A highly sensitive, rapid screening method for the detection of antibodies directed against HLA class I and II antigens

Abstract

Screening of potential transplant recipients for antibodies that can cause graft rejection is an essential part of the pre-transplant monitoring carried out by tissue typing laboratories. This is a time-consuming process and the rapid reporting of results is dependent on the maintenance of frozen cell panels. The usual procedure of screening against a panel of random cells takes up to 6 weeks. In this study we have used flow cytometric analysis of pooled chronic lymphatic leukaemia (CLL) cells to detect antibodies directed against HLA antigens. We show that FACS screening of pooled cells can accurately and rapidly detect these antibodies and that the method is suitable for routine use. An estimate of the degree of patient panel reactivity can be determined within a few hours. In addition, the technique is more sensitive than those conventionally used, an advantage that may be of importance in preventing graft damage.

Recurrent transfusion‐related acute lung injury

“…Sera were screened by a LIFT procedure based on that described by Harmer et al 5 Pooled lymphocytes were incubated with test serum for 40 minutes at 37°C. After washing in 1‐percent (vol/vol) FCS in PBS, 50 μL of a 1‐in‐100 dilution of FITC‐conjugated anti‐human IgG was added and the test was incubated at 22°C for an additional 30 minutes.…”

Mothers without HLA antibodies before transplantation have a low risk of alloimmunization post‐transplantation

“…However, this technique requires a panel of lymphocytes, which may vary from one assay to another and may detect only HLA Abs that induce lymphocyte lysis through complement activation. More sensitive and more specific techniques for HLA Ab screening have been developed such as purified HLA antigens that are coated on a plastic plate, enzyme‐linked immunosorbent assay (11) or, more recently, flow cytometry techniques such as the Luminex technique using purified HLA antigens attached to microbeads (12, 13). This means that certain results obtained with older techniques should be reconsidered using more recent technology.…”

Characteristics, Detection, and Clinical Relevance of Alloantibodies in Kidney Transplantation

“…FC cell-based technology has been introduced in two variations: donor-specific CM (Garovoy et al, 1983) and anti-HLA antibody screening using a pool of Epstein-Barr virus (EBV)-transformed B lymphoblastoid cells or peripheral blood lymphocytes (Harmer et al, 1993;Shroyer et al, 1995). This assay uses a laser beam to detect alloantibodies.…”