RT @fcaprioli76: Will it ever be possible to find a definitive cure for #inflammatoryboweldiseases? Find out most recent developments towar….

RT @stratifnc: Our work on the sexual dimorphism of colorectal cancer is finally out in Oncoimmunology!. A huge thanks to @GeorgiaLattanzi….

Pure joy is when your collaborators receive recognition for their work 🎉.Chiara Amoroso @fcaprioli76 and the @LoungeTcell team best award for our #fmt study in #IBD at the 238 Congress of the Falk Foundation @policlinicoMI

Oops we did it again! #iNKT cells are functionally shut down by bugs (here Porphiromonas gingivalis) in #colorectal #cancer. Soon a summary of the study, now Thanks to @epiangie @GeorgiaLattanzi @stratifnc , the whole @LoungeTcell and @AIRC_it .

Lab moving ongoing .✔️-4 done.✔️-80 done.✔️-20 in progress.✔️10 years of reagents, lab books, various stuff (some of not certain utility).✔️car packed.✔️some tears .✔️new beginnings ahead at @BtBsUNIMIB .✔️@LoungeTcell alive&kicking!

In the rush of the return home from #UEGWeek we did not celebrate the achievement of @dnoviello_93, Who won the Best National Abstract award. @fcaprioli76 clinical and myself research super-proud mentors! Grande Daniele🌟

Fun and inspiration, plus cool science and rigorous clinical science #UEGWeek2024 .We proudly presented the work of @federicaperillo and jointly coordinated with @stratifnc on CAC patients and the consequences of deregulated T cells activation

Finally, this paper is dedicated to the memory of our dear colleague Nicolas Barnich who recently passed away. His untimely departure is a profound loss to the scientific community and to all who had the privilege of knowing him.

A big thanks goes to all the people involved in the study, from @gabrijejuni , to X-less Moira Paroni, to @fcaprioli76 @dnoviello_93 @policlinicoMI @LaStatale @unimib for their huge effort on this exciting project. We also thank @FondCariplo that financially supported us.

Summary: our findings indicated that the transdifferentiation of pTh17 cell is strongly driven in CD patients by AIEC determinants, such as rfaP or ybaT , generated within AIEC-infected DCs, and presented to cTh17 cells together with very high IL-23 levels.

Clinical Significance: 1) YbaT is ubiquitously found across all E. coli pathotypes, differently from other virulence factors like FimH. 2) Targeting of ybaT and rfaP inhibits pTh17 generation without affecting protective Th17 cells.

rfaP is a LPS core heptose(I) kinase modifying the LPS inner core.Its absence alters structure and stability of the outer membrane (👉different antigens!). ybaT is an aminoacid permease involved in the glutamate dependent copper (Cu) tolerance (👉less stress resistance!).

To exclude putative polar effects of transposon insertion in our mutants, we generated individual isogenic deletion mutants in rfaP and ybaT genes, confirming our data with DC (persistence and IL23 secretion) and Th17 (transdifferentiation)

Functional validation with DCs and Th17 cells. Six mutants impairedd IL23 secretin, but ΔybaT (T59.F12) and to a lesser extent ΔrfaP (T12.G9 and T45.G12) mutants drastically impaired also the skewing of cTh17 into pTh17 cells.

The DNA sequencing after rescue cloning revealed that the transposon insertions in these 13 selected LF82 mutants occurred within genes involved in a few specific pathways including metabolic processes, stress response, LPS and capsule biosynthesis.

After 3 rounds of progressively more restrictive screenings, we identified 22 mutants further tested on CD-derived Dendritic cells, At the end, 13 mutants showed different parameters (intracellular bacterial load, capability to persist, IL23 secretion) in CD-vs HD- derived DCs.

But still we had not discovered how AIEC was capable of hacking DCs and transdifferentiating cTh17 cells in pTh17 cells. @gabrijejuni generated a library of 10,058 (!!!) AIEC mutants and tested all of them for their capability to induce IL23 and iL1b secretion by human DC

Third, when we exposed cTh17 to DCs isolated from CD patients only AIEC-infected DCs supported the transdifferentiation in vitro of cTh17 cells into pathogenic pTh17, whose differentiation was blocked by anti IL23 (and anti IL1b) antibodies.

Second, AIEC persist longer in CD-derived DC as compared to those derived from healthy donors, promoting a more sustained (in time) production of IL23 and IL1b.

AIEC, how did you manage to seduce Th17 cells? We turned to old school immunology: we isolated immune cells from CD patients and we put them with LF82. AIEC was allying with dendritic cells (DC), inducing them to secrete f IL23 and IL1b, required for Th17 differentiation.