Serologic and molecular studies to identify neonatal alloimmune neutropenia in a cohort of 10,000 neonates

Br J Haematol. 2021 Feb;192(4):778-784. doi: 10.1111/bjh.17295. Epub 2021 Feb 2.

Abstract

Neonatal alloimmune neutropenia (NAIN) is caused by maternal alloimmunisation to fetal human neutrophil antigens (HNAs). This study investigated maternal HNA/HLA alloantibodies involved with NAIN and identified the frequency of NAIN in Brazilian neonates. Neonatal neutropenia (neutrophil count < 1.5 × 109 /L) was investigated in samples from 10,000 unselected neonates, resulting in 88 neutropenic newborns (NBs) and their 83 mothers. Genotyping was performed by PCR-SSP (HNA-1/-4) and PCR-RFLP (HNA-3/-5). Serologic studies were performed by GAT (granulocyte agglutination test), Flow-WIFT (white blood cells immunofluorescence test) and LABScreen-Multi-HNA-Kit (OneLambda®) (LSM). Neonatal neutropenia was identified in 88/10,000 (0·9%) NBs. Genotyping revealed 60·2% maternal-fetal HNA incompatibilities (31·8% for HNA-1; 14·8% for HNA-3; 15·9% for HNA-4; 21·6% for HNA-5). Serologic studies revealed 37·3% of mothers with positive results with at least one technique. The detected anti-HNA specificities were confirmed in eight positive cases related to HNA-1/-3 systems. In cases with maternal-fetal HNA-4/-5 incompatibility, no specific neutrophil alloantibodies were found but anti-HLA I/II were present. Anti-HNA-2 was not identified. This is a large Brazilian study which involved the investigation of antibodies against all five HNA systems in neutropenia cases and showed a frequency of NAIN in 8/10,000 neonates. Among the HNA antibodies identified, we highlight the anti-HNA-1d and anti-HNA-3b, antibodies unusual in alloimmunised women, and rarely related to NAIN cases.

Keywords: Anti-HNA-1d; Anti-HNA-3b; maternal-fetal HNA incompatibility; neonatal alloimmune neutropenia; neutrophil alloimmunisation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Brazil / epidemiology
  • Female
  • Genotype
  • Humans
  • Infant, Newborn
  • Infant, Newborn, Diseases / blood
  • Infant, Newborn, Diseases / diagnosis*
  • Infant, Newborn, Diseases / epidemiology
  • Infant, Newborn, Diseases / genetics
  • Isoantibodies / blood
  • Isoantibodies / genetics
  • Isoantibodies / immunology
  • Leukocyte Count
  • Male
  • Neutropenia / blood
  • Neutropenia / diagnosis*
  • Neutropenia / epidemiology
  • Neutropenia / genetics
  • Neutrophils / immunology

Substances

  • Isoantibodies