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Microarray analysis of immediate-type allergy in KU812 cells in response to fulvic acid

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Abstract

Fulvic acid (FA) is class of compounds of humic substances formed through the degradation of organic substances by chemical and biological processes. FA has been utilized in traditional Chinese medicine and possesses various pharmacological properties. Previously, we reported that FA extracted from solubilized excess sludge (SS-FA) had an inhibitory effect on β-hexosaminidase release in human leukemia basophilic (KU812) cells. In this study, we investigated the effects of SS-FA on the immediate-type allergic reaction and studied its possible mechanisms of action in KU812 cells following activation with phorbol myristate acetate (20 nmol L−1) plus calcium ionophore A23187 (1 μmol L−1) (PMACI). The inhibitory effect of SS-FA on degranulation in PMACI-stimulated KU812 cells was examined using histamine release assay. SS-FA significantly decreased the histamine release in KU812 cells at concentrations of 0.1–10.0 μg mL−1. To gain more information regarding the mechanism of the suppression of degranulation following SS-FA treatment, microarray was conducted to determine which genes were differentially expressed in response to SS-FA in PMACI-activated KU812 cells. From a total of 201 genes in the DNA chip, 28 genes were up-regulated and 173 genes were down-regulated in cells pretreated with SS-FA for 15 min and stimulated with PMACI. From the 71 genes that showed more than two fold change in expression, 16 genes were significantly down-regulated that were subjected to hierarchical clustering. SS-FA affected the expression of genes that were involved in the following pathways: signal transduction, cytokine–cytokine receptor interaction, immune response, cell adhesion molecules and IgE receptor β subunit response.

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Abbreviations

DNP-BSA:

Dinitrophenylated bovine serum albumin

FA:

Fulvic acid

FBS:

Fetal bovine serum

KU812:

Human leukemia basophilic

PMA:

Phorbol 12-myristate 13-acetate

PMACI:

PMA plus calcium ionophore A23187

RBL-2H3:

Rat basophilic leukemia

SLE:

Systemic lupus erythematosus

SS-FA:

Fulvic acid extracted from solubilized sludge

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Acknowledgments

This research was partially supported by the Mitsui & Co., Ltd. Environment Fund.

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Authors and Affiliations

  1. Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8572, Japan

    Hideko Motojima, Myra O. Villareal, Junkyu Han & Hiroko Isoda

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  1. Hideko Motojima
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  2. Myra O. Villareal
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  3. Junkyu Han
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  4. Hiroko Isoda
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Correspondence to Hiroko Isoda.

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Motojima, H., O. Villareal, M., Han, J. et al. Microarray analysis of immediate-type allergy in KU812 cells in response to fulvic acid. Cytotechnology 63, 181–190 (2011). https://doi.org/10.1007/s10616-010-9333-6

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  • DOI: https://doi.org/10.1007/s10616-010-9333-6

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