Journal Article

Mesenteric Lymph Node Transplantation in Mice to Study Immune Responses of the Gastrointestinal Tract

Shaikh, Haroon, Mosca, Josefina Peña, Mokhtari, Zeinab, Graf, Caroline, Büttner-Herold, Maike, Le, Duc-Dung, Huehn, Jochen, Vargas, Juan Gamboa, Pezoldt, Joern, Rosenwald, Andreas, Jarick, Katja J., Ulbrich, Maria, Viera, Estibaliz Arellano, Heinze, Katrin G., Beilhack, Andreas

2021

Overview

Mesenteric lymph nodes (mLNs) are sentinel sites of enteral immunosurveillance and immune homeostasis. Immune cells from the gastrointestinal tract (GIT) are constantly recruited to the mLNs in steady-state and under inflammatory conditions resulting in the induction of tolerance and immune cells activation, respectively. Surgical dissection and transplantation of lymph nodes (LN) is a technique that has supported seminal work to study LN function and is useful to investigate resident stromal and endothelial cell biology and their cellular interactions in experimental disease models. Here, we provide a detailed protocol of syngeneic mLN transplantation and report assays to analyze effective mLN engraftment in congenic recipients. Transplanted mLNs allow to study T cell activation and proliferation in preclinical mouse models. Donor mLNs proved viable and functional after surgical transplantation and regenerated blood and lymphatic vessels. Immune cells from the host completely colonized the transplanted mLNs within 7-8 weeks after the surgical intervention. After allogeneic hematopoietic cell transplantation (allo-HCT), adoptively transferred allogeneic CD4 + T cells from FVB/N (H-2 q ) mice homed to the transplanted mLNs in C57BL/6 (H-2 b ) recipients during the initiation phase of acute graft-versus-host disease (aGvHD). These CD4 + T cells retained full proliferative capacity and upregulated effector and gut homing molecules comparable to those in mLNs from unmanipulated wild-type recipients. Wild type mLNs transplanted into MHCII deficient syngeneic hosts sufficed to activate alloreactive T cells upon allogeneic hematopoietic cell transplantation, even in the absence of MHCII + CD11c + myeloid cells. These data support that orthotopically transplanted mLNs maintain physiological functions after transplantation. The technique of LN transplantation can be applied to study migratory and resident cell compartment interactions in mLNs as well as immune reactions from and to the gut under inflammatory and non-inflammatory conditions.

• Publisher: Frontiers Media SA,Frontiers Media S.A
• Subject: Acute Disease / acute graft-versus host disease / Adoptive Transfer / Allografts / alloreactive T cells / Animal models / Animals / Antibodies / Antigens / CD11c antigen / CD4 antigen / Cell activation / Cell Differentiation / Cell Proliferation / Digestive system / Disease Models, Animal / Endothelial cells / Gastrointestinal tract / Graft Survival / Graft vs Host Disease - immunology / Graft vs Host Disease - metabolism / Graft-versus-host reaction / Hematopoietic Stem Cell Transplantation / Hematopoietic stem cells / Histocompatibility antigen H-2 / Histocompatibility Antigens Class II - immunology / Homeostasis / Homing behavior / Immune response / Immunological tolerance / Immunology / Immunosurveillance / Inflammation / Laboratories / lymph node stromal cells / lymph node transplantation / Lymph nodes / Lymph Nodes - immunology / Lymph Nodes - metabolism / Lymph Nodes - transplantation / Lymphatic system / Lymphocyte Activation / Lymphocytes / Lymphocytes T / mesenteric lymph node / Mesentery / Mice / Mice, Inbred C57BL / mouse models / Myeloid cells / Phenotype / Shaving & shavers / Stem cell transplantation / Surgery / Syngeneic grafts / T-Lymphocytes - immunology / T-Lymphocytes - metabolism / T-Lymphocytes - transplantation / Transplantation Tolerance / Transplantation, Isogeneic