A whole blood alternative to traditional methods for CD4+ T lymphocyte determination.

Flow cytometry, the method of choice for determining subset cell populations, requires sophisticated instrumentation and highly trained operators. An alternative method for subpopulation determinations is presented that is fast, simple, and readily interpreted for absolute subset counts. This method uses antibody labeled microspheres added to whole blood and after lysing, read on a hemocytometer using an ordinary light microscope.

A simple and rapid method for removal of specific cell populations from whole blood.

A method is presented in which specific cells can be rapidly separated from whole blood by the use of antibody labelled dense particles utilizing gravity as a means of separation. This method specifically removes the targeted cells with minimal non-specific cell loss. The method is simple to perform and rapid, requiring less than 10 min for cell/particle binding and separation.

Lymphocyte subset determination using a hematology analyzer.

Anti-CD4 antibody (T4)-coated microspheres were used to label CD4 cells in whole blood. The mixture was lysed and analyzed by a modified Coulter VCS hematology analyzer, which differentiated microsphere-labeled cells by a change in Coulter volume, conductance, and light scatter. %CD3+/CD4+ fluorescent values from a profile were compared to %CD4 values using the VCS-microsphere method.

A rapid and reliable assay to enumerate CD4+ T lymphocytes in whole blood.

We compared the performance of a rapid and simple anti-CD4 antibody-coated microsphere assay with flow cytometry and immunofluorescence for quantitation of absolute count of CD4+ T lymphocytes. A longitudinal evaluation of CD4+ T lymphocytes by flow cytometry and microsphere assay in 10 human immunodeficiency virus (HIV)-seronegative and 59 HIV-seropositive individuals was conducted over a period of 9 months. Standard flow cytometry analysis was performed to establish the absolute CD4+ T-lymphocyte count.

A rapid manual method for CD4+ T-cell quantitation for use in developing countries.

Objective: To evaluate a manual method (Cytosphere) for quantifying CD4+ T-cell numbers.

Design: Cross-sectional study of HIV-1-seronegative and HIV-1-seropositive individuals evaluated for absolute CD4 counts by both standardized flow cytometric measurements and manual Cytosphere technology using a hemacytometer.

Setting: University research hospitals in both the United States and Africa.

Molecular comparison of mouse Thy-1 and its human homologue.

Physicochemical techniques were used to verify previous immunochemical studies showing homology of the human Thy-1 (formerly p25) antigen and the murine Thy-1 or theta antigen. Peptide maps and amino acid compositions showed close similarity between these two proteins; however, they were not identical. These data confirm that the p25 antigen is the human homologue of mouse Thy-1.

Isolation and partial characterization of the human homologue of Thy-1.

A glycoprotein of 25,000 daltons was isolated from a human T lymphoblastoid cell line (MOLT-3). A monovalent antiserum raised against this antigen precipitated mouse Thy-1.1 antigen as well as a 25,000-dalton antigen from human blood T cells.

Expression of two differentiation antigens on normal and cultured human T cells.

An antiserum specific for human T lymphocytes (AMT) was used to examine patterns of T cell surface antigen expression and to isolate their reactive membrane antigens. By a quantitative adsorption assay, different plateaus of AMT reactivity with blood T cells were observed after serial adsorptions with individual T cell lines. MOLT-3 cells removed 95% of AMT activity to blood T cells whereas MOLT-4 removed 70% and HSB-2 removed only 30%.

Isolation and characterizaiton of murine cell surface components. I. Purification of milligram quantities of Thy-1.1.

The Thy-l.1 molecule was isolated from the BW5147 murine lymphoblastoid cell line. The initial step in purification was the preparation of a crude plasma membrane fraction followed by acetone precipitation.

Growth properties and alloantigenic expression of murine lymphoblastoid cell lines.

Murine lymphoblastoid cell lines were evaluated for their expression of Thy-1 and thymus leukemia (TL) differentiation alloantigens. Two culture conditions were shown to affect this expression. Cells grown in fetal bovine serum (FBS)-enriched medium expressed up to 15 times the amount of TL as cells grown in horse serum (HS)-enriched medium.