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Characterization of surface markers of bovine gut mucosal leukocytes using monoclonal antibodies
1Department of Veterinary Microbiology, Western College of Veterinary Medicine, University of Saskatchewan, Saskatoon, Canada.
Insights
Bovine intestinal leukocytes differ significantly across locations. Intraepithelial lymphocytes (IEL) show distinct T cell, B cell, and MHC phenotypes compared to lamina propria (LPL) and Peyer's patch (PPL) cells.
Area of Science:
- Immunology
- Gastroenterology
- Veterinary Science
Background:
- The bovine intestinal immune system's leukocyte populations are crucial for mucosal defense.
- Understanding the phenotypic differences of leukocytes in various intestinal compartments is essential for immune research.
Purpose of the Study:
- To characterize and compare the surface phenotypes of bovine leukocytes from the intraepithelium (IEL), lamina propria (LPL), and Peyer's patches (PPL) of the small intestine.
- To identify significant differences in leukocyte populations across these distinct intestinal locations.
Main Methods:
- Isolation of leukocytes from bovine small intestinal mucosa (IEL, LPL, PPL).
- Analysis of cell surface phenotypes using laser flow cytometry (LFC) and monoclonal antibodies (mAb).
- Comparison of cell populations based on T cell, B cell, monocyte, and MHC antigen expression.
Main Results:
- Intraepithelial lymphocytes (IEL) had significantly lower percentages of pan T cells, B cells, and T helper (Th) cells compared to LPL and PPL.
- IEL exhibited a higher proportion of T cytotoxic/suppressor (Tc/s) cells relative to LPL.
- Significant differences in MHC class I and class II expression were observed between IEL and PPL, with IEL showing lower expression.
Conclusions:
- Bovine IEL possess a distinct leukocyte phenotype compared to LPL and PPL, characterized by a higher proportion of "null" cells and different T cell subset distributions.
- LPL and PPL present phenotypically similar leukocyte populations, contrasting with the unique profile of IEL.
- These findings highlight the specialized immune functions of different bovine intestinal compartments.
Abstract:
The surface phenotypes of bovine intestinal leukocytes isolated from the intraepithelium (IEL), lamina propria (LPL) and Peyer's patches (PPL) of the small intestinal mucosa of normal adult cows were determined using monoclonal antibodies (mAb) specific to adult bovine peripheral blood leukocytes (PBL). Laser flow cytometric (LFC) analysis demonstrated that IEL contained significantly (P less than 0.1 to 0.02) fewer cells (26%) expressing the pan T cell phenotype in comparison to LPL (38%) and PPL (44%). Similarly, significantly (P less than 0.01 to 0.001) lower numbers of B cells were observed among IEL (10%) compared to LPL (28%) and PPL (33%). While approximately equal numbers of B7A1+ "null" cells (10%) and DH59B+ "Ia+ monocytes/granulocytes" (16.5%) were observed among the three intestinal cell populations, IEL contained significantly (P less than 0.1 to 0.05) lower numbers (19%) of T helper (Th) cells in comparison to LPL (44%) and PPL (38%). In contrast, lymphocytes with the T cytotoxic/suppressor (Tc/s) phenotype were significantly lower (P less than 0.01 to 0.001) among LPL (14.5%) compared to IEL (25%) and PPL (23%). While the numbers of cells expressing class I major histocompatibility complex (MHC) surface antigens (H58A+) were approximately equal among LPL (79%) and PPL (87%), a significant difference (P less than 0.02) was observed between IEL (71%) and PPL. Similarly, while approximately equal numbers of cells expressing the MHC class II surface phenotype were observed among LPL (42%) and PPL (46%), IEL contained significantly (P less than 0.01) fewer (31%) MHC class II cells in comparison to PPL. Enrichment for T cells by plastic adherence and Sephadex G-10/nylon wool fractionation revealed a significant (P less than 0.01) and proportional increase in T lymphocyte subsets expressing pan T, Th and Tc/s phenotypes among the three cell populations. Similarly, enrichment for B cells by the same techniques showed a significant (P less than 0.01) and proportional increase in cells expressing the panB cell phenotype among LPL and PPL. Marked differences in cell size distribution and cell surface density were observed when the three intestinal leukocyte populations were compared by LFC using monoclonal antibodies directed at various cell surface markers. Furthermore, considerable quantitative variations of each cell surface marker were observed among the individual animals tested. The results of this study indicate that bovine IEL, which contain a high percentage of cells (greater than 30%) with no known phenotype are significantly different from LPL and PPL which are phenotypically similar cell populations.(ABSTRACT TRUNCATED AT 400 WORDS)
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