Comparison of different IVIg preparations on IL-17 production by human Th17 cells Mohan S. Maddur a,b,c , Srini V. Kaveri a,b,c,d, ⁎, Jagadeesh Bayry a,b,c,d, ⁎ a Unité 872, Institut National de la Santé et de la Recherche Médicale Paris, F-75006, France b Centre de Recherche des Cordeliers, Equipe 16, Immunopathology and Therapeutic Immunointervention, Université Pierre et Marie Curie - Paris 6, UMR S 872, 15 rue de l'Ecole de Médicine, Paris, F-75006, France c Université Paris Descartes, UMR S 872, Paris, F-75006, France d International Associated Laboratory IMPACT, Institut National de la Santé et de la Recherche Médicale -France and Indian Council of Medical Research - India, National Institute of Immunohaemotology, Mumbai, India abstract article info Available online 2 March 2011 Variations in intravenous immunoglobulin (IVIg) products are a common feature. In this report, we compared the effect of different IVIg products on IL-17 production by human Th17 cells. We found that irrespective of products, IgGs of all IVIg preparations were equally effective in inhibiting the production of IL-17A from Th17 cells. © 2011 Elsevier B.V. All rights reserved. Intravenous immunoglobulin (IVIg) is a therapeutic preparation of pooled normal polyspecific IgG obtained from plasma pools of thousands of healthy blood donors. In addition to patients with primary immunodeficiencies, IVIg is used as a therapeutic agent in large number of autoimmune, allergic and inflammatory diseases [1-3]. Several mutually nonexclusive mechanisms have been described for the therapeutic efficacy of IVIg [1,2,4-6]. Unlike recombinant proteins, variations in IVIg products are a common feature. A recent article by Ahmed R et al., elegantly outlines the differences between commercially available IVIg products with respect to various parameters including stabilizing agents and formula- tion [7]. The variations in the source of plasma and IgG purification methods have been shown to influence the immunoglobulin content to infectious agents and the outcome of infection rate in immunodeficiency patients [8-10]. However, much data is not available on comparison of immunoregulatory properties of IVIg in the context of autoimmune and inflammatory conditions. Since, IgG molecules are the principal compo- nents of IVIg that mediate therapeutic effect, we compared the effect of IgG molecules of different IVIg products on IL-17A production by human Th17 cells. Th17 cells, a recently identified subset of Th cells that produce IL-17, mediate the clearance of extracellular pathogens such as Klebsiella and Candida. In addition, several reports also demonstrate that Th17 cells and IL-17 play a crucial role in the pathogenesis of various autoimmune, allergy and inflammatory conditions [11-17]. Since IVIg is beneficial in diseases such as Kawasaki disease, systemic lupus erythematosus, anti- neutrophil cytoplasmic antibody (ANCA)-associated vasculitis, derma- tomyositis and pemphigus, where Th17 cells are also implicated in the pathogenesis [1,18,19], we aimed at comparing the effect of various IVIg preparations on the production of IL-17A by Th17 cells. CD4 + Th cells were purified from peripheral blood mononuclear cells of healthy blood donors. For differentiation of Th17 cells, CD45RA + CD25 - naïve Th cells were stimulated in X-VIVO serum-free medium with anti- CD3 and anti-CD28 monoclonal antibodies and cytokines TGF-β (5 ng/ ml) and IL-21 (25 ng/ml) for 6 days. For amplification of Th17 cells, CD45RO + memory Th cells were stimulated in X-VIVO serum-free medium with anti-CD3 and anti-CD28 monoclonal antibodies and cytokines IL-1β (12.5 ng/ml) and IL-6 (25 ng/ml) for 6 days [20]. Various IVIg preparations (Sandoglobulin, Gamunex, Privigen, Octagam, and Tegeline) were dialyzed in large volumes of PBS and RPMI-1640 medium and were added at a concentration of 25 mg/ml/0.25 million cells after 12 h initiation of culture as described previously [20]. Sandoglobulin and Tegeline are freeze-dried IVIg products while others are liquid prepara- tions. The stabilizing agents are sucrose in Sandoglobulin, maltose in Octagam, L-proline in Privigen, glycine in Gamunex and saccharose in Tegeline. As shown in Fig. 1A and B, we found that irrespective of products, IgGs of all IVIg preparations were equally effective in inhibiting the production of IL-17A either from differentiating or expanding Th17 cells. The effect of IVIg was more prominent on inhibition of IL-17 from differentiating Th17 cells. Together, our data indicate that under ex-vivo experimental conditions, the IVIg products are similar in their mechanisms. However, data from patients are warranted to corroborate these experimental results that will help to formulate the guidelines for the use of specific IVIg preparations in various autoimmune diseases. Acknowledgements Supported by grants from the Institut National de la Santé et de la Recherche Médicale (INSERM), Centre National de la Recherche Autoimmunity Reviews 10 (2011) 809–810 ⁎ Corresponding authors at: INSERM U 872, Equipe 16-Centre de Recherche des Cordeliers, 15 rue de l'Ecole de Médicine, Paris, F-75006, France. Tel.: +33 1 55 42 82 66; fax: +33 1 55 42 82 62. E-mail addresses: srini.kaveri@crc.jussieu.fr (S.V. Kaveri), jagadeesh.bayry@crc.jussieu.fr (J. Bayry). 1568-9972/$ – see front matter © 2011 Elsevier B.V. All rights reserved. doi:10.1016/j.autrev.2011.02.007 Contents lists available at ScienceDirect Autoimmunity Reviews journal homepage: www.elsevier.com/locate/autrev