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Correlation of Circulating CD64+/CD163+ Monocyte Ratio and stroma/peri-tumoral CD163+ Monocyte Density with Human Papillomavirus Infected Cervical Lesion Severity

  • Original Article
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Cancer Microenvironment

Abstract

HPV infected cervical cells secrete mediators that are gradually changed and have influence on infiltrating M2 phenotypic monocytes in cervical lesions. However, profiles of circulating immune cells in women with cervical lesions and M2 phenotypic monocyte activity in HPV infected cervical lesions are limited. This study aimed to investigate circulating monocyte populations correlated with M2 phenotype density and its activity in HPV infected cervical lesions. HPV DNA was investigated in cervical tissues using PCR. High risk HPV E6/E7 mRNA was detected using in situ hybridization. CD163 immunohistochemical staining was performed for M2 macrophage. CD163 and Arg1 mRNA expression were detected using real-time PCR. Circulating monocyte subpopulations were analyzed using flow cytometry. CD163 and Arg1 mRNA expression were increased according to cervical lesion severity and corresponding with density of M2 macrophage in HSIL and SCC in stroma and peri-tumoral areas. Additionally, the relationship between M2 macrophage infiltration and high risk HPV E6/E7 mRNA expression was found and corresponded with cervical lesion severity. Circulating CD14+CD16+ and CD14+CD163+ monocytes were elevated in No-SIL and cervical lesions. Interestingly, CD14+CD64+ monocyte was greatly elevated in HSIL and SCC, whereas intracellular IL-10+ monocytes were not significantly different between cervical lesions. The correlation between increasing ratio of circulating CD64+/CD163+ monocyte and density of infiltrating CD163+ monocytes was associated with severity of HPV infected cervical lesions. The elevated circulating CD64+/CD163+ monocyte ratio correlates to severity of HPV infected cervical lesions and might be a prognostic marker in cervical cancer progression.

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Abbreviations

APC:

Antigen presenting cell

Arg1:

Arginase 1

CD:

Cluster of differentiation

DapB:

Bacterial dihydrodipicolinate reductase

DNA:

Deoxyribonucleic acid

FFPE:

Formalin-fixed paraffin-embedded tissue

FITC:

Fluorescein isothiocyanate

GAPDH:

Glyceraldehyde 3-phosphate dehydrogenase

HR-HPV:

High-risk human papillomavirus

HSIL:

High-grade squamous intraepithelial lesion

IHC:

Immunohistochemical staining

IL-10:

Interleukin 10

ISH:

In situ hybridization

LSIL:

Low-grade squamous intraepithelial lesion

mRNA:

Messenger ribonucleic acid

No-SIL:

No squamous intraepithelial lesion

PBMC:

Peripheral blood mononuclear cell

PCR:

Polymerase chain reaction

PE:

Phycoerythrin

PPIB:

Human peptidylpropyl isomerase B

SA-HRP:

Streptavidin horseradish peroxidase

SCC:

Squamous cell carcinoma

TMA:

Tissue microarray

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Acknowledgements

We would like to acknowledge Prof. David Blair for editing the MS via Publication Clinic KKU.

Author information

Authors and Affiliations

  1. Department of Microbiology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Piyawut Swangphon, Chamsai Pientong & Tipaya Ekalaksananan

  2. HPV & EBV and Carcinogenesis Research Group, Khon Kaen University, Khon Kaen, 40002, Thailand

    Piyawut Swangphon, Chamsai Pientong, Nuchsupha Sunthamala, Sureewan Bumrungthai & Tipaya Ekalaksananan

  3. Department of Biology, Faculty of Science, Mahasarakham University, Mahasarakham, 44150, Thailand

    Nuchsupha Sunthamala

  4. Graduate School, Khon Kaen University, Khon Kaen, Thailand

    Nuchsupha Sunthamala

  5. Division of Microbiology and Parasitology, School of Medical Sciences, University of Phayao, Phayao, 56000, Thailand

    Sureewan Bumrungthai

  6. Department of Molecular Immunology, Tokyo Medical and Dental University, Tokyo, 113-0034, Japan

    Miyuki Azuma

  7. Department of Obstetrics and Gynecology, Faculty of Medicine, Khon Kaen University, Khon Kaen, 40002, Thailand

    Pilaiwan Kleebkaow

  8. Department of Obstetrics and Gynecology, Khon Kaen Central Hospital, Khon Kaen, 40000, Thailand

    Thumwadee Tangsiriwatthana & Ussanee Sangkomkamhang

  9. Department of Pharmacology, Faculty of Medicine, Khon Kaen University, Khon Kaen, Thailand

    Bunkerd Kongyingyoes

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  1. Piyawut Swangphon
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  2. Chamsai Pientong
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Corresponding author

Correspondence to Tipaya Ekalaksananan.

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Conflict of Interest

The authors declare that they have no conflict of interest for this study.

Ethical Approval

Ethical approval of this study was obtained from the Khon Kaen University Ethics Committee for Human Research (KKUEC) and the Khon Kaen Central Hospital Ethics Committee for Human Research with reference numbers HE531387 and No. 20/04/2554, respectively.

Funding

Budgets for chemicals, reagents and sample collection processes were supported by Khon Kaen University (Grant numbers 551603, 564103 and 573003) and Faculty of Medicine, Khon Kaen University (Grant number I54141). PS was supported by the research fund for supporting lecturer to admit high potential student to study and research on his expert program from Graduate School (Grant number 511H1201). NS was supported by the Post-doctoral Program from Research Affairs and Graduate School, Khon Kaen University (Grant number 58222). The funders have no role in study design, data collection and analysis, preparation of the manuscript, and decision to publish.

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Swangphon, P., Pientong, C., Sunthamala, N. et al. Correlation of Circulating CD64+/CD163+ Monocyte Ratio and stroma/peri-tumoral CD163+ Monocyte Density with Human Papillomavirus Infected Cervical Lesion Severity. Cancer Microenvironment 10, 77–85 (2017). https://doi.org/10.1007/s12307-017-0200-2

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