J Korean Med Sci.  2006 Aug;21(4):627-632. 10.3346/jkms.2006.21.4.627.

Granulocyte Antibodies in Korean Neonates with Neutropenia

• Han TH
• Chey MJ
• Han KS

• KMID: 2157812
• DOI: http://doi.org/10.3346/jkms.2006.21.4.627

Abstract

Neonatal alloimmune neutropenia (NAN) is a disease that can cause severe and prolonged neutropenia in neonates. However, no report is available on the incidence of granulocyte antibody in neonates, the target antigen of this antibody, and the estimated incidence of NAN in Korea. Among a total of 856 neonates admitted to a neonatal intensive care unit (NICU) over a five year period, a total of 105 neonates with neutropenia were enrolled in this study. Positive reactions were observed in the sera of six neonates (5.7%, 6/105) by mixed passive hemagglutination assay (MPHA). To confirm the presence of NAN, MPHA and granulocyte antigen typing (HNA-1a, -1b, -2a, -4a, and -5a) were performed on neonatal and maternal blood. To differentiate granulocyte antibody and HLA antibody, MPHA was also performed using HLA antibody adsorbed serum. We confirmed three cases (2.9%, 3/105) of NAN among neonates with neutropenia in which granulocyte antibody specificities (two anti-HNA-1b and one anti-HNA-1a) and fetomaternal granulocyte antigen mismatches were identified. In this study, the estimated incidence of NAN was 0.35% (3/856) among neonates admitted to NICUs in Korea.

Keyword

Neutropenia; Neonatal Alloimmune Neutropenia; Granulocyte Antibodies

MeSH Terms

Polymerase Chain Reaction/methods
Neutropenia/blood/diagnosis/*immunology
Korea
Isoantigens/genetics/immunology
Isoantibodies/*immunology
Intensive Care Units, Neonatal/statistics & numerical data
Infant, Newborn
Humans
Hemagglutination Tests
HLA Antigens/immunology
Granulocytes/*immunology
Genotype
Antibody Specificity/immunology

Figure

• 

  Fig. 1 HNA-1a, HNA-1b, HNA-4a genotyping by PCR-SSP. Lane 9 shows a DNA ladder marker (Bioneer, Daejeon, Korea). The amplification products (439 bp) of the internal control (HGH gene) are present in each lane. Lanes 1, 3, 5, and 7 are positive controls for HNA-1a (141 bp), HNA-1b (219 bp), HNA-4a-positive (124 bp), and HNA-4a-negative (124 bp), respectively. Lanes 2, 4, 6, and 8 are negative controls for HNA-1a, HNA-1b, HNA-4a+, and HNA-4a-, respectively. Lanes 10-13 contain amplification products of HNA-1a, HNA-1b, HNA-4a+, and HNA-4a-, respectively from a DNA sample that is a HNA-1-heterozygote (HNA-1a/HNA-1b) and a HNA-4a-heterozygote (HNA-4a+/HNA-4a-).

• 

  Fig. 2 HNA-5a genotyping by Bsp1,286 I allele-specific restriction enzyme analysis (ASRA). Lane 1 shows a DNA ladder marker (Bioneer, Daejeon, Korea); lane 2 shows an undigested 709 bp PCR product of the αL chain of β2integrin cDNA; lane 3 shows an HNA-5a+ homozygote sample (297 bp, 217 bp, and 195 bp); lane 4 shows a HNA-5a heterozygote samples (412 bp, 297 bp, 217 bp, and 195 bp); and lane 5 shows a HNA-5a- homozygote sample (412 bp, and 297 bp).

• 

  Fig. 3 Granulocyte antibody test using the mixed passive hemagglutination assay (MPHA). The neonates' and mothers' sera were tested using extracted granulocyte antigen-coated microplates (from six donors) as a solid phase and sheep RBCs coated with rabbit F(ab')2 anti-human IgG as indicator cells. To differentiate granulocyte antibody from HLA antibody, sera (left half) with positive reactions were compared with HLA antibody adsorbed sera (right half). In the sera of neonate 1 (N1) and her mother (M1), anti-HNA-1a; in the sera of neonate 2 (N2) and his mother (M2), anti-HNA-1b; in the sera of neonate 3 (N3) and his mother (M3), anti-HNA-1b and anti-HLA. P, pasitive control; N, negative control.

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