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

. 2022 Aug 18;14(1):2112881. doi: 10.1080/19490976.2022.2112881

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

© 2022 The Author(s). Published with license by Taylor & Francis Group, LLC.

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

PMC Copyright notice

Figure 3. — Abiotic bacterial culture supernatant modulates NK cell anti-PDAC activity in vitro. Stool was collected from germ-free (GF) and SPF-Rag1^−/− mice and cultured anaerobically for 3 days. Abiotic culture supernatant was prepared after filtration and NK-92MI cells treated with 1% of this supernatant for 3 days. After exposure, NK-92MI cells were harvested and then co-cultured with the human PDAC cell line L3.6pl or BxPC3 for 4 hours and flow cytometry performed to determine cytotoxicity as described (a). NK cells exposed to abiotic culture supernatant from SPF-derived stool had decreased cytotoxicity to L3.6pl and BxPC3 compared to GF-derived abiotic supernatant in vitro (b). Furthermore, NK-92MI cells had a decreased migratory ability in a Boyden chamber assay when pre-treated with abiotic culture supernatant from SPF-Rag1^−/− stool (c). These findings of decreased cytotoxicity and decreased migration correlated with decreased activation of NK-92MI cells as measured by flow cytometry for IFNγ(+) NK-92MI cells after exposure to abiotic culture supernatant from SPF-Rag1^−/− stool (d). To interrogate pathways responsible for altered cytoxicity, qPCR of treated NK-92MI cells again demonstrated decreased NK cell activation (IFNγ+) but also decreased Perforin expression when exposed to abiotic culture supernatant from SPF stool (e). Furthermore, decreased NK cell cytotoxicity and migration pathways were found via qPCR array as well as increased expression of inhibitory NK cell pathways in NK-92MI cells treated with abiotic culture supernatant from SPF stool (f). (*) p < .05, (**) p < .01.