Isolation and phenotypic characterization of colonic macrophages
| العنوان: | Isolation and phenotypic characterization of colonic macrophages |
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| المؤلفون: | Martin Hausmann, Reinhard Andreesen, Gerhard Rogler, V. Gross, Daniela Vogl, E. Aschenbrenner, Jürgen Schölmerich, Werner Falk, T. Andus |
| المصدر: | Clinical and Experimental Immunology. 112:205-215 |
| بيانات النشر: | Oxford University Press (OUP), 1998. |
| سنة النشر: | 1998 |
| مصطلحات موضوعية: | Phagocyte, Colon, CD14, Sialic Acid Binding Ig-like Lectin 3, Immunology, Antigen-Presenting Cells, Antigens, Differentiation, Myelomonocytic, chemical and pharmacologic phenomena, Cell Separation, Biology, CD16, Th2 Cells, Intestinal mucosa, Antigens, CD, medicine, Humans, Immunology and Allergy, Macrophage, Intestinal Mucosa, Receptors, Immunologic, Antigen-presenting cell, Immunity, Mucosal, Lamina propria, Membrane Glycoproteins, Tumor Necrosis Factor-alpha, Macrophages, hemic and immune systems, Original Articles, Th1 Cells, Flow Cytometry, Phenotype, medicine.anatomical_structure, Tumor necrosis factor alpha, B7-2 Antigen, Biomarkers, Interleukin-1 |
| الوصف: | SUMMARY Macrophages play an important role in the intestinal mucosal immune system. However, they are a poorly defined cell population. We therefore determined their phenotype in normal colonic mucosa. Macrophages were isolated from colonic biopsies and surgical specimens by collagenase digestion. Colonic macrophages were positively sorted by anti-CD33 magnetic beads. Flow cytometric triple fluorescence analysis was applied to study CD14, CD16, CD33, CD44, CD11b, CD11c, CD64, HLA-DR, CD80, CD86 and CD3/CD19 expression. CD33 was evaluated as a positive marker for intestinal macrophages. CD33+ cells isolated from normal colonic mucosa showed co-expression of the established intracellular macrophage marker CD68 in FACS analysis. CD33+ cells were capable of phagocytosis. Isolation of this cell population by magnetic anti-CD33 beads and culture resulted in a 4·2–40-fold increase in IL-1β and 4·5–44-fold increase in tumour necrosis factor-alpha (TNF-α) secretion compared with unsorted lamina propria mononuclear cells (LPMC). Of the CD33+ cells, 90·9 ± 6·9% (mean ± s.d.) were CD44+. However, macrophages from colonic mucosa showed only a low expression of CD14 (10·5 ± 3·8%), CD16 (10·1 ± 3·9%), HLA-DR (27·3 ± 9·2%), CD11b (17·4 ± 6·8%), CD11c (17·8 ± 10·4%). Furthermore, expression of CD80 (9·2 ± 4·2%) and CD86 (15·1 ± 7·3%) was low, suggesting a low ability of normal intestinal macrophages to activate T cells and T cell-mediated immune responses. We conclude that CD33 is useful for the isolation and flow cytometric characterization of colonic macrophages. These cells exhibit a single phenotype in normal mucosa (CD33++, CD44++, CD14−, CD16−, CD11b−, CD11c−, HLA-DRlow, CD80−, CD86−) lacking lipopolysaccharide (LPS) receptor and costimulatory molecules. |
| تدمد: | 1365-2249 0009-9104 |
| DOI: | 10.1046/j.1365-2249.1998.00557.x |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::c458a1d4ad33ee649b0679d68f620ee4 https://doi.org/10.1046/j.1365-2249.1998.00557.x |
| Rights: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....c458a1d4ad33ee649b0679d68f620ee4 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 13652249 00099104 |
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| DOI: | 10.1046/j.1365-2249.1998.00557.x |