Comparison of tube and gel red blood cell agglutination techniques in detecting chimeras after major ABO-mismatched allogeneic hematopoietic stem cell transplantation.

We compared the ability of tube and gel red blood cell (RBC) agglutination techniques to follow erythroid engraftment in a patient who received a major ABO-mismatched peripheral blood stem cell transplant and bone marrow transplant. Tube and gel RBC agglutination techniques were used to detect mixed-field reactivity in cell mixtures containing A/O and c+/c- RBCs and the ability of these two technologies to detect RBC chimeras were compared. We detected c+ RBCs in c+/c- RBC populations microscopically at 1% by the tube RBC agglutination technique, but not until 10% by the gel technique.

Detection of granulocyte antibodies by flow cytometry without the use of pure granulocyte isolates.

Established methods used to detect serum antibodies to granulocytes require the isolation of granulocytes. Flow cytometric analysis of granulocytes with monoclonal antibodies eliminates the need for granulocyte isolation. The purpose of this study was to develop a method to evaluate reactions of antibodies to granulocytes without separating granulocytes from other leukocytes.

Comparison of DATs using traditional tube agglutination to gel column and affinity column procedures.

Background: Detection of immunoglobulin or complement bound to RBCs by using the DAT is valuable in the diagnosis of immune-mediated hemolytic anemia. Traditionally, the DAT has been performed by tube agglutination using anti-IgG or anti-C3d. The purpose of this study was to compare the tube agglutination DAT to gel microcolumn, affinity microcolumn, and flow cytometric DATs on RBCs coated in vitro and on patient RBC samples.

Detection of HLA antibodies by using flow cytometry and latex beads coated with HLA antigens.

Background: Detection of HLA class I antibodies in sera is needed in various clinical situations. The standard method for detecting HLA class I antibodies is the complement-dependent lymphocytotoxicity (CDC) assay, but solid-phase assays are now available.

Study Design And Methods: This study assessed the ability of a flow cytometric assay using latex beads coated with HLA class I antigens to detect HLA class I-specific antibodies.

Granulocyte storage and antigen stability.

Background: Current methods for the detection of granulocyte antibodies require panels of freshly isolated cells. This makes these assays time-consuming, costly, and technically difficult.

Study Design And Methods: The immunofluorescence method of detecting the binding of antibodies to granulocytes was modified for use with a flow cytometer, and methods were tested to store granulocytes for use in that assay.