Oregon Health & Science University

State of Oregon

Gregory P Lunstrum

Douglas R Keene

M Peter Marinkovich

Robert E Burgeson

Patricia Rousselle

Patricia Rouselle

Robnert E Burgeson

Improving adhesion of epidermis to an underlying dermal substrate including the step of providing an amount of purified kalinin between the epidermis and dermal substrate wherein the kalinin provides adhesion between the human dermis and epidermis wherein said kalinin is a trimeric molecule having the following three nonidentical chains: a chain which includes the BM165 epitope, either a 155 kilodalton chain or a 105 kilodalton chain, and a 140 kilodalton chain.

Products and methods for improving keratinocyte adhesion to the dermis

The isolated proteins kalinin and k-laminin are disclosed to provide adhesion between epidermal keratinocytes and the underlying dermis. Purified kalinin localizes to the anchoring filaments of basement membranes of human subepithelial skin, trachea, esophagus, cornea and amnion when such areas are probed with BM165 monoclonal antibody after localization. The protein has a molecular weight of approximately 410-495 kDa and exists in a cell-associated form (about 495 kDa) and two medium-associated forms (about 460 and 410 kDa, respectively). The BM165 epitope is located on the 165-kDa and 200 kDa subunits. Kalinin has a rotary-shadow image revealing an asymmetric rod 107-nm long having two globules at a first end and a single globule at an opposing end. The k-laminin adhesion molecule is an isolated heterotrimeric laminin variant that has a molecular weight of about 650 kDa and separates on western blots into first and second fragments that are similar to the B1 and B2 fragments of laminin. K-laminin also includes a third fragment of 190 kDa that is not immunoreactive with monoclonal antibodies 1F5, 11D5 and 4C7, but is immunoreactive with a monoclonal antibody BM165. Kalinin and k-laminin are preferably covalently associated. A method is also disclosed for improving adhesion of transplanted keratinocytes to an underlying substrate, such as the human dermis, by optimizing production of kalinin from cultured keratinocytes, or by providing an exogenous source of kalinin between the keratinocytes and substrate.

A purified protein kalinin is disclosed that provides adhesion between epidermal keratinocytes and the underlying dermis. Purified kalinin localizes to the anchoring filaments of basement membranes or human subepithelial skin, trachea, esophagus, cornea and amnion when such areas are probed with BM165 monoclonal antibody after localization. The protein has a molecular weight of approximately 400-460 kDa and exists in a cell-associated form (about 460 kDa) and two medium-associated forms (about 440 and 400 kDa, respectively). The cell-associated form comprises a 200-, a 155- and a 140-kDa subunit, all normally held together by disulflde bonds. The cell-associated form is subjected to extracellular processing to produce the two medium-associated forms, wherein, in the 440-kDa form, the 200-kDa subunit has been processed to a 165-kDa subunit and, in the 400-kDa form, the 155-kDa subunit has been processed to a 105-KDa subunit. The BM165 epitope is located on the 165-kDa subunit. Kalinin has a rotary-shadow image revealing an asymmetric rod 107-nm long having two globules at a first end and a single globule at an opposing end. A method is also disclosed for improving adhesion of transplanted keratinocytes to an underlying substrate, such as the human dermis, by optimizing production of kalinin from cultured keratinocytes, or by providing an exogenous source of kalinin between the keratinocytes and substrate.

Growing community of inventors

Portland, OR, United States of America

Gregory P Lunstrum