Human C3 Deficiency Associated with Impairments in Dendritic Cell Differentiation, Memory B Cells, and Regulatory T Cells This information is current as of September 27, 2021. Arije Ghannam, Martine Pernollet, Jean-Luc Fauquert, Nicole Monnier, Denise Ponard, Marie-Bernadette Villiers, Josette Péguet-Navarro, Arlette Tridon, Joel Lunardi, Denis Gerlier and Christian Drouet J Immunol 2008; 181:5158-5166; ; Downloaded from doi: 10.4049/jimmunol.181.7.5158 http://www.jimmunol.org/content/181/7/5158 http://www.jimmunol.org/ References This article cites 47 articles, 17 of which you can access for free at: http://www.jimmunol.org/content/181/7/5158.full#ref-list-1 Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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The Journal of Immunology Human C3 Deficiency Associated with Impairments in Dendritic Cell Differentiation, Memory B Cells, and Regulatory T Cells1 Arije Ghannam,* Martine Pernollet,† Jean-Luc Fauquert,‡ Nicole Monnier,§ Denise Ponard,¶ Marie-Bernadette Villiers,ʈ Josette Pe´guet-Navarro,# Arlette Tridon,‡ Joel Lunardi,§ Denis Gerlier,* and Christian Drouet2** Primary C3 deficiency, a rare autosomal inherited disease (OMIM 120700), was identified in a 2-year-old male suffering from recurrent pyogenic infections from early infancy with undetectable total complement hemolytic activity (CH50) and C3 values. The nonconsanguineous parents and the two patients’ two siblings had 50% normal serum C3 concentration. The molecular abnormality associated a paternal allele coding C3 with the missense mutation p.Ser550Pro and an apparently null maternal allele, Downloaded from with production of a defective protein that could no longer be secreted. Vaccination of the child did not induce a long-term Ab response. Accordingly, switched memory IgD؊CD27؉ B cells were barely detected, amounting to only 2.3% of peripheral blood -CD19؉ cells. Cells were significantly defective in stimulating alloreactive responses. The in vitro development of immature den dritic cells and their maturation capacity were greatly impaired, with decreased CD1a expression and IL-12p70 secretion ability. These cells were unable to induce autologous B cell proliferation and Ig secretion in the presence of CD40L and C3. Finally, the /regulatory T cell development ability of CD4؉ T cells after CD3 and CD46 activation in the presence of IL-2 was significantly http://www.jimmunol.org impaired. Thus, the association of important functional defects of dendritic cells, acquisition of B cell memory, and regulatory T cells with human C3 deficiency strongly supports a major role for C3 in bridging innate and adaptive immunity in humans. The Journal of Immunology, 2008, 181: 5158–5166. he activation of C3 and subsequent production of various 120700) was recognized as being associated with a striking sus- proteolytic fragments are crucial and a strategic function ceptibility to pyogenic infections (1, 5). The rare cases of complete T for both innate and adaptive immune defense. The liver is C3 deficiency are associated with large deletions, microdeletions, the main site of C3 synthesis (1), which is increased during acute or mutations in splice sequences (for review, see Ref. 4). Het- by guest on September 27, 2021 inflammation. Diffusible amounts are also produced by activated erozygous individuals with missense mutations are at risk for non- monocytes, macrophages (2), and dendritic cells (DCs)3 (3, 4). infectious diseases, such as membranoproliferative glomerulone- Human complete C3 deficiency with two affected alleles (OMIM phritis (5), age-related macular degeneration (6), and atypical hemolytic and uremic syndrome (7). Evidence that C3 is involved in adaptive immunity was demonstrated in C3–/– animals, which *VirPatH, Universite´de Lyon, Universite´Lyon1, Centre National de la Recherche exhibited a substantial reduction in germinal center (GC) numbers, † Scientifique FRE3011, Lyon, France; Etablissement Franc¸ais du Sang, Grenoble, low IgG Ab titers after virus infection (8), and impaired T-inde- France; ‡Centre Hospitalier Universitaire Clermont-Ferrand, Universite´d’Auvergne, Clermont-Ferrand, France; §Centre Hospitalier Universitaire Grenoble, Institut Na- pendent and T-dependent responses (9–12). The activation and the tional de la Sante´et de la Recherche Me´dicale, Unite´836, Universite´Joseph Fourier, covalent attachment of C3 to an Ag enhance the specific B cell Grenoble, France; ¶Laboratoire d’Immunologie, Centre Hospitalier Universitaire Grenoble, Grenoble, France; ʈInstitut National de la Sante´et de la Recherche Me´di- response by at least two mechanisms. On one hand, coligation of cale, Unite´823, Universite´Joseph Fourier, Grenoble, France; #EA 41-69, Universite´ the CD21/CD19/CD81 coreceptors lowers the threshold for B cell Lyon1, Hoˆpital E. Herriot, Lyon, France; and **GREPI/TIMC-IMAG Centre Na- activation both in vivo and ex vivo (13, 14), and the ligation of tional de la Recherche Scientifique Unite´Mixte de Recherche 5525, Universite´Joseph Fourier, Centre Hospitalier Universitaire Grenoble, Grenoble, France CD21 is required for the survival of activated B cells within the Received for publication May 27, 2008. Accepted for publication August 1, 2008. follicles and GCs (10). In contrast, the complement receptor-de- pendent retention of Ag on follicular DCs provides an Ag source The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance for the clonal selection within the GC (10, 15, 16). The direct with 18 U.S.C. Section 1734 solely to indicate this fact. coupling of C3d or C3b to a protein Ag lowers the amount of 1 This work was partly supported by Grant PHRC 2002 from the French Ministry of protein required for an optimal Ab response by as much as 10,000- Health (to C.D.). A.G. is a recipient of a fellowship from Tishreen University, Syria. fold (17, 18) and leads to more stable IgG production and better J.-L.F. took care of the family; A.G., M.P., N.M., and D.P. performed experiments; memory stimulation (19), resulting in an improved, long-lasting A.G., M.-B.V., J.P.-N., A.T., J.L., and D.G. analyzed results; A.G., D.G., and C.D. made the figures; and A.G., D.G., and C.D. designed the research and wrote the paper. response (20). Similarly, the coupling of C3d to the influenza hem- Part of this work was presented as an oral communication at the 11th European agglutinin in a DNA vaccine enhances the humoral response (21). Meeting on Complement in Human Diseases (Cardiff, U.K.), with the abstract iden- C3 also appears to be critical for optimal Ag presentation in allo- tification: Ghannam et al. 2007. Absence of memory B cells and unbalanced T cell responses in a C3 deficient patient. Mol. Immunol. 44: 3910 (Abstr.). reactive responses (3, 22, 23). Considering the importance of C3 in 2 Address correspondence and reprint requests to Dr. Christian Drouet, GREPI/ TIMC-IMAG CNRS UMR5525, Universite´Joseph Fourier, CHU Grenoble, PO Box complement-induced regulatory T cell; GC, germinal center; PPD, tuberculin purified 217, Grenoble, F-38043 France. E-mail address: [email protected] derivative; SNP, single nucleotide polymorphisms; Treg, regulatory T cell. 3 Abbreviations used in this paper: DC, dendritic cell; ACM, Ag preparation from Candida albicans; AT, tetanus toxoid; CH50, complement hemolytic activity; cTreg, Copyright © 2008 by The American Association of Immunologists, Inc. 0022-1767/08/$2.00 www.jimmunol.org The Journal of Immunology 5159 the recovery from primary infection, the costimulation of B cells Ag proliferation assays via the CD21/CD19/CD81 coreceptors to recruit leukocytes at the PBMCs were prepared by density-gradient centrifugation over lymphocyte infection site has been suggested as a minor pathway (24). In con- separation medium (Eurobio). A total of 50 ␮g/ml tetanus toxoid (AT; ϩ ϩ trast, the ability of C3 to enhance specific CD4 and CD8 T cell Aventis Pasteur), 2500 UI/ml tuberculin purified derivative (PPD; Aventis responses can be critical in mediating antiviral protection (24, 25). Pasteur), or 25 ␮g/ml Ag preparation from Candida albicans (ACM; Bio- ϫ 5 ␮ All of these findings may have implications in vaccine Rad) was added to 1 10 PBMCs in triplicate in 96-well plates (200 l; BD Falcon). Mitogen (0.5 ␮g/ml)-activated cells and nonstimulated cells development. were used as positive mitogenic and negative control, respectively. The In the present study, we characterized a complete C3 deficiency proliferation was monitored after an 8-h [3H]thymidine (1.0 ␮Ci/ml) in- in a nonconsanguineous French family. In addition to a new mis- corporation at day 6 for each Ag. Tests were conducted in triplicate, and sense C3 mutant, our results identify several new immune dys- the results were expressed as mean net cpm Ϯ SD. Each experiment was conducted twice, and the experiments shown are representative of all functions (4). An important defect of memory B cells was found, the data. associated with the impairment of vaccine Ab production. The in vitro differentiation of myeloid DCs was greatly impaired. The Alloreactive mixed culture complement-induced regulatory T cells (Tregs) were lacking in the Responder and allogeneic irradiated (30 Gy) stimulator PBMCs were young patient and his heterozygous parents.