PD-L1/PD-1 and CTLA-4 Expression in Equine Penile Squamous Cell Carcinomas
Abstract
:Simple Summary
Abstract
1. Introduction
2. Materials and Methods
2.1. Case Selection
- confirmed penile localization of the lesions;
- and availability of >0.5 cm2 of FFPE tumor tissue evaluated on section.
2.2. DNA Extraction and EcPV2 Detection
2.3. RNA Extraction and Real-Time PCR
2.4. Western Blotting
2.5. Immunohistochemistry
3. Results
3.1. Case Selection and Histology
3.2. DNA Extraction and EcPV2 Detection
3.3. PD-1 and PD-L1 Gene Expression
3.4. Western Blotting
3.5. Immunohistochemistry
4. Discussion
5. Conclusions
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Acknowledgments
Conflicts of Interest
References
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Gene | Primer Pairs Sequences | Amplicon Length | Accession and References |
---|---|---|---|
EcPV2-L1 | F-5′- TTGTCCAGGAGAGGGGTTAG-3′ | 81 | NC_012123 [7] |
R-5′- TGCCTTCCTTTTCTTGGTGG-3′ | |||
pEcPV2-L1 | FAM-CGTCCAGCACCTTCGACCACCA-TAMRA | ||
EcPV2-E6 | F-5′-CGTTGGCCTTCTTTGCATCT-3′ | 81 | NC_012123 [8] |
R-5′-AGGTTCAGGTCTGCTGTGTT-3′ | |||
p-EcPV2-E6 | FAM- CCGTGTGGCTATGCTGATGACATTTGG-TAMRA | ||
Ec-B2M DNA detection | F-5′-CTGATGTTCTCCAGGTGTTCC-3′ | 114 | NM_001082502 [7] |
R-5′-TCAATCTCAGGCGGATGGAA-3′ | |||
p-B2M | FAM-ACTCACGTCACCCAGCAGAGA-TAMRA | ||
Ec-PDCD1 (PD1) | F-5′-GCCTGTGTCCTGACCACC-3′ | 150 | XM_023642815 |
R-5′-CTCCGGGGTCTTCTCTCG-3′ | |||
Ec-CD274 (PDL1) | F-5′-GTGCTGACTACAAGCGGATT-3′ | 120 | XM_001492842 |
R-5′-GGTAACCCTCAGCCTGACAT-3′ | |||
Ec-B2M cDNA | F-5′-TCCTGCTCGGGCTACTCTC-3′ | 83 | NM_001082502 [7] |
R-5′-TGCTGGGTGACGTGAGTAAA-3′ |
Case ID | Histological Diagnosis | DNA | cDNA | Normalized Expression | ||||
---|---|---|---|---|---|---|---|---|
B2M | L1 | E6 | L1 | E6 | PDCD1 | CD274 | ||
1 | SCC | + | +++ | ++ | + | ++ | Negative | ND |
2 | SCC | + | + | + | ND | ND | ND | 5.3 ± 7.3 |
3 | CIS | + | +++ | ++ | + | + | ND | Negative |
4 | SCC | + | ++++ | +++ | + | + | ND | ND |
6 | P | + | + | + | ND | ND | Negative | ND |
7 | SCC | + | + | + | ND | ND | Negative | ND |
8 | SCC | + | ++++ | +++ | + | + | Negative | ND |
9 | SCC | + | ++++ | +++ | + | + | Negative | Negative |
10 | SCC | + | ++++ | +++ | + | + | Negative | ND |
11 | SCC | + | + | + | ND | ND | Negative | ND |
12 | SCC | + | ++++ | +++ | + | + | Negative | ND |
13 | SCC | + | + | + | ND | ND | Negative | ND |
14 | SCC | + | ++ | + | + | ND | ND | ND |
15 | SCC | + | ++++ | +++++ | ++ | + | Negative | ND |
17 | SCC | + | ++++ | +++ | + | + | Negative | ND |
18 | SCC | + | ++ | ++ | ND | ND | Negative | 0.6 ± 0.7 |
20 | SCC | + | ND | ND | ND | ND | Negative | Negative |
21 | CIS | + | ++++ | +++ | ++ | ++ | ND | 9.3 ± 8.4 |
23 | SCC | + | +++ | ++++ | + | + | Negative | ND |
24 | SCC | + | ND | ND | ND | ND | Negative | ND |
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Porcellato, I.; Mecocci, S.; Brachelente, C.; Cappelli, K.; Armando, F.; Tognoloni, A.; Chiaradia, E.; Stefanetti, V.; Mechelli, L.; Pepe, M.; Gialletti, R.; Passeri, B.; Ghelardi, A.; Razzuoli, E. PD-L1/PD-1 and CTLA-4 Expression in Equine Penile Squamous Cell Carcinomas. Animals 2021, 11, 2121. https://doi.org/10.3390/ani11072121
Porcellato I, Mecocci S, Brachelente C, Cappelli K, Armando F, Tognoloni A, Chiaradia E, Stefanetti V, Mechelli L, Pepe M, Gialletti R, Passeri B, Ghelardi A, Razzuoli E. PD-L1/PD-1 and CTLA-4 Expression in Equine Penile Squamous Cell Carcinomas. Animals. 2021; 11(7):2121. https://doi.org/10.3390/ani11072121
Chicago/Turabian StylePorcellato, Ilaria, Samanta Mecocci, Chiara Brachelente, Katia Cappelli, Federico Armando, Alessia Tognoloni, Elisabetta Chiaradia, Valentina Stefanetti, Luca Mechelli, Marco Pepe, Rodolfo Gialletti, Benedetta Passeri, Alessandro Ghelardi, and Elisabetta Razzuoli. 2021. "PD-L1/PD-1 and CTLA-4 Expression in Equine Penile Squamous Cell Carcinomas" Animals 11, no. 7: 2121. https://doi.org/10.3390/ani11072121