Detection Of Cell Surface Markers Research Articles

A combined method for detection of cell surface marker expression and bromodeoxyuridine (BrdU) uptake by epidermal cells in suspension

A rapid and convinient immunocytochemical method is presented for the simultaneous detection of surface antigen expression and S-phase cells using monoclonal antibodies and cells in suspension. The first reagent permits the nature of the cells to be determined while the second immunostaining stage reveals which of these cells are proliferating (BrdU-positive) cells. Staining and fixation conditions have been defined for the detection of cell surface markers by red fluorescence and BrdU labelling by green fluorescence. All the steps were carried out on cells in suspension and the membrane antigen distributon was not affected by the BrdU staining protocol. The method has been applied to two human epidermal cell subpopulations: Langerhans' cells and basal keratinocytes. This technique should have general applicability to the study of the cytokinetic characteristics of phenotypically defined cells in heterogeneous populations.

Flow cytometric analysis of cell proliferation dynamics in the B cell compartment of the mouse

Using a method which allows simultaneous flow cytometric detection of cell surface markers and 5-bromo-2'-deoxyuridine (BrdU) incorporation, the distribution and proliferative behavior of B lineage subpopulations was studied in intact adult mice. In the bone marrow we could define two subsets of B cells on the basis of differential expression of the pan-B cell marker B220 and of membrane-associated mu and delta immunoglobulin heavy chains. B220dull mu+ delta- B cells were found to emerge from rapidly dividing cells and probably represent B cells recently generated from B220dull mu- pre-B cells. In contrast, only few, if any, of the B220bright mu+ delta+ B cells were labeled with BrdU after a period of 8 days, suggesting that these cells represent long-lived B cells residing in the bone marrow. Analysis of BrdU-incorporation into splenic B cells showed that only 20% of these cells had gone through cell division during the preceding 8 days. Almost none of the B cells in the peritoneum, a large fraction of which belongs to the Ly1 B subset, were labeled with BrdU over a period of 7 days in 8-month-old animals.

Lymphocyte Functions of Child Patients with ALL (Acute Lymphoblastic Leukemia) in Remission

Functional activities of various subsets of peripheral blood lymphocytes from patients with acute lymphoblastic leukemia in remission were studied. In the detection of cell surface markers and the responsiveness to mitogens, no significant abnormalities were found. However, impaired functional activities of various subsets of lymphocytes such as helper T cells and B cells in an in vitro pokeweed mitogen-stimulated immunoglobulin-producing system, and natural killer cells and cells responsible for antibody-dependent cellular cytotoxicity in 51Cr release assays were found. The results of the induction of specific killer T cells varied. The impairment of lymphocyte functions seemed to be related to the defective defense mechanism which even in patients in remission leads to serious life-threatening infections.

7] Rosetting techniques to detect cell surface markers on mouse and human lymphoreticular cells

Publisher Summary This chapter explores wide variety of techniques that have been used to detect cell surface antigens and receptors on subpopulations of lymphoreticular cells. Among these are rosetting techniques which involve the specific binding of indicator cells or immunoabsorbent beads to distinct cell populations to form so called “rosettes.” It discusses that rosetting assays are simple, rapid, well established, and provide a read-out which is visible under the light microscope. They are easily amenable to double labeling technique. It provides an amplification of the signal in that an intact indicator cell or bead binds via a ligand to a specific “acceptor” molecule on the cell surface. They offer a convenient means of separating rosetting cells by density or sedimentation velocity. It discusses detailed description of immunobead indicator methods and antigen-specific rosettes. The chapter also provides a brief discussion of lymphocyte-tumor cell conjugates, and an evaluation of separation techniques.