Characteristics of carrbohydrate determinants of murine lymphoma NK/Ly cell surface by binding of native lectins with subsequent their detection by indirect immunochemical method
M. Lutsyk, V. Kovalyshyn, A. Yashchenko
Carbohydrate determinants on surface of murine lymphoma NK/Ly cells were characterized using indirect immunocytochemical detection of bound lectins (Con A, LcL, PNA, WGA) with the aid of light and electron microscopy. Prefixed cells were treated with native lectins, thereafter bound lectins were detected by specific antilectin antibodies (or horsåradish peroxidase for Con A) labelled with colloidal gold. For light microscopy gold particles were visualized by procedure of physical development with silver acetate. On the surface of NK/Ly cells high affinity WGA-binding determinants were maximally exposed indicating on significant amount of sialic acids in glycans of N- and especially O-type. Lentil lectin detected the lowest number of determinants with low binding affinity. Relatively high percentage of PNA+ cells indicates on significant proportion of asialo-O-glycans, thus exposing T-F antigen. The advantages of indirect immunochemical method for detection of lectin-binding sites as compared with direct methods in which are used label modified lectins are discussed.
Download full text in PDF format
|
