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METHOD FOR DETERMINING SUBPOPULATION COMPOSITION OF SKIN CELLS AND OBTAINING SKIN CYTOIMMUNOGRAM

FIELD: medicine.

SUBSTANCE: method for determining the subpopulation composition of skin cells and obtaining the skin cytoimmunogram comprises sampling a skin biopsy specimen to a depth of 2 mm, homogenizing the tissue in a 0.9% aqueous solution of sodium chloride at a temperature of +23…+25°C, extracting the homogenate, filtering the homogenate through an inert filtering partition with a pore diametre of 20 mcm, centrifuging the homogenate at 400 g for 5 minutes at a temperature +23…+25°C. After centrifugation, the viability of the skin cells is determined, the skin cells are incubated for 20 minutes with monoclonal antibodies fluorochrome-labeled for flow cytometry, phenotyping of the skin cells is carried out, the number of skin cells of a specific phenotype - skin cytoimmunogram - are determined: keratinocytes CD49f+, among them activated CD49f+HLA-DR+; fibroblasts (fibrocytes) CD45-CD14-CD44+, among them activated CD45-CD14-CD44+CD80+; mast cells CD249+, among them activated CD249+CD63+; monocytes (macrophages) CD45+CD14+, among them activated CD45+CD14+HLA-DR+; intra-epidermal macrophages CD207+, among them activated CD207+CD80+, CD207+HLA-DR+, CD207+CD80+HLA-DR+; endothelial cells CD146+, among them activated CD146+CD34+, CD146+HLA-DR+, CD146+CD54+, CD146+CD54+HLA-DR+; lymphocyte populations: T-lymphocytes CD45+CD3+, T-helpers CD45+CD3+CD4+CD8-, T-suppressors CD45+CD3+CD4-CD8+, B-lymphocytes CD45+CD3-CD19+, NK-lymphocytes CD45+CD3-CD16+CD56+.

EFFECT: application of this method allows to make a quantitative and functional characteristic of all the skin constituent elements for use in diagnosis.

METHOD FOR DETERMINING THE SUBPOPULATION COMPOSITION OF SKIN CELLS AND OBTAINING A SKIN CYTOIMMUNOGRAM

Patent holder: Goltsov Sergey Viktorovich
Type: patent for an invention
Patent number: RU 2630607 C1
Patent office: Russia
Publication year: 2017
Application number: 2016121997
Registration date: 02.06.2016
Publication date: 11.09.2017

eLIBRARY ID: 38270421

INTERNATIONAL PATENT CLASSIFICATION:
G01N 33/53 Research or analysis of materials by special methods not classified in groups 1/00 - 31/00 / biological materials, e.g. blood, urine; devices for counting and measuring blood cells (hemocytometers) / chemical analysis of biological materials, e.g. blood, urine; tests based on the binding of biospecific ligands; immunological tests / immunological analysis; analysis of biospecific binding; materials for this purpose

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