A new look at an old case: An auto-anti-P with pseudo-LKE activity

Abstract

ABSTRACT

A hemolytic anti‐LKE associated with a rare LKE‐negative, “weak P” red blood cell phenotype: alloanti‐LKE and alloanti‐P recognize galactosylgloboside and monosialogalactosylgloboside (LKE) antigens

“…Four examples of alloanti‐P showed activity against Gb5. PCH serum was specific for Gb4, as was a historical low‐titer “anti‐LKE” (UM1314, Table ) …”

“…LKE typing was performed with MoAb MC813‐70 as described . Samples agglutinating at IS were typed as LKE‐S.…”

“…For immunostaining, HPTLC plates were coated in 0.2% poly(iso)butyl‐methacrylate in hexane for 1 minute and dried for at least 1 hour at room temperature (RT) . For staining with human antisera, plates were initially blocked for 1.5 hours with phosphate‐buffered saline (PBS)‐3% bovine serum albumin (BSA) and then overlaid with antisera diluted 1:20 in PBS‐5% BSA for 2 hours.…”

“…For staining with human antisera, plates were initially blocked for 1.5 hours with phosphate‐buffered saline (PBS)‐3% bovine serum albumin (BSA) and then overlaid with antisera diluted 1:20 in PBS‐5% BSA for 2 hours. After washing, bound antibody was detected by sequential incubation with biotinylated anti‐human immunoglobulin (IgM, IgG), streptavidin‐linked alkaline phosphatase, and black alkaline phosphatase substrate . To estimate relative antibody binding, the amount of bound antibody was divided by total GSL as determined by scanning densitometry (antibody staining [area]/GSL quantity [area by charring]).…”

“…LKE typing was performed with MoAb MC813-70 as described. 4,18 Samples agglutinating at IS were typed as LKE-S. Negative samples were tested by the IAT using antimouse IgG.…”