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Updated: Jun 27, 2026
Isolating And Immunostaining Lymphocytes and Dendritic Cells from Murine Peyer's Patches
Published on: March 17, 2013
Isolating mucosal lymphocytes from biopsy tissue for cellular immunology assays
Barbara L Shacklett1, J William Critchfield, Donna Lemongello
1Department of Medical Microbiology, and Division of Infectious Diseases, School of Medicine, University of California, Davis, CA, USA.
Insights
Researchers developed a minimally invasive method to analyze T-cells from rectal biopsies, crucial for understanding gastrointestinal tract HIV-1 transmission and pathogenesis.
Area of Science:
- Immunology
- Virology
- Gastroenterology
Background:
- Mucosal tissues in the gastrointestinal and genitourinary tracts are key entry points for HIV-1.
- Understanding human mucosal T-cell responses is vital for HIV-1 pathogenesis research.
- Current knowledge of mucosal T-cell immunity, particularly in the gut, is limited.
Purpose of the Study:
- To present a reproducible method for isolating and analyzing mucosal T-lymphocytes.
- To enable the study of T-cell responses in sites not easily accessible.
- To facilitate research into HIV-1 transmission and pathogenesis at mucosal sites.
Main Methods:
- Isolation and culture of T-lymphocytes from gastrointestinal biopsy tissue.
- Utilizing flexible sigmoidoscopy for minimally invasive rectal biopsy acquisition.
- Analysis of CD4+ and CD8+ T-cells from rectal biopsies.
Main Results:
- Established a well-accepted, minimally invasive method for obtaining human mucosal tissue.
- Successfully isolated and prepared mucosal T-cells for analysis.
- Demonstrated the applicability of these methods to various lymphoid tissues.
Conclusions:
- The described method allows for the study of human mucosal T-cell responses.
- This technique provides access to critical tissues for HIV-1 research.
- The approach is adaptable for analyzing T-cells from different mucosal sites.
Abstract:
Mucosal tissues of the gastrointestinal and genitourinary tracts serve as major portals of HIV-1 transmission, and recent literature has highlighted the important role of these tissues in pathogenesis. However, our understanding of human mucosal T-cell responses remains limited. We have previously reported methods for isolating, culturing and analyzing mucosal T-lymphocytes obtained from gastrointestinal biopsy tissue. This method of acquiring tissue is minimally invasive and well accepted by patients, and allows sampling of sites that would not otherwise be accessible without surgical intervention. This chapter summarizes the approach currently in use in our laboratory to isolate and study CD4+ and CD8+ T-cells from rectal biopsies obtained through flexible sigmoidoscopy. These methods are also applicable, with minor modifications, to small tissue samples obtained from other lymphoid tissues.
