Abstract
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 KIT. 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 myelocytomatosis 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.
Highlights
Mast cells are known to play a major role in innate and acquired immunity
All cell lines are available for study of preclinical human mast cell biology in lieu of primary mast cell cultures derived from bone marrow, or peripheral or cord blood precursors [5]
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Summary
Mast cells are known to play a major role in innate and acquired immunity. Since 1988, a number of cell lines have been described, including HMC-1 (Human Mast Cell leukemia-1), LAD2 (Laboratory of Allergic Diseases 2), LUVA (Laboratory of University of VirginiA), and ROSA cell lines [1,2,3,4]. HMC-1 and LAD2 cell lines were derived from patients with mast cell leukemia. LADR cells share some similarities to LAD2 cells while differing in some important aspects of degranulation, surface receptor expression, protease content, We firgsetneexexpparnesdsieodn,aannddstuhsecenptcibhialirtyactoteinrifzecetdionL. WanhdeLnAiDnf2eccetellds wweirtehinHitIiVallwy iitnhcudbiaffteerdent tropisms, HIV-BAL, HIV-LAV, and HIV-TYBE infectivity, as measured by p24 assay, were detected in LADR cells earlier (day 8) than LAD2 cells (day 12), consistent with their differential surface expression of CD184 (Figure 3C). When infected with HIV with different tropisms, HIV-BAL, HIV-LAV, and HIV-TYBE infectivity, as measured by p24 assay, were Int.dJ.eMteocl.teSdci.i2n01L9,A2D0, R552c0ells earlier (day 8) than LAD2 cells (day 12), consistent with their differenti4aol f 9 surface expression of CD184 (Figure 3C).
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