Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2018 Apr 9;9:688. doi: 10.3389/fimmu.2018.00688

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

Copyright © 2018 Becker, Nagarkatti and Nagarkatti.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

PMC Copyright notice

Locked nucleic acid (LNA) reduces intragraft effector cells and cytokines. Female C57BL/6 mice were given either syn (BL6) or allo (C₃H) tail skin grafts. Mice receiving allografts were given either LNA-466 (10 mg/kg) or LNA-Ctrl (10 mg/kg) intraperitoneally 1 day before skin transplantation and every third day after that until termination of the study. Upon rejection, grafts were aseptically excised, minced, and enzymatically digested to dislodge graft-infiltrating cells (GICs). GICs were spun down, culture supernatants were collected, and live cells were used for flow cytometric analysis. Representative dot plots displaying naïve (CD62L^Low, CD44^Neg), memory (CD62L⁺, CD44^HI), and effector (CD62L^Low, CD44^HI) cell types gated on CD8⁺ [(A), upper row] and CD4⁺ [(A), lower row] cells. Percentages for CD8⁺ and CD4⁺ are quantified in (B,C), respectively. Dot plots of GICs double-positive for FoxP3 and CD62L, data are gated on CD4⁺ cells (D), and quantified in (E); n = 7 (allogeneic + Ctrl) or 8 (allogeneic + LNA). GIC supernatants were collected and subjected to enzyme-linked immunosorbent assay for the interrogation of effector cytokines TNFα (F) and IFNγ (G), as well as anti-inflammatory cytokines transforming growth factor (TGF)-β1 (H) and TGF-β2 (I). Data are presented as mean ± SEM. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001 by ANOVA with Tukey’s multiple comparisons test or a Student’s t-test.

Locked nucleic acid (LNA) reduces intragraft effector cells and cytokines. Female C57BL/6 mice were given either syn (BL6) or allo (C₃H) tail skin grafts. Mice receiving allografts were given either LNA-466 (10 mg/kg) or LNA-Ctrl (10 mg/kg) intraperitoneally 1 day before skin transplantation and every third day after that until termination of the study. Upon rejection, grafts were aseptically excised, minced, and enzymatically digested to dislodge graft-infiltrating cells (GICs). GICs were spun down, culture supernatants were collected, and live cells were used for flow cytometric analysis. Representative dot plots displaying naïve (CD62L^Low, CD44^Neg), memory (CD62L⁺, CD44^HI), and effector (CD62L^Low, CD44^HI) cell types gated on CD8⁺ [(A), upper row] and CD4⁺ [(A), lower row] cells. Percentages for CD8⁺ and CD4⁺ are quantified in (B,C), respectively. Dot plots of GICs double-positive for FoxP3 and CD62L, data are gated on CD4⁺ cells (D), and quantified in (E); n = 7 (allogeneic + Ctrl) or 8 (allogeneic + LNA). GIC supernatants were collected and subjected to enzyme-linked immunosorbent assay for the interrogation of effector cytokines TNFα (F) and IFNγ (G), as well as anti-inflammatory cytokines transforming growth factor (TGF)-β1 (H) and TGF-β2 (I). Data are presented as mean ± SEM. *P < 0.05, **P < 0.005, ***P < 0.001, ****P < 0.0001 by ANOVA with Tukey’s multiple comparisons test or a Student’s t-test.