Subclass of Donor-specific Antibody as a Risk Factor of Antibody Mediated Rejection in Renal Transplantation

Study Overview
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Brief Summary
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Antibody-mediated rejection is now recognized as the first cause of long-term kidney transplant loss. This type of rejection is mediated by the presence of graft-specific antibodies, usually directed against HLA (Human Leukocyte Antigens), called DSA (Donor Specific Antibody). De novo DSA (ie, post-transplantation) is detected in approximately 20% of transplant recipients in the first five years, and is a major risk factor for antibody mediated rejection and graft loss. All anti-HLA antibodies therefore do not have the same pathogenicity. Some teams have shown that the detection of IgG3 anti-HLA by cytometry is associated with a higher risk of humoral rejection but these results have not been confirmed by others. One of the limitations of the cytometry by Luminex technique is that it only informs the detection of each subclass but does not allow analysis of the distribution of the different subclasses of a DSA. A method has been developed for the relative detection and quantification of different subclasses of the DSA using the mass spectrometry technique and will be tested during this study. This new quantification method therefore opens up the prospect of studying whether, not only the presence but especially the distribution of IgG subclasses, in particular IgG3, could constitute a reliable and robust marker of humoral rejection.

Detailed Description
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Antibody-mediated rejection is now recognized as the first cause of long-term kidney transplant loss. This type of rejection is mediated by the presence of graft-specific antibodies, usually directed against HLA (Human Leukocyte Antigens), called DSA (Donor Specific Antibody). It results from the interaction between DSA and HLA present on the surface of graft endothelial cells, complement activation, endothelial cell activation and recruitment of inflammatory cells within the renal microcirculation leading to a graft dysfunction. De novo DSA (ie, post-transplantation) is detected in approximately 20% of transplant recipients in the first five years, and is a major risk factor for antibody mediated rejection and graft loss. However, the presence of a DSA is not always associated with humoral rejection. All anti-HLA antibodies therefore do not have the same pathogenicity. The antibody titre (assessed by the fluorescence average (MFI) on Luminex beads) is involved in the risk of rejection but is far from explaining the disparities in clinical evolution. DSA are mainly IgG of different subclasses whose distribution could have a major impact on their pathogenicity. Indeed, complement-dependent cytotoxicity (CDC) and antibody-dependent cellular cytotoxicity (ADCC) functions differ according to IgG subclass. IgG3 is the subclass that has the greatest potential for complement activation followed by IgG1. IgG3 and IgG1 are also the two subclasses with the best affinities for the FcγRIIIa activating receptor for ADCC mediated by Natural Killer (NK) cells. Some teams have shown that the detection of IgG3 anti-HLA by cytometry is associated with a higher risk of humoral rejection but these results have not been confirmed by others. One of the limitations of the cytometry by Luminex technique is that it only informs the detection of each subclass but does not allow analysis of the distribution of the different subclasses of a DSA. A method has been developed for the relative detection and quantification of different subclasses of the DSA using the mass spectrometry technique and will be tested during this study. This new quantification method therefore opens up the prospect of studying whether, not only the presence but especially the distribution of IgG subclasses, in particular IgG3, could constitute a reliable and robust marker of humoral rejection.

Official Title
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Characterization of the Subclass of Donor-specific Antibody Determined in Mass Spectrometry as a Risk Factor of Antibody Mediated Rejection in Renal Transplantation

Conditions
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Antibody-mediated Rejection

Participation Criteria
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Eligibility Criteria
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Inclusion Criteria: - Patients over 18 years old Renal transplant patients followed at the University Hospital of Montpellier presenting a DSA de novo during follow-up. Affiliation or beneficiary of a social security scheme. Inclusion criteria specific to patients whose graft biopsy has already been performed Patients who consented to the collection of a plasma sample on graft biopsy day and use in future programs (Collection DC-2014-2328) Collection of non-opposition to participate in the study Inclusion criteria specific to patients whose biopsy of the graft has not already been performed: Eligible for graft biopsy for humoral-mediated rejection (according to Banff 2015 classification) (Appendix 1) Collection of non-opposition to participate in the study Signature of informed consent to participate in the collection of a plasma sample on graft biopsy day and use in future programs (Collection DC-2014-2328) Exclusion Criteria: Pregnant or lactating women according to Article L1121 5 of the health public Code Vulnerable persons according to article L1121-6 of the health public Code Major persons placed under guardianship or curator

Ages Eligible for Study
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Minimum Age: 18 Years

Sexes Eligible for Study
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All

Accepts Healthy Volunteers
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No

Study Plan
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How is the study designed?
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Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| patient with kidney transplant<br>Blood sample will be took from subjects during this research | |

What is the study measuring?
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Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| distribution of DSA IgG subclasses | the performance of the distribution of DSA IgG subclasses for the diagnosis of humoral rejection in renal transplant patients with Donor Specific Antibodies | day 0 (inclusion visit) | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| histological criteria for humoral rejection | Link between the distribution of DSA IgG subclasses and histological criteria for humoral rejection (according to the Banff International Classification 2015) | day 0 (inclusion visit) | 
| degradation of graft function | Link between the distribution of DSA IgG subclasses and degradation of graft function (glomerular filtration rate loss of more than 25%) | 1 year after inclusion of subjects | 
| graft loss | Link between the distribution of DSA IgG subclasses and graft loss | 1 year after inclusion of subjects | 

 Terms related to the study
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Keywords Provided by Centre Hospitalier Valida
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Renal transplantation, antibody-mediated rejection, DSA, mass spectrometry technique