Significance of Qualitative and Quantitative Evaluations of Anti-Hla Antibodies in Kidney Transplantation

Ishida, Hideki; Shimizu, Tomokazu; Shirakawa, Hiroki; Omoto, Kazuya; Tanabe, Kiyokatsu

doi:10.1097/00007890-201007272-00516

Cited by 8 publications

(13 citation statements)

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“…47,86,87 DSA with a Negative XM: Defining Risk Remains Challenging The increased sensitivity of SAB allows DSA to be detected even with a negative XM. 80,88,89 AMR rates of 20%-55% are reported under these circumstances, 22,23,83,88,90,91 with AMR negatively affecting the impact of DSA on graft survival in some studies 90 but not all. 83 In series where DSA are associated with worse allograft survival, 82,83,88,[92][93][94][95] DSA MFI is an imperfect discriminator of outcomes.…”

Section: Utility Of Hla Antibody Testing Pretransplantmentioning

confidence: 94%

Utility of HLA Antibody Testing in Kidney Transplantation

Konvalinka¹,

Tinckam

2015

Journal of the American Society of Nephrology

165

135

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HLA antigens are polymorphic proteins expressed on donor kidney allograft endothelium and are critical targets for recipient immune recognition. HLA antibodies are risk factors for acute and chronic rejection and allograft loss. Solid-phase immunoassays for HLA antibody detection represent a major advance in sensitivity and specificity over cell-based methods and are widely used in organ allocation and pretransplant risk assessment. Post-transplant, development of de novo donorspecific HLA antibodies and/or increase in donor-specific antibodies from pretransplant levels are associated with adverse outcomes. Although single antigen bead assays have allowed sensitive detection of recipient HLA antibodies and their specificities, a number of interpretive considerations must be appreciated to understand test results in clinical and research contexts. This review, which is especially relevant for clinicians caring for transplant patients, discusses the technical aspects of single antigen bead assays, emphasizes their quantitative limitations, and explores the utility of HLA antibody testing in identifying and managing important pre-and post-transplant clinical outcomes.

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Section: Utility Of Hla Antibody Testing Pretransplantmentioning

confidence: 94%

Utility of HLA Antibody Testing in Kidney Transplantation

Konvalinka¹,

Tinckam

2015

Journal of the American Society of Nephrology

165

135

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“…We recently demonstrated the relationship between quantitative assessment of DSAs using the molecules of equivalent soluble fluorochrome (MESF) assay and the rate of graft rejection after transplantation. 11 However, in human clinical renal transplantation, there are various immunological backgrounds and immunosuppressive agents, and obtaining multiple biopsies from a patient has risks and is costly.…”

mentioning

confidence: 99%

Sequential analysis of donor-specific antibodies and pathological findings in acute antibody-mediated rejection in a rat renal transplantation model

Kohei

Tanabe

Horita

et al. 2013

Kidney International

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Alloantibodies contribute significantly to renal transplant rejection by activation of complement and various cytokines with a variety of effector cells, and are a major cause of allograft loss. Although there is clinical evidence of antibody- and complement-mediated injury in renal transplantation, the mechanism of antibody-mediated rejection remains largely unknown. In order to understand the sequential production of antibodies and complement components, we presensitized recipient rats by skin transplantation. Anti-donor-specific IgG levels reached a maximum 2 weeks following presensitization after which the rats underwent renal transplantation from the same donor strain. We then evaluated sequential pathological findings based on the Banff classification and several factors related to graft rejection. In this presensitized model, peritubular capillaries were already dilated and stained for C4d. Neutrophil and mononuclear cell infiltration in these capillaries was detected beginning 2 h after transplantation. Donor-specific antibody IgG levels decreased rapidly and anti-IgG antibody stained glomerular and peritubular capillaries in the grafts beginning 2 h after transplantation. Additionally, several cytokines and complement components showed marked changes in the presensitized group. Thus, in the donor-specific presensitized recipient, alloantibodies and complement were activated immediately after transplant. C4d deposition in peritubular capillaries appears to be a key factor for the diagnosis of antibody-associated rejection.

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“…There is a strong trend towards the employment of multiplexed diagnostic methods in various clinical demands, such as detection of HLA antigens for transplants, chronic leukemias, microbiological diagnostic, and allergies as well as detection of hepatitis C virus [25, 28, 29]. The combination of microspheres and flow cytometry as a tool for diagnosis has not been solely used in the identification of human diseases.…”

Section: Discussionmentioning

confidence: 99%

“…Nowadays, QFCM is applied in instrument calibration and linearity verification [22], as well as in immunotherapy studies [18, 37], evaluation of membrane antigens expression in HIV [38] or leukemia patients [24, 39, 40], anti-HLA antibodies for kidney transplantation [25] and platelet-derived microparticles analysis [23]. There are different applications and manufacturers of microspheres for QFCM studies, as well as different types of evaluation that can be performed [22].…”

Section: Discussionmentioning

confidence: 99%

“…MESF studies are based on the equivalence between the number of fluorochromes in two solutions, where one can be a suspension of labeled microspheres and the other an unknown sample [21]. The applications of QFCM include calibration and linearity verification of instruments [22] as well as research and diagnostic studies [23–25]. Despite the different techniques proposed for this quantitative evaluation, there is a consensus on the requirement of efficient standardization of the methodology with respect to instruments and reagents (antibodies and microspheres), since at least for reagents the limit of detection will be necessarily fixed at the highest possible population of the MESF kits applied [26, 27].…”

Section: Introductionmentioning

confidence: 99%

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Flow Cytometry as a Tool for Quality Control of Fluorescent Conjugates Used in Immunoassays

et al. 2016

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The use of antibodies in immunodiagnostic kits generally implies the conjugation of these proteins with other molecules such as chromophores or fluorochromes. The development of more sensitive quality control procedures than spectrophotometry is essential to assure the use of better fluorescent conjugates since the fluorescent conjugates are critical reagents for a variety of immunodiagnostic kits. In this article, we demonstrate a new flow cytometric protocol to evaluate conjugates by molecules of equivalent soluble fluorochromes (MESF) and by traditional flow cytometric analysis. We have coupled microspheres with anti-IgG-PE and anti-HBSAg-PE conjugates from distinct manufactures and/or different lots and evaluated by flow cytometry. Their fluorescence intensities were followed for a period of 18 months. Our results showed that there was a great difference in the fluorescence intensities between the conjugates studied. The differences were observed between manufactures and lots from both anti-IgG-PE and anti-HBSAg-PE conjugates. Coefficients of variation (CVs) showed that this parameter can be used to determine better coupling conditions, such as homogenous coupling. The MESF analysis, as well as geometric mean evaluation by traditional flow cytometry, showed a decrease in the values for all conjugates during the study and were indispensable tools to validate the results of stability tests. Our data demonstrated the feasibility of the flow cytometric method as a standard quality control of immunoassay kits.

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Significance of Qualitative and Quantitative Evaluations of Anti-Hla Antibodies in Kidney Transplantation

Cited by 8 publications

References 0 publications

Utility of HLA Antibody Testing in Kidney Transplantation

Utility of HLA Antibody Testing in Kidney Transplantation

Sequential analysis of donor-specific antibodies and pathological findings in acute antibody-mediated rejection in a rat renal transplantation model

Flow Cytometry as a Tool for Quality Control of Fluorescent Conjugates Used in Immunoassays

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