Identification of Non-HLA Target Antigens Recognized After Lung Transplantation

Sequential analysis by immunoprecipitation-MALDI-TOF: A novel method for detection and identification of alloantibody specificities

“…The lysate of cells with bound antibodies is then incubated with protein G beads, which precipitate antigen-immunoglobulin (Ig)G-complexes. The precipitated proteins are eluted from the beads, separated on an SDS-PAGE gel, and visualized by silver Using the SEREX method for a study of lung-transplanted patients, Otten et al [13] identified antibodies reacting with intracellular gene products (LOC284058, XP_931864, PMSC4, and ZNF33A), the secreted protein PLUNC and the cell-surface protein F3. The SEREX method includes the construction of a cDNA library from target tissue, which is tested with a serum for reactivity against library-encoded antigens.…”

“…Because native human cells are used in our test, disadvantages associated with artificial expression systems such as those mentioned earlier can be avoided. At the same time, unlike some previous reports on detection of antibodies against intracellular structures [12,13], our method detects only proteins that are expressed on the cell surface and, therefore, are more likely to be relevant in the clinical transplantation setting.…”

“…In the literature, 2 methods have been used for characterization of previously unknown antigens recognized by antibodies in the sera of patients after organ transplantation [12,13]. One method is the serologic analysis of antigens by recombinant expression cloning (SEREX) [13,14].…”

“…One method is the serologic analysis of antigens by recombinant expression cloning (SEREX) [13,14]. The other method is based on protein arrays [12].…”

Significance of histocompatibility in organ transplantation

“…Proteomic technologies offer the best opportunities to search for endothelial and tissue antigens to which patients may produce antibodies. One preliminary study [36] of 11 lung recipients, for example, identified six non-HLA antigens from a library of 30 000 gene expression products derived from donor trachea, some of which were recognized by antibodies from different patients. Unfortunately there was no opportunity to correlate the presence of these antibodies with bronchiolitis obliterans or any adverse events following detection of these antibodies.…”