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Volume 12
Issue 8
10.3390/v12080787
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Article

Comparison of Multiplexed Immunofluorescence Imaging to Chromogenic Immunohistochemistry of Skin Biomarkers in Response to Monkeypox Virus Infection

by
Anup Sood
1,
Yunxia Sui
1,
Elizabeth McDonough
1,
Alberto Santamaría-Pang
1,
Yousef Al-Kofahi
1,
Zhengyu Pang
1,
Peter B. Jahrling
2,
Jens H. Kuhn
2,* and
Fiona Ginty
1,*
1
GE Research, 1 Research Circle, Niskayuna, NY 12309, USA
2
Integrated Research Facility at Fort Detrick, National Institute of Allergy and Infectious Diseases, National Institutes of Health, B-8200 Research Plaza, Frederick, MD 21702, USA
*
Authors to whom correspondence should be addressed.
Viruses 2020, 12(8), 787; https://doi.org/10.3390/v12080787
Submission received: 23 May 2020 / Revised: 15 July 2020 / Accepted: 19 July 2020 / Published: 23 July 2020
(This article belongs to the Special Issue Smallpox and Emerging Zoonotic Orthopoxviruses 2.0: What Is Coming Next?)
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Abstract

Over the last 15 years, advances in immunofluorescence-imaging based cycling methods, antibody conjugation methods, and automated image processing have facilitated the development of a high-resolution, multiplexed tissue immunofluorescence (MxIF) method with single cell-level quantitation termed Cell DIVETM. Originally developed for fixed oncology samples, here it was evaluated in highly fixed (up to 30 days), archived monkeypox virus-induced inflammatory skin lesions from a retrospective study in 11 rhesus monkeys to determine whether MxIF was comparable to manual H-scoring of chromogenic stains. Six protein markers related to immune and cellular response (CD68, CD3, Hsp70, Hsp90, ERK1/2, ERK1/2 pT202_pY204) were manually quantified (H-scores) by a pathologist from chromogenic IHC double stains on serial sections and compared to MxIF automated single cell quantification of the same markers that were multiplexed on a single tissue section. Overall, there was directional consistency between the H-score and the MxIF results for all markers except phosphorylated ERK1/2 (ERK1/2 pT202_pY204), which showed a decrease in the lesion compared to the adjacent non-lesioned skin by MxIF vs an increase via H-score. Improvements to automated segmentation using machine learning and adding additional cell markers for cell viability are future options for improvement. This method could be useful in infectious disease research as it conserves tissue, provides marker colocalization data on thousands of cells, allowing further cell level data mining as well as a reduction in user bias.
Keywords: immunohistochemistry; IHC; monkeypox; monkeypox virus; MPXV; multiplexed immunofluorescence; MxIF; Orthopoxvirus; Poxviridae; poxvirus immunohistochemistry; IHC; monkeypox; monkeypox virus; MPXV; multiplexed immunofluorescence; MxIF; Orthopoxvirus; Poxviridae; poxvirus

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MDPI and ACS Style

Sood, A.; Sui, Y.; McDonough, E.; Santamaría-Pang, A.; Al-Kofahi, Y.; Pang, Z.; Jahrling, P.B.; Kuhn, J.H.; Ginty, F. Comparison of Multiplexed Immunofluorescence Imaging to Chromogenic Immunohistochemistry of Skin Biomarkers in Response to Monkeypox Virus Infection. Viruses 2020, 12, 787. https://doi.org/10.3390/v12080787

AMA Style

Sood A, Sui Y, McDonough E, Santamaría-Pang A, Al-Kofahi Y, Pang Z, Jahrling PB, Kuhn JH, Ginty F. Comparison of Multiplexed Immunofluorescence Imaging to Chromogenic Immunohistochemistry of Skin Biomarkers in Response to Monkeypox Virus Infection. Viruses. 2020; 12(8):787. https://doi.org/10.3390/v12080787

Chicago/Turabian Style

Sood, Anup, Yunxia Sui, Elizabeth McDonough, Alberto Santamaría-Pang, Yousef Al-Kofahi, Zhengyu Pang, Peter B. Jahrling, Jens H. Kuhn, and Fiona Ginty. 2020. "Comparison of Multiplexed Immunofluorescence Imaging to Chromogenic Immunohistochemistry of Skin Biomarkers in Response to Monkeypox Virus Infection" Viruses 12, no. 8: 787. https://doi.org/10.3390/v12080787

APA Style

Sood, A., Sui, Y., McDonough, E., Santamaría-Pang, A., Al-Kofahi, Y., Pang, Z., Jahrling, P. B., Kuhn, J. H., & Ginty, F. (2020). Comparison of Multiplexed Immunofluorescence Imaging to Chromogenic Immunohistochemistry of Skin Biomarkers in Response to Monkeypox Virus Infection. Viruses, 12(8), 787. https://doi.org/10.3390/v12080787

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