Open AccessCarbohydrate‐Labelled Glycoproteins as Markers of Human Lymphocyte Subsets Stimulated with Mitogen and Alloantigen
Author(s) -
EIJK Ronald V. W.,
MUHLRADT Peter F.
Publication year - 1981
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1981.tb06192.x
Subject(s) - concanavalin a , pokeweed mitogen , spleen , glycoprotein , lymphocyte , biology , population , thymocyte , microbiology and biotechnology , thymidine , mitogen activated protein kinase , immunology , biochemistry , t cell , immune system , medicine , in vitro , environmental health
Human lymphocytes from thymus, spleen, and blood were stimulated by concanavalin A, phytohemagglutinin or pokeweed mitogen to incorporate radioactive thymidine and various carbohydrates. Peripheral‐blood lymphocytes were also activated by alloantigen in a mixed lymphocyte reaction. Incorporation of thymidine and carbohydrates was parallel in stimulated cells. Carbohydrate‐labelled cells were extracted with Triton X‐100 buffer and the extracts subjected to sodium dodecylsulfate/polyacrylamide gel electrophoresis and subsequent autoradiography. From a comparison of the resulting glycoprotein patterns the following conclusions could be drawn. (a) The same population of thymocytes responds to all three mitogens. (b) Pokeweed‐mitogen‐stimulated blood lymphocytes are similar to this mitogen‐responsive thymocyte population. (c) The glycoprotein pattern of pokeweed‐mitogen‐responsive spleen cells shows two characteristic glycoproteins, GP 50 and GP 78, and resembles that of murine B cells. (d) Whereas concanavalin A and phytohemagglutinin activate identical subpopulations of T cells in the spleen, this is not the case in the blood. (e) Concanavalin‐A‐activated and alloantigen‐activated blood lymphocytes express a glycoprotein GP 185 which is not found on phytohemagglutinin‐responsive cells. (f) The kinetic relationship during alloantigen stimulation in a mixed lymphocyte reaction between the appearance of glycoprotein GP 185 and subsequent maximal killing activity in an assay using release of 51 Cr makes it likely that glycoprotein GP 185 is a marker for cytotoxic T lymphocytes.