Abstract
Recent evidence has indicated the importance of lysosomes in natural killing (NK). The lysosomotropic dye neutral red, at concentrations greater than 2 X 10(-5) M, inhibits NK activity of large granular lymphocytes (LGL). Inhibition is at the level of effector cell activity rather than target cell susceptibility. Neutral red inhibits NK activity without affecting the viability of the effector cell population or the response of lymphocytes to the mitogen, phytohaemagglutinin (PHA). While the mechanism of this inhibition is unknown, neutral red-treated peripheral blood lymphocytes (PBL) do not suppress untreated effector cell populations. Furthermore, neutral red does not influence the binding of effector cells to target cells. Since natural killing by neutral red-treated PBL recovers slowly upon incubation of the cells at 37 degrees, inhibition is not due to selective toxocity. Trypan blue, another frequently used lysosomotropic vital stain, has no effect when effector cells are pretreated with 10(-3) M dye for 30 min or less. However, effector cells first treated with neutral red lose red granular staining following treatment with trypan blue for as little as 10 min at room temperature. Natural killing by neutral red pretreated PBL also recovers more quickly following brief incubation with trypan blue. Treatment of effector cells with trypan blue and neutral red simultaneously prevents accumulation of neutral red in cytoplasmic granules and subsequent inhibition of NK.
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Selected References
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- Aterman K. The development of the concept of lysosomes. A historical survey, with particular reference to the liver. Histochem J. 1979 Sep;11(5):503–541. doi: 10.1007/BF01012536. [DOI] [PubMed] [Google Scholar]
- Beck F., Lloyd J. B., Griffiths A. Lysosomal enzyme inhibition by trypan blue: a theory of teratogenesis. Science. 1967 Sep 8;157(3793):1180–1182. doi: 10.1126/science.157.3793.1180. [DOI] [PubMed] [Google Scholar]
- Bulychev A., Trouet A., Tulkens P. Uptake and intracellular distribution of neutral red in cultured fibroblasts. Exp Cell Res. 1978 Sep;115(2):343–355. doi: 10.1016/0014-4827(78)90288-4. [DOI] [PubMed] [Google Scholar]
- Cameron D. J., Churchill W. H. Macrophage mediated cytotoxicity in man: role of hydrocortisone, trypan blue, chloroquine and prednisolone. Int J Immunopharmacol. 1981;3(1):77–85. doi: 10.1016/0192-0561(81)90047-3. [DOI] [PubMed] [Google Scholar]
- Carpen O., Virtanen I., Saksela E. Ultrastructure of human natural killer cells: nature of the cytolytic contacts in relation to cellular secretion. J Immunol. 1982 Jun;128(6):2691–2697. [PubMed] [Google Scholar]
- Cudkowicz G., Hochman P. S. Carrageenan-induced decline of natural killer activity. I. In vitro activation of adherent non-T-suppressor cells. Cell Immunol. 1980 Aug 1;53(2):395–404. doi: 10.1016/0008-8749(80)90340-8. [DOI] [PubMed] [Google Scholar]
- Grossi C. E., Cadoni A., Zicca A., Leprini A., Ferrarini M. Large granular lymphocytes in human peripheral blood: ultrastructural and cytochemical characterization of the granules. Blood. 1982 Feb;59(2):277–283. [PubMed] [Google Scholar]
- Haliotis T., Roder J., Klein M., Ortaldo J., Fauci A. S., Herberman R. B. Chédiak-Higashi gene in humans I. Impairment of natural-killer function. J Exp Med. 1980 May 1;151(5):1039–1048. doi: 10.1084/jem.151.5.1039. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Hibbs J. B., Jr Heterocytolysis by macrophages activated by bacillus Calmette-Guérin: lysosome exocytosis into tumor cells. Science. 1974 Apr 26;184(4135):468–471. doi: 10.1126/science.184.4135.468. [DOI] [PubMed] [Google Scholar]
- Nathan C. F., Silverstein S. C., Brukner L. H., Cohn Z. A. Extracellular cytolysis by activated macrophages and granulocytes. II. Hydrogen peroxide as a mediator of cytotoxicity. J Exp Med. 1979 Jan 1;149(1):100–113. doi: 10.1084/jem.149.1.100. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Ohkuma S., Poole B. Cytoplasmic vacuolation of mouse peritoneal macrophages and the uptake into lysosomes of weakly basic substances. J Cell Biol. 1981 Sep;90(3):656–664. doi: 10.1083/jcb.90.3.656. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Pappenheimer A. M. EXPERIMENTAL STUDIES UPON LYMPHOCYTES : I. THE REACTIONS OF LYMPHOCYTES UNDER VARIOUS EXPERIMENTAL CONDITIONS. J Exp Med. 1917 May 1;25(5):633–650. doi: 10.1084/jem.25.5.633. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Press J. L. H-2 haplotype-specific conversion of the Ir gene phenotype expressed by low-responder mice immunized with a p-azophenylarsonate conjugate of (T,G)-A--L. J Immunol. 1982 May;128(5):2236–2242. [PubMed] [Google Scholar]
- Roder J. C., Argov S., Klein M., Petersson C., Kiessling R., Andersson K., Hansson M. Target-effector cell interaction in the natural killer cell system. V. Energy requirements, membrane integrity, and the possible involvement of lysosomal enzymes. Immunology. 1980 May;40(1):107–116. [PMC free article] [PubMed] [Google Scholar]
- Roder J. C., Helfand S. L., Werkmeister J., McGarry R., Beaumont T. J., Duwe A. Oxygen intermediates are triggered early in the cytolytic pathway of human NK cells. Nature. 1982 Aug 5;298(5874):569–572. doi: 10.1038/298569a0. [DOI] [PubMed] [Google Scholar]
- Roder J. C., Lohmann-Matthes M. L., Domzig W., Wigzell H. The beige mutation in the mouse. II. Selectivity of the natural killer (NK) cell defect. J Immunol. 1979 Nov;123(5):2174–2181. [PubMed] [Google Scholar]
- Ruiz-Argüelles A., Seroogy K. B., Ritts R. E., Jr In vitro effect of cimetidine on human cell-mediated cytotoxicity. I. Inhibition of natural killer cell activity. Cell Immunol. 1982 May 1;69(1):1–12. doi: 10.1016/0008-8749(82)90045-4. [DOI] [PubMed] [Google Scholar]
- Scornik J. C., Ruiz P., Hoffman E. M. Suppression of cell-mediated tumor cell lysis and complement-induced cytotoxicity by trypan blue. J Immunol. 1979 Sep;123(3):1278–1284. [PubMed] [Google Scholar]
- Shau H., Dawson J. R. Regulation of human natural killing by levamisole. Cancer Immunol Immunother. 1982;13(1):24–29. doi: 10.1007/BF00200196. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Shau H., Koren H. S., Dawson J. R. Human natural killing against ovarian carcinoma. Br J Cancer. 1983 May;47(5):687–695. doi: 10.1038/bjc.1983.107. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Timonen T., Ortaldo J. R., Herberman R. B. Characteristics of human large granular lymphocytes and relationship to natural killer and K cells. J Exp Med. 1981 Mar 1;153(3):569–582. doi: 10.1084/jem.153.3.569. [DOI] [PMC free article] [PubMed] [Google Scholar]
- Tracey D. E., Adkinson N. F., Jr Prostaglandin synthesis inhibitors potentiate the BCG-induced augmentation of natural killer cell activity. J Immunol. 1980 Jul;125(1):136–141. [PubMed] [Google Scholar]
- Verhoef J., Sharma S. D. Inhibition of human natural killer activity by lysosomotropic agents. J Immunol. 1983 Jul;131(1):125–131. [PubMed] [Google Scholar]
- de Duve C., de Barsy T., Poole B., Trouet A., Tulkens P., Van Hoof F. Commentary. Lysosomotropic agents. Biochem Pharmacol. 1974 Sep 15;23(18):2495–2531. doi: 10.1016/0006-2952(74)90174-9. [DOI] [PubMed] [Google Scholar]
