154 interaction promotes human B lymphocytes differentiation into plasma cells. Currently, CD138 is the hallmark marker enabling the detection of human plasma cells, both in vitro and in vivo; its presence can be monitored by flow cytometry using a specific antibody. We have developed a culture system allowing for the differentiation of memory B lymphocytes. In order to detect the newly formed plasma cells, we have compared their staining using five anti-CD138 monoclonal antibodies (mAbs). As a reference, we also tested human cell lines, peripheral blood mononuclear cells, and bone marrow samples. The five anti-CD138 mAbs stained RPMI-8226 cells (>98%) with variable stain index (SI). The highest SI was obtained with B-A38 mAb while the lowest SI was obtained with DL-101 and 1D4 mAbs. However, the anti-CD138 mAbs were not showing equivalent CD138+ cells frequencies within the generated plasma cells. B-A38, B-B4, and MI-15 were similar (15–25%) while DL-101 mAb stained a higher proportion of CD138-positive cells (38–42%). DL-101 and B-A38 mAbs stained similar populations in bone marrow samples but differed in their capacity to bind to CD138high and CD138lo cell lines. In conclusion, such cellular fluctuations suggest heterogeneity in human plasma cell populations and/or in CD138 molecules. |
url |
http://dx.doi.org/10.1155/2014/536482 |
work_keys_str_mv |
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