PMC9483160_01
Residual SARS-CoV-2 detected in the appendix and skin tissues of patient 1 using multiplex immunohistochemistry and RNAscope (A, D) Representative images of the (A) appendix and (D) skin tissues stained with hematoxylin and eosin, with differentiated staining of nuclear (hematoxylin) and cytoplasmic (eosin) components. (B, E) Representative images of the (B) appendix and (E) skin tissues stained for DAPI (blue), CK/EpCAM (red), CD45 (cyan), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (yellow). (C, F) Representative images of the (C) appendix and (F) skin tissues stained for DAPI (blue), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (red). (G-I) Representative images of the (G, H) appendix tissue obtained from patient 1 and (I) normal colon tissue obtained from patients not affected by COVID-19, subjected to RNAscope in situ hybridization with a nuclear component (hematoxylin) counterstained. SARS-CoV-2 spike RNAs are labelled as green dots, examples of the green dots are marked by black arrows. (A-F) Scale bar, 100 mum. (G-I) Scale bar, 20 mum.
fimmu-13-939989-g001.jpg
PMC9483160_02_fimmu-13-939989-g001.jpg
f1
With the same techniques used for patient 1, viral NP and spike protein was detected and observed in the tumor-adjacent area ( Figures 2A, B ,  Supplementary Figure 1 ).
Residual SARS-CoV-2 detected in the breast tissue of patient 2 using multiplex immunohistochemistry and RNAscope (A) Representative images of the breast tissue stained with hematoxylin and eosin, with differentiated staining of nuclear (hematoxylin) and cytoplasmic (eosin) components. (B) Representative images of the breast tissue stained for DAPI (blue), CK/EpCAM (red), CD45 (cyan), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (yellow). (C) Representative images of the breast tissue stained for DAPI (blue), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (red). (D-F) Representative images of (D, E) the breast tissue obtained from patient 2 and (F) normal breast tissue obtained from patients not affected by COVID-19, subjected to RNAscope in situ hybridization with a nuclear component (hematoxylin) counterstained. SARS-CoV-2 spike RNAs are labelled as green dots, examples of the green dots are marked by black arrows. (A-C) Scale bar, 100 mum. (D-F) Scale bar, 20 mum.
fimmu-13-939989-g002.jpg
PMC9483160_02_fimmu-13-939989-g002.jpg
f2
With the same techniques used for patient 1, viral NP and spike protein was detected and observed in the tumor-adjacent area ( Figures 2A, B ,  Supplementary Figure 1 ). These viral antigens also co-localized with myeloid and macrophage markers CD68 ( Figure 2C ), CD14, CD206, and CD169 ( Supplementary Figure 3 ). Viral RNA was also detected in the breast, within both the extracellular space of the tissue ( Figure 2D ) and within the cells ( Figure 2E ). The specificity of the used probe was tested on normal breast tissue obtained from an independent cohort in 2018 or earlier ( Figure 2F ).
PMC9483160_02
Residual SARS-CoV-2 detected in the appendix and skin tissues of patient 1 using multiplex immunohistochemistry and RNAscope (A, D) Representative images of the (A) appendix and (D) skin tissues stained with hematoxylin and eosin, with differentiated staining of nuclear (hematoxylin) and cytoplasmic (eosin) components. (B, E) Representative images of the (B) appendix and (E) skin tissues stained for DAPI (blue), CK/EpCAM (red), CD45 (cyan), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (yellow). (C, F) Representative images of the (C) appendix and (F) skin tissues stained for DAPI (blue), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (red). (G-I) Representative images of the (G, H) appendix tissue obtained from patient 1 and (I) normal colon tissue obtained from patients not affected by COVID-19, subjected to RNAscope in situ hybridization with a nuclear component (hematoxylin) counterstained. SARS-CoV-2 spike RNAs are labelled as green dots, examples of the green dots are marked by black arrows. (A-F) Scale bar, 100 mum. (G-I) Scale bar, 20 mum.
fimmu-13-939989-g001.jpg
PMC9483160_02_fimmu-13-939989-g001.jpg
f1
With the same techniques used for patient 1, viral NP and spike protein was detected and observed in the tumor-adjacent area ( Figures 2A, B ,  Supplementary Figure 1 ).
Residual SARS-CoV-2 detected in the breast tissue of patient 2 using multiplex immunohistochemistry and RNAscope (A) Representative images of the breast tissue stained with hematoxylin and eosin, with differentiated staining of nuclear (hematoxylin) and cytoplasmic (eosin) components. (B) Representative images of the breast tissue stained for DAPI (blue), CK/EpCAM (red), CD45 (cyan), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (yellow). (C) Representative images of the breast tissue stained for DAPI (blue), SARS-CoV-2 nucleocapsid protein (COVID NP; green), and CD68 (red). (D-F) Representative images of (D, E) the breast tissue obtained from patient 2 and (F) normal breast tissue obtained from patients not affected by COVID-19, subjected to RNAscope in situ hybridization with a nuclear component (hematoxylin) counterstained. SARS-CoV-2 spike RNAs are labelled as green dots, examples of the green dots are marked by black arrows. (A-C) Scale bar, 100 mum. (D-F) Scale bar, 20 mum.
fimmu-13-939989-g002.jpg
PMC9483160_02_fimmu-13-939989-g002.jpg
f2
With the same techniques used for patient 1, viral NP and spike protein was detected and observed in the tumor-adjacent area ( Figures 2A, B ,  Supplementary Figure 1 ). These viral antigens also co-localized with myeloid and macrophage markers CD68 ( Figure 2C ), CD14, CD206, and CD169 ( Supplementary Figure 3 ). Viral RNA was also detected in the breast, within both the extracellular space of the tissue ( Figure 2D ) and within the cells ( Figure 2E ). The specificity of the used probe was tested on normal breast tissue obtained from an independent cohort in 2018 or earlier ( Figure 2F ).