Isolation of murine intrahepatic immune cells employing a modified procedure for mechanical disruption and functional characterization of the B, T and natural killer T cells obtained
| Autor: | M Rahman Qazi, Joseph W. DePierre, K G Blom, J B Noronha Matos, B D Nelson, Manuchehr Abedi-Valugerdi |
|---|---|
| Rok vydání: | 2008 |
| Předmět: |
Male
Pathology medicine.medical_specialty Cell Survival Immunology B-Lymphocyte Subsets Cell Separation Streptamer Biology Immunophenotyping Interferon-gamma Mice Interleukin 21 Immune system T-Lymphocyte Subsets medicine Animals Immunology and Allergy Interferon gamma Cells Cultured Cell Proliferation Immunobiology Natural killer T cell Molecular biology In vitro Mice Inbred C57BL Immunoglobulin M Liver Concanavalin A biology.protein Natural Killer T-Cells Immunocompetence medicine.drug |
| Zdroj: | Clinical and Experimental Immunology. 155:320-329 |
| ISSN: | 1365-2249 0009-9104 |
| DOI: | 10.1111/j.1365-2249.2008.03815.x |
| Popis: | SummaryIntrahepatic immune cells (IHIC) are known to play central roles in immunological responses mediated by the liver, and isolation and phenotypic characterization of these cells is therefore of considerable importance. In the present investigation, we developed a simple procedure for the mechanical disruption of mouse liver that allows efficient isolation and phenotypic characterization of IHIC. These cells are compared with the corresponding cells purified from the liver after enzymatic digestion with different concentrations of collagenase and DNase. The mechanical disruption yielded viable IHIC in considerably greater numbers than those obtained following enzymatic digestion. The IHIC isolated employing the mechanical disruption were heterogeneous in composition, consisting of both innate and adaptive immune cells, of which B, T, natural killer (NK), NK T cells, granulocytes and macrophages were the major populations (constituting 37·5%, 16·5%, 12·1%, 7·9%, 7·9% and 7·5% of the total number of cells recovered respectively). The IHIC obtained following enzymatic digestion contained markedly lower numbers of NK T cells (1·8%). The B, T and NK T cells among IHIC isolated employing mechanical disruption were found to be immunocompetent, i.e. they proliferated in vitro in response to their specific stimuli (lipopolysaccharide, concanavalin A and α-galactosylceramide respectively) and produced immunoglobulin M and interferon-γ. Thus, the simple procedure for the mechanical disruption of mouse liver described here results in more efficient isolation of functionally competent IHIC for various types of investigation. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|
Plný text