Variant RHCE alleles are found mainly in Afro-descendant individuals, as well as in patients with sickle cell disease (SCD). The most common variants are related to the RHCE*ce allele, which can generate partial e and c antigens. Although RHCE variant alleles have been extensively studied, defining their clinical significance is a difficult task. We evaluated the risk of RhCE alloimmunization as a consequence of partial antigens in patients with a positive phenotype transfused with red blood cell (RBC) units with the corresponding antigen. A retrospective study was performed with Brazilian patients, evaluating the number of antigen-positive transfused RBC units (incompatible due to partial antigen) in 27 patients with SCD carrying RHCE variant alleles who did not develop antibodies as well as evaluating the variants present in 12 patients with partial phenotype and correlated antibody (one patient with SCD and 11 patients with other pathologies). Two patients showed variant alleles with molecular changes that had not yet been described. Variant RHCE alleles were identified in a previous study using molecular methods. RHCE*ceVS.01 was the most frequent allele found among the patients without antibodies. Six patients with partial c antigen had a mean of 3.8 c+ RBC units transfused, and 10 patients with partial e antigen were exposed for a mean of 7.2 e+ RBC units. Among the variant alleles found in alloimmunized patients, the most frequent was RHCE*ceAR, which was found in five patients; the antibodies developed were anti-hrS and/or anti-c. Our results showed that RHCE*ceVS.01 is indeed the most frequent variant allele in our cohort of patients with SCD, but the partial antigens that were identified have low risk of alloimmunization. RHCE*ceAR is the most impactful variant in the Brazilian population with high risk of alloimmunization and clinically significant anti-hrS formation.
The MAM blood group system (International Society of Blood Transfusion [ISBT] 041) consists of one high-prevalence antigen to date, first detected in a 31-year-old woman during her third pregnancy. Epithelial membrane protein 3 (EMP3) was recently identified as the gene coding the MAM antigen. Six unique genetic variants have been described in EMP3 in 11 MAM- individuals. EMP3 is an 18-kDa glycoprotein with a large extracellular domain containing at least one N-glycosylation site. The normal function of EMP3 is still unclear, but ex vivo culture of erythropoietic progenitor cells from MAM- individuals shows an increased yield of reticulocytes, suggesting that EMP3 acts as a brake during normal erythropoiesis. EMP3 is abundant on different cell types, including many epithelial tissues and blood cells. Interestingly, EMP3 expression has been suggested as a prognostic marker for a number of cancer types, both for good and poor prognoses. EMP3 may act as a tumor suppressor or an oncogene in different cancer contexts. The protein appears to interact with other cell surface receptors and affects the downstream signaling and function of these proteins. MAM- red blood cells express low levels of CD44 and, consequently, the antigens of the Indian blood group system are only weakly expressed. Clinically, the MAM blood group antigen is important with regard to blood transfusion and pregnancy. Anti-MAM can cause severe hemolytic disease of the fetus and newborn in some pregnancies but have little to no effect in other pregnancies. Cases are typically not detected until problems occur during pregnancy, making the availability of compatible blood a challenge.
Proteolytic enzymes are used to treat red blood cells (RBCs) to aid in complex antibody identification. Although there are many enzymes that can be used, for the purpose of this method review, enzyme-treated RBCs refers only to RBCs treated with ficin or papain. Ficin and papain can increase the sensitivity of antibody detection by modifying the RBC membrane. Enzyme treatment and test methods can be performed using one-stage or two-stage procedures. Enzyme treatment is especially useful for the differentiation of multiple antibodies, enhancement of detection of weak antibodies, and adsorption methods. In all cases, quality control is required to ensure adequate treatment of RBCs before additional testing. Ficin and papain are useful tools for both immunohematology reference laboratories and transfusion services.