Antibodies against Lutheran blood group antigens have been observed during first-time pregnancy. Samples from a woman of African descent were tested in our immunohematology laboratory on several occasions since 2001. Her samples were phenotyped as Lu(a+b-), and anti-Lub was suspected but not identified. She was asked to make autologous donations in preparation for her delivery, which she did. In 2010, two antibodies were identified: anti-Lea and -Lub. Six years later, a third investigation was requested. This time, an antibody directed at a high-prevalence Lutheran antigen was found in addition to the anti-Lea and -Lub previously observed. Her serum was compatible with three out of five Lu(a-b-) reagent red blood cells (RBCs). One of the incompatible Lu(a-b-) reagent RBCs was known to be In(Lu) (KLF1 mutation). The genetic background of the other reagent RBC was unknown. The LU cDNA sequence analysis revealed the presence of the c.230G>A (Lua), c.679C>T (LU:-16), and a silent polymorphism c.1227G>T. Anti-Lu16 was highly suspected. This would be the fifth case of LU:-16 with antibodies reported, all within women of African heritage with the Lu(a+b-) phenotype. Hemolytic disease of the fetus and newborn was not noted in these cases.