Dataset Information

Gene expression profiles of heat shock proteins 70 and 90 from Empoasca onukii (Hemiptera: Cicadellidae) in response to temperature stress.

ABSTRACT: Empoasca onukii Matsuda is a worldwide pest that causes great economic loss in tea growing areas and is significantly affected by temperatures. Heat shock protein (Hsp) genes are important in insects' response to temperature stress. In this study, two full-length Hsp genes, Eohsp90 and Eohsp70, were cloned from E. onukii using rapid amplification of complementary DNA ends. The open reading frames of Eohsp90 and Eohsp70 were 2,172?bp and 2,016?bp in length, respectively. Their deduced amino acid sequences of Eohsp90 and Eohsp70 showed high homology with other species. Subsequently, the transcriptional expression of Eohsp90 and Eohsp70 in E. onukii adults exposed to various temperatures (-5, 0, 10, 15, 20, 25, 30, 35, 38, 41 and 44°C) for 1?h, and at extreme temperatures (0°C and 41°C) for various time duration (0, 20, 40, 60, 80, 100, and 120?min) were investigated via real-time quantitative polymerase chain reaction. The relative expression levels of both Eohsp90 and Eohsp70 in E. onukii adults were upregulated as the temperature rises or falls over time, except in the -5°C or 44°C temperature groups. Moreover, the expression level in the temperature elevated groups was higher than that of the lower temperature groups. In addition, the Eohsp70 generally demonstrated a higher transcriptional level than Eohsp90, and both genes had a higher expression profile in female adults compared with the males. The expression profiles indicated that Eohsp90 and Eohsp70 may play important roles in E. onukii adult responses to ecologically relevant environmental temperature threat.

SUBMITTER: Qiao L

PROVIDER: S-EPMC4535498 | biostudies-literature | 2015

REPOSITORIES: biostudies-literature

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Gene expression profiles of heat shock proteins 70 and 90 from Empoasca onukii (Hemiptera: Cicadellidae) in response to temperature stress.

Qiao Li L Wu Jun X JX Qin Dao Z DZ Liu Xiang C XC Lu Zhao C ZC Lv Li Z LZ Pan Zi L ZL Chen Hao H Li Guang W GW

Journal of insect science (Online) 20150417

Empoasca onukii Matsuda is a worldwide pest that causes great economic loss in tea growing areas and is significantly affected by temperatures. Heat shock protein (Hsp) genes are important in insects' response to temperature stress. In this study, two full-length Hsp genes, Eohsp90 and Eohsp70, were cloned from E. onukii using rapid amplification of complementary DNA ends. The open reading frames of Eohsp90 and Eohsp70 were 2,172 bp and 2,016 bp in length, respectively. Their deduced amino acid ...[more]

PMID: 25888707

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Project description:BackgroundHeat shock proteins (HSPs) are found in all prokaryotes and most compartments of eukaryotic cells. Members of the HSP family mediate immune responses to tissue damage or cellular stress. However, little is known about the immune response induced by the oriental liver fluke, Clonorchis sinensis, even though this organism is carcinogenic to humans. We address this issue in the present study in mouse bone marrow dendritic cells (mBMDCs), using recombinant HSP70 and 90 from C. sinensis (rCsHSP70 and rCsHSP90).MethodsrCsHSP70 and rCsHSP90 were produced in an E. coli system. Purified recombinant proteins were treated in BMDCs isolated from C57BL/6 mice. T cells were isolated from Balb/c mice and co-cultured with activated mBMDCs. Expression of surface molecules was measured by flow cytometry and cytokine secretion was quantified using ELISA. C57BL/6 mice were divided into four groups, including peptide alone, peptide/Freund's adjuvant, peptide/CsHSP70, peptide/CsHSP90, and were immunized intraperitoneally three times. Two weeks after final immunization, antibodies against peptide were measured using ELISA.ResultsBoth proteins induced a dose-dependent upregulation in major histocompatibility complex and co-stimulatory molecule expression and increased secretion of pro-inflammatory cytokines including interleukin (IL)-1β, -6, and -12p70 and tumor necrosis factor-α in mBMDCs. Furthermore, when allogenic T cells were incubated with mBMDCs activated by rCsHSP70 and rCsHSP90, the helper T cell (Th)1 cytokine interferon-γ was up-regulated whereas the level of the Th2 cytokine IL-4 was unchanged. These results indicate that rCsHSPs predominantly induce a Th1 response. Over and above these results, we also demonstrated that the production of peptide-specific antibodies can be activated after immunization via in vitro peptide binding with rCsHSP70 or rCsHSP90.ConclusionThis study showed for the first time that the HSP or HSP/peptide complexes of C. sinensis could be considered as a more effective vaccine against C. sinensis infection as results of the activator of host immune response as well as the adjuvant for antigenic peptide conjugate to induce peptide-specific antibody response in mice.

Dataset Information

Gene expression profiles of heat shock proteins 70 and 90 from Empoasca onukii (Hemiptera: Cicadellidae) in response to temperature stress.

Publications

Gene expression profiles of heat shock proteins 70 and 90 from Empoasca onukii (Hemiptera: Cicadellidae) in response to temperature stress.

Similar Datasets

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