SLAMF4 Is a Negative Regulator of Expansion of Cytotoxic Intraepithelial CD8+ T Cells That Maintains Homeostasis in the Small Intestine

Gastroenterology. 2015 May;148(5):991-1001.e4. doi: 10.1053/j.gastro.2015.02.003. Epub 2015 Feb 10.

Abstract

Background & aims: Intraepithelial T lymphocyte cells (IEL) are the first immune cells to respond to pathogens; they help maintain the integrity of the epithelial barrier. We studied the function of the mouse glycoprotein Signaling Lymphocyte Activation Molecule Family receptor (SLAMF) 4 (encoded by Slamf4) on the surface of CD8αβ αβ T-cell receptor (TCR)(+) IELs, and the roles of these cells in homeostasis of the small intestine in mice.

Methods: SLAMF4(-) CD8(+) αβTCR(+) cells isolated from spleens of OT-I Rag1(-/-) mice were induced to express gut-homing receptors and transferred to C57BL/6J mice; levels of SLAMF4(+) cells were measured in small intestine tissues. After administration of anti-CD3 or antigen, with or without anti-SLAM4, to C57BL/6J and Slamf4(-/-) mice, CD8αβ αβTCR(+) IELs were collected; cytokine production and cytotoxicity were measured. Depletion of CX3CR1(+) phagocytes was assessed in mice by live-cell confocal imaging or by cytofluorometry; small intestine tissues were analyzed by histology and inflammation was quantified.

Results: Splenic CD8(+) αβTCR(+) cells began to express SLAMF4 only after migrating to the small intestine. Injection of C57BL/6J mice with anti-SLAMF4 and anti-CD3 increased levels of interleukin 10 and interferon gamma secretion by IEL, compared with injection of anti-CD3 only. Similarly, the number of granzyme B(+) cytotoxic CD8(+) αβTCR(+) IELs increased in Slamf4(-/-) mice after injection of anti-CD3 and anti-SLAMF4, administration of antigen, or injection of anti-CD3. Surprisingly, in vivo activation of CD8αβ(+) IELs with anti-CD3 or antigen caused transient depletion of CX3CR1(+) phagocytes, which was prolonged by co-injection with anti-SLAMF4 or in Slamf4(-/-) mice. Anti-CD3 aggravated inflammation in the small intestines of Slamf4(-/-) mice and Eat2a(-/-)Eat2b(-/-) mice, indicated by flattened villi and crypt hyperplasia.

Conclusions: In mice, the intestinal environment induces SLAMF4 expression and localization to the surface of CD8(+) αβTCR(+) IELs. Signaling via SLAMF4 controls expansion of cytotoxic CD8αβ(+) IELs, which regulate the reversible depletion of lamina propria phagocytes and inflammation in the small intestine.

Keywords: Immune Regulation; Immunity; Intestinal Epithelium; T-Cell Development.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Adaptor Proteins, Signal Transducing / deficiency
  • Adaptor Proteins, Signal Transducing / genetics
  • Animals
  • Antigens, CD / genetics
  • Antigens, CD / metabolism*
  • CX3C Chemokine Receptor 1
  • Cell Movement
  • Cell Proliferation*
  • Cells, Cultured
  • Cytokines / immunology
  • Cytokines / metabolism
  • Dendritic Cells / immunology
  • Dendritic Cells / metabolism
  • Green Fluorescent Proteins / genetics
  • Green Fluorescent Proteins / metabolism
  • Homeodomain Proteins / genetics
  • Homeodomain Proteins / metabolism
  • Homeostasis
  • Hyperplasia
  • Intestinal Mucosa / immunology
  • Intestinal Mucosa / metabolism*
  • Intestinal Mucosa / pathology
  • Intestine, Small / immunology
  • Intestine, Small / metabolism*
  • Intestine, Small / pathology
  • Lymphocyte Activation*
  • Mice, Inbred C57BL
  • Mice, Knockout
  • Phagocytes / immunology
  • Phagocytes / metabolism
  • Receptors, Antigen, T-Cell, alpha-beta / immunology
  • Receptors, Antigen, T-Cell, alpha-beta / metabolism
  • Receptors, Chemokine / genetics
  • Receptors, Chemokine / metabolism
  • Receptors, Immunologic / deficiency
  • Receptors, Immunologic / genetics
  • Receptors, Immunologic / metabolism*
  • Signal Transduction
  • Signaling Lymphocytic Activation Molecule Family
  • Spleen / immunology
  • Spleen / metabolism
  • T-Lymphocytes, Cytotoxic / immunology
  • T-Lymphocytes, Cytotoxic / metabolism*

Substances

  • Adaptor Proteins, Signal Transducing
  • Antigens, CD
  • CX3C Chemokine Receptor 1
  • Cd244a protein, mouse
  • Cx3cr1 protein, mouse
  • Cytokines
  • Eat2a protein, mouse
  • Eat2b protein, mouse
  • Homeodomain Proteins
  • Receptors, Antigen, T-Cell, alpha-beta
  • Receptors, Chemokine
  • Receptors, Immunologic
  • Signaling Lymphocytic Activation Molecule Family
  • RAG-1 protein
  • Green Fluorescent Proteins