Analysis of innate lymphoid cells during the follow-up of intestinal transplant patients:a preliminary study

PUCCI MOLINERIS M; RUMBO C; NACHMAN F; FUXMAN C; RUMBO M; GONDOLESI G; MEIER D

Simposio; International Small Bowel Transplant Symposium 2015; 2015

Intestinal Transplant Association

Intestinal transplantation (ITx) presents the highest risk of graft lossamong solid organ transplants due to the enormous amount of immunecells and intraluminal commensal bacteria. ITx patients are exposedto an aggressive immunosuppressive therapy, which raisessusceptibility to infections and might dysbalance the intestinal homeostasisof the microflora and the immune system. Recently, it hasbeen shown in a mouse model that the dysfunction of the innate lymphoidcell subgroup 3 (ILC3) defined as Lin- CD127+ CD117+NKp44+ CD56+/- disturbs the intestinal homeostasis inducing inflammation.The ILC3s secreting IL-22 and IL17-A by IL-23 induction formpart of a newly described group of immune cells lacking B and T cellreceptors and differ from conventional natural killer cells (cNK). Inhumans, an increased number of ILC3s was detected in patients withCrohn´s disease (CD). Our aim was to elucidate whether the integrityof ILC3s is disturbed by the immunosuppressive regimen in the ITxsetting. We isolated lamina propria cells from biopsies obtained duringthe follow up of ITx patients divided by the presence of rejection(mild rejection (n=2) and normal group (n=11)); from one patient withrefractory Crohn?s disease (n=1); and from non-immunosuppressedpatients as control samples (n=3). For the enrichment of ILCs, wedepleted CD4+ cells by magnetic-activated cell sorting. The resultingCD4- cells were stained with fluorochrome-conjugated anti-CD3,CD20, CD127, CD117, NKp44 and CD56 antibodies and analyzedby flow cytometry. Activated cNK cells were determined as CD3-CD56+ CD117- NKp44+ cells. Our results show a 4-fold decrease inpercentage of ILCs (p