Summary
The pathway of interleukin 1 (IL-1) secretion from the cell remains unclear. IL-1β is the major form produced by human monocytes, and is synthesized as a precursor of 35kDa which is processed to the extracellular biologically active 17kDa form. We have examined the intracellular localization of IL-1β in lipopolysaccharide (LPS)-stimulated human peripheral blood monocytes, by immunocytochemistry and immunoprecipitation of subcellular fractions. LPS treatment slightly damaged the cells. Unstimulated cells showed very little immunolabelling. In contrast, there was heavy immunolabelling on LPS stimulated cells. Immunolabelling occured within the cytoplasm, nucleus and mitochondria. There was no immunolabelling on the membranous secretory organelles and the plasma membrane. Blebs of cytoplasm budding from the cell surface were immunolabelled, suggesting an alternative route of secretion of IL-1β from the cell. Immunoprecipitation studies confirmed these results.
Similar content being viewed by others
References
Auron, P. E., Webb, A. C., Rosenwasser, L. J., Mucci, S. F., Rich, A., Wolff, S. M. &Dinarello, C. (1984). Nucleotide sequence of human monocyte Interleukin-1 precursor mRNA.Proc. Natl. Acad. Sci. USA 81, 7907–11.
Bakouche, O. Brown, D. C. &Lachman, L. B. (1987) Subcellular localisation of human monocytes Interleukin-1: Evidence for an inactive precursor molecule and a possible mechanism for IL1 release.J. Immunol. 138, 4249–55.
Bomford, R., Abdulla, E., Hughes-Jenkins,C., Simpkin, D. &Schmidt, J. (1987) Antibodies to Interleukin-1 raised with synthetic peptides: identification of external sizes and analysis of interleukin-1 synthesis in stimulated human peripheral blood monocytes.Immunology,62, 543–9.
Clarke, B. D., Collins, K. L., Gandy, M. S., Webb, A. C. &Auron, P. E. (1986) Genomic sequence for human pro-interleukin-1 beta: possible evolution from a reverse transcribed pro-interleukin-1-alpha gene.Nuc. Acids Res. 14, 7897–914.
Deshates, R. J., Koch, B. D., Werner-Wasbourne, M., Craig, E. A. &Schekman, R. (1988) A subfamily of stress proteins facilitates translocation of secretory and mitochondrial precursor polypeptides.Nature (Lond.) 332, 800–5.
Dinarello, C. (1988). Biology of interleukin-1.FASEB J. 2, 108–15.
Geri, I. &Lepe-zuniga, J. C. (1983) Interleukin 1: uniqueness of its production and spectrum of activities. InLymphokines (edited byPick, E.) Vol. 9, pp. 109–26. Orlando FA: Academic Press.
Hazuda, D. J., Lee, J. C. &Young, P. R. (1988) The kinetics of interleukin-1 secretion for activated moncytes. Differences between interleukin-1α and interleukin-1β.J. Biol. Chem. 263, 8473–9.
Hurt, E. C. &van Loon, A. P. G. (1986) Low proteins find mitochondria and intramitochondrial compartments.T.J.B.S. 11, 204–7.
Kronheim, S., March, C., Erb, S., Conlon, P., Mochizuki, D. &Hopp, T. (1985) Human interleukin-1: purification to homogeneity.J. Exp. Med. 161, 490–502.
Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4.Nature (Lond.) 227, 680–5.
March, C. J., Mosley, B., Larsen, A., Ceretti, D. P., Braedi, G., Price, V., Gillis, S., Henney, C. S., Kronheim, S. R., Grabstein, K., Conlon, P. J., Hopp, T. P. &Cosman, D. (1985) Cloning, sequence and expression of two distinct human interleukin-1 complementary DNA's.Nature (Lond.),315, 641–7.
Matsushima, K. T., Copeland, K., Onozaki, K. &Oppenheim, J. J. (1986a) Purification and biochemical characteristics of two distinct human interleukin-1′s from the myelomonocytic THP-1 cell line.Biochemistry 25, 3429.
Matsushima, K., Taguchi, M., Kovacs, E. J., Young, H. A. &Oppenheim, J. J. (1986b) Intracellular localisation of human monocyte associated interleukin-1 (IL-1) activity and release of biologically active IL-1 from monocytes by trypsin and plasmin.J. Immunol. 136, 2883–91.
Mosley, B., Urdal, D. L., Prickett, K. S., Larson, A., Cosman, D., Conlon, P. J., Gillis, S. &Dower, S. K. (1987) The interleukin-1 receptor binds the human interleukin-1α precursor, but not the interleukin-1β precursor.J. Biol. Chem. 262, 2941–4.
Oppenheim, J. J., Kovacs, E. J., Matsushima, K. &Durrum, S. K. (1986) There is more than one interleukin-1.Immunol. Today 7, 45–56.
Schmidt, J. (1984) Purification of human interleukin-1.J. Exp. Med. 160, 772–87.
Singer, I., Scott, S., Hall, G. L., Limjuco, G., Chin, J. &Schmidt, J. (1988) Interleukin-1β is localised in the cytoplasmic ground substance but is largely absent from the Golgi apparatus and the plasa membranes of stimulated human monocytes.J. Exp. Med. 167, 389–407.
Tokuyasu, K. T. (1986) Immunocryoultramicrotomy.J. Microsc. 143, 139–49.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Beesley, J.E., Bomford, R. & Schmidt, J.A. Ultrastructural localization of Interleukin 1 in human peripheral blood monocytes; evidence for IL-1β in mitochondria. Histochem J 22, 234–244 (1990). https://doi.org/10.1007/BF02386010
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF02386010