In-situ-topoproteome analysis of cutaneous lymphomas: perspectives of assistance for dermatohistologic diagnostics by Multi Epitope Ligand Cartography (MELC).
BACKGROUND
Immunophenotyping is essential for diagnostics of cutaneous lymphomas. In this regard we present a skin tissue-adapted application platform of MELC technology.


METHODS
This topoproteome analysis allows the subcellular colocalization 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 x 450 nm2, 20x 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 lymphomas a MELC-library of 23 markers was established. According to an inaugurative detailed procedure the CMP frequency was determined in a normalization to 100 microm horizontal skin width. By a TopoMiner strategy mycosis fungoides could be separated from the other states with a maximum of significance (p < or = 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.
Bonnekoh
Bernd
B
Clinic for Dermatology and Venereology, Otto-von-Guericke-University, Magdeburg, Germany. bernd.bonnekoh@medizin.unimagdeburg.de
Pommer
Ansgar J
AJ
Böckelmann
Raik
R
Philipsen
Lars
L
Hofmeister
Henning
H
Gollnick
Harald
H
D014408:Biomarkers, Tumor / Q000032:analysis*; D018604:Epitope Mapping / Q000379:methods; D020869:Gene Expression Profiling / Q000379:methods*; D006801:Humans; D016410:Lymphoma, T-Cell, Cutaneous / Q000175:diagnosis* / Q000378:metabolism*; D009363:Neoplasm Proteins / Q000032:analysis*; D020543:Proteome; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
D014408:Biomarkers, Tumor; D009363:Neoplasm Proteins; D020543:Proteome