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 Previous Article | Table of Contents | Next Article 
Blood, Vol. 91 No. 2 (January 15), 1998:
pp. 673-679
Fc RIIIB Gene Duplication: Evidence for Presence and
Expression of Three Distinct FcRIIIB Genes in
NA(1+,2+)SH(+) Individuals
By
Harry R. Koene,
Marion Kleijer,
Dirk Roos,
Masja de Haas, and
Albert E.G.Kr. Von dem Borne
From the Central Laboratory of the Netherlands Red Cross Blood
Transfusion Service and Laboratory for Clinical and Experimental
Immunology, Academic Medical Center, University of Amsterdam,
Amsterdam, The Netherlands and the Department of Hematology, Academic
Medical Center, Amsterdam, The Netherlands.
 |
ABSTRACT |
Recently, a new alloantigen on IgG Fc receptor type IIIb
(Fc RIIIb), SH, was described (Bux et al, Blood 89:1027,
1997). We identified three healthy individuals whose neutrophils
reacted positively with the SH antiserum. The neutrophil antigen (NA) phenotype of all three donors was NA(1+,2+). Analysis of genomic DNA showed that the three donors were positive for the described SH-encoding mutation in the NA2-FcRIIIB gene,
266C A. However, NA(1,2) genotyping and nucleotide sequencing
of an NA2-specific fragment amplified from the genomic DNA
fragment showed that these individuals carried three FcRIIIB
genes, namely, NA1-FcRIIIB, NA2-FcRIIIB, and
SH-FcRIIIB, encoding NA1-FcRIIIb, NA2-FcRIIIb, and
SH-FcRIIIb, respectively. Southern blot analysis confirmed these
findings. Furthermore, all three transcripts were isolated from
neutrophil mRNA. To investigate whether the presence of three FcRIIIB genes resulted in a higher membrane expression of
FcRIIIb, we measured the reactivity of neutrophils from
NA(1+,2+)SH(+) individuals with a panel of CD16 monoclonal
antibodies (MoAbs) in comparison with neutrophils from
NA(1+,2+)SH( ) controls. Reactivity of four different
anti-pan-FcRIII MoAbs and NA2-specific MoAb GRM1 was higher with
SH(+) neutrophils compared with controls, whereas that of
NA1-specific MoAbs was similar, which is in concordance with the
results from the genomic analysis. We observed that reactivity with
NA2-specific CD16 MoAb PEN1 was sixfold higher in SH(+) individuals compared with controls. Apparently, the 60AlaAsp substitution in SH-FcRIIIb influences the epitope recognized by PEN1. In
conclusion, we identified three NA(1+,2+)SH(+) individuals
carrying three FcRIIIB genes and observed a clear
gene-dosage effect on the level of expression of neutrophil FcRIIIb.
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INTRODUCTION |
NEUTROPHIL ANTIGENS (NAs) are involved in
several clinical conditions, such as blood transfusion reactions and
immune-mediated neutropenia.1 The NA system has been
extensively investigated and is located on IgG Fc receptor type IIIb
(FcRIIIb; CD16).2,3 NA1- and NA2-FcRIIIb differ by
four amino acids in the membrane-distal Ig-like domain.4
These differences lead to distinct glycosylation patterns of NA1- and
NA2-FcRIIIb.2,3 Although the amino acid differences are
not located in the membrane-proximal, ligand-binding domain of the
receptor, the FcRIIIb-NA(1,2) isoforms differ in their capacity to
interact with IgG.5-8 Alloantibodies against NA1-FcRIIIb
and, to a lesser extent, NA2-FcRIIIb can be detected in a large
proportion of patients suffering from neonatal immune-mediated neutropenia.9
Recently, Bux et al10 described four cases of alloimmune
neutropenia in which alloantibodies recognizing a thus far unknown antigen on FcRIIIb were identified. The newly identified alloantigen was termed SH and has a gene frequency of 4% in the German
population.10 Nucleotide sequence analysis showed that the
SH-FcRIIIB gene differed from NA2-FcRIIIB by a
single base pair (266CA), encoding an AlaAsp
substitution at amino acid position 60. The authors concluded that
SH-FcRIIIb is the product of an NA2-FcRIIIB
polymorphism.10
In this report, we analyzed three individuals whose neutrophils were
phenotyped as NA(1+,2+)SH(+). Genomic analysis showed that in these
individuals, an FcRIIIB gene encoding SH-FcRIIIb exists
alongside an NA1- and an NA2-FcRIIIB gene. All three
FcRIIIB genes were transcribed into mRNA. Furthermore, in
all three donors we observed a clear gene-dosage effect with regard to
the neutrophil membrane expression of FcRIIIb, attributable to the
equal expression of the three isoforms.
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MATERIALS AND METHODS |
Antibodies and antisera.
Anti-pan-FcRIII (CD16) monoclonal antibodies (MoAbs) used were
CLBFcRgran1 (mIgG2a), 3G8 (mIgG1), BW209/2 (mIgG2a), and MEM154 (mIgG1). CLBgran11 (mIgG2a) and MG38 (mIgG1) recognize NA1-FcRIIIb, whereas GRM1 (mIgG2a) recognizes NA2-FcRIIIb and FcRIIIa. PEN1 (mIgG2a) reacts with a small oligosaccharide moiety of NA2-FcRIIIb and with FcRIIIa.11 BW209/2 was a generous gift from Dr
Kurrle (Behring Werke, Marburg, Germany) and GRM1 was provided by Dr Garrido (Hospital des Nieves, Granada, Spain). CLBFcRgran1, CLBgran11, and irrelevant control MoAbs were from our own institute. The other
CD16 MoAbs were obtained via the Fifth International Workshop on
Leukocyte Antigens (Boston, MA; November 1993). A human antiserum recognizing SH-FcRIIIb was obtained via the Second International Granulocyte Serology Workshop (Helsinki, Finland; May 1996). Sera from
healthy AB-positive individuals were used as controls. Fluorescein isothiocyanate (FITC)-labeled F(ab )2 fragments of
goat-antimouse-Ig and FITC-goat-antihuman-Ig from our institute were
used to detect MoAb and human antibody binding, respectively.
Isolation of cells.
EDTA-anticoagulated blood was centrifuged over a Ficoll-Hypaque
gradient with a specific gravity of 1.076 g/mL (Pharmacia Fine
Chemicals, Uppsala, Sweden). Mononuclear cells were harvested from the
interphase for DNA isolation and the pellet was treated with ice-cold
NH4Cl solution (155 mmol/L NH4Cl, 10 mmol/L
KHCO3, 0.1 mmol/L EDTA, pH 7.4) to lyse erythrocytes. The
remaining cells were more than 95% neutrophils.
Flow cytometry.
Neutrophils were tested for reactivity with a panel of CD16 MoAbs and
with the SH antiserum by indirect immunofluorescence.12 Briefly, neutrophils were fixed with 1% paraformaldehyde (PFA; wt/vol)
and were incubated with MoAb or human antiserum for 30 minutes at room
temperature. After washing with phosphate-buffered saline (PBS)
containing 0.2% bovine serum albumin (wt/vol) the cells were stained
with FITC-labeled F(ab)2 fragments of
goat-antimouse-Ig or FITC-goat-antihuman-Ig. Binding of the
conjugate was assessed with a FACScan flowcytometer (Becton Dickinson,
San Jose, CA).
Soluble FcRIII enzyme-linked immunosorbent assay (ELISA).
The plasma level of sFcRIII was measured by a sandwich ELISA,
essentially as previously described.13 Briefly, ELISA
plates were incubated with CD16 MoAb CLBFcRgran1 and blocked with PBS containing 2% (vol/vol) milk. The plates were incubated with plasma that was diluted in High Performance ELISA (HPE) buffer (CLB, Amsterdam, The Netherlands). Subsequently, a biotinylated polyclonal rabbit-antihuman-FcRIII antibody diluted in HPE buffer was added. After incubation with horse radish peroxidase-labeled streptavidin, a
substrate was added to measure the amount of bound antibody. Plasma
from 90 healthy individuals was pooled and used to construct a
calibration curve. The level of sFcRIII in this pool was set at 100 arbitrary units (AU).
FcRIIIB-NA(1,2) genotyping assays.
Genotyping for the FcRIIIB-NA(1,2) polymorphism was
performed as described before.14 In brief, two sets of
primers specifically annealing to either an NA1-FcRIIIB or
an NA2-FcRIIIB fragment were used. NA1-FcRIIIB-
and NA2-FcRIIIB-specific fragments were separately
amplified from gDNA in a Perkin Elmer Cetus Cycler (Norwalk, CT) in a
total volume of 50 µL. Amplification of a fragment of the
p22-phox (CYBA) gene served as an internal control in
each polymerase chain reaction (PCR). The nucleotide sequence of
FcRIIIB-NA2-specific fragments was determined by cycle
sequencing of purified PCR products with 33P-labeled
terminators with the Thermo Sequenase kit, according to the
manufacturer's instructions (Amersham Life Sciences, Cleveland, OH).
Southern blot-based restriction fragment length polymorphism (RFLP)
assay.
Southern blot-based restriction fragment analysis was performed as
previously described.14 Briefly, 10 µg of genomic DNA was
digested overnight with BamH1 and EcoR1 (Promega,
Madison, WI). After gel electrophoresis and transfer to
nylon sheets, the blot was hybridized with 32P-labeled
pGP5, a probe that contains the entire coding region of NA1-FcRIIIb
(Dr G. Peltz, DNAX Research Institute of Molecular and Cellular
Biology, Palo Alto, CA).15 This probe hybridizes to both
isoforms of FcRIIIB, as well as to FcRIIIA. The
double digestion resulted in an FcRIIIB- and an
FcRIIIA-specific fragment of 2.0 and 5.5 kb, respectively.
The labeling intensities of the two fragments were measured and
compared by phospho-imaging (Fuji, London, UK). NA(1+,2+) genotyped
donors and known hemizygous FcRIIIB gene-deficient donors
served as controls.
Sequence analysis of FcRIIIB-encoding cDNA.
Messenger RNA was isolated from purified neutrophils with a
CsCl2 gradient and was reverse transcribed into cDNA. The
entire coding region of FcRIIIB was amplified by
PCR,16 after which the products were cloned into a pGEM-T
vector according to the manufacturer's instructions (Promega). After
transformation of Escherichia coli, inserts were amplified by
PCR and sequenced with 32P-end-labeled primers (Amersham)
with the BRL cycle sequencing kit (BRL, Gaithersburg, MD).
| |
RESULTS |
Genomic analysis.
In a population of 55 healthy laboratory workers, we identified three
healthy donors whose neutrophils reacted positively with the SH
antiserum giving a phenotype frequency of approximately 5%, in
concordance with previous findings.10 Neutrophils from all
three donors were phenotyped as NA(1+,2+)SH(+).
Figure 1 shows a representative experiment
with neutrophils from an SH() and an SH(+) donor. To determine
whether the NA2-FcRIIIB gene contained the reported
266CA mutation,10 we specifically amplified a fragment of the NA2-FcRIIIB gene from gDNA by an
allele-specific primer-annealing (ASPA) PCR. As shown in
Fig 2A to C, direct sequencing of these
products showed that the individuals were heterozygous at nucleotide
position 266. At this position, a C as well as an A were detected,
suggesting that two different fragments were amplified with the
NA2-FcRIIIB-specific primer set. Sequence analysis of
NA2-FcRIIIB-specific fragments amplified from genomic DNA from
two NA(1+,2+)SH() controls showed a single band at position 266 (Fig 2D to E).
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| Fig 1.
Reactivity of neutrophils from an NA(1+,2+)SH(+)
donor and an NA(1+,2+)SH() control with human anti-NA1- and
anti-NA2-FcRIIIb antisera and the SH antiserum. AB serum was used
as control.
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| Fig 2.
Nucleotide sequence (GATC) of
FcRIIIB-NA2-specific genomic DNA fragments. (A to C) Fragments
from three NA(1+2+)SH(+) individuals. Two bands (A and C) are
visible at nucleotide position 266, indicating the presence of a normal
(266C) as well as of a mutated (266A) FcRIIIB-NA2 gene. (D
and E) Sequences of NA(1+2+)SH() controls.
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Additionally, NA2-FcRIIIB-specific PCR products were
digested with SfaNI, which recognizes a 5-GATGC-3
sequence present only in the SH-FcRIIIB-derived
fragment.10 Figure 3 shows the results of this digestion. Lane 1 shows SfaNI treatment of a
fragment amplified from a plasmid containing SH-FcRIIIb-encoding
cDNA, resulting in a single band of 137 bp, confirming that complete digestion is obtained by SfaNI analysis. Lanes 2 to 4 contain SfaNI-treated DNA fragments from NA(1+,2+)SH() controls.
Only the undigested fragment of 168 bp is present. In lanes 5 to 7 DNA
fragments from three NA(1+,2+)SH(+) individuals are shown. Digested
(137 bp) and undigested (168 bp) fragments are visible, indicating the
presence of an SH-FcRIIIB and an NA2-FcRIIIB sequence.
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| Fig 3.
SfaNI digestion of the 168-bp
FcRIIIB-NA2-specific gDNA fragment containing the SH
mutation at nucleotide position 266. SfaNI recognizes a
5-GATGC-3 sequence, which is only present in the FcRIIIB-SH gene. Lane 1 shows digestion of a fragment
amplified from a plasmid containing SH-FcRIIIb-encoding cDNA,
confirming that complete digestion is obtained by Sfa NI
treatment. DNA fragments from NA(1+,2+)SH() individuals were not
digested (lanes 2 through 4), whereas an additional band of 137 bp is
visible after digestion of fragments from SH(+) individuals (lanes 5 through 7).
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To confirm the hypothesis that more than two FcRIIIB genes
are present in the genome of NA(1+,2+)SH(+) individuals, a Southern blot-based RFLP assay was performed. The number of FcRIIIB
genes was determined by comparing the labeling intensities of an
FcRIIIB-specific and an FcRIIIA-specific
fragment. Genomic DNA from two of the three individuals was available
for testing. Table 1 shows the quantitative results of
the Southern blot, obtained with a phospho-imager. For these two
NA(1+,2+)SH(+) individuals, the ratio between the FcRIIIB-
and FcRIIIA-specific band is 1.16 and 1.14, respectively. This is approximately three times higher than the ratio obtained for three individuals with only one FcRIIIB gene (0.37 ± 0.17), and 1.5-fold the ratio found for six NA(1+,2+)SH()
controls (0.69 ± 0.12).
cDNA analysis.
Messenger RNA was isolated from purified neutrophils from one
NA(1+,2+)SH(+) individual and reversely transcribed into cDNA to
investigate whether all three FcRIIIB genes were
transcribed. The entire coding region of FcRIIIB was
amplified by PCR and cloned into E coli. Figure
4, panel 1, shows part of the sequence of an NA1 transcript,
whereas the nucleotide sequence shown in panel 2 is derived from an
NA2 transcript. The NA1-NA2 difference at nucleotide
position 227 is depicted (AG), and all other described nucleotide differences were normally present (not shown).4 The sequence shown in panel 3 only differed from a normal NA2 sequence at nucleotide position 266 (CA). This
substitution predicts the AlaAsp substitution in
SH-FcRIIIb, described by Bux et al.10 Sequence analysis
of the complete coding region showed no other nucleotide substitutions.
In a total of 30 sequenced clones with an FcRIIIb-encoding insert
from this NA(1+,2+)SH(+) individual, 6 were found to carry the
NA1-FcRIIIB insert, 21 carried the NA2-FcRIIIB
insert, and 3 carried the SH-FcRIIIB insert.
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| Fig 4.
Sequence analysis of cDNA, derived from
NA(1+2+)SH(+) neutrophils. The nucleotide differences between the
three transcripts are shown in the box below the figure. Panel 1 shows
the normal NA1-FcRIIIB-derived sequence as indicated by the
presence of an A at nucleotide position 226. The sequence shown in
panel 2 is derived from an NA260Ala-FcRIIIB
clone (226G and 266C). The third distinct transcript is derived from an
NA2SH-FcRIIIB gene. The sequence is identical to
the NA260Ala-FcRIIIB sequence except for the A
at nucleotide position 266, encoding an Asp at amino acid position
60.
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Neutrophil FcRIIIb expression.
We determined the reactivity of NA(1+,2+)SH(+) neutrophils with a panel
of CD16 MoAbs in comparison with neutrophils from donors who were
NA(1+,2+)SH() in three separate experiments
(Table 2). Neutrophils from SH() and either
homozygous NA1- or NA2-positive individuals were simultaneously
analyzed in two of these experiments. Compared with neutrophils from
NA(1+,2+)SH() individuals, SH(+) neutrophils showed a higher
reactivity with anti-pan-FcRIII MoAbs CLBFcRgran1, 3G8, MEM154, and
BW209/2 (170%, 119%, 140%, and 163%, respectively). The
NA(1+,2+)SH(+) neutrophils reacted approximately twice as strong
with the NA2-specific MoAb GRM1 as did control neutrophils (Table 2).
However, the reactivity of the NA(1+,2+)SH(+) neutrophils was
comparable to that of NA(1,2+)-genotyped donors, who carried two
NA2 genes, as determined by Southern blot
analysis,13 with mean fluorescent intensities
(mfis) of 1,480 and 1,585 ± 568, respectively.
However, reactivity of the NA2-specific MoAb PEN1 was about sixfold
higher for NA(1+,2+)SH(+) neutrophils compared with
NA(1+,2+)SH() neutrophils and threefold higher compared with
NA(1,2+)SH() neutrophils. Reactivity with NA1-specific MoAbs
CLBFcRgran11 and MG38 was similar between SH() and SH(+) NA(1+,2+) neutrophils, suggesting an equal membrane
expression of NA1-FcRIIIb (Table 2). Figure 5 shows
the results of a representative experiment.
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| Fig 5.
Staining pattern of NA(1+2+)SH() (dashed lines)
and NA(1+2+)SH(+) neutrophils (continuous lines) with a panel of
CD16 MoAbs. One representative experiment of three is shown. (A and B)
The reactivity of SH(+) neutrophils with anti-pan-FcRIII MoAbs
(CLBFcRgran1 and 3G8) was higher than that of SH() neutrophils. (C
and D) The reactivity of both types of neutrophils with
NA1-FcRIIIb-specific MoAbs CLBFcRgran11 and MG38 was comparable.
(E) Reactivity of NA2-FcRIIIb-specific MoAb GRM1 was approximately
twice as high with SH(+) neutrophils compared with SH()
neutrophils. (F) NA2-FcRIIIb-specific MoAb PEN1 showed a reactivity
with SH(+) neutrophils that was sixfold higher compared with the
reactivity of SH() neutrophils.
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Because the level of soluble (s)FcRIII in plasma correlates with the
number of FcRIIIB genes,13 we determined the
amount of sFcRIII in plasma from the three SH(+) individuals. In
concordance with the high membrane expression levels, we found plasma
sFcRIII levels of 197, 279, and 137 AU (mean, 204 ± 71 AU).
However, this value was not significantly different from the mean
sFcRIII level in 24 NA(1+,2+)SH() individuals (106 ± 24 AU13; Welch's approximate t-test, P = .07).
| |
DISCUSSION |
Recently, Bux et al identified a new alloantigen on
FcRIIIb.10 Sequence analysis showed that this antigen,
termed SH, is encoded by a point mutation in the NA2-FcRIIIB
gene. We amplified both a normal NA2-encoding sequence and an NA2
sequence containing the SH mutation from genomic DNA from three
NA(1+,2+)SH(+) individuals by means of an
NA2-FcRIIIB-specific PCR. Digestion of these
NA2-FcRIIIB-specific fragments with SfaNI confirmed
the presence of two NA2 genes. Furthermore, a Southern
blot-based RFLP assay showed that three FcRIIIB genes are
present in the genome of these NA(1+,2+)SH(+) individuals. All three
FcRIIIB genes were transcribed, because three distinct
transcripts encoding either NA1-, NA2-, or SH-FcRIIIb were isolated
from neutrophil mRNA. These data indicate that these three donors each
carry three FcRIIIB genes, namely, NA1-FcRIIIB, NA2-FcRIIIB, and SH-FcRIIIB. Six of 14 SH(+)
individuals described by Bux et al were phenotyped as NA(1,2+),
and the remaining eight were NA(1+,2+).10 Moreover, three
NA(1+,2+)SH(+) donors were reanalyzed and were found to carry three
FcRIIIB genes as well (J. Bux, personal communication). Only
confirmation by PCR and/or Southern blotting can settle the
question as to whether any SH positivity without gene duplication
exists. However, with these methods the possibility of one
FcRIIIB gene-deficient chromosome and two FcRIIIB
genes on the other one cannot be ruled out.14 If we
assume that the SH-FcRIIIB gene is only present in
association with a second FcRIIIB gene on the same
chromosome, then the presence of NA(1,2+)SH(+) phenotyped donors
in the study of Bux et al might indicate that NA2-FcRIIIB is
located on the same chromosome as SH-FcRIIIB. It is
conceivable that before or after the SH mutation occurred, an unequal
crossing-over event between two chromosomes carrying FcRIIIB
genes has led to the supposed NA2-SH allele. The counterpart of
this unequal crossing-over is an FcRIIIB gene deletion,
which has been described.14,17-20 The genotype frequencies
of SH-FcRIIIB and FcRIIIB gene deletion, being
4% and 3% to 9%, respectively, are not contradicting this
theory.10,18,19 Theoretically, SH() donors with
three FcRIIIB genes (gene duplication without mutation)
should exist. Southern blot analysis of gDNA from a large group of
donors could settle this question.
Neutrophils from healthy individuals carry 100,000 to 300,000 copies of
FcRIIIb per cell.21 Previously, we described that the
amount of FcRIIIb on the neutrophil membrane correlates with the
number of FcRIIIB genes.13 Individuals who are
hemizygous FcRIIIB-gene-deficient have approximately half
the neutrophil FcRIIIb expression and half the plasma soluble
(s)FcRIII level of individuals carrying two FcRIIIB
genes. Moreover, reactivity of neutrophils from NA-homozygous
individuals with NA-specific MoAbs is twice as high compared with
NA(1+,2+) neutrophils.13 The three NA(1+,2+)SH(+)
individuals that we tested had a higher neutrophil FcRIIIb
expression as measured with anti-pan-FcRIII MoAbs, with the
exception of 3G8. Furthermore, the reactivity with the NA2-specific
MoAb GRM1 was approximately twice as high compared with SH()
neutrophils, whereas the reactivity with NA1-specific MoAbs was
similar. MoAb 3G8 recognizes an epitope in the membrane-proximal domain
of FcRIII,22 whereas binding of GRM1 is dependent on the
presence of 47Ser.22,23 Whether the 60AlaAsp
substitution influences the affinity of 3G8 will have to be
investigated in studies with SH-FcRIIIb-transfected cells or with
neutrophils from donors at the genomic level proven to carry only the
SH-FcRIIIB gene and no NA2-FcRIIIB. Our data
suggest that the substitution of the hydrophobic alanine to the
negatively charged aspartic acid in SH-FcRIIIb influences the
epitope recognized by PEN1, which might be located in the
membrane-distal Ig-like domain because of its NA2
specificity.11 Finally, SH(+) individuals seemed to have
higher levels of plasma sFcRIII compared with NA(1+,2+)SH() control donors. This suggests that the gene-dosage effect
observed for plasma sFcRIII levels also holds for individuals with
three FcRIIIB genes. Therefore, our findings imply that the
large interindividual variation in neutrophil FcRIIIb expression and
plasma levels of sFcRIII might be partly caused by differences in
the number of FcRIIIB genes. One could hypothesize that the
number of FcRIIIb copies on the neutrophil membrane influences the
effector functions of the cell. However, hemizygous and homozygous
FcRIIIB gene deficiency does not seem to be associated with
an increased infection risk.14 Further experiments should
elucidate whether the neutrophil response to opsonized particles
correlates with the number of FcRIIIB genes.
It should be further investigated whether the SH mutation influences
the ligand-binding capacity of the receptor. The NA1-FcRIIIb and
NA2-FcRIIIb isoforms have been shown to interact differently with
IgG, although the amino acid differences are all in the membrane-distal domain.5-8 Moreover, mutations in the membrane-distal
domain of FcRIIa affected the ligand-binding capacity as
well.24
The Second International Granulocyte Serology Workshop agreed for the
time being to term the new antigen SH. For the nomenclature of this
geno/phenotype it is important to elucidate whether SH positivity will
always be accompanied by NA2 positivity, either because of the close
homology between the SH and NA2 isoforms, or because of genetic linkage
between the two genes. Thus far, only the SH antiserum, genomic
analysis, and possibly CD16 MoAb PEN1 can distinguish between NA2- and
NA2SH-FcRIIIb. To underline the close connection of the
SH antigen with the NA(1,2) system and to emphasize the similarity to
NA2-FcRIIIB, we propose that the antigen be termed
NA2SH-FcRIIIb or NA3.
In conclusion, we detected the presence of three FcRIIIB
genes in three individuals whose neutrophils were phenotyped as NA(1+,2+)SH(+). These three genes, NA1-FcRIIIB,
NA2-FcRIIIB, and SH-FcRIIIB, were all transcribed and
a clear gene-dosage effect regarding neutrophil FcRIIIb expression
was observed. Our data indicate that it may be possible that a
chromosomal locus exists on which NA2-FcRIIIB is located in
tandem with SH-FcRIIIB.
| |
FOOTNOTES |
Submitted April 22, 1997;
accepted September 17, 1997.
Supported by Grant No. 900-012-092 of the Netherlands Organization for
Scientific Research (NWO).
Address reprint requests to Albert E.G.Kr. Von dem Borne, Central
Laboratory of the Netherlands Red Cross Blood Transfusion Service,
Department of Experimental Immunohematology, Plesmanlaan 125, 1066 CX
Amsterdam, The Netherlands.
The publication costs of this article were defrayed in part by page
charge payment. This article must therefore be hereby marked
"advertisement" in accordance with 18 U.S.C. section 1734 solely
to indicate this fact.
| |
ACKNOWLEDGMENT |
We thank the Department of Leukocyte and Thrombocyte Serology of the
CLB for neutrophil phenotyping and R. Dee for helpful technical
suggestions.
| |
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Abstract
Introduction
Abstract