Profiling Immune Cells in HPV16+ vHSIL

Formalin-fixed paraffin-embedded biopsies of 24 HPV16⁺ vulvar high-grade squamous intraepithelial lesions (vHSIL) patients, treated with an experimental vaccine containing 13 SLPs covering the entire amino acid sequence of HPV16 oncoproteins E6 and E7, were cut in 4 um thick sections. These samples were stained with two multiplex immunofluorescence panels,20 (link) one for T cells and one for myeloid cells. The slides were then scanned with the Vectra multispectral imaging system (PerkinElmer), and cells were, after manual training, automatically phenotyped and counted with inForm advanced image analysis software (PerkinElmer).

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Beyranvand Nejad E., Labrie C., Abdulrahman Z., van Elsas M.J., Rademaker E., Kleinovink J.W., van der Sluis T.C., van Duikeren S., Teunisse A.F., Jochemsen A.G., Oosting J., de Miranda N.F., Van Hall T., Arens R, & van der Burg S.H. (2020). Lack of myeloid cell infiltration as an acquired resistance strategy to immunotherapy. Journal for Immunotherapy of Cancer, 8(2), e001326.

Publication 2020

Vectra multispectral imaging system inForm advanced image analysis software

Amino acid sequence Biopsies Cells Embedded paraffin Formalin High grade squamous intraepithelial lesions Hpv16 Hpv16 e6 Immunofluorescence Myeloid cells Oncoproteins Patients T cells Vaccine Vectra Vulvar

Corresponding Organization : Leiden University Medical Center

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Variable analysis

independent variables

Experimental vaccine containing 13 SLPs covering the entire amino acid sequence of HPV16 oncoproteins E6 and E7

dependent variables

Phenotyping and counting of cells in the formalin-fixed paraffin-embedded biopsies of 24 HPV16+ vulvar high-grade squamous intraepithelial lesions (vHSIL) patients using multiplex immunofluorescence panels and Vectra multispectral imaging system

control variables

Formalin-fixed paraffin-embedded biopsies of 24 HPV16+ vulvar high-grade squamous intraepithelial lesions (vHSIL) patients
Staining with two multiplex immunofluorescence panels, one for T cells and one for myeloid cells
Scanning of slides with the Vectra multispectral imaging system (PerkinElmer)
Automated phenotyping and counting of cells using inForm advanced image analysis software (PerkinElmer)

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