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In‐situ‐topoproteome analysis of cutaneous lymphomas: Perspectives of assistance for dermatohistologic diagnostics by Multi Epitope Ligand Cartography (MELC)
Author(s) -
Bonnekoh Bernd,
Pommer Ansgar J.,
Böckelmann Raik,
Philipsen Lars,
Hofmeister Henning,
Gollnick Harald
Publication year - 2008
Publication title -
jddg: journal der deutschen dermatologischen gesellschaft
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.463
H-Index - 60
eISSN - 1610-0387
pISSN - 1610-0379
DOI - 10.1111/j.1610-0387.2008.06754.x
Subject(s) - mycosis fungoides , cutaneous t cell lymphoma , immunophenotyping , pathology , medicine , in situ , lymphoma , dermatology , chemistry , flow cytometry , immunology , organic chemistry
Summary Background: Immunophenotyping is essential for diagnostics of cutaneous lymphomas. In this regard we present a skin tissue‐adapted application platform of MELC technology. Patients and Methods: This topoproteome analysis allows the subcellular colo‐calization of at least n = 100 epitopes in situ. For this purpose the specimen is processed by a Toponome Imaging Cycler ® for a n‐fold repetition of the following cycle: 1) staining with a fluorophore‐labeld antibody, 2) fluorescence‐imaging, and 3) photobleaching. Overlay and binarization of fluorescence images lead to combinatorial molecular phenotypes (CMP), which relate to a pixel or microtopographic unit (450 × 450 nm 2 , 20× objective). Skin biopsies were derived from patients with mycosis fungoides (patch/plaque lesions), psoriasis, atopic eczema and from healthy skin donors. Results: In orientation to the WHO‐EORTC‐classification of cutaneous lym‐phomas a MELC‐library of 23 markers was established. According to an inaugurative detailed procedure the CMP frequency was determined in a normalization to 100 μm horizontal skin width. By a TopoMiner strategy mycosis fungoides could be separated from the other states with a maximum of significance (p ≤ 0.03) by at least 10‐fold overexpression of the following tumor cell‐representative CMP‐motif: CD3+/CD4+/CD1a‐/CD7‐/CD8‐/CD45R0+/CD45RA‐/CD11a+. Conclusions: The skin tissue‐adapted MELC‐application‐platform extends substantially conventional lymphoma diagnostics by an unprecedented dimension of in‐situ‐analysis of marker combinatorics including its exact quantification and visualization.