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Mast Cells in Leprosy Skin Lesions J Clin Pathol: First Published As 10.1136/Jcp.43.3.196 on 1 March 1990

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Mast Cells in Leprosy Skin Lesions J Clin Pathol: First Published As 10.1136/Jcp.43.3.196 on 1 March 1990 196 J Clin Pathol 1990;43:196-200 Mast cells in leprosy skin lesions J Clin Pathol: first published as 10.1136/jcp.43.3.196 on 1 March 1990. Downloaded from I A Cree, G Coghill, J Swanson Beck Abstract prominent, there is an influx of lymphocytes, The variability ofmast cell density within fibroblast proliferation is evident, and giant cell and between leprosy skin lesions was formation occurs.7 Patients of the same classi- examined as a basis for future studies, fication on the Ridley-Jopling scale vary in and whether the number of mast cells in their susceptibility to reversal reactions. the lesion was determined by local or It has long been known that mast cells are systemic factors was evaluated. The mast intimately concerned with the production of cell density in the granuloma, skin oedema, one ofthe features ofreversal reaction. appendages, and intervening dermis was Their possible role in the pathogenesis of such assessed by counting mast cells in glycol reactions was investigated by Chowdhury and methacylate sections stained with Ghosh,5 who found a lower density ofmast cells Giemsa stain and relating these counts to in tuberculoid lesions in reaction compared area measurements obtained by plani- with non-reactive tuberculoid lesions. There is metry. In biopsy specimens taken from also some evidence of mast cell degranulation the edge of established lesions the density in erythema nodosum leprosum (ENL, type II of mast cells within the granulomata was leprosy reactions).9 O The aim ofthis investiga- considerably higher than that in the tion was to examine the variability of mast cell intervening dermis and was comparable density within and between leprosy skin lesions with that found in the appendages. No as a basis for future studies, and to ascertain major differences in mast cell density whether the number of mast cells in the lesion were found between unaffected skin and is determined by local or systemic factors. the centre or edge of individual lesions. Three methods of staining mast cells are Mast cell densities in biopsy specimens currently in use: (a) metachromatic staining of from the edge of different lesions on the their granules"; (b) histochemical demonstra- same patient were also similar, suggest- tion of enzymes12; and (c) immunoperoxidase http://jcp.bmj.com/ ing that the mast cell density within the staining using monoclonal antibodies.'"" As granulomata is independent of the site of methods (b) and (c) both require frozen sec- the lesion and is determined systemically. tions metachromatic staining was the only option available for this study. The method developed by Vogt et al" using glycol meth- The skin lesions of leprosy contain many if not acrylate sections stained with Giemsa stain was all of the cell types which occur in normal skin, chosen for this study because it combined the on September 28, 2021 by guest. Protected copyright. in addition to those which migrate into the excellent morphological detail of plastic lesions from peripheral blood as part of the embedding with a reliable method of showing granulomatous process. The contribution of the presence of metachromatic granules. cells normally present in the dermis to granu- loma formation and progression is uncertain. Mast cells have received little attention in leprosy recently, but evidence linking them Methods with the development of delayed hypersen- Two groups of 4 mm skin punch biopsy sitivity reactions' raises the possibility that they specimens were obtained under local anaes- might be of some importance in leprosy lesions. thesia with 10O lignocaine from patients with Mast cells are heterogeneous with respect to leprosy in India and Bangladesh. All of the their morphology, biochemistry, and func- patients were untreated or had received treat- tion.2" The differences between them appear to ment (WHO multiple drug therapy regimen) on the tissue and species from which for less than one month. Multiple biopsy Department of depend Pathology, Ninewells they are derived, so that is is not possible to specimens were taken from individual patients; Hospital and Medical apply the results of studies of mast cell dis- for the examination of intralesional variation in School, Dundee, tribution or function in one site or animal to mast cell numbers, specimens were taken from Scotland I A Cree another with any certainty. the centre of the lesion, the edge of the lesion, G Coghill Type 1 reversal reactions are thought to and from the apparently unaffected skin out- J Swanson Beck reflect increased delayed hypersensitivity to side the lesion at a point 2 cm from the nearest Correspondence to: Mycobacterium leprae and account for much of identifiable margin of the lesion in 22 patients Dr I A Cree, Department of of interlesional Pathology, Ninewells the nerve damage which occurs in borderline (biopsy series 1); for the study Hospital and Medical leprosy patients.5 Clinically there is inflamma- variation specimens were taken from the edge School, Dundee, DD1 9SY, same in Scotland. tion of both skin lesions and affected nerves, of two different lesions on the patient was made to Accepted for publication with pain, swelling, and tendemess on pal- 35 cases (biopsy series 2). An effort 31 August 1989 pation.6 Histologically, intercellular oedema is choose lesions of disparate size and appearance Mast cells in leprosy skin lesions 197 Table 1 Age and sex distribution ofpatients and Ridley-Jopling classification BIOPSY SERIES 1 The average mast cell density in biopsy Age Sex Ridley-Jopling classification specimens from unaffected skin was 17 6 per ratio Mean (SD) (M:F) Idt TT BT BB BL LL Total mm2. Specimens taken from the centre of the J Clin Pathol: first published as 10.1136/jcp.43.3.196 on 1 March 1990. Downloaded from lesion had an average of 18 7 mast cells per Series 1 36 (13 0) 16:6 1 0 14 0 3 4 22 mm2, while at the edge of the lesion the average Series 2 38 (15 1) 28:7 0 1 19 2 1 12 35 mast cell density was 19 5 per mm2. There was Idt = indeterminate; TT = tuberculoid; BT = borderline-tubeculoid; BB = borderline; no significant difference between the densities BL = borderline-lepromatous; LL = lepromatous. in either of these sites, suggesting that the total number of mast cells in the skin does not where possible to determine the maximum change greatly within the lesion and is similar variation which might occur. The number, age, to that in normal skin. When the component and sex distribution, and Ridley-Jopling clas- mast cell densities from different sites in the sification of the patients in each series is shown same lesion (granuloma, appendages, and in table 1. intervening dermis) were compared in each After being transported to Dundee in 4% biopsy specimen there was no discernible pat- buffered formaldehyde each biopsy specimen tern in either paucibacillary or multibacillary was bisected: one half was embedded in paraf- patients. fin wax, the other in glycol methacrylate." Paraffin wax sections (5 gim) were stained with Adikim haematoxylin and eosin and by the Wade-Fite 10oorIr ..de r Aa. W..Awam method for diagnostic purposes, while 2 gm 7vr A.. sections were cut from the glycol-methacrylate ... tow embedded half of each biopsy specimen and a stained for mast cell quantitation using the Giemsa method developed by Vogt et al." 4Jt ..S The dermal area and the areas of the section 5 occupied by granuloma and by skin append- ages (sebaceous glands, sweat glands, and hair follicles) were measured by planimetry.'6 Mast i cell counts were performed on non-overlap- I ping fields at x 400 magnification by scanning I the whole section using a 25 square eyepiece A._ grid. To avoid overestimation of mast cell density cells were counted only if part of the nucleus could be seen, and anucleate fragments were ignored. Mast cell counts were expressed for each Auk -Aftl http://jcp.bmj.com/ component (dermis, granuloma, and append- sV.w_ age) as cells per mm.2 The results for paucibacillary and multibacillary cases were Figure I Part of an epithelioid cellgranuloma in a compared using the Mann-Whitney U test for biopsy specimen takenfrom a patient with borderline- non-parametric data. As lesion selection may tuberculoid leprosy. Two mast cells (arrows) within the have occurred when taking biopsy specimens gramuloma show prominent nucleoli and many granules. (Giemsa.) on September 28, 2021 by guest. Protected copyright. from different lesions (biopsy series 2) the results in this group were randomised before the relation between mast cell densities in -e: -e individual patients was assessed by linear regression analysis. Results The appearance of the mast cells in leprosy lesions is very variable. Several mor- phologically distinct variants were present: "active mast cells" with large ovoid nuclei, prominent nucleoli, and extensive cytoplasm with many granules; "fusiform mast cells" with elongated dark-staining fusiform nuclei and little cytoplasm; and "resting mast cells" with small dark nuclei, scanty cytoplasm, and few granules. These cell types were most com- monly found in inflammatory foci (fig 1), applied to sebaceous or sweat glands (fig 2), and in the unaffected dermis (fig 2), respectively. There were too many cells of intermediate Figure 2 A fieldfrom the unaffected dermis in a case of morphology, however, to allow these types of borderline-tuberculoid leprosy showing a dermal mast cell. to be differentiated and counted (arrow) with dense granules and an elongated mast cell mast cell applied to the side of a capillary adjacent to a sweat separately on morphological grounds alone. gland (arrowhead). (Giemsa.) 198 Cree, Coghill, Swanson Beck Table 2 Average mast cell densities (per mm2) in skin 400- and component parts of specimens takenfrom edge of leprosy lesions 350- J Clin Pathol: first published as 10.1136/jcp.43.3.196 on 1 March 1990.
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