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

2017 ◽  
Vol 49 (9) ◽  
pp. 2031-2035 ◽  
Author(s):  
M.A. Moreno Gonzales ◽  
D.G. Mitema ◽  
B.H. Smith ◽  
C.A. Schinstock ◽  
M.D. Stegall ◽  
...  
Keyword(s):  
Class I ◽  
Hla Antibodies ◽  
Single Antigen

C1q-Binding HLA Antibodies Do Not Predict Platelet Transfusion Failure in TRAP Study Participant

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1562-1562
Author(s):  
Rachael P. Jackman ◽  
Douglas Bolgiano ◽  
Mila Lebedeva ◽  
Sherrill J. Slichter ◽  
Philip J. Norris
Keyword(s):  
Conflicts Of Interest ◽  
Fluorescent Antibody ◽  
Study Participant ◽  
Class I ◽  
Detection Methods ◽  
Antibody Detection ◽  
Hla Antibodies ◽  
Luminex Assay ◽  
Single Antigen ◽  
Antibody Levels

Abstract Introduction: In the Trial to Reduce Alloimmunization to Platelets (TRAP) study, 101 of the 530 subjects became clinically refractory (CR) to platelet transfusions in the absence of detectable antibodies against HLA as measured by the lymphocytotoxicity assay (LCA). Using more sensitive bead-based detection methods we have previously demonstrated that while many of these LCA- patients do have anti-HLA antibodies, that these low to moderate level antibodies do not predict refractoriness. In addition to being less sensitive then bead based methods, the LCA screen only detects complement-binding antibodies. As these antibodies could be important for platelet rejection, we assessed if previously undetected complement-binding antibodies could account for some of the refractoriness seen in LCA- patients. Methods: 169 LCA- (69 CR, 100 non-CR) and 20 LCA+ (10 CR, 10 non-CR) subjects were selected from the TRAP study. Anti-class I HLA IgG and C1q binding antibodies were measured in serum or plasma using two different multi-analyte, semi-quantitative, bead-based fluorescent antibody detection assays: the LabScreen mixed Luminex assay, and the LabScreen single antigen class I assay with or without added EDTA (One Lambda). Groups were compared using an unpaired t-test, a=0.05, and correlation between variables was also assessed. Results: New measurements of anti-class I HLA IgG antibodies reliably reproduced earlier data with a strong correlation between the old and new measurements (R2=0.9736, p<0.0001). While some of the LCA- subjects did have detectable C1q-binding anti-class I HLA antibodies, and some LCA+ subjects did not, levels of these antibodies were significantly higher among LCA+ subjects (p<0.0001). Levels of C1q-binding anti-class I HLA antibodies were not significantly different between CR and non-CR among either the LCA- or LCA+ subjects. Conclusions: While complement-binding anti-class I HLA antibody levels were higher in the LCA+ subjects, higher levels of these antibodies were not seen in CR LCA+ patients as compared with non-CR LCA+ patients. Complement-binding anti-class I HLA antibodies do not account for refractoriness seen among the LCA- TRAP subjects. This work confirms that low to mid level anti-class I antibodies do not drive refractoriness to platelets, and suggests that antibody-independent mechanisms cause refractoriness in patients lacking higher levels of anti-HLA antibodies. Disclosures No relevant conflicts of interest to declare.

ADSORPTION OF CYTOTOXIC ANTI-HLA ANTIBODIES WITH HLA CLASS I IMMUNOSORBANT BEADS

1990 ◽  
Vol 49 (5) ◽  
pp. 925-930 ◽  
Author(s):  
LYNN D. DEVITO ◽  
HANS W. SOLLINGER ◽  
WILLIAM J. BURLINGHAM
Keyword(s):  
Hla Class I ◽  
Class I ◽  
Hla Antibodies

Reassessment of T Lymphocytes Crossmatches Results Prediction With Luminex Class I Single Antigen Flow Beads Assay

2017 ◽  
Vol 101 (3) ◽  
pp. 624-630 ◽  
Author(s):  
Jonathan Visentin ◽  
Thomas Bachelet ◽  
Cécile Borg ◽  
Nicolas Franchini ◽  
Thoa Nong ◽  
...  
Keyword(s):  
T Lymphocytes ◽  
Class I ◽  
Single Antigen

Anti-HLA antibodies interfere in the detection of soluble HLA class I molecules

1999 ◽  
Vol 60 (5) ◽  
pp. 414-423 ◽  
Author(s):  
Carin A Koelman ◽  
Wilma Ensink ◽  
Arend Mulder ◽  
Janneke Tanke ◽  
Ilias I.N Doxiadis ◽  
...  
Keyword(s):  
Hla Class I ◽  
Class I ◽  
Hla Antibodies ◽  
Hla Class I Molecules ◽  
Soluble Hla

scholarly journals Sex-Specific Differences in HLA Antibodies after Pneumococcal Vaccination in Kidney Transplant Recipients

Vaccines ◽  
2019 ◽  
Vol 7 (3) ◽  
pp. 84 ◽  
Author(s):  
Lindemann ◽  
Oesterreich ◽  
Wilde ◽  
Eisenberger ◽  
Muelling ◽  
...  
Keyword(s):  
Kidney Transplant ◽  
De Novo ◽  
Pneumococcal Vaccination ◽  
Human Leukocyte ◽  
Hla Class I ◽  
Current Data ◽  
Class I ◽  
Transplant Recipients ◽  
Kidney Transplant Recipients ◽  
Hla Antibodies

In transplant recipients vaccination against Streptococcus pneumoniae is recommended to reduce mortality from invasive pneumococcal disease. It is still debated if vaccination in transplant recipients triggers alloresponses. Therefore, it was our aim to define if vaccination with Prevenar 13®, a 13-valent, conjugated pneumococcal vaccine (Pfizer, New York, NY, USA) that acts T cell dependently, induces human leukocyte antigen (HLA) antibodies in clinically stable kidney transplant recipients. Forty-seven patients were vaccinated once with Prevenar 13® and HLA antibodies were determined prior to vaccination and at month 1 and 12 thereafter. In parallel, pneumococcal IgG antibodies were measured. Using Luminex™ Mixed Beads technology (One Lambda/Thermo Fisher, Canoga Park, CA, USA) we observed overall no change in HLA antibodies after vaccination. Pneumococcal antibodies increased significantly at month 1 (p < 0.0001) and remained elevated at month 12 (p < 0.005). A more detailed analysis of HLA antibodies showed that in 18 females HLA class I and II antibodies increased significantly at month 1 and 12 (p < 0.05); whereas in 29 males HLA class I and II antibodies tended to decrease. Using Luminex™ Single Antigen Beads assay, no de novo donor-specific HLA antibodies were detected after vaccination. In conclusion, the current data indicate that females may be more susceptible to the induction of (non-specific) HLA antibodies after vaccination.

DETECTION OF AUTOANTIBODIES AND ALLOANTIBODIES AGAINST PLATELET-ASSOCIATED ANIGENS

1987 ◽  
Author(s):  
R McMillan ◽  
P Tani ◽  
P Berchtold ◽  
F Millard
Keyword(s):  
Hla Antigens ◽  
Class I ◽  
High Dose ◽  
Hla Antibodies ◽  
Negative Results ◽  
Single Donor ◽  
Prognostic Group ◽  
Successful Outcomes ◽  
Well Assay ◽  
High Degree

Although autoantibodies against the platelet glycoproteins (GP) and a 1loantibodies toward class I HLA antigens have been demonstrated using various methods, practical techniques for their detection have not been available.We studied 59 patients with chronic immune thrombocytopenic purpura (ITP) where platelet-associated and plasma autoantibodies against the GP Ilb/IIIa complex and GP lb were measured using a newly developed imraunobead assay and a previously described microtiter well assay. The specificity of both assays depends on the monoclonal antibody employed (e.g., GPIIb/IIIa, lb or HLA). Platelet-associated autoantibody was detected using the immunobead assay in 21 of 28 patients (13 with anti-GPIIb/IIIa and 8 with anti-GPIb). Plasma autoantibodies were noted in 34 of 59 patients (21 with anti-GPIIb/IIIa, 11 with anti-GPIb and 2 with both). Positive results were noted in 30 of 59 ITP patients using the immunobead assay and in only 14 of 59 using the microtiter well assay, suggesting that solubilization prior to antibody addition, as in the microtiter well assay, alters epitope stability. However, the microtiter well assay appears to define a poor prognostic group. Of 11 patients with positive well assays who underwent splenectomy, only one achieved a complete remission, 3 have died and all require continuous high-dose steroids or immunosuppressants. Of the 31 thrombocytopenic controls studied, all gave negative results using both assays.The immunobead assay was adapted to specifically measure anti-HLA antibodies and was used to evaluate 51 single donor platelet transfusions given to 7 patients refractory to random platelets. Twenty-nine of 33 (88%) transfusions associated with a negative assay had successful outcomes (1 hr increment >7500) while only 2 of 18 (11.1%) episodes associated with a positive test had successful outcomes. Only 1 unsuccessful transfusion episode was associated with a negative immunobead and a positive antiglobulin test suggesting that transfusion-related alloantibodies, other than to class I HLA antigens, are an uncommon cause of refractoriness.We conclude that these two clinically adaptable assays are capable of measuring both autoantibodies and anti-HLA alloantibodies with a high degree of sensitivity.

scholarly journals The Kinetics of Anti-HLA Antibodies in the First Year after Kidney Transplantation: In Whom and When Should They Be Monitored?

2018 ◽  
Vol 2018 ◽  
pp. 1-6 ◽  
Author(s):  
Maria Cristina Ribeiro de Castro ◽  
Erick A. Barbosa ◽  
Renata P. Souza ◽  
Fabiana Agena ◽  
Patrícia S. de Souza ◽  
...  
Keyword(s):  
Acute Rejection ◽  
De Novo ◽  
First Year ◽  
Sharp Drop ◽  
Hla Antibodies ◽  
Single Antigen ◽  
The Impact ◽  
Kinetics Of ◽  
Time Point ◽  
Rejection Rates

The impact of the kinetics of the anti-HLA antibodies after KTx on the occurrence of acute rejection as well as the better time-point to monitor anti-HLA Abs after transplantation is not completely defined. This prospective study followed 150 patients over 12 months after transplantation. Serum IgG anti-HLA Abs were detected by single antigen beads after typing donors and recipients for loci A, B, C, DR, and DQ. Before KTx, 89 patients did not present anti-HLA Abs and 2% developed “de novo” Abs during the 1st year, 39 patients were sensitized without DSAs, and 13% developed DSA after surgery; all of them presented ABMR. Sensitized patients presented higher acute rejection rates (36.4% versus 13.5%, p<0.001), although 60% of the patients did not present ABMR. Patients, in whom DSA-MFI decreased during the first two weeks after surgery, did not develop ABMR. Those who sustained their levels presented a rate of 22% of ABMR. 85% of patients developed ABMR when MFIs increased early after transplantation (which occurred in 30% of the DSA positive patients). In the ABMR group, we observed an iDSA-MFI sharp drop on the fourth day and then an increase between the 7th and 14th POD, which suggests DSA should be monitored at this moment in sensitized patients for better ABMR prediction.

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