Cookies on this website

We use cookies to ensure that we give you the best experience on our website. If you click 'Accept all cookies' we'll assume that you are happy to receive all cookies and you won't see this message again. If you click 'Reject all non-essential cookies' only necessary cookies providing core functionality such as security, network management, and accessibility will be enabled. Click 'Find out more' for information on how to change your cookie settings.

An immunofluorescent staining method using specific monoclonal antibodies was used to detect IL-1 alpha and IL-1 beta in individual cells in stimulated human peripheral blood. No staining was seen in unstimulated cells but intense, maximal staining of approximately 5% of the cells was seen 20-24 h after activation with PHA/PMA. The large irregularly shaped stained cells were surrounded by smaller unstained cells with lymphocyte-like morphology. By 44 h post activation a few cells only showed weak staining. The staining pattern was different for the two molecules studied, with a granular pattern for IL-1 alpha staining and diffuse cytoplasmic staining for IL-1 beta. Staining post-activation could be abolished by preincubation of the monoclonal antibody with the appropriate recombinant IL-1, but not by pre-incubation with the other IL-1 type. When both anti-IL-1 alpha and anti-IL-1 beta were used together two populations of cells were identified; one had the granular staining as seen with anti-IL-1 alpha alone and the other had the diffuse staining pattern as seen with anti-IL-1 beta. The percentage of cells showing bright staining with anti-IL-1 alpha and anti-IL-1 beta together was approximately equal to the sum of the percentage of cells staining for IL-1 alpha or IL-1 beta alone. This study demonstrates a method for the detection of individual IL-1 alpha- and IL-1 beta- producing cells and suggests that in activated human peripheral blood IL-1 alpha and IL-1 beta are produced by separate populations of cells.

Type

Journal article

Journal

J immunol methods

Publication Date

21/06/1989

Volume

120

Pages

277 - 283

Keywords

Antibodies, Monoclonal, Cell Separation, Cytoplasm, Fluorescent Antibody Technique, Humans, Interleukin-1, Kinetics, Leukocytes, Mononuclear, Lymphocyte Activation, Phytohemagglutinins, Tetradecanoylphorbol Acetate