Reactivity of Twenty-two Cytotoxic Human Monoclonal HLA Antibodies Towards Soluble HLA Class I in an Enzyme-Linked Immunosorbent Assay (PRA-STAT®)

Mulder, Arend; Kardol, M. J.; Regan, John F.; Buelow, Roland; Claas, Frans H.J.

doi:10.1016/s0198-8859(97)00146-8

Cited by 41 publications

(38 citation statements)

References 15 publications

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“…This study was performed using IgG-and IgM-type mAbs derived from Caucasoid women (Dutch) who became sensitized during pregnancy by paternal antigens [32][33][34]. All were supernatants of cloned hybridomas generated from Epstein-Barr virus-transformed B cells.…”

Section: Human Monoclonal Antibodiesmentioning

confidence: 99%

“…For the 5 eplets in this study, we have calculated from the models in Fig. 1 that eplets and their corresponding 15-Å 2 surface areas have an average of 33 residue locations (range [25][26][27][28][29][30][31][32][33][34][35][36][37][38][39][40]. These numbers are higher than the general 15 to 25 range of contact residues in structural epitopes of experimentally tested protein antigens.…”

Section: Molecular Modeling Of the Topography Of Structural Epitopesmentioning

confidence: 99%

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Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies

Duquesnoy¹,

Marrari²,

Mulder³

et al. 2012

Human Immunology

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Section: Human Monoclonal Antibodiesmentioning

confidence: 99%

Section: Molecular Modeling Of the Topography Of Structural Epitopesmentioning

confidence: 99%

Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies

Duquesnoy¹,

Marrari²,

Mulder³

et al. 2012

Human Immunology

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“…HLA antibody-producing EBV cell lines were stabilized by electrofusion and rigorous cloning. [22][23][24][25] The HLA specificity of mAbs was determined by testing with a large (n Ͼ 240) panel of HLA-typed peripheral blood mononuclear cell suspensions in the conventional complement-dependent microcytotoxicity (CDC) assay. For flow cytometry application, HLA hu-mAbs were used as undiluted hybridoma culture supernatants.…”

Section: Monoclonal and Polyclonal Antibodiesmentioning

confidence: 99%

Down-regulation of HLA-A and HLA-Bw6, but not HLA-Bw4, allospecificities in leukemic cells: an escape mechanism from CTL and NK attack?

Demanet

Mulder

Deneys

et al. 2004

Blood

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“…the assays, but antibody specificities and test sensitivities vary in detail due to test-specific characteristics (19,29,34,35). The IPIm detects a higher percentage of panel reactivity in sera from high-risk patients awaiting a kidney than the CDC, and this is in good agreement with graft failure (14).…”

Section: Discussionmentioning

confidence: 94%

“…The main conclusion is that the ELISA method is a good alternative, with an 80% agreement between the CDC and ELISA (15,19,32,34), and an 80% agreement between flow cytometry and ELISA (30). But the problem of IgM reactivity of the CDC and of nonreactivity with non-complement-fixing antibodies remains.…”

Section: Discussionmentioning

confidence: 99%

HLA‐antibody testing: The immune phagocytosis inhibition test is superior to the PRA‐STAT and NIH lymphocytotoxic test with respect to specificity

Flesch¹,

Philipp²,

Cassens

et al. 2001

Clinical Laboratory Analysis

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We compared the specificity and sensitivity of four different methods for the detection of antibodies specific for HLA antigens. The NIH version of the complement-dependent cytotoxic test (CDC) was used as the gold standard to which we compared two Fcgamma receptor (FcgammaR)-dependent immune phagocytosis inhibition tests (IPI) and one commercial enzyme-labelled immunosorbent assay (ELISA) with soluble HLA class I-antigen preparations bound to the plate (PRA-STAT). Both IPI tests are based on the fact that HLA-antibodies specifically bind to antigens on the monocyte surface via their Fab portion, and in so doing block a neighbouring FcgammaR with their Fc region. This blockade prevents phagocytosis of IgG-coated red blood cells (RBCs), which can be measured either microscopically (IPIm) or photometrically (IPIp). The four assays were used in blind tests on 20 human alloantisera or monoclonal antibodies with known HLA-antigen reactivities. Additionally, two monoclonal antibodies and one human serum were titrated to elucidate the sensitivity of each test. After all tests were completed, the identities of the samples were disclosed. Both IPI methods detected and identified all clinically relevant HLA class I and class II specific antibodies. In contrast, the CDC was not able to detect noncytotoxic HLA-antibodies and HLA class II specific antibodies; however, it detected clinically insignificant IgM lymphocytotoxins. The PRA-STAT assay enabled identification of all cytotoxic and noncytotoxic IgG antibodies with specificity for HLA-class I antigens. With respect to sensitivity, the CDC and the IPI methods were superior to the PRA-STAT. These facts demonstrate the advantage of IPI methods in the detection of clinically relevant HLA-antibodies.

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Reactivity of Twenty-two Cytotoxic Human Monoclonal HLA Antibodies Towards Soluble HLA Class I in an Enzyme-Linked Immunosorbent Assay (PRA-STAT®)

Cited by 41 publications

References 15 publications

Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies

Structural aspects of human leukocyte antigen class I epitopes detected by human monoclonal antibodies

Down-regulation of HLA-A and HLA-Bw6, but not HLA-Bw4, allospecificities in leukemic cells: an escape mechanism from CTL and NK attack?

HLA‐antibody testing: The immune phagocytosis inhibition test is superior to the PRA‐STAT and NIH lymphocytotoxic test with respect to specificity

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