The Use of Magnetic Beads Coated With Soluble Hla Class I or Class Ii Proteins in Antibody Screening and for Specificity Determination of Donor-Reactive Antibodies1

Sumitran-Karuppan, S; Möller, Erna

doi:10.1097/00007890-199605270-00022

Cited by 35 publications

(15 citation statements)

References 7 publications

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“…However, our absorption experiments demonstrated that this assay was capable to detect antibodies directed against specific HLA-DR antigens and was not interfered by other molecules. Recently, antigen-specific immunotests using purified antigens coated in microtiter plate [30] or beads [31,32] were developed for the characterization of HLA class II antibodies in ELISA or flow cytometry technique, respectively. However, some epitopes could be destroyed by the isolation processes which might impair the binding of human alloantibodies.…”

Section: Discussionmentioning

confidence: 99%

Association of HLA-DR Antibodies and Graft Rejection in Renal Transplantation: Posttransplant Analysis Using the Monoclonal Antibody-Specific Immobilization of Leukocyte Antigens Assay (MAILA)

Renner¹,

Santoso²,

Müller‐Steinhardt³

et al. 2004

Transfus Med Hemother

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Background: The clinical impact of HLA-DR alloantibodies in renal transplantation is still controversial. This may be partly due to the lack of specificity and sensitivity of current detection methods, e.g. complement-dependent cytotoxicity test employing B lymphocytes as target antigens. Patients and Methods: In this study, we analyzed the incidence and specificity of HLA-DR antibodies after renal transplantation by use of the antigen-specific capture assay MAILA. 53 primary cadaver kidney recipients were recruited for a prospective study and were retrospectively assigned to the following groups: 20 patients with unsuspicious clinical course within the first 3 months after transplantation (control group), 20 patients undergoing histologically proven acute rejection within the first 3 months after transplantation, and 13 patients with chronic rejection 1 year or later after transplantation. 105 patients’ sera collected prospectively were analyzed by MAILA using 13 homozygous B-lymphoblastoid cell lines (B-LCL) carrying the common HLA-DR specificities. Results: In the control group, only 1 of 19 patients (5.3%) without acute rejection revealed HLA-DR antibodies after transplantation. In contrast, HLA-DR alloantibodies could be detected post transplant in 36.8% (7/19) and 69.2% (9/13) of patients with acute and chronic rejection, respectively. The frequency of HLA-DR antibodies present in both study groups was significantly higher in comparison to the control group. Of the 17 patients who received a graft across a donor recipient HLA-DR mismatch, 5 patients exhibited donor-specific alloantibodies. However, the majority of antibodies detected was not specific for mismatched HLA-DR alloantigens. Conclusion: HLA-DR antibodies were significantly associated with transplant rejection in first cadaver renal transplantation, especially in patients undergoing chronic rejection. Although donor specificity was not observed in the majority of patients, monitoring of HLA-DR antibodies might support diagnosis of graft rejection.

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Section: Discussionmentioning

confidence: 99%

Association of HLA-DR Antibodies and Graft Rejection in Renal Transplantation: Posttransplant Analysis Using the Monoclonal Antibody-Specific Immobilization of Leukocyte Antigens Assay (MAILA)

Renner¹,

Santoso²,

Müller‐Steinhardt³

et al. 2004

Transfus Med Hemother

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“…However, compared to other reliable techniques for HLA–Ab screening, i.e. ELISA [11]and fluorescence–activated cell sorting [12], which are not adversely affected by therapeutically applied LyAb, this new method is both cost effective and simple.…”

Section: Discussionmentioning

confidence: 99%

An Extended Lymphocytotoxicity Test for Patients Treated with Lymphocytotoxic Antibodies

2000

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Background and Objectives: After organ transplantation, HLA antibodies (HLA–Ab) stimulated by transplant or transfusion are usually indistinguishable from passive therapeutic lymphocytotoxic antibodies (LyAb), e.g. antilymphocyte globulin (ALG), antithymocyte globulin (ATG) and OKT3, by standard lymphocytotoxicity tests (LCTs). Herein, an extended technique capable of distinguishing between these two types of antibodies is described. Materials and Methods: Serum samples from 11 patients who received therapeutic antibodies were screened for HLA–Ab by LCTs. The administered LyAb were murine OKT3 (n = 2), equine ALG (n = 3), rabbit ATG (n = 2), and combinations of ALG and OKT3 (n = 1), ATG and ALG (n = 1), and ATG and OKT3 (n = 2). To discriminate between therapeutically applied antibodies and active HLA alloimmunization, the sera were preincubated with species–specific anti–Fc IgG, thus blocking the therapeutic antibodies. Results: The sera of all patients treated showed strong positive lymphocytotoxic reactions [panel reactive antibodies (PRA) 50–100%]. After incubation with the corresponding Fc antibodies, in 9 of the 11 samples the amount of PRA decreased to the level present before transplantation (0–18%). In the remaining cases, new–onset alloimmunization was detected. Conclusion: This extended LCT allows the screening of serum samples for active HLA–Ab even in the presence of therapeutic LyAb.

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“…We described previously a new and quick method for the isolation of HLA class I and class II antigens using paramagnetic microbeads. 25 Microbeads coated with pooled HLA class I or II antigens were used for the removal of HLA antibodies from patient sera. The absorbed sera were retested for binding to LSECs.…”

Section: Methodsmentioning

confidence: 99%

A novel mechanism of liver allograft rejection facilitated by antibodies to liver sinusoidal endothelial cells†

Sumitran–Holgersson

Karrar

et al. 2004

Hepatology

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The Use of Magnetic Beads Coated With Soluble Hla Class I or Class Ii Proteins in Antibody Screening and for Specificity Determination of Donor-Reactive Antibodies1

Cited by 35 publications

References 7 publications

Association of HLA-DR Antibodies and Graft Rejection in Renal Transplantation: Posttransplant Analysis Using the Monoclonal Antibody-Specific Immobilization of Leukocyte Antigens Assay (MAILA)

Association of HLA-DR Antibodies and Graft Rejection in Renal Transplantation: Posttransplant Analysis Using the Monoclonal Antibody-Specific Immobilization of Leukocyte Antigens Assay (MAILA)

An Extended Lymphocytotoxicity Test for Patients Treated with Lymphocytotoxic Antibodies

A novel mechanism of liver allograft rejection facilitated by antibodies to liver sinusoidal endothelial cells†

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