HIV Type 1 Glycoprotein 120 Inhibits Human B Cell Chemotaxis to CXC Chemokine Ligand (CXCL) 12, CC Chemokine Ligand (CCL)20, and CCL21 This information is current as of October 6, 2021. Gamal Badr, Gwenoline Borhis, Dominique Treton, Christiane Moog, Olivier Garraud and Yolande Richard J Immunol 2005; 175:302-310; ; doi: 10.4049/jimmunol.175.1.302 http://www.jimmunol.org/content/175/1/302 Downloaded from References This article cites 64 articles, 39 of which you can access for free at: http://www.jimmunol.org/content/175/1/302.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! Every submission reviewed by practicing scientists • Fast Publication! 4 weeks from acceptance to publication by guest on October 6, 2021 *average Subscription Information about subscribing to The Journal of Immunology is online at: http://jimmunol.org/subscription Permissions Submit copyright permission requests at: http://www.aai.org/About/Publications/JI/copyright.html Email Alerts Receive free email-alerts when new articles cite this article. Sign up at: http://jimmunol.org/alerts The Journal of Immunology is published twice each month by The American Association of Immunologists, Inc., 1451 Rockville Pike, Suite 650, Rockville, MD 20852 Copyright © 2005 by The American Association of Immunologists All rights reserved. Print ISSN: 0022-1767 Online ISSN: 1550-6606. The Journal of Immunology HIV Type 1 Glycoprotein 120 Inhibits Human B Cell Chemotaxis to CXC Chemokine Ligand (CXCL) 12, CC Chemokine Ligand (CCL)20, and CCL211 Gamal Badr,* Gwenoline Borhis,* Dominique Treton,* Christiane Moog,† Olivier Garraud,‡ and Yolande Richard2* We analyzed the modulation of human B cell chemotaxis by the gp120 proteins of various HIV-1 strains. X4 and X4/R5 gp120 inhibited B cell chemotaxis toward CXCL12, CCL20, and CCL21 by 40–50%, whereas R5 gp120 decreased inhibition by 20%. This gp120-induced inhibition was strictly dependent on CXCR4 or CCR5 and lipid rafts but not on CD4 or VH3-expressing BCR. Inhibition did not impair the expression or ligand-induced internalization of CCR6 and CCR7. Our data suggest that gp120/ CXCR4 and gp120/CCR5 interactions lead to the cross-desensitization of CCR6 and CCR7 because gp120 does not bind CCR6 ␤ and CCR7. Unlike CXCL12, gp120 did not induce the activation of phospholipase C 3 and PI3K downstream from CXCR4, Downloaded from whereas p38 MAPK activation was observed. Similar results were obtained if gp120-treated cells were triggered by CCL21 and CCL20. Our results are consistent with a blockade restricted to signaling pathways using phosphatidylinositol-4,5-bisphosphate as a substrate. X4 and X4/R5 gp120 induced the cleavage of CD62 ligand by a mechanism dependent on matrix metalloproteinase ␣ 1 and 3, CD4, CXCR4, G i, and p38 MAPK, whereas R5 gp120 did not. X4 and X4/R5 gp120 also induced the relocalization of cytoplasmic CD95 to the membrane and a 23% increase in CD95-mediated apoptosis. No such effects were observed with R5 gp120. The gp120-induced decrease in B cell chemotaxis and CD62 ligand expression, and increase in CD95-mediated B cell http://www.jimmunol.org/ apoptosis probably have major deleterious effects on B cell responsiveness during HIV infection and in vaccination trials. The Journal of Immunology, 2005, 175: 302–310. uman immunodeficiency virus-1 infection is associated a superantigen to conserved VH3 framework regions, activating with strong polyclonal B cell activation, increasing the and depleting these cells (17, 18). As VH3 genes are the key de- H percentage of activated B cells in the periphery (1–5), terminants of Abs specific for bacterial polysaccharide Ags, the and with strong, sustained follicular hyperplasia in secondary lym- depletion of VH3-expressing B cells may contribute to the observed phoid organs during the asymptomatic phase of the disease (6–8). increase in the incidence of secondary infections in HIV-infected pa- The B cells of HIV-infected patients spontaneously secrete Ig but tients (19). Scamurra et al. (10) recently showed in vivo that the de- by guest on October 6, 2021 are unable to mount a T cell-dependent B cell response (1, 9, 10). pletion of peripheral VH3-expressing memory B cells, but not of naive Antiretroviral therapy decreases HIV-1-driven B cell hyperactiv- B cells, in HIV-infected patients involves gp120 binding. ity, polyclonal B cell activation, and Ig production in patients (11). Lymphocyte recirculation, which is critical for effective immu- These findings strongly suggest that the sustained replication of nity, is tightly regulated by the expression of adhesion molecules HIV-1 affects differentiation in lymphoid tissue. The mechanisms and chemoattractant receptors on lymphocytes, combined with the by which HIV-1 impairs the humoral response may result from spatial and temporal expression of ligands for these receptors by a intrinsic B cell defects or dysfunctional dialogue between T and B variety of tissue cells (20–23). Human B cells express several cells. Soluble Tat and gp120, biologically active extracellular pro- chemokine receptors, including CXCR4, CXCR5, CCR6, and teins released by HIV-1-infected cells, induce intrinsic defects in CCR7, and respond to their cognate ligands CXCL12, CXCL13, human B cells (12–15). We previously showed that Tat selectively CCL20, and CCL21 and CCL19, respectively. Triggering of the inhibits the BCR-mediated proliferation of naive and memory B BCR, CD40 and the IL-4R modulates chemokine receptor expres- cells, and the production of cytokines and Ig. In contrast, Tat dou- sion and chemotaxis in B cells (24–26). Chemokines can bind to 3 ␣ bles the germinal center cell proliferation induced by CD40 mAb various pertussis toxin (PTX) -sensitive (G i) and PTX-insensi- ␣ ␣ ␣ plus IL-4 (16). The HIV-1 envelope protein gp120 may bind like tive (G q and G 15/16)G proteins, but chemotaxis is only ob- ␣ served upon activation of G i protein-coupled receptors (22). There is strong evidence that G␤␥, rather than G␣ subunits initiate *Institut National de la Sante´et de la Recherche Medicale, Unite´131, Institut Paris- i Sud sur les Cytokines, Clamart, France; †Universite´ Louis Pasteur, Strasbourg, the chemotactic response (27). We recently showed that human B France; and ‡Groupe Immunite´ des Muqueuses et Agents Pathoge`nes, Faculte´de cell chemotaxis to CXCL12, CXCL13, CCL21, and CCL20 de- Me´decine, Universite´Jean Monnet, Saint-Etienne, France pends on PI3K, phospholipase C (PLC)␤3, protein kinase C Received for publication November 1, 2004. Accepted for publication April 20, 2005. (PKC), RhoA, and NF-␬B. Although activated by chemokine re- The costs of publication of this article were defrayed in part by the payment of page ceptor-ligand interactions, neither p38 MAPK nor ERK1/2 are in- charges. This article must therefore be hereby marked advertisement in accordance ␤␥ with 18 U.S.C. Section 1734 solely to indicate this fact. volved in B cell chemotaxis (28). G stimulation also activates G 1 This work was supported by grants from the Institut National de la Sante´etdela Recherche Me´dicale, Universite´Paris-Sud, and Sidaction. 2 Address correspondence and reprint requests to Dr. Yolande Richard, Institut 3 Abbreviations used in this paper: PTX, pertussis toxin; PLC, phospholipase C; PKC, National de la Sante´et de la Recherche Me´dicale, Unite´131, Universite´Paris- protein kinase C; MFI, mean fluorescence intensity; CD62L, L-selectin; MARCKS, myr- Sud, 32 rue des Carnets, 92 140 Clamart, France. E-mail address: yolande.richard istoylated alanine-rich C kinase substrate; M␤CD, methyl-␤-cyclodextrin; MMP, matrix @ipsc.u-psud.fr metalloproteinase; PIP2, phosphatidylinositol-4,5-bisphosphate. Copyright © 2005 by The American Association of Immunologists, Inc. 0022-1767/05/$02.00 The Journal of Immunology 303 protein-coupled receptor kinases, which phosphorylate chemokine For in vitro culture assays, B cells (2 ϫ 106 cells/ml) were cultured in receptors, inducing their binding to ␤-arrestin (29) and their rapid RPMI 1640 (Invitrogen SARL) supplemented with 10 mM HEPES, 2 mM ␮ internalization (30). L-glutamine, 100 U/ml penicillin, 100 g/ml streptomycin, 1 mM sodium pyruvate, and 10% heat-inactivated FCS (complete medium) for 2 h, with We analyzed the effect of recombinant CXCR4-binding (X4), or without recombinant 10 nM gp120 (unless otherwise indicated). Pre- CCR5-binding (R5), or dual-binding (X4/R5) gp120 on the che- liminary experiments showed that B cell chemotaxis was maximally in- motactic response of human primary B cells. We found that gp120 hibited after2hofincubation with 10 nM gp120 from the various HIV-1 strongly inhibited the chemotactic response to CCL20 and CCL21, strains (data not shown). without inducing spontaneous apoptosis or impairing the expres- Flow cytometry sion and ligand-induced internalization of CCR6 and CCR7. B cell chemotaxis was inhibited more strongly by X4 and X4/R5 gp120 Chemokine receptor expression was analyzed by flow cytometry, using than by R5 gp120, consistent with CXCR4 being the more strongly PE-conjugated anti-CCR6 (clone 53103.111, IgG2b), anti-CCR5 (clone 45531.111, IgG2b), anti-CCR7 (clone 150503, IgG2a), and anti-CXCR4 expressed of these two receptors. The use of Abs blocking (clone 44717.111, IgG2b) mAbs from R&D Systems. PE-conjugated CD19 CXCR4, CCR5, or gp120 totally prevented the gp120-induced in- (IgG1) and CD95 (IgG1) mAbs and FITC- and PE-conjugated mouse iso- hibition of B cell chemotaxis whereas soluble CD4 slightly in- type-matched control IgG were purchased from BD Biosciences. FITC- creased it. In contrast to what was observed with CXCL12, we conjugated CD62L (clone FMC46, IgG2a) was purchased from Diaclone. To detect cytoplasmic CD95 expression, we treated B cells with the Cyto- found that X4 or X4/R5 gp120 did not activate the CXCR4- fix/Cytoperm kit (BD Biosciences) before staining with PE-conjugated dependent phosphorylation of PLC␤3, PI3K/AKT, ERK1/2, and CD95 mAb or mouse IgG1.