Ultrastructural analysis of human skin biopsy specimens from patients receiving recombinant human stem cell factor: Subcutaneous injection of rhSCF induces dermal mast cell degranulation and granulocyte recruitment at the injection site  Ann M. Dvorak, MD, John J. Costa, MD, Rita A. Monahan-Earley, BA, Patricia Fox, BA, Stephen J. Galli, MD  Journal of Allergy and Clinical Immunology  Volume 101, Issue 6, Pages 793-806 (June 1998) DOI: 10.1016/S0091-6749(98)70404-X Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 1 A, Normal skin blood vessel in a control biopsy site obtained 6 days after the initiation of rhSCF dosing. Thin endothelial cells appear intact. B, Vessel in rhSCF injection site obtained on first day of rhSCF dosing. Vessel exhibits swollen, electron-lucent, necrotic endothelial cells (E). Lumen is packed with portions of cytoplasm from damaged cells and remnants of neutrophils. Numerous polymorphonuclear neutrophilic granulocytes (arrows) have migrated through vessel and reside beneath vascular basal lamina. A, magnification ×6000; B, ×4500. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 2 Biopsy specimen of rhSCF injection site obtained on day 6 of rhSCF dosing. Papillary dermis shows dense aggregates of fibrin, some of them bound to collagen (in A) and necrotic eosinophils and neutrophils among collagen bundles (in B). Extracellular eosinophil and neutrophil granules retain their granule membranes, indicating their origin from cells undergoing necrosis rather than degranulation. A, magnification ×14,000; B, ×9500. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 3 Mast cell (M) in rhSCF injection site (obtained on day 6 of rhSCF dosing) has extruded most of its granules (arrows). The mast cell is in close contact with a fibroblast (F), which has greatly expanded cisterns of rough endoplasmic reticulum (arrowhead). Magnification ×14,000. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 4 Blood vessels and granulocytes in rhSCF injection site biopsy specimen obtained on day 13 of rhSCF dosing. Blood vessels exhibit perivascular neutrophils (N), eosinophils (E), and basophils (B). Basophils (e.g., B in A and B) exhibit extensive PMD, characterized by presence of numerous empty granules in their cytoplasm. Some eosinophils exhibit alterations in their secondary granules, involving both matrix and core compartments. A, magnification ×7000; B, ×13,500. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig 5 Biopsy specimen of rhSCF injection site obtained on day 13 of rhSCF dosing contains some mast cells that have many secretory granules that exhibit no evidence of AND. Many developing granules show central dense foci of condensation (arrows) superimposed on underlying granule patterns. These patterns include crystals (closed arrowhead in B) and scrolls (open arrowhead in B). A, magnification ×16,000; B, ×36,000. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 6 Montage of cytoplasmic granules from mast cells in rhSCF injection site biopsy specimen (obtained on day 13 of rhSCF dosing) illustrating different ultrastructural patterns and processes. In A, granule contains dense particles. In B, regular crystalline array fills large portion of this granule, which exhibits mixed structural pattern. Mixed granule in C contains prominent scrolls at periphery of homogeneously dense material. D, Two granules with central condensing nucleoids (arrows), finding typical of mast cell granule building. E, Granule that exhibits focal, irregular electron-lucent areas within otherwise electron-dense granule matrix is characteristic of PMD (arrows). A, magnification ×84,000; B, ×90,000; C, ×78,000; D, ×52,000; E, ×61,000. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 7 Mast cell in papillary dermis of rhSCF injection site biopsy specimen (obtained on day 13 of rhSCF dosing) reveals morphology typical of AND. Altered, swollen granules are present in intracytoplasmic degranulation chambers that open to cell surface, and swollen, membrane-free, extruded granules (arrows) are present in interstitium at distance from probable mast cell of origin. These extruded granules rest in amorphous, slightly dense pools of interstitial edema that separate adjacent bands of collagen. Electron-dense fibrin fragments are also visible, some of which (open arrowhead) are attached to mast cell surface. Magnification ×13,000. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions

Fig. 8 High-magnification micrograph of rhSCF injection site biopsy specimen, which was obtained on day 13 of rhSCF dosing, illustrating extracellular, membrane-free mast cell granules in close proximity to surfaces of elongated, narrow cell processes of dermal dendrocytes. Dermal dendrocyte at top illustrates intermediate filament-packed process of dermal dendrocyte in continuity with dendrocyte cell body. Banded interstitial collagen fibers are plentiful. Magnification ×37,000. Journal of Allergy and Clinical Immunology 1998 101, 793-806DOI: (10.1016/S0091-6749(98)70404-X) Copyright © 1998 Mosby, Inc. Terms and Conditions