PMID- 30609079 OWN - NLM STAT- MEDLINE DCOM- 20200729 LR - 20220325 IS - 1600-0625 (Electronic) IS - 0906-6705 (Print) IS - 0906-6705 (Linking) VI - 28 IP - 3 DP - 2019 Mar TI - Single cell analyses reveal specific distribution of anti-bacterial molecule Perforin-2 in human skin and its modulation by wounding and Staphylococcus aureus infection. PG - 225-232 LID - 10.1111/exd.13870 [doi] AB - Perforin-2 (P-2) is a recently described antimicrobial protein with unique properties to kill intracellular bacteria. We investigated P-2 expression pattern and cellular distribution in human skin and its importance in restoration of barrier function during wound healing process and infection with the common wound pathogen Staphylococcus aureus. We describe a novel approach for the measurement of P-2 mRNA within individual skin cells using an amplified fluorescence in situ hybridization (FISH) technique. The unique aspect of this approach is simultaneous detection of P-2 mRNA in combination with immune-phenotyping for cell surface proteins using fluorochrome-conjugated antibodies. We detected P-2 transcript in both hematopoietic (CD45(+) ) and non-hematopoietic (CD45(-) ) cutaneous cell populations, confirming the P-2 expression in both professional and non-professional phagocytes. Furthermore, we found an induction of P-2 during wound healing. P-2 overexpression resulted in a reduction of intracellular S. aureus, while infection of human wounds by this pathogen resulted in P-2 suppression, revealing a novel mechanism by which S. aureus may escape cutaneous immunity to cause persistent wound infections. CI - (c) 2019 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd. FAU - Strbo, Natasa AU - Strbo N AUID- ORCID: 0000-0003-2152-2433 AD - Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida. FAU - Pastar, Irena AU - Pastar I AUID- ORCID: 0000-0003-0197-6198 AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Romero, Laura AU - Romero L AD - Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida. FAU - Chen, Vivien AU - Chen V AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Vujanac, Milos AU - Vujanac M AD - Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida. FAU - Sawaya, Andrew P AU - Sawaya AP AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Jozic, Ivan AU - Jozic I AUID- ORCID: 0000-0001-5114-9524 AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Ferreira, Andrea D F AU - Ferreira ADF AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Wong, Lulu L AU - Wong LL AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Head, Cheyanne AU - Head C AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Stojadinovic, Olivera AU - Stojadinovic O AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Garcia, Denisse AU - Garcia D AD - Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida. FAU - O'Neill, Katelyn AU - O'Neill K AD - Department of Microbiology and Immunology, University of Miami Miller School of Medicine, Miami, Florida. FAU - Drakulich, Stefan AU - Drakulich S AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Taller, Seth AU - Taller S AD - Plastic Surgery, University of Miami Miller School of Medicine, Miami, Florida. FAU - Kirsner, Robert S AU - Kirsner RS AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. FAU - Tomic-Canic, Marjana AU - Tomic-Canic M AUID- ORCID: 0000-0002-9341-0193 AD - Department of Dermatology and Cutaneous Surgery, Wound Healing and Regenerative Medicine Research Program, University of Miami Miller School of Medicine, Miami, Florida. LA - eng GR - R01 NR015649/NR/NINR NIH HHS/United States PT - Journal Article PT - Research Support, N.I.H., Extramural DEP - 20190212 PL - Denmark TA - Exp Dermatol JT - Experimental dermatology JID - 9301549 RN - 0 (Pore Forming Cytotoxic Proteins) RN - 0 (perforin 2) RN - EC 3.1.3.48 (Leukocyte Common Antigens) RN - EC 3.1.3.48 (PTPRC protein, human) SB - IM MH - Animals MH - Cell Membrane/metabolism MH - Endothelial Cells/immunology/metabolism MH - Fibroblasts/metabolism MH - HEK293 Cells MH - Humans MH - Immunophenotyping MH - In Situ Hybridization, Fluorescence MH - Intraepithelial Lymphocytes/immunology/metabolism MH - Keratinocytes/immunology/metabolism MH - Leukocyte Common Antigens/metabolism MH - Mice MH - Pore Forming Cytotoxic Proteins/*metabolism MH - Single-Cell Analysis/*methods MH - Skin/*metabolism/microbiology MH - Staphylococcal Infections/*metabolism MH - Staphylococcus aureus MH - *Wound Healing PMC - PMC7461719 MID - NIHMS1005243 OTO - NOTNLM OT - Perforin-2 OT - fish-flow RNA assay OT - intracellular bacteria OT - skin OT - wound COIS- Conflict of interest Statement Authors declare no conflict of interest. EDAT- 2019/01/05 06:00 MHDA- 2020/07/30 06:00 PMCR- 2020/09/01 CRDT- 2019/01/05 06:00 PHST- 2018/05/04 00:00 [received] PHST- 2018/12/03 00:00 [revised] PHST- 2019/01/02 00:00 [accepted] PHST- 2019/01/05 06:00 [pubmed] PHST- 2020/07/30 06:00 [medline] PHST- 2019/01/05 06:00 [entrez] PHST- 2020/09/01 00:00 [pmc-release] AID - 10.1111/exd.13870 [doi] PST - ppublish SO - Exp Dermatol. 2019 Mar;28(3):225-232. doi: 10.1111/exd.13870. Epub 2019 Feb 12.