Isolation and purification of natural killer cells subpopulations using mononuclear cells
Natural Killer (NK) cells are the main lymphocyte population expressing P75 B chain of the IL-2 receptor (IL-2R). Consequently, incubation of peripheral blood lymphocytes with IL-2 induce selective activation of NK cells and results in NK activity and generation of Lymphokine activated killer (LAK)...
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| Format: | Article |
| Language: | fas |
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Tehran University of Medical Sciences
1999-06-01
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| Series: | Tehran University Medical Journal |
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| Online Access: | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/5688.pdf&manuscript_id=5688 |
| _version_ | 1818993123666165760 |
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| author | Mosaffa N Labibi F |
| author_facet | Mosaffa N Labibi F |
| author_sort | Mosaffa N |
| collection | DOAJ |
| description | Natural Killer (NK) cells are the main lymphocyte population expressing P75 B chain of the IL-2 receptor (IL-2R). Consequently, incubation of peripheral blood lymphocytes with IL-2 induce selective activation of NK cells and results in NK activity and generation of Lymphokine activated killer (LAK) cells activity and proliferation. One of the early events during IL-2 activation of peripheral blood lymphocyte in both rodents and humans is adherence of some NK cells to plastic surface. The cells adherence to plastic after 24 hr of culture with IL-2 are almost exclusively CD56+, have the morphology large granular cells to yield a highly entiched population of activated NK cells that have been used for systemic adoptive immunotherapy. To test these hypothesis, we used highly purified population of human peripheral NK cells through the biological and nonimmunclogical phenotyping technique. Blood mononuclear cells were separated by centrifugation of ficol-hypaque gradient from normal blood donor (20-30 years age). We depleted after purification of nonadherent cells with nylonwool. We collected with rosette technique to remove cells with high affinity SRBC receptors. These cells separate in two parts A-NK and NA-NK by mononuclear celss activated supernatant media. The main objective results of this study show that the subpopulation of human NK cell which develope early adherent to plastic surface in the presence of supernatant mononuclear celss activation media was functionally more cytotoxic and killed K562 targets in single cell sytotoxicity manner and LDH activity assay than nonadherent NK cells and resting NK cells |
| first_indexed | 2024-12-20T20:37:03Z |
| format | Article |
| id | doaj.art-61349bfcad0446899814252d6c412242 |
| institution | Directory Open Access Journal |
| issn | 1683-1764 1735-7322 |
| language | fas |
| last_indexed | 2024-12-20T20:37:03Z |
| publishDate | 1999-06-01 |
| publisher | Tehran University of Medical Sciences |
| record_format | Article |
| series | Tehran University Medical Journal |
| spelling | doaj.art-61349bfcad0446899814252d6c4122422022-12-21T19:27:12ZfasTehran University of Medical SciencesTehran University Medical Journal1683-17641735-73221999-06-01571915Isolation and purification of natural killer cells subpopulations using mononuclear cellsMosaffa NLabibi FNatural Killer (NK) cells are the main lymphocyte population expressing P75 B chain of the IL-2 receptor (IL-2R). Consequently, incubation of peripheral blood lymphocytes with IL-2 induce selective activation of NK cells and results in NK activity and generation of Lymphokine activated killer (LAK) cells activity and proliferation. One of the early events during IL-2 activation of peripheral blood lymphocyte in both rodents and humans is adherence of some NK cells to plastic surface. The cells adherence to plastic after 24 hr of culture with IL-2 are almost exclusively CD56+, have the morphology large granular cells to yield a highly entiched population of activated NK cells that have been used for systemic adoptive immunotherapy. To test these hypothesis, we used highly purified population of human peripheral NK cells through the biological and nonimmunclogical phenotyping technique. Blood mononuclear cells were separated by centrifugation of ficol-hypaque gradient from normal blood donor (20-30 years age). We depleted after purification of nonadherent cells with nylonwool. We collected with rosette technique to remove cells with high affinity SRBC receptors. These cells separate in two parts A-NK and NA-NK by mononuclear celss activated supernatant media. The main objective results of this study show that the subpopulation of human NK cell which develope early adherent to plastic surface in the presence of supernatant mononuclear celss activation media was functionally more cytotoxic and killed K562 targets in single cell sytotoxicity manner and LDH activity assay than nonadherent NK cells and resting NK cellshttp://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/5688.pdf&manuscript_id=5688Plastic adherence suppernatant culture |
| spellingShingle | Mosaffa N Labibi F Isolation and purification of natural killer cells subpopulations using mononuclear cells Tehran University Medical Journal Plastic adherence suppernatant culture |
| title | Isolation and purification of natural killer cells subpopulations using mononuclear cells |
| title_full | Isolation and purification of natural killer cells subpopulations using mononuclear cells |
| title_fullStr | Isolation and purification of natural killer cells subpopulations using mononuclear cells |
| title_full_unstemmed | Isolation and purification of natural killer cells subpopulations using mononuclear cells |
| title_short | Isolation and purification of natural killer cells subpopulations using mononuclear cells |
| title_sort | isolation and purification of natural killer cells subpopulations using mononuclear cells |
| topic | Plastic adherence suppernatant culture |
| url | http://journals.tums.ac.ir/PdfMed.aspx?pdf_med=/upload_files/pdf/5688.pdf&manuscript_id=5688 |
| work_keys_str_mv | AT mosaffan isolationandpurificationofnaturalkillercellssubpopulationsusingmononuclearcells AT labibif isolationandpurificationofnaturalkillercellssubpopulationsusingmononuclearcells |