Flow-Cytometric Analysis of HNA-2 Expression and Phenotypes Among Thai Blood Donors

Nathalang, Oytip; Siriphanthong, Kanokpol; Petvises, Sawang; Jeumjanya, Nichapa

doi:10.3343/alm.2018.38.4.362

Cited by 6 publications

(14 citation statements)

References 28 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Regarding our recent study of HNA‐2 antigen expression in 220 unrelated Thai blood donors by flow cytometry, only one donor had the HNA‐2 null phenotype . In the first cohort, 3 SNPs including SNP c.787A>T (rs201821720) in exon 7, c.1291G>A (rs78718189) and c.1254G>A (rs188387562) in exon 9 were observed in 4 samples.…”

Section: Resultsmentioning

confidence: 96%

“…Regarding our recent study of HNA-2 antigen expression in 220 unrelated Thai blood donors by flow cytometry, only one donor had the HNA-2 null phenotype. 10 In the first cohort, 3 SNPs including (Table 2).…”

Section: Resultsmentioning

confidence: 99%

“…Study subjects consisting of 49 blood donors with known percentage of CD177 expression (%HNA‐2 antigen expression) from the first cohort based on the fluorescence intensity of different neutrophil populations were categorised into four groups including 22 subjects with increased expression (≥60%), 16 with intermediate expression (40.0 to 59.9%), 10 with weak expression (5.0 to 39.9%) and 1 null expression (<5.0%). Additionally, the second cohort of 771 DNA samples was obtained from unrelated central Thai blood donors with unknown CD177 expression.…”

Section: Methodsmentioning

confidence: 99%

“…CD177 expression on neutrophils was measured by flow cytometry according to a related study . Briefly, polymorphonuclear cells (PMNs) were isolated from EDTA blood samples using Polymorphprep (AXIS‐SHIELD; Alere Technologies AS, Oslo, Norway) to investigate HNA‐2 expression.…”

Section: Methodsmentioning

confidence: 99%

“…Individuals who have HNA‐2 antigen expression of less than 5% are classified as HNA‐2 null. The frequency of the HNA‐2 null phenotype has been reported to be approximately 0.5% to 5% among Thai and other populations . HNA‐2 isoantibodies produced by HNA‐2 null individuals are implicated in transfusion‐related acute lung injury (TRALI) and immune neutropenia …”

Section: Introductionmentioning

confidence: 99%

See 4 more Smart Citations

A novel nonsense mutation found in the CD177 gene of Thai individuals with the HNA‐2 null phenotype

Siriphanthong

Petvises

Thanongsaksrikul

et al. 2019

Transfusion Medicine

Self Cite

View full text Add to dashboard Cite

Summary Objectives The aim of this study is to explore the molecular basis and to develop a simple sequence‐specific primer polymerase chain reaction (PCR‐SSP) technique for screening genotypes associated with the human neutrophil antigen‐2 (HNA‐2) null phenotype among Thai blood donors. Background Single‐nucleotide polymorphisms (SNPs) c.787A>T of the CD177 gene is well known to be primarily demonstrated as a genetic determinant for HNA‐2 deficiency. Methods The SNPs in the CD177 gene (exons 7 and 9) of 49 Thai blood donors with the known percentage of CD177 expression by flow cytometry including 48 HNA‐2 positive and 1 HNA‐2 null individuals were identified by long‐range PCR amplification and sequencing. Moreover, screening for the c.1254G>A mutation was developed using an in‐house PCR‐SSP technique and tested among 771 unrelated donor samples. Results A HNA‐2 null sample from the first cohort was heterozygous for c.787A/T and homozygous for c.1291G/G, namely, a 787A‐1291G/787T‐1291G (AG/TG) genotype. Interestingly, we could identify SNP c.1254G>A (rs188387562, p. Trp418Ter) that caused a nonsense mutation of the CD177 gene in exon 9. This individual might have the 787A‐1254A‐1291G/787T‐1254G‐1291G genotype. From the second cohort (771 unrelated donors), the 1254GG homozygote was the most common (96.37%), followed by the 1254GA heterozygote (3.50%) and 1254AA homozygote (0.13%). Blood samples of two individuals with 787AT‐1254GA‐1291GG and 787AA‐1254AA‐1291GG genotypes were tested and the HNA‐2 antigen expressions were 0.03% and 0.16% in rank. Conclusions The c.787A>T is a primary genetic hallmark to determine the HNA‐2 null phenotype. Additional screening of the novel c.1254G>A in combination with c.787A>T is a suitable, convenient and effective diagnosis among Thais.

show abstract

Section: Resultsmentioning

confidence: 96%

Section: Resultsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

See 3 more Smart Citations

A novel nonsense mutation found in the CD177 gene of Thai individuals with the HNA‐2 null phenotype

Siriphanthong

Petvises

Thanongsaksrikul

et al. 2019

Transfusion Medicine

Self Cite

View full text Add to dashboard Cite

show abstract

Human neutrophil antigens: Nature, clinical significance and detection

Browne

Dearman

Poles

2020

Int J Immunogenetics

View full text Add to dashboard Cite

Within granulocyte immunology literature, the terms 'granulocyte' and 'neutrophil' are often used interchangeably. As neutrophils constitute >99% of the granulocyte population in homeostasis, the glycoproteins expressed by granulocytes are referred to collectively as the human neutrophil antigens (HNAs). This review will discuss the general characteristics of granulocytes and their contribution towards the most frequently associated clinical disorders/complications. An overview of the described HNA is provided with the most recent nomenclature as defined by the International Society of Blood Transfusion (ISBT). Current approaches to HNA characterization and antibody detection will also be outlined before finally discussing limitations and possible improvements. 1.1 | What are granulocytes? Granulocytes are granulocytic, polymorphonuclear white blood cells (WBC) produced in the bone marrow which are subdivided into neutrophils, basophils and eosinophils by the different staining properties of their granules. Neutrophils are the most abundant WBC (1.8-7.5 × 10 9 /L) in a healthy individual, whereas basophils and eosinophils are considerably less numerous (0.

show abstract

Human neutrophil antigen 2 sequence‐based typing: Joining the hunt for the CD177 answer

et al. 2021

View full text Add to dashboard Cite

Background and Objectives: Isoantibodies to human neutrophil antigen 2 (CD177) have been associated with several clinical conditions but to date the molecular basis for altered or non-expression has not been determined. Reliance on phenotyping and crossmatch to investigate these neutropenic clinical cases are inconvenient for the patients and demanding of resources within the laboratory. Therefore, a molecular approach has been introduced to address both issues. Materials and Methods:A DNA panel of 100 randomly selected blood donors were collected and supplemented with 18 DNA samples from blood donors previously shown to be CD177 null. All DNA samples were sequence-based typed for all exons and observed polymorphisms recorded. The DNA from two families previously investigated for neonatal alloimmune neutropenia due to CD177 isoantibodies were also analysed. Results:The incidence of CD177 null could be associated with a known exon 7 single-nucleotide polymorphism in 16/21 known CD177 null samples, which is consistent with previously published findings. Two additional mutations that may lead to null expression were also identified, of which one may be novel. In both family investigations, this same mutation could also be observed in the maternal DNA sample. Conclusion:Based on these observations, introduction of CD177 genotyping into routine use would identify null expression in over 75% (16/21) of associated cases.In turn, this could significantly reduce the need for supplementary testing and associated inconvenience to patients while permitting increased efficiency of laboratory testing. An added benefit would potentially elucidate other clinically relevant mutations and associated antigenic targets.

show abstract

Flow-Cytometric Analysis of HNA-2 Expression and Phenotypes Among Thai Blood Donors

Cited by 6 publications

References 28 publications

A novel nonsense mutation found in the CD177 gene of Thai individuals with the HNA‐2 null phenotype

A novel nonsense mutation found in the CD177 gene of Thai individuals with the HNA‐2 null phenotype

Human neutrophil antigens: Nature, clinical significance and detection

Human neutrophil antigen 2 sequence‐based typing: Joining the hunt for the CD177 answer

Contact Info

Product

Resources

About