Abstract
A simple and rapid method is reported here for enriching murine spleen cells with natural killer (NK) function as assessed by short-term cytolysis assay of 51Cr-labelled YAC-1 lymphoma target cells. The established method used for the enrichment of NK reactive cells, including large granular lymphocytes (LGL) from human and rat peripheral blood lymphocytes, does not substantially enrich for mouse splenic NK cell activity. A reproducible procedure for enriching mouse splenic NK cells has been developed using a four- or five-step discontinuous Percoll gradient in the density range of 1.062 g/ml (top) to 1.092 g/ml (bottom) and osmolarity (310-340 mOsm/kg) nearer to that of mouse blood and tissue. A four- to 25-fold (usually about nine-fold) increase in NK cell activity, consisting of 50-100% of the recovered lytic unit activity, is found in which are the cells forming band 3, approximately 10% of the recovered cell number. This cell population with enriched NK cell activity has a characteristic density less than or equal to 1.077 g/ml, but more than 1.070 g/ml when centrifuged under appropriate conditions. Similar enrichment was obtained with a four-step gradient at an uniform osmolarity of 320 mOsm/kg throughout. Although the lymphocytes in band 3 show relatively little heterogeneity in appearance, only a minor population of the cells contain granules.
Full text
PDF








Images in this article
Selected References
These references are in PubMed. This may not be the complete list of references from this article.
- Herberman R. B., Holden H. T. Natural cell-mediated immunity. Adv Cancer Res. 1978;27:305–377. doi: 10.1016/s0065-230x(08)60936-7. [DOI] [PubMed] [Google Scholar]
- Itoh K., Suzuki R., Umezu Y., Hanaumi K., Kumagai K. Studies of murine large granular lymphocytes. II. Tissue, strain, and age distributions of LGL and LAL. J Immunol. 1982 Jul;129(1):395–405. [PubMed] [Google Scholar]
- Julius M. H., Simpson E., Herzenberg L. A. A rapid method for the isolation of functional thymus-derived murine lymphocytes. Eur J Immunol. 1973 Oct;3(10):645–649. doi: 10.1002/eji.1830031011. [DOI] [PubMed] [Google Scholar]
- Kiessling R., Wigzell H. An analysis of the murine NK cell as to structure, function and biological relevance. Immunol Rev. 1979;44:165–208. doi: 10.1111/j.1600-065x.1979.tb00270.x. [DOI] [PubMed] [Google Scholar]
- Kumagai K., Itoh K., Suzuki R., Hinuma S., Saitoh F. Studies of murine large granular lymphocytes. I. Identification as effector cells in NK and K cytotoxicities. J Immunol. 1982 Jul;129(1):388–394. [PubMed] [Google Scholar]
- Luini W., Boraschi D., Alberti S., Aleotti A., Tagliabue A. Morphological characterization of a cell population responsible for natural killer activity. Immunology. 1981 Aug;43(4):663–668. [PMC free article] [PubMed] [Google Scholar]
- Reynolds C. W., Timonen T., Herberman R. B. Natural killer (NK) cell activity in the rat. I. Isolation and characterization of the effector cells. J Immunol. 1981 Jul;127(1):282–287. [PubMed] [Google Scholar]
- Silverstein E., Sokoloff L., Mickelsen O., Jay G. E. Primary Polydipsia and Hydronephrosis in an Inbred Strain of Mice. Am J Pathol. 1961 Feb;38(2):143–159. [PMC free article] [PubMed] [Google Scholar]
- Tagliabue A., Befus A. D., Clark D. A., Bienenstock J. Characteristics of natural killer cells in the murine intestinal epithelium and lamina propria. J Exp Med. 1982 Jun 1;155(6):1785–1796. doi: 10.1084/jem.155.6.1785. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tagliabue A., Luini W., Soldateschi D., Boraschi D. Natural killer activity of gut mucosal lymphoid cells in mice. Eur J Immunol. 1981 Nov;11(11):919–922. doi: 10.1002/eji.1830111112. [DOI] [PubMed] [Google Scholar]
- Timonen T., Reynolds C. W., Ortaldo J. R., Herberman R. B. Isolation of human and rat natural killer cells. J Immunol Methods. 1982;51(3):269–277. doi: 10.1016/0022-1759(82)90393-3. [DOI] [PubMed] [Google Scholar]
- Timonen T., Saksela E. Isolation of human NK cells by density gradient centrifugation. J Immunol Methods. 1980;36(3-4):285–291. doi: 10.1016/0022-1759(80)90133-7. [DOI] [PubMed] [Google Scholar]