Detection of Gov system antibodies by MAIPA reveals an immunogenicity similar to the HPA‐5 alloantigens

Berry, JS; Murphy, Chris; Smith, Graham; Ranasinghe, E.; Finberg, Robert W.; Walton, J.; Brown, Julianne R.; Navarrete, C.; Metcalfe, Paul; Ouwehand, Willem H.

doi:10.1046/j.1365-2141.2000.02170.x

Cited by 94 publications

(121 citation statements)

References 30 publications

(46 reference statements)

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“…1,11,12 Until recently, however, the importance of the Gov antigens in alloantibody-mediated platelet destruction has not been defined. We have recently shown that the incidence of Gov alloantibodies, especially in PR patients, is high with an incidence exceeded only by those directed against the human platelet antigen 1 (HPA-1) system antigens.…”

Section: Introductionmentioning

confidence: 99%

“…We have recently shown that the incidence of Gov alloantibodies, especially in PR patients, is high with an incidence exceeded only by those directed against the human platelet antigen 1 (HPA-1) system antigens. 11 Unfortunately, Gov phenotyping by serologic techniques is problematic for several reasons. First, the polyclonal antisera that are required are in limited supply and of variable quality, and often contain HLA, and sometimes HPA, alloantibodies as well.…”

Section: Introductionmentioning

confidence: 99%

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A tyrosine703serine polymorphism of CD109 defines the Gov platelet alloantigens

Schuh

Watkins

Nguyen

et al. 2002

Blood

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The biallelic platelet-specific Gov antigen system-implicated in refractoriness to platelet transfusion, neonatal alloimmune thrombocytopenia, and posttransfusion purpura-is carried by the glycosylphosphatidylinositol (GPI)-linked protein CD109. The recent identification of the human CD109 complementary DNA (cDNA) has allowed the molecular nature of the Gov alleles to be elucidated. By using reverse transcriptase-polymerase chain reaction (RT-PCR) to amplify CD109 cDNAs from 6 phenotypically homozygous Gov aa and Gov bb individuals, we have determined that the Gov alleles differ by an A to C single nucleotide polymorphism (SNP) at position 2108 of the cod-

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

A tyrosine703serine polymorphism of CD109 defines the Gov platelet alloantigens

Schuh

Watkins

Nguyen

et al. 2002

Blood

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“…3,12,13 Previously undefined, 10,16,17 the immunogenicity of the Gov antigens is now known to be similar to that of the HPA-5 antigens and is exceeded only by that of the HPA-1 antigens. 18 CD109 is also expressed by activated T cells. After T-cell activation in vitro, CD109 becomes detectable on CD4 ϩ and CD8 ϩ T cells by day 1, peaks by day 6, and thereafter decreases in the absence of additional interleukin-2.…”

Section: Introductionmentioning

confidence: 99%

Cell surface antigen CD109 is a novel member of the α2 macroglobulin/C3, C4, C5 family of thioester-containing proteins

Lin

Sutherland

Horsfall

et al. 2002

Blood

166

163

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Cell surface antigen CD109 is a glycosylphosphatidylinositol (GPI)-linked glycoprotein of approximately 170 kd found on a subset of hematopoietic stem and progenitor cells and on activated platelets and T cells. Although it has been suggested that T-cell CD109 may play a role in antibodyinducing T-helper function and it is known that platelet CD109 carries the Gov alloantigen system, the role of CD109 in hematopoietic cells remains largely unknown. As a first step toward elucidating the function of CD109, we have isolated and characterized a human CD109 cDNA from KG1a and endothelial cells. The isolated cDNA comprises a 4335 bp open-reading frame encoding a 1445 amino acid (aa) protein of approximately 162 kd that contains a 21 aa Nterminal leader peptide, 17 potential Nlinked glycosylation sites, and a C-terminal GPI anchor cleavage-addition site. We report that CD109 is a novel member of the ␣2 macroglobulin (␣2M)/C3, C4, C5 family of thioester-containing proteins, and we demonstrate that native CD109 does indeed contain an intact thioester. Analysis of the CD109 aa sequence suggests that CD109 is likely activated by proteolytic cleavage and thereby becomes capable of thioester-mediated covalent binding to adjacent molecules or cells. In addition, the predicted chemical reactivity of the activated CD109 thioester is complementlike rather than resembling that of ␣2M proteins. Thus, not only is CD109 potentially capable of covalent binding to carbohydrate and protein targets, but the t ½ of its activated thioester is likely extremely short, indicating that CD109 action is highly restricted spatially to the site of its activation. IntroductionTo identify new surface antigens expressed by primitive hematopoietic stem and progenitor cells, we raised a series of monoclonal antibodies (mAbs) against the primitive CD34 ϩ acute myeloid leukemia cell line, KG1a. 1-3 Four of these mAbs-8A3, 7D1, 8A1, and 7C5-recognized a novel glycoprotein of approximately 170 kd that was expressed in a restricted pattern in the hematopoietic compartment and by endothelial cells. 4 Subsequently found to be identified by a number of additional mAbs, this antigen was designated CDw109 in 1993 5 and CD109 in 1996. 6 Antibodies to CD109 recognize monomeric polypeptides of about 170 kd and 150 kd in lysates of KG1a cells, T-cell lines, and activated T lymphoblasts, endothelial cells, and activated platelets. 3,[7][8][9][10][11] Peptide mapping and amino acid (aa) analysis indicate that the 150-kd form is likely derived proteolytically from the 170-kd form. 3,10 An additional band of about 120 kd that is occasionally observed arises through calcium-dependent proteolysis of the larger forms. 3,10 CD109 contains several N-linked endoglycosidase H-sensitive hybrid-type glycans but no O-linked glycans. 3,10 Consistent with this finding, ABH blood group antigens have recently been shown to be carried by platelet CD109. 12 KG1a CD109 is susceptible to cleavage with phosphatidylinositolspecific phospholipase C (PI-PLC), indicating that CD109 is bo...

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“…For instance, we included a monoclonal antibody against Fcc receptor II (CD32) in a mixture of mAb to detect the nonspecific binding of immunoglobulins or immune complexes (Koksch et al, 1995). With regard to the sensitivity of the SASPA, antibodies against HPA-5 and -15 antigens, which are weakly expressed on the platelets with approximately 2000 binding sites per platelet Santoso et al, 1989;Sutherland et al, 1991;Haregewoin et al, 1994;Berry et al, 2000), were also investigated. In this evaluation, anti-HPA-15b and -HPA-5a, as well as -HPA-5b, could be detected at higher dilution steps than MAIPA, and one serum containing IgG anti-HPA-5b could be only recognized by SASPA.…”

Section: Discussionmentioning

confidence: 99%

A novel method for simultaneous analysis of specific platelet antibodies: SASPA

et al. 2004

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Anti-platelet antibodies cause several thrombocytopenic disorders such as autoimmune or alloimmune thrombocytopenia, post-transfusion purpura, drug-induced thrombocytopenia and refractoriness to platelet transfusion by eliminating platelets from the circulation. These antibodies can target specific platelet glycoproteins (GP), human leucocyte antigen (HLA) class I antigens or phospholipids (Kurata et al, 1993;Alcorta et al, 1996;Macchi et al, 1996Macchi et al, , 1997Cuadrado et al, 1997;Panzer et al, 1997;Kiefel et al, 2001;Ranasinghe et al, 2001;Cines & Blanchette, 2002). The detection of these antibodies with high sensitivity and characterization of their specificity is important for diagnosis and specific therapy.The platelet immunofluorescence test (PIFT) has been widely used as a screening method for the detection of platelet antibodies (von dem Borne et al, 1978;Christopoulos et al, 1993;Porcelijn and von dem Borne, 1998;Hagenström et al, 2000). In order to differentiate the detected antibodies, the monoclonal antibody-specific immobilization of platelet antigen (MAIPA) assay has been mostly used as the reference assay (Kiefel et al, 1987). Several other antibody-detection systems, such as immunoblotting, solid-phase radioimmunoassay and modified antigen-capture enzyme-linked immunosorbent assay (MACE), have been reported (Tijhuis et al, 1991;Hou et al, 1995). However, the drawbacks of these tests SummaryGlycoprotein (GP)-specific platelet antibodies can cause allo-immune and auto-immune thrombocytopenia. The specific detection of relevant antibodies is a prerequisite for diagnosis and treatment. Here, we describe an improved method based on simultaneous detection of various plateletspecific immunoglobulin G (IgG) and IgM antibodies. Bead populations with distinct fluorescence intensities, coated with monoclonal antibodies specific for mouse heavy chain isotypes, were used for the simultaneous immobilization of platelet-GP [IIb/IIIa, Ib/IX, human leucocyte antigen (HLA) class I, or Ia/IIa, CD32, GPIV or CD109, Ib/IX, HLA class I]. In order to detect human IgG and IgM antibodies simultaneously, phycoerythrin-and fluorescein isotiocyanate-conjugated goat anti-human IgG and IgM were added. On this basis, the abundance of six distinct antibodies (three anti-GP, each with subclasses IgG and IgM) were simultaneously analysed without cross-reaction by flow cytometry. For evaluation, sera and platelets from 169 patients with platelet-binding and/or platelet-associated antibodies were investigated. The monoclonal antibody-specific immobilization of platelet antigen (MAIPA) assay was performed in parallel as reference test. The simultaneous analysis of platelet-specific antibodies (SASPA) assay was able to detect all platelet-specific IgG and IgM that were also recognized by MAIPA with a comparable sensitivity. SASPA proved to be a rapid and reliable assay that required less platelets than other methods. This method has the potential to pave the way for new investigations of platelet-specific antibodies.

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Detection of Gov system antibodies by MAIPA reveals an immunogenicity similar to the HPA‐5 alloantigens

Cited by 94 publications

References 30 publications

A tyrosine703serine polymorphism of CD109 defines the Gov platelet alloantigens

A tyrosine703serine polymorphism of CD109 defines the Gov platelet alloantigens

Cell surface antigen CD109 is a novel member of the α2 macroglobulin/C3, C4, C5 family of thioester-containing proteins

A novel method for simultaneous analysis of specific platelet antibodies: SASPA

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