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Updated: Aug 13, 2026
Isolation and Flow Cytometric Characterization of Murine Small Intestinal Lymphocytes
Published on: May 8, 2016
Isolation of functionally active intraepithelial lymphocytes and enterocytes from human small and large intestine
C Lundqvist1, M L Hammarström, L Athlin
1Department of Immunology, University of Umeå, Sweden.
Insights
A new method isolates human intraepithelial lymphocytes (IEL) and enterocytes from the same person. This allows for studying their functions and interactions in local immunity.
Area of Science:
- Gastroenterology
- Immunology
- Cell Biology
Background:
- Human intraepithelial lymphocytes (IEL) and enterocytes play crucial roles in gut immunity and function.
- Efficient isolation methods are needed to study these cells and their interactions.
Purpose of the Study:
- To develop a mild and effective method for simultaneously isolating human IEL and enterocytes from the same tissue.
- To enable further research into the functions and interactions of these cells in local immunity.
Main Methods:
- Mechanical disruption, reducing agent treatment, and Percoll gradient centrifugation were used.
- Magnetic beads with anti-epithelial (BerEP4) and anti-leukocyte (CD45) antibodies purified IEL and enterocytes, respectively.
- The method was applied to jejunal, ileal, and colonic tissue specimens.
Main Results:
- High yields of viable IEL and enterocytes were recovered from various intestinal segments.
- Purified IEL and enterocyte fractions demonstrated high purity (>96% and >98%, respectively).
- Both cell types were viable and capable of in vitro expansion.
Conclusions:
- A novel, mild purification technique allows for the simultaneous isolation of pure, viable human IEL and enterocytes.
- This method facilitates the study of IEL-enterocyte interactions and their roles in local immunity.
- The isolation procedure is applicable to different regions of the human intestine.
Abstract:
A mild purification method has been developed for the isolation of human intraepithelial lymphocytes (IEL) and enterocytes from the same individual. The isolation procedure includes mechanical disruption of the mucosal layer, treatment with reducing agent and sedimentation followed by Percoll gradient centrifugation. Finally, epithelial cells are removed from the IEL fraction using magnetic beads coated with the anti-epithelial antigen monoclonal antibody (mAb) BerEP4. Leucocytes are removed from the enterocyte fraction using magnetic beads coated with mAbs directed against common leucocyte antigen (CD45). Using this procedure IEL and enterocytes have been isolated from apparently normal jejunal, ileal and colonic tissue specimens. Recoveries of IEL were 7 x 10(5), 4 x 10(5) and 1 x 10(5)/cm2 mucosa from jejunum, ileum and colon respectively. 1-2 x 10(6) enterocytes/cm2 mucosa were recovered from small intestine while the corresponding value for colonic biopsies was approximately 2 x 10(5) enterocytes/cm2. The IEL fraction was pure as judged by the low percentages of B cells, macrophages and BerEP4 positive cells (less than 4%) present in the purified fraction. The enterocyte fraction contained less than 2% CD45+ cells. The two cell fractions were viable and expanded in vitro. Enterocytes expanded spontaneously while IEL required initial stimulation with mitogens. The isolation procedure described here will make it possible to study the function of human IEL, interactions between IEL and enterocytes and the role of both cell types in local immunity.
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