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Characterization of HLA Antibodies by C3d Assay and IgG Subclasses in Sensitized Kidney Transplant Patients

A. Casillas-Abundis, J. M. Arreola-Guerra, J. Alberu, M. I. Solis-G, L. Llorente, J. Granados, M. Vilatoba, R. Cruz-Martinez, L. Morales-Buenrostro, A. G. Contreras

Instituto Nacional de Ciencias Medicas y Nutricion SZ, Mexico City, Mexico

Meeting: 2019 American Transplant Congress

Abstract number: A155

Keywords: Highly-sensitized, HLA antibodies, IgG, Monoclonal antibodies

Session Information

Date: Saturday, June 1, 2019

Session Name: Poster Session A: Biomarkers, Immune Monitoring and Outcomes

Session Time: 5:30pm-7:30pm

 Presentation Time: 5:30pm-7:30pm

Location: Hall C & D

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  • Comparison Between IgG, C1q and C3d Single Antigen Bead Assays in Detecting Class I Complement-Binding Anti-HLA Antibodies.

*Purpose: To determine the detection rate of IgG subclasses (IgGsc) and C3d-binding HLA-antibodies (abs) in a cohort of highly sensitized kidney transplant patients.

*Methods: We evaluated, in cryopreserved sera from 90 potential renal transplant recipients, HLA-abs Class I and Class II using single antigen beads (SAB). C3d was detected by Luminex (Lifecodes®), IgG 1-4 subclasses were evaluated using monoclonal abs for each IgGsc (Luminex).

*Results: The SAB were able to reveal 1,684 abs, additionally 63 were only detected by C3d assay and 24 by IgGsc (n=1771).The IgGsc assay showed 367 abs with at least one IgGsc, the most common was IgG1 alone or associated with other subclasses (351, 75.6%); only 7 IgG2 and 9 IgG3 were found isolated form IgG1. IgG4 was identified in 17 abs, always in combination with IgG1. A MFI >4000 of the HLA-SAB was directly related with the detection of one or more IgGsc (*p=<0.001). Figure 1.The C3d assay exposed 216 complement-binding abs; 128 of these were identified in the IgGsc as well. Table 1 depict a more detailed analysis of each Class I and Class II category, its IgG detection and C3d-binding capacity.A multivariate analysis using different models showed that the best predictors of IgGsc detection were: finding an HLA-DQ ab and the positivity of the C3d assay (OR, (CI 95%) 1.72 (1.18-2.51), p= 0.004 and 7.41 (5.4-10.1), p= <0.001). Comparatively, finding ≥2 IgGsc was the best predictor for a positive C3d.

*Conclusions: The ability of the IgG monoclonal abs to detect IgGsc depends on the MFI of the HLA-SAB; a high MFI improves the assessment of IgGsc. The HLA-DQ abs that were C3d positive had a higher probability of detection of IgGsc than any other combination. The classification of abs as one or more IgGsc increases the risk of these being C3d positive.

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Table 1. Results of IgGsc detection and C3d-binding.

All Abs

(n=1771)

IgG subclasses Detection (n=367)Positive C3d assay (n=216)
HLA-Ab Class II, n (%)636 (35.91)116 (31.6)161 (74.5)
HLA A, n (%)291 (16.43)69 (18.8)23 (10.6)
HLA B, n (%)566 (31.96)145 (39.51)22 (10.1)
HLA C, n (%)278 (15.7)37 (10.08)10 (4.63)
HLA DP, n (%)156 (8.81)1 (0.27)12 (5.56)
HLA DQ, n (%)170 (9.6)63 (17.17)52 (24.07)
HLA DR, n (%)310 (17.5)52 (14.17)97 (44.9)

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To cite this abstract in AMA style:

Casillas-Abundis A, Arreola-Guerra JM, Alberu J, Solis-G MI, Llorente L, Granados J, Vilatoba M, Cruz-Martinez R, Morales-Buenrostro L, Contreras AG. Characterization of HLA Antibodies by C3d Assay and IgG Subclasses in Sensitized Kidney Transplant Patients [abstract]. Am J Transplant. 2019; 19 (suppl 3). https://atcmeetingabstracts.com/abstract/characterization-of-hla-antibodies-by-c3d-assay-and-igg-subclasses-in-sensitized-kidney-transplant-patients/. Accessed December 5, 2019.

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