Summary
The subcellular localization of lactoferrin in human neutrophils was studied by an electron-microscopic immunoperoxidase method. This molecule was detected in small granules of blood polymorphonuclear leukocytes. A morphometrical analysis showed that there was no significant difference in the mean size between lactoferrin-positive and myeloperoxidase-negative granules. In contrast, the mean size of myeloperoxidase-positive granules was significantly larger than that of lactoferrin-positive granules. This indicates that lactoferrin is contained in the myeloperoxidase-negative, secondary, granules of human neutrophils. In immature bone marrow mononuclear neutrophils, lactoferrin was present in cytoplasmic granules of somewhat larger size than lactoferrin-positive granules of polymorphonuclear leucocytes. A morphometrical study showed that the mean size of lactoferrin-positive granules was significantly greater in immature bone marrow cells than in polymorphonuclear leucocytes. This indicates that lactoferrin-positive granules decrease in size as the cells mature. Besides cytoplasmic granules, lactoferrin was demonstrated in the Golgi complex and a part of the rough endoplasmic reticulum of immature bone marrow neutrophils, probably myelocytes and early metamyelocytes. These results show that lactoferrin is synthesized and packed into secondary granules in immature bone marrow neutrophils and therefore that the secondary granules are a type of secretory granule.
Similar content being viewed by others
References
Ambruso DR, Johnston RB (1981) Lactoferrin enhances hydroxyl radical production by human neutrophils, neutrophil particulate fraction and an enzymatic generating system. J Clin Invest 67:352–360
Baggiolini M, Bretz U, Dewald B, Feigenson ME (1978) The polymorphonuclear leukocyte. Agents Actions 8:3–10
Bainton DF, Ullyot JL, Farquhar MG (1971) The development of neutrophilic polymorphonuclear leukocytes in human bone marrow. J Exp Med 134:907–934
Borgers M, Thone F, DeCree J, DeCock W (1978) Alkaline phosphatase activity in human polymorphonuclear leukocytes. Histochem J 10:31–43
Boxer LA, Haak RA, Yang H-H, Wolach JB, Whitcomb JA, Butterick CJ, Baehner RL (1982) Membrane-bound lactoferrin alters the surface properties of polymorphonuclear leucocytes. J Clin Invest 70:1049–1057
Breton-Gorius J, Mason DY, Buriot D., Vilde J-E, Griscelli C (1980) Eactoferrin deficiency as a consequence of a lack of specific granules in neutrophils from a patient with recurrent infections: detection by immunoperoxidase staining for lactoferrin and cytochemical electron microscopy. Am J Pathol 99:413–428
Bretz U, Baggiolini M (1974) Biochemical and morphological characterization of azurophil and specific granules of human neutrophilic polymorphonuclear leukocytes. J Cell Biol 63:251–269
Coates TD, Oseas RS, Yang HH, Baehner RE, Boxer EA (1980) Polymorphonuclear leukocyte aggregation promoted by lactoferrin mediated reduction in cell surface charge. Clin Res [Abstr] 28:307A
Cramer E, Pryzwansky KB, Villeval J-E, Testa U, Breton-Gorius J (1985) Ultrastructural localization of lactoferrin and myeloperoxidase in human neutrophils by immunogold. Blood 65:423–432
Gallin JI, Fletcher MP, Seligmann BE, Hoffstein S, Cehrs K, Mounessa N (1982) Human neutrophil-specific granule deficiency: a model to assess the role of neutrophil-specific granule in the evolution of the inflammatory response. Blood 59:1317–1329
Kane SP, Peters TJ (1975) Analytical subcellular fractionation of human granulocytes with reference to the localization of vitamine B12-binding proteins. Clin Sci Mol Med 49:171–182
Klebanoff SJ, Clark RA (1978) The neutrophil: function and clinical disorders. North-Holland Publ Comp, Amsterdam, p 29
Mason DY, Farrell C, Taylor CR (1975) The detection of intracellular antigens in human leucocytes by immunoperoxidase staining. Br J Haematol 31:361–370
Miyauchi J, Watanabe Y, Enomoto Y, Takeuchi K (1983) Eactoferrin-deficient neutrophil polymorphonuclear leucocytes in leukaemias: a semiquantitative and ultrastructural cytochemical study. J Clin Pathol 36:1397–1405
Miyauchi J (1984) Distribution and subcellular localization of lactoferrin in human tissues with special reference to the submandibular gland. Acta Histochem Cytochem 17:77–89
Miyauchi J, Sasadaira H, Watanabe K, Watanabe Y (1985) Ultrastructural immunocytochemical localization of lysozyme in human monocytes and macrophages. Cell Tissue Res 242:269–277
Murphy G, Reynolds JJ, Bretz U, Baggiolini M (1977) Collagenase is a component of the specific granules of human neutrophil leukocytes. Biochem J 162:195–197
Nakane PK, Pierce GB Jr (1967) Enzyme-labeled antibodies for the light and electron microscopic localization of tissue antigens. J Cell Biol 33:307–318
Parmley RT, Takagi M, Barton JC, Boxer LA, Austin RL (1982) Ultrastructural localization and iron-binding protein in human neutrophils and rabbit heterophils. Am J Pathol 109:343–358
Pryzwansky KB, Rausch PG, Spitznagel JK, Herion JC (1979) Immunocytochemical distinction between primary and secondary granule formation in developing human neutrophils: Correlations with Romanovsky stains. Blood 53:179–185
Snedecor GW, Cochran WG (1980) Statistical method. The Iowa State University Press, Iowa, USA, p 96–98
Spitznagel JK, Dalldorf FG, Leffell MS, Folds JD, Welsh IRH, Cooney MH, Martin LE (1974) Character of azurophil and specific granules purified from human polymorphonuclear leukocytes. Lab Invest 30:774–785
Watanabe Y (1979) Ultrastructure of human neutrophils and leukemic cells. Transactiones Societatis Pathologicae Japonicae 68:3–34 (in Japanese)
Wilson MB, Nakane PK (1978) Recent development in the periodate method of conjugating horseradish peroxidase (HRPO) to antibodies. In: Knapp W, Holubar K, Wick G (eds) Immunofluorescence and related staining techniques. Elsevier/North-Holland Biomedical Press, Amsterdam, p 215–224
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Miyauchi, J., Watanabe, Y. Immunocytochemical localization of lactoferrin in human neutrophils. Cell Tissue Res. 247, 249–258 (1987). https://doi.org/10.1007/BF00218306
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00218306