Description and Characterization of a Novel Human Mast Cell Line for Scientific Study
Background: Laboratory of allergic diseases 2 (LAD2) human mast cells were developed over 15 years ago and have been distributed worldwide for studying mast cell proliferation, receptor expression, mediator release/inhibition, and signaling. LAD2 cells were derived from CD34+ cells following marrow...
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doaj-1567b502fcae42dea856319532f89c132020-11-24T21:33:38ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-11-012022552010.3390/ijms20225520ijms20225520Description and Characterization of a Novel Human Mast Cell Line for Scientific StudyArnold S. Kirshenbaum0Yuzhi Yin1J. Bruce Sundstrom2Geethani Bandara3Dean D. Metcalfe4Mast Cell Biology Section, Laboratory of Allergic Diseases, Bethesda, MD 20892, USAMast Cell Biology Section, Laboratory of Allergic Diseases, Bethesda, MD 20892, USAAIDS Research Review Branch/SRP, National Institute of Allergy and Infectious Diseases, National Institutes of Health, Bethesda, MD 20892, USAMast Cell Biology Section, Laboratory of Allergic Diseases, Bethesda, MD 20892, USAMast Cell Biology Section, Laboratory of Allergic Diseases, Bethesda, MD 20892, USABackground: Laboratory of allergic diseases 2 (LAD2) human mast cells were developed over 15 years ago and have been distributed worldwide for studying mast cell proliferation, receptor expression, mediator release/inhibition, and signaling. LAD2 cells were derived from CD34+ cells following marrow aspiration of a patient with aggressive mastocytosis with no identified mutations in <i>KIT</i>. Another aspiration gave rise to a second cell line which has recently been re-established (LADR). We queried whether LADR had unique properties for the preclinical study of human mast cell biology. Methods: LADR and LAD2 cells were cultured under identical conditions. Experiments examined proliferation, beta-hexosaminidase (β-hex) release, surface receptor and granular protease expression, infectivity with HIV, and gene expression. Results: LADR cells were larger and more granulated as seen with Wright−Giemsa staining and flow cytometry, with cell numbers doubling in 4 weeks, in contrast to LAD2 cells, which doubled every 2 weeks. Both LADR and LAD2 cells released granular contents following aggregation of FcεRI. LADR cells showed log-fold increases in FcεRI/CD117 and expressed CD13, CD33, CD34, CD63, CD117, CD123, CD133, CD184, CD193, and CD195, while LAD2 cells expressed CD33, CD34, CD63, CD117, CD133, CD193 but not CD13, CD123, CD184, or CD195. LADR tryptase expression was one-log-fold increased. LADR cell and LAD2 cell chymase expression were similar. Both cell lines could be infected with T-tropic, M-tropic, and dual tropic HIV. Following monomeric human IgE stimulation, LADR cells showed greater surface receptor and mRNA expression for CD184 and CD195. Expression arrays revealed differences in gene upregulation, especially for the suppressor of cytokine signaling (SOCS) family of genes with their role in JAK2/STAT3 signaling and cellular myelo<b>c</b>ytomatosis oncogene (c-MYC) in cell growth and regulation. Conclusions: LADR cells are thus unique in that they exhibit a slower proliferation rate, are more advanced in development, have increased FcεRI/CD117 and tryptase expression, have a different profile of gene expression, and show earlier infectivity with HIV-BAL, LAV, and TYBE when compared to LAD2 cells. This new cell line is thus a valuable addition to the few FcεRI+ human mast cell lines previously described and available for scientific inquiry.https://www.mdpi.com/1422-0067/20/22/5520mast celllad cellshivallergyfcεr1stem cell factorc-<i>kit</i>socs<i>myc</i> |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Arnold S. Kirshenbaum Yuzhi Yin J. Bruce Sundstrom Geethani Bandara Dean D. Metcalfe |
spellingShingle |
Arnold S. Kirshenbaum Yuzhi Yin J. Bruce Sundstrom Geethani Bandara Dean D. Metcalfe Description and Characterization of a Novel Human Mast Cell Line for Scientific Study International Journal of Molecular Sciences mast cell lad cells hiv allergy fcεr1 stem cell factor c-<i>kit</i> socs <i>myc</i> |
author_facet |
Arnold S. Kirshenbaum Yuzhi Yin J. Bruce Sundstrom Geethani Bandara Dean D. Metcalfe |
author_sort |
Arnold S. Kirshenbaum |
title |
Description and Characterization of a Novel Human Mast Cell Line for Scientific Study |
title_short |
Description and Characterization of a Novel Human Mast Cell Line for Scientific Study |
title_full |
Description and Characterization of a Novel Human Mast Cell Line for Scientific Study |
title_fullStr |
Description and Characterization of a Novel Human Mast Cell Line for Scientific Study |
title_full_unstemmed |
Description and Characterization of a Novel Human Mast Cell Line for Scientific Study |
title_sort |
description and characterization of a novel human mast cell line for scientific study |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1422-0067 |
publishDate |
2019-11-01 |
description |
Background: Laboratory of allergic diseases 2 (LAD2) human mast cells were developed over 15 years ago and have been distributed worldwide for studying mast cell proliferation, receptor expression, mediator release/inhibition, and signaling. LAD2 cells were derived from CD34+ cells following marrow aspiration of a patient with aggressive mastocytosis with no identified mutations in <i>KIT</i>. Another aspiration gave rise to a second cell line which has recently been re-established (LADR). We queried whether LADR had unique properties for the preclinical study of human mast cell biology. Methods: LADR and LAD2 cells were cultured under identical conditions. Experiments examined proliferation, beta-hexosaminidase (β-hex) release, surface receptor and granular protease expression, infectivity with HIV, and gene expression. Results: LADR cells were larger and more granulated as seen with Wright−Giemsa staining and flow cytometry, with cell numbers doubling in 4 weeks, in contrast to LAD2 cells, which doubled every 2 weeks. Both LADR and LAD2 cells released granular contents following aggregation of FcεRI. LADR cells showed log-fold increases in FcεRI/CD117 and expressed CD13, CD33, CD34, CD63, CD117, CD123, CD133, CD184, CD193, and CD195, while LAD2 cells expressed CD33, CD34, CD63, CD117, CD133, CD193 but not CD13, CD123, CD184, or CD195. LADR tryptase expression was one-log-fold increased. LADR cell and LAD2 cell chymase expression were similar. Both cell lines could be infected with T-tropic, M-tropic, and dual tropic HIV. Following monomeric human IgE stimulation, LADR cells showed greater surface receptor and mRNA expression for CD184 and CD195. Expression arrays revealed differences in gene upregulation, especially for the suppressor of cytokine signaling (SOCS) family of genes with their role in JAK2/STAT3 signaling and cellular myelo<b>c</b>ytomatosis oncogene (c-MYC) in cell growth and regulation. Conclusions: LADR cells are thus unique in that they exhibit a slower proliferation rate, are more advanced in development, have increased FcεRI/CD117 and tryptase expression, have a different profile of gene expression, and show earlier infectivity with HIV-BAL, LAV, and TYBE when compared to LAD2 cells. This new cell line is thus a valuable addition to the few FcεRI+ human mast cell lines previously described and available for scientific inquiry. |
topic |
mast cell lad cells hiv allergy fcεr1 stem cell factor c-<i>kit</i> socs <i>myc</i> |
url |
https://www.mdpi.com/1422-0067/20/22/5520 |
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