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The genome of polymorphonuclear neutrophils maintains normal coding sequences.

ABSTRACT: Genetic studies often use genomic DNA from whole blood cells, of which the majority are the polymorphonuclear myeloid cells. Those cells undergo dramatic change of nuclear morphology following cellular differentiation. It remains elusive if the nuclear morphological change accompanies sequence alternations from the intact genome. If such event exists, it will cause a serious problem in using such type of genomic DNA for genetic study as the sequences will not represent the intact genome in the host individuals. Using exome sequencing, we compared the coding regions between neutrophil, which is the major type of polymorphonuclear cells, and CD4+ T cell, which has an intact genome, from the same individual. The results show that exon sequences between the two cell types are essentially the same. The minor differences represented by the missed exons and base changes between the two cell types were validated to be mainly caused by experimental errors. Our study concludes that genomic DNA from whole blood cells can be safely used for genetic studies.

SUBMITTER: Xiao F

PROVIDER: S-EPMC3826757 | biostudies-literature | 2013

REPOSITORIES: biostudies-literature

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The genome of polymorphonuclear neutrophils maintains normal coding sequences.

Xiao Fengxia F Kim Yeong C YC Wen Hongxiu H Luo Jiangtao J Chen Peixian P Cowan Kenneth K Wang San Ming SM

PloS one 20131108 11

Genetic studies often use genomic DNA from whole blood cells, of which the majority are the polymorphonuclear myeloid cells. Those cells undergo dramatic change of nuclear morphology following cellular differentiation. It remains elusive if the nuclear morphological change accompanies sequence alternations from the intact genome. If such event exists, it will cause a serious problem in using such type of genomic DNA for genetic study as the sequences will not represent the intact genome in the h ...[more]

PMID: 24250807

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Project description:Oral polymorphonuclear neutrophils (oPMNs) are essential for the maintenance of oral health. Functionally active oPMNs have been described in oral health, gingivitis and periodontitis. Interestingly, recent evidence showed that oPMNs from chronic periodontitis patients have a similar functional and reactivity pattern compared to oPMNs from healthy subjects. However, it remains unknown whether oPMNs respond to the initiation and resolution of gingival inflammation. Therefore, this study aimed to evaluate the oPMNs’ transcriptional responses during the initiation and resolution of an experimentally induced gingivitis. Oral rinses were obtained from 8 subjects at 4 different time points during the experiment for the isolation of oPMNs; day 0 (baseline), day 9 (during gingivitis challenge), day 14 (end of the challenge) and day 21 (resolution). Transcriptome of oPMNs was determined by RNAsequencing. Differentially expressed genes (DEGs) was selected at P<0.01 level, and evaluated for pathway regulation at different time points using Ingenuity Pathway Analysis suite. During the gingivitis challenge, dental plaque levels and gingival bleeding levels were significantly increased at day 14 compared to baseline. Transcriptome analysis indicate four major clusters of DEGs, which consisted of a group of 256 initial response DEGs (day 9 only), 221 late response DEGs (day 14 only), 53 persistent response DEGs (consistent at day 9 and day 14), and a group of 524 DEGs showing response only after the gingivitis challenge (day 21 only). Pathway analysis of the initial response DEGs showed their involvement in many immune regulatory pathways (interleukin signaling, reactive oxygen species processes and granulocyte adhesion and diapedesis). Late responding DEGs mainly showed activation of signaling pathways and phagosome formation as part of the phagocytosis process, whereas DEGs at day 21 were involved in epithelial adherence signaling and other miscellaneous related signaling pathways. This study showed that oPMNs have specific expression profiles during the development and resolution of experimental gingivitis. Even though, the specific observed pathways were not identical within each time point, the altered pathways indicate PMN responses in general and suggests an immunomodulatory role for the oPMNs. The oPMNs may therefore play a co-controlling part in the initiation and resolution of gingival inflammation.

Dataset Information

The genome of polymorphonuclear neutrophils maintains normal coding sequences.

Publications

The genome of polymorphonuclear neutrophils maintains normal coding sequences.

Similar Datasets

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