Please enter verification code
Characterization of the Anti-HLA Class I and II IgG Antibodies in Moroccan IVIg Using Regular Beads and Ibeads in Luminex Multiplex Single Antigen Immunoassay
International Journal of Immunology
Volume 5, Issue 4, August 2017, Pages: 53-65
Received: May 15, 2017; Accepted: May 23, 2017; Published: Jul. 18, 2017
Views 2865      Downloads 213
Fatiha EL Hilali, Department of Biology, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Vadim Jucaud, Terasaki Foundation Laboratory, California, USA
Hajar EL Hilali, Department of Biology, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Mohammed Hassan Bhuiyan, Department of Biology, College of Staten Island, City University of New York, New York, USA
Andrew Mancuso, Department of Biology, College of Staten Island, City University of New York, New York, USA
Nancy LiuSullivan, Department of Biology, College of Staten Island, City University of New York, New York, USA
Abdeslem Elidrissi, Department of Biology, College of Staten Island, City University of New York, New York, USA
Hamid Mazouz, Department of Biology, Faculty of Sciences, Moulay Ismail University, Meknes, Morocco
Article Tools
Follow on us
Therapeutic Immunoglobulin Intravenous (IVIg), approved to treat a wide range of autoimmune and primary immunodeficiency diseases, contain mixture of polyreactive and polyclonal IgG purified from the pooled plasma of thousands of donors. The aim of this study is to characterize the profiles of anti- Human Leukocyte Antigen (HLA) class-I and class-II IgG antibodies in four lots of Moroccan IVIg preparations using Luminex Multiplex Single Antigen Bead Immunoassay and to compare it with the unique high frequency HLA types found in the Moroccan population. Anti-HLA class I IgG profiles were assessed using regular (Labscreen) Beads and iBeads. The regular beads are coated with all conformational and structural variants of HLA-I (HLA heavy chain (HC) with β2-microglobulin (β2m) with or without peptides, β2m-free HC with or without peptides or HC only), quite contrast to iBeads, which contained only native tissue-associated HLA HC with β2m and with or without peptides. The level of antibodies was measured as Mean Fluorescent Intensity (MFI). The reactivity of anti-HLA-I IgG antibodies to different alleles of HLA-I loci differed in their recognition of native HLA-I and other structural variants of the HLA-I. High MFI IgG antibodies in the IVIg corresponded with several high frequency HLA-I alleles (B*0801, B*5001, Cw*0602 and Cw*0702) and HLA-II haplotypes (DQA1*0201-DQB1*0201/DRB1*0301), which accounted for 50% of the total gene frequencies in the Moroccan population. HLA-I reactivity of the IVIg with iBeads confirms that the IgG reacting to normal tissue associated with peptide -associated or -free β2mHC. These findings caution the use of high dose IVIg for the carriers of the high frequency HLA types for it may cause tissue injury. The β2m-free-HC reactivity of IVIg indicates the potential of IVIg to bind to activated T and B cells that express these variants, to suppress antibody production. Such an immunomodulation by IVIg renders benefit for patients with autoimmune diseases and organ transplantation.
Intravenous Immunoglobulin, HLA, Antibodies, Moroccan IVIg, Beads, Ibeads, MFI
To cite this article
Fatiha EL Hilali, Vadim Jucaud, Hajar EL Hilali, Mohammed Hassan Bhuiyan, Andrew Mancuso, Nancy LiuSullivan, Abdeslem Elidrissi, Hamid Mazouz, Characterization of the Anti-HLA Class I and II IgG Antibodies in Moroccan IVIg Using Regular Beads and Ibeads in Luminex Multiplex Single Antigen Immunoassay, International Journal of Immunology. Vol. 5, No. 4, 2017, pp. 53-65. doi: 10.11648/j.iji.20170504.11
Copyright © 2017 Authors retain the copyright of this article.
This article is an open access article distributed under the Creative Commons Attribution License ( which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
A. A. Bousfina, L. Jeddane, A. Conicno-Nato, Primary Immunodeficiencies in developing countries. In: Primary Immunodeficiency disorders. A historic and Scientific Perspective. (Eds.), Etzioni A, Ochs HD, Elsevier Inc. USA, 2014, pp. 65-76.
F. El Hilali, H. El Hilali, M. F. Belahsen, A. El Idrissi, H. Mazouz, “Uses of Intravenous Immunoglobulin: A 13-Year Evaluation of Guillain–Barré Syndrome in Fez, Morocco,” J Biol Med, vol. 1, pp. 001-005, 2017.
HLA nomenclature., 2017 (accessed 06.05.17).
F. Choukri, A. Chakib, H. Himmich, H. Raissi, S. Caillat-Zucman, “HLA class I polymorphism in a Moroccan population from Casablanca,” Eur J Immunogenet, vol. 29, pp. 205-11, 2002.
D. Piancatelli, A. Canossi, A. Aureli, K. Oumhani, T. Del Beato, M. Di Rocco, G. Liberatore, A. Tessitore, K. Witter, R. El Aouad, D. Adorno, “Human leukocyte antigen-A, -B, and -Cw polymorphism in a Berber population from North Morocco using sequence-based typing,” Tissue Antigens, vol. 63, pp. 158-72, 2004.
H. Izaabel, H. J. Garchon, S. Caillat-Zucman, G. Beaurain, O. Akhayat, J. F Bach, A. Sanchez-Mazas, “HLA class II DNA polymorphism in a Moroccan population from the Souss, Agadir area,” Tissue Antigens, vol.51, pp. 106-10, 1998.
E. Gómez-Casado, P. del Moral, J. Martínez-Laso, A. García-Gómez, L. Allende, C. Silvera-Redondo, J. Longas, M. González-Hevilla, M. Kandil, J. Zamora, A. Arnaiz-Villena, “HLA genes in Arabic-speaking Moroccans: close relatedness to Berbers and Iberians,” Tissue Antigens, vol. 55, pp. 239-49, 2000.
K. Oumhani, A. Canossi, D. Piancatelli, M. Di Rocco, T. Del Beato, G. Liberatore, A. Aureli, A. Benjoaud, R. El Aouad, D. Adorno, C. U. Casciani, “Sequence-Based analysis of the HLA-DRB1 polymorphism in Metalsa Berber and Chaouya Arabic-speaking groups from Morocco,” Hum Immunol, vol. 63, pp. 129-38, 2002.
M. Kabbaj, M. Oudghiri, A. Naya, H. Naamane, S. Bennani, “Polymorphism of human leukocyte antigen-A, -B, and -DRB1 in a Moroccan population from Casablanca: study of the allelic and the haplotypic frequencies,” Ann Biol Clin (Paris), vol. 69, pp. 295-301, 2011.
C. Brick, O. Atouf, M. Essakalli, “The HLA system in the Moroccan population: General review,” Transfus Clin Biol, vol. 22, pp. 299-311, 2015.
Z. V. Collins, P. F. Arnold, F. Peetoom, G. S. Smith, R. L. Walford, “A naturally occurring monospecific anti-HL-A8 isoantibody,” Tissue Antigens, vol. 3, pp. 358–363, 1973.
V. Lepage, L. Degos, J. Dausset, “A “natural” anti-HLA-A2 antibody reacting with homozygous cells,” Tissue Antigens, vol. 8, pp.139–142, 1976.
M. M. Tongio, A. Falkenrodt, Y. Mitsuishi, A. Urlacher, J. P. Bergerat, M. L. North, S. Mayer, “Natural HLA antibodies,” Tissue Antigens, vol. 26, pp. 271–285, 1985.
F. Ameglio, F. Saba, A. Bitti, A. Aceti, N. Tanigaki, R. Sorrentino, A. Dolei, R. Tosi, “Antibody reactivity to HLA classes I and II in sera from patients with hydatidosis,” J. Infect. Dis, vol. 156, pp. 673–676, 1987.
A. Ma´jsky´, “Natural HLA-A, B and DR antibodies in the serum of nonimmunized men,” Vnitr. Lek, vol. 35, pp. 467–471, 1989.
M. A. Luscher, G. Choy, J. E. Embree, N. J. Nagelkerke, J. J. Bwayo, S. Njenga, F. A. Plummer, B. H. Barber, K. S. Mac Donald, “Anti-HLA alloantibody is found in children but does not correlate with a lack of HIV type 1 transmission from infected mothers,” AIDS Res. Hum. Retroviruses, vol.14, pp. 99–107, 1998.
B. Zhou, S. Saito, Y. Nakazawa, N. Kobayashi, M. Matsuda, Y. Matsumoto, T. Hosoyama, K. Koike, “Existence of an immunoglobulin G component of naturally occurring HLA class I antibodies that are not directed against self-antigens in human serum,” Tissue Antigens, vol. 72, pp. 98–104, 2008.
L. E. Morales-Buenrostro, P. I. Terasaki, L. A. Marino-Va´zquez, J. H. Lee, N. El-Awar, J. Alberu´, ““Natural” human leukocyte antigen antibodies found in nonalloimmunized healthy males,” Transplantation, vol. 86, pp. 1111–1115, 2008.
M. H. Ravindranath, H. Kaneku, N. El-Awar, L. E. Morales-Buenrostro, P. I. Terasaki, “Antibodies to HLA-E in nonalloimmunized males: pattern of HLA-Ia reactivity of anti-HLA-E-positive sera,” J Immunol, vol. 185, pp. 1935-48, 2010.
M. H. Ravindranath, P. I. Terasaki, C. Y. Maehara, V. Jucaud, S. Kawakita, T. Pham, W. Yamashita, “Immunoglobulin (Ig)G purified from human sera mirrors intravenous Ig human leucocyte antigen (HLA) reactivity and recognizes one's own HLA types, but may be masked by Fab complementarity-determining region peptide in the native sera,” Clin Exp Immunol, vol. 179, pp. 309-3282015.
M. H. Ravindranath, V. Jucaud, N. Banuelos, M. J. Everly, J. Cai, A. Nguyen, P. I. Terasaki, Nature and Clonality of the Fluoresceinated Secondary Antibody in Luminex Multiplex Bead Assays Are Critical Factors for Reliable Monitoring of Serum HLA Antibody Levels in Patients for Donor Organ Selection, Desensitization Therapy, and Assessment of the Risk for Graft Loss. J Immunol. 2017, DOI:
D. W. King, E. Reed, N. Suciu-Foca, “Complexes of soluble HLA antigens and anti-HLA autoantibodies in human sera: possible role in maintenance of self-tolerance,” Immunol Res, vol. 8, pp. 249-62, 1989.
K. Zeki, F. Shirakawa, T. Fujihira, M. Kanatani, K. Watanabe, H. Suzuki, S. Eto, “Circulating monocyte (macrophage)-specific antibodies in patients with autoimmune thyroid diseases,” Clin Endocrinol (Oxf), vol. 31, pp. 1-13, 1989.
M. C. Dooren, W. H. Ouwehand, A. J. Verhoeven, et al., “Adult respiratory distress syndrome after experimental intravenous gamma-globulin concentrate and monocyte-reactive IgG Abs,” Lancet, vol. 352, pp. 1601-1602, 1998.
A. Rizk, K. C. Gorson, L. Kenney, R. Weinstein, “Transfusion-related acute lung injury after the infusion of IVIG,” Transfusion, vol. 41, pp. 264-268, 2001.
P. V. Voulgari, S. Paschou, E. Svarna, et al, « Images in rheumatology. Transfusion-related acute lung injury during intravenous immunoglobulin treatment,” Journal of Rheumatology, vol. 37, pp. 190-19, 2010.
V. Gupta, P. Gupta, T. P. Yadav, “Transfusion related acute lung injury with intravenous immunoglobulin,” Indian Pediatrics, vol. 48, pp. 807-808, 2011.
D. R. Reddy, P. K. Guru, M. M. Blessing, J. R. Stubbs, A. A. Rabinstein, E. F. Wijdicks, “Transfusion-Related Acute Lung Injury after IVIG for Myasthenic Crisis,” Neurocrit Care, vol. 23, pp. 259-261, 2015.
R. Kumar, M. J. Sedky, S. J. Varghese, O. E. Sharawy, “Transfusion Related Acute Lung Injury (TRALI): A Single Institution Experience of 15 Years,” Indian J. Hematol. Blood Transfusion, vol. 32, pp. 320-327, 2016.
P. M. Kopko, T. G. Paglieroni, M. A. Popovsky, “TRALI: correlation of antigen-Ab and monocyte activation in donor-recipient pairs,” Transfusion, vol. 43, pp. 177–184, 2003.
P. M. Kopko, P. V. Holland, “Transfusion-related acute lung injury,” British Journal of Haematology, vol. 105, pp. 322–329, 2011.
U. J. Sachs, “A threshold model for the susceptibility to transfusion-related acute lung injury,” Transfusion Clinical Biology, vol. 19, pp. 109–116, 2012.
D. Glotz, J. P. Haymann, N. Sansonetti, et al, “Suppression of HLA-specific alloAbs by high dose intravenous immunogloubulins (IVIg),” Transplantation, vol. 56, pp. 335-337, 1993.
D. Glotz, C. Antoine, J. P. Haymann, P. Julia, A. Doboust, J. Bariety, “Intravenous immunogloubulins and Kidney transplantation in patients with anti-HLA antibodies,” Adv. Nephrol. Necker Hosp, vol. 30, pp. 221-233, 2000.
N. El-Awar, A. Nikaein, M. Everly, J. Hopefield, A. Nguyen, “A Novel HLA Class I Single Antigen Bead Preparation Eliminates False Positive Reactions Attributed to Natural Antibodies – in the Sera of Normal Males and Pre-Transplant Patients,” Hum Immunol, vol. 71 (2010) S26. DOI: 10.1016/j.humimm.2010.06.060.
J. Szenczi, J. Kardos, A. Gyorgy, P. Závodszky, “The effect of solvent environment on the conformation and stability of human polyclonal IgG in solution,” Biologicals, vol. 34, pp. 5–14, 2006.
W. K. Bleeker, J. L. Teeling, A. J. Verhoeven, G. M Rigter, J, Agterberg, A. T. Tool, A. H. Koenderman, T. W. Kuijpers, C. E. Hack, “Vasoactive side effects of intravenous immunoglobulin preparations in a rat model and their treatment with recombinant platelet-activating factor acetylhydrolase,” Blood, vol. 95, pp. 1856–61, 2000.
United States Patent Number: 5,871,736. Bruegger et al., Date of Patent‘. Feb. 16, 1999. Liquid immunoglobulin foreign patent documents formulations.
D. Fenyo, Q. Wang, J. De Grasse, J. C. Padova, M. Cadene, B. T. Chait, MALDI Sample Preparation: the Ultra Thin Layer Method. JoVE. e192. 2007.
Maldi-sample-preparation-the-ultra-thin-layer-method. maldi-sample-preparation-the-ultra-thin-layer-method. 2017 (accessed 06.05.17).
M. H. Ravindranath, P. I. Terasaki, T. Pham, V. Jucaud, S. Kawakita, “Therapeutic preparations of IVIg contain naturally occurring anti-HLA-E antibodies that react with HLA-Ia (HLA-A/-B/-Cw) alleles,” Blood, vol. 121, pp. 2013-2028, 2013.
J. Visentin, G. Guidicelli, T. Nong, M. J. Moreau. P. Merville, C. Lionel, J. Lee, J. Taupin, “Evaluation of the iBeads assay as a tool for identifying class I HLA antibodies,” Human Immunol, vol. 76, pp. 651-6, 2015.
V. Jucaud V, M. H. Ravindranath, P. I. Terasaki, “Conformational Variants of the Individual HLA-I antigens on Luminex Single Antigen Beads used in Monitoring HLA Antibodies: Problems and Solutions”, Transplantation, vol. 101, pp. 764- 77, 2017.
A. Zerrouki, S. Ouadghiri, N. Benseffaj, R. Razine, M. Essakalli, “High background in Luminex® assay for HLA antibody screening: Interest of Adsorb Out™,” Transpl Immunol, vol. 36, pp. 20-24, 2016.
E. Schnabl, H. Stockinger, O. Majdic, H. Gaugitsch, I. J. Lindley, D. Maurer, A. Hajek Rosenmayr, W. Knapp, “Activated human T lymphocytes express MHC class I heavy chains not associated with beta 2-microglobulin,” J Exp Med, vol. 171, pp. 1431-1442, 1990.
J. Madrigal, A. Belichm, M. P. Benjamin, R. J, Little, A. M, Hildebrand, W. H, Mann, D. L. Parham, “Molecular definition of a polymorphic antigen (LA45) of free HLA-A and -B heavy chains found on the surfaces of activated B and T cells,” J Exp Med. 174, 1085-1095, 1991.
S. Demaria, R. Schwab, Y. Bushkin, “The origin and fate of beta 2m-free MHC class I molecules induced on activated T cells,” Cell Immunol, vol. 142, pp. 103-113, 1992.
D. Zhu, M. H. Ravindranath, P. I. Terasaki, T. Miyazaki, T. Pham, V. Jucaud, “Suppression of allo-human leucocyte antigen (HLA) Abs secreted by B memory cells in vitro: intravenous immunoglobulin (IVIg) versus a monoclonal anti-HLA-E IgG that mimics HLA-I reactivities of IVIg,” Clin. Exper. Immunol, vol. 177, pp. 464–477, 2014.
M. H. Ravindranath, P. I. Terasaki, C. Y. Maehara et al.,“Immunoglobulin (Ig)G purified from human sera mirrors intravenous Ig human leucocyte antigen (HLA) reactivity and recognizes one’s own HLA types, but may be masked by Fab complementarity determining region peptide in the native sera,” Clin. Exper. Immunol, vol. 179, pp. 309–328, 2015.
Powerpoint presentation of The ALPHA-TRAXTM Program.
J. S. Finlayson, B. L. Armstrong, A. M. Young, “Reversibility of human immunoglobulin G dimerization,” Acta Radiol Suppl, vol. 310, pp. 114-23, 1971.
E. M. Yoo, L. A. Wims, L. A. Chan, S. L. Morrison, “Human IgG2 can form covalent dimers,” J Immunol, vol. 170, pp. 3134-8, 2003.
A. McAuley, J. Jacob, C. G. Kolvenbach, K. Westland, H. J. Lee, S. R. Brych, D. Rehder, G. R. Kleemann, D. N. Brems, M. Matsumura, “Contributions of a disulfide bond to the structure, stability, and dimerization of human IgG1 antibody CH3 domain,” Protein Sci, vol. 17, pp. 95-106, 2008.
Science Publishing Group
1 Rockefeller Plaza,
10th and 11th Floors,
New York, NY 10020
Tel: (001)347-983-5186