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Demonstration of Charcot-Leyden crystals in eosinophilic cystitis.

To the Editor.--The presence and diagnostic relevance of Charcot-Leyden crystals (CLCs) in eosinophilic cystitis (EC) was reported in detail recently. (1) Although their association with the disease is well established, CLCs are still poorly studied. (2-4) Appreciation of CLCs on standard hematoxylin-eosin-stained tissue sections is not always straightforward, especially with the background of apoptotic changes accompanied by different degrees of necrosis seen in the acute phase of EC. (1,5) The CLCs stain faintly with eosin and do not polarize on examination under polarized light microscopy.

Chemical knowledge of CLCs was limited until 1980 when it was discovered that they contain cationic proteins that stain satisfactorily with hematoxylin-basic fuchsin-picric acid in brilliant dark-pink coloration. (2)

Charcot-Leyden crystals consist of a 13 000-molecular weight acidic protein whose amino acid composition distinguished it from the 9300-molecular weight major basic protein, the principal constituent of eosinophil large granules; this protein is identical to lysophospholipase (3) and is stained by basic fuchsin in the Ziehl-Neelsen method.

We suggest 2 helpful although not so popular diagnostic methods--autofluorescence and acid-fast stain--for easy detection of CLCs. (6,7)

We performed a study on the incidence of CLCs in EC focusing precisely on these 2 diagnostic features (autofluorescence and acid-fast stain) comparing them with standard hematoxylin-eosin-stained tissue sections. Our study examined 15 cases with EC. On our hematoxylin-eosinstained sections (Ref. No. 676500; Thermo Scientific Shandon, Pittsburgh, Pennsylvania), CLCs were readily seen in only 4 of 15 cases (Figure, a). The CLCs stained red with Ziehl-Neelsen (Ref. No. 1.09215.0500; Merck, Darmstadt, Germany) in 6 of 15 cases and exhibited yellow-green autofluorescence (8 of 15 cases) when formalin-fixed, paraffin-embedded, hematoxylin-eosin-stained sections were viewed with fluorescent microscope (Zeiss) with an IVFI condenser using filters FT 510 and LP 520 (Figure, b and c). Moreover, fluorescence made detection of CLCs fast and easy especially in hypercellular areas versus the time-consuming screening for them with conventional light microscopy. The remaining Ziehl-Neelsen-negative and fluorescencenegative cases presented with a severe degree of necrosis, which in our opinion was the reason for dye impermeability and noninformative fluorescence.




Although formalin-induced autofluorescence and acid-fast stain of CLCs are probably nonspecific phenomena, use of the fluorescent microscope and the Ziehl-Neelsen stain can successfully help in the diagnosis of acute-phase EC, especially in samples with massive tissue necrosis.

Diana Staribratova, MD, PhD

Vesselin Belovejdov, MD, PhD

Dimitri Staikov, MD

Department of Pathology

Medical University of Plovdiv

4000 Plovdiv, Bulgaria

Dorian Dikov, MD

Service d'Anatomie et de Cytologie Pathologiques

Centre Hospitalier de Lagny Marne-la-Valkee

Lagny-sur-Marne, France

(1.) Popescu OE, Landas SK, Haas GP. The spectrum of eosinophilic cystitis in males: case series and literature review. Arch Pathol Lab Med. 2009;133(2):289-294.

(2.) Dincsoy HP, Burton TJ, van der Bel-Kahn JM. Circulating Charcot-Leyden crystals in the hypereosinophilic syndrome. Am J Clin Pathol. 1981; 75(2):236-243.

(3.) Weller PF, Goetzel EJ, Austen F. Identification of human eosinophil lysophospho-lipase as the constituent of Charcot-Leyden crystals (phospholipase B). Med Sci. 1980;77(12):7440-7443.

(4.) Goodman TR, Connolly B, Taylor G. Eosinophilic cystitis following an urachal remnant. Pediatr Radiol. 1999;29(6):487-488.

(5.) Staribratova D, Staikov D, Dikov D, et al. Eosinophilic cystitis: clinico-morphologic aspects. Virchows Archiv. 2003;443(3):432.

(6.) Lo JW, Fung CHK. Autofluorescent Charcot-Leyden crystals. Arch Pathol Lab Med. 1992; 116(11):1101.

(7.) Bu XM, Yao LQ, Zheng ZY, Xiong XS. Demonstration of Charcot-Leyden crystals by acid-fast stains applied on tissues. Zhonghua Bing Li Xue Za Zhi. 2006;35(1):47.

The authors have no relevant financial interest in the products or companies described in this article.
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Author:Staribratova, Diana; Belovejdov, Vesselin; Staikov, Dimitri; Dikov, Dorian
Publication:Archives of Pathology & Laboratory Medicine
Article Type:Letter to the editor
Date:Oct 1, 2010
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