Surface Plasmon Resonance Analysis Shows an IgG-Isotype-Specific Defect in ABO Blood Group Antibody Formation in Patients with Common Variable Immunodeficiency

Abstract

BackgroundCommon variable immunodeficiency (CVID) is the most common clinically severe primary immunodeficiency and comprises a heterogeneous group of patients with recurrent severe bacterial infections due to the failure to produce IgG antibodies after exposure to infectious agents and immunization. Diagnostic recommendations for antibody failure include assessment of isoagglutinins. We have readdressed this four decades old but still accepted recommendation with up to date methodology.MethodsAnti-A/B IgM- and IgG-antibodies were measured by Diamed-ID Micro Typing, surface plasmon resonance (SPR) using the Biacore® device and flow cytometry.ResultsWhen Diamed-ID Micro Typing was used, CVID patients (n = 34) showed IgG- and IgM-isoagglutinins that were comparable to healthy volunteers (n = 28), while all XLA patients (n = 8) had none. Anti-A/B IgM-antibodies were present in more than 2/3 of the CVID patients and showed binding kinetics comparable to anti-A/B IgM-antibodies from healthy individuals. A correlation could be found in CVID patients between levels of anti-A/B IgM-antibodies and levels of serum IgM and PnP–IgM-antibodies. In contrast in CVID patients as a group ABO antibodies were significantly decreased when assessed by SPR, which correlated with levels of switched memory, non-switched memory and naïve B cells, but all CVID patients had low/undetectable anti-A/B IgG-antibodies.ConclusionThese results indicate that conventional isoagglutinin assessment and assessment of anti-A/B IgM antibodies are not suited for the diagnosis of impaired antibody production in CVID. Examination of anti-A/B IgG antibodies by SPR provides a useful method for the diagnosis of IgG antibody failure in all CVID patients studied, thus indicating an important additional rationale to start immunoglobulin replacement therapy early in these patients, before post-infectious sequelae develop.

Pneumococcal Polysaccharide Vaccination Elicits IgG Anti-A/B Blood Group Antibodies in Healthy Individuals and Patients with Type I Diabetes Mellitus

“…Therefore, we used SPR to measure anti-A/B antibodies as previously described (6, 11, 12). Amine-labeled blood group A and B trisaccharides (Dextra Laboratories Ltd., Reading, UK) were immobilized on two-channel sensor chips CM5 (General Electric Health Care, Freiburg, Germany) using standard protocols (31, 32).…”

“…Amine-labeled blood group A and B trisaccharides (Dextra Laboratories Ltd., Reading, UK) were immobilized on two-channel sensor chips CM5 (General Electric Health Care, Freiburg, Germany) using standard protocols (31, 32). In brief, the CM5 sensor chip surface was activated with 100 μl 0.05 mol/l N -hydroxy-succinimide and 0.2 mol/l N -ethyl- N ′-dimethylamino-propylcarbodiimide injected into the buffer stream of HBS-EP (0.01 mol/l HEPES buffer, pH 7.4, containing 0.15 mol/l NaCl, 3 mmol/l EDTA, and 0.005% surfactant P20) as described previously (6). The buffer stream was passed through FC-1 and FC-2 of the two-channel Biacore ® X instrument (GE Health Care) at a flow rate of 5 μl/min for 20 min.…”

“…Antibodies to carbohydrate epitopes share certain characteristics with natural antibodies, e.g., the absence of extensive class switching and affinity maturation due to absent T-cell help (5). Isoagglutinins, i.e., antibodies against carbohydrate epitopes that form the AB0 antigens on red blood cells (RBCs), are considered prototypic natural antibodies that were shown to be compromised in patients with immunodeficiency (1, 4, 6). Their appearance can be explained by inapparent stimulation particularly from intestinal bacteria because antigens cross-reactive with blood group polysaccharides are widely distributed in the environment (1, 7, 8).…”

“…In the present study, we investigated whether vaccination with Pneumo 23 Vaccine “Pasteur Merieux” (Pn23) has an effect on blood group ABO antibodies in healthy individuals and in patients with DM I. To screen for blood group anti-A/B antibodies, we used the commercially available DiaMed-ID Micro Typing System based on erythrocyte agglutination (6). To measure blood group-specific anti-A/B antibodies in healthy individuals and in patients with DM I in an isotype-specific manner, we used surface plasmon resonance (SPR) technology and synthetic A/B trisaccharides bound via amine-coupling to the CM5 chip (6, 11, 12).…”

“…To screen for blood group anti-A/B antibodies, we used the commercially available DiaMed-ID Micro Typing System based on erythrocyte agglutination (6). To measure blood group-specific anti-A/B antibodies in healthy individuals and in patients with DM I in an isotype-specific manner, we used surface plasmon resonance (SPR) technology and synthetic A/B trisaccharides bound via amine-coupling to the CM5 chip (6, 11, 12). SPR using blood group-specific A/B synthetic trisaccharides is the ideal technology to investigate a potential IgM to IgG isotype shift and to investigate changes in binding characteristics of the relevant blood group anti-A/B antibodies under near to physiological conditions in real time.…”