Herpes Simplex Virus Type 1 Targets the MHC Class II Processing Pathway for Immune Evasion This information is current as Jürgen Neumann, Anna Maria Eis-Hübinger and Norbert of October 2, 2021. Koch J Immunol 2003; 171:3075-3083; ; doi: 10.4049/jimmunol.171.6.3075 http://www.jimmunol.org/content/171/6/3075 Downloaded from References This article cites 53 articles, 23 of which you can access for free at: http://www.jimmunol.org/content/171/6/3075.full#ref-list-1 http://www.jimmunol.org/ Why The JI? Submit online. • Rapid Reviews! 30 days* from submission to initial decision • No Triage! 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The Journal of Immunology Herpes Simplex Virus Type 1 Targets the MHC Class II Processing Pathway for Immune Evasion1 Ju¨rgen Neumann,* Anna Maria Eis-Hu¨binger,† and Norbert Koch2* HSV type 1 (HSV-1) has evolved numerous strategies for modifying immune responses that protect against infection. Important targets of HSV-1 infection are the MHC-encoded peptide receptors. Previous studies have shown that a helper T cell response and Ab production play important roles in controlling HSV-1 infection. The reduced capacity of infected B cells to stimulate CD4؉ T cells is beneficial for HSV-1 to evade immune defenses. We investigated the impact of HSV-1 infection on the MHCII processing pathway, which is critical to generate CD4؉ T cell help. HSV-1 infection targets the molecular coplayers of MHC class II processing, HLA-DR (DR), HLA-DM (DM), and invariant chain (Ii). HSV-1 infection strongly reduces expression of Ii, which impairs formation of SDS-resistant DR-peptide complexes. Residual activity of the MHC class II processing pathway is diminished by viral envelope glycoprotein B (gB). Binding of gB to DR competes with binding to Ii. In addition, we found gB associated with DM molecules. Both, gB-associated DR and DM heterodimers are exported from the endoplasmic reticulum, as indicated by Downloaded from carbohydrate maturation. Evaluation of DR, DM, and gB subcellular localization revealed abundant changes in intracellular distribution. DR-gB complexes are localized in subcellular vesicles and restrained from cell surface expression. The Journal of Immunology, 2003, 171: 3075–3083. he immune system is highly efficient in defending against heterodimers. B cells play a vital role in the humoral defense of viral infection. The virus-host relationship, however, is HSV-1 infection and might also be a target for virus evasion strat- http://www.jimmunol.org/ T balanced by virus evasion strategies (1). Infected cells egies (5). The transfer of mAb against glycoprotein D to T cell display viral sequences that are bound to MHC receptors at the cell subset-depleted mice demonstrated that Abs provide protection surface and are recognized by T cells. CD8ϩ T cells detect viral against HSV-1-induced disease (6). To counteract the host immune peptides derived from the biosynthesis of infected cells. Hence, system, HSV-1 might influence the presentation of viral sequences biosynthesis of viral proteins is directly linked to formation of by MHCII polypeptides. MHC class I (MHCI)3-peptide complexes exposed on cell mem- Unlike MHCI molecules that use cytoplasmic degradation as a branes. HSV type 1 (HSV-1) infections elicit both cytotoxic and source for peptides, MHCII heterodimers acquire their antigenic Ab responses. One mechanism employed by the virus to prevent peptides in endosomal/lysosomal compartments. Maturation of cytotoxic responses to HSV-1 infection is early inhibition of MHCII molecules along their biosynthetic route involves degra- by guest on October 2, 2021 MHCI molecule transport to cell surfaces (2, 3). Binding of the dation and release of the associated invariant chain (Ii) and renders HSV-1 polypeptide ICP47 to the TAP transporter inhibits peptide the heterodimer susceptible to peptide acquisition (7, 8). A final translocation from the cytosol to the endoplasmic reticulum (ER) step in activation is achieved by the nonclassical MHCII polypep- lumen and prevents presentation of viral sequences by MHCI mol- tide DM. DM heterodimers do not appear to bind peptides (9). ecules. Such presentation is important to elicit a CD8ϩ T cell re- Their task is to release a fragment of Ii, which is lodged in the sponse (3, 4). In addition to a cytotoxic response, an Ab response MHCII groove (10, 11). In addition, DM molecules edit peptides and additional defense mechanisms, such as the release of IFNs, for stable MHCII complexes and for other structural factors (12). provide protection against virus dissemination and reinfection. HSV-1 has been shown to inhibit the ability of lymphoblastoid MHC class II (MHCII) is one key to developing a pathogen-spe- B cells to stimulate CD4ϩ T cells (13). By interfering with specific cific immune response. Generation of CD4ϩ T cell help for Ab steps in the biosynthetic MHCII processing pathway, HSV-1 production requires stimulation by virus peptide-loaded MHCII polypeptides might manipulate MHCII presentation. Recently, we discovered that the HSV-1 glycoprotein B (gB), transiently ex- pressed in COS-7 cells, binds to HLA-DR molecules (14). *Section of Immunobiology, Institute for Molecular Physiology, University of Bonn, The impact of HSV-1 infection on the MHCII processing path- Bonn, Germany; and †Institute for Medical Microbiology and Immunology, Univer- sity of Bonn Medical Center, Bonn, Germany way and the role of the virus envelope protein gB was investigated Received for publication April 17, 2003. Accepted for publication July 14, 2003. in this study. We demonstrate that HSV-1 infection strongly de- creases the amount of Ii in B lymphoblastoid cells. gB binds to DR 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 and to DM heterodimers, thereby manipulating the MHCII pro- with 18 U.S.C. Section 1734 solely to indicate this fact. cessing pathway. Our results suggest that HSV-1 uses at least two 1 This work was supported by a Grant Ko 810/6-1 from the Deutsche distinct mechanisms to down-regulate a CD4 T cell response: 1) Forschungsgemeinschaft. interruption of MHCII Ag processing by reduction of Ii expression 2 Address correspondence and reprint requests to Dr. Norbert Koch, Universita¨t Bonn, and 2) interaction of gB with DM and DR polypeptides. Abteilung Immunbiologie, Institut fu¨r Molekulare Physiologie Ro¨merstrasse 164, 53117 Bonn, Germany. E-mail address: norbert.koch@uni-bonn.de 3 Abbreviations used in this paper: MHCI and II, MHC class I and class II; HSV-1, Materials and Methods HSV type 1; Ii, invariant chain; ER, endoplasmic reticulum; gB, glycoprotein B; Biochemicals and Abs NP40 NonidetP-40; EndoH, endoglycosidase H; PGNaseF, N-glycosidase F; CatB, cathepsin B; moi, multiplicity of infection; PVDF, polyvinylidene difluoride; HCMV, Bu-45 is a mouse mAb directed against the luminal domain of human Ii human CMV; LC, Langerhans cell. (15) and was used for immunoprecipitation. Immunoprecipitation of DR Copyright © 2003 by The American Association of Immunologists, Inc. 0022-1767/03/$02.00 3076 MHC CLASS II IS TARGETED BY HSV-1 Downloaded from http://www.jimmunol.org/ by guest on October 2, 2021 FIGURE 1. MHC and HSV-1 protein expression by infected cells. A, JESTHOM cells were infected with HSV-1 for 24 h. Infected cells were MACS selected with mAb 2c/2 against gB, surface stained for HSV-1 proteins, and assayed by flow cytometry (upper panel). Uninfected cells incubated with anti-HSV-1 Abs are displayed as a dark curve. MHCII (middle panel) and MHCI (lower panel) surface molecules were stained with fluorescein-conjugated mAbs L243 and W6/32 (dark curve, uninfected cells; white curve, infected cells). B, Lysates from cells exposed to HSV-1 for 18, 40, and 64 h; MACS selected for gB and from uninfected cells were immunoblotted and stained for gB (mAb 2c/2), actin, and Ii (mAbs AC40 and Bu-43), DR␣ (mAb 1B5) MHCI H chain (mAb HC10), and DM␣ (mAb 5C1). The relative amount of Ii was estimated by densitometric scanning (data not shown). C, JESTHOM cells were infected with HSV-1. After 8 h, only a small fraction of the cells display gB on the cell surface. gB-expressing cells were separated by MACS. Cell lysates from infected and uninfected cells were immunoblotted and stained with mAbs for actin and Ii. D, JESTHOM cells were infected with HSV-1 for 72 h followed by MACS selection. Infected and uninfected cells were starved, metabolically labeled with [35S]methionine, and chased for up to 180 min. Ii was immunoprecipitated from cell lysates with mAb Bu-45. SDS-PAGE-separated immunoprecipitates were exposed to x-ray filmsfor4h(upper panels). The gel shown in the upper left panel in addition was exposed for 12 h (middle left). The intensity of Ii bands was densitometrically scanned (lower panels). The estimated t1/2 of Ii in infected and uninfected cells is indicated. was performed with mAb ISCR3 (16) while FITC-conjugated mAbs L243 class I was performed with FITC-conjugated mAb W6/32 (18), and stain- (17), Tu¨39-FITC (BD PharMingen, San Diego, CA), and I251 were used ing of cell surface-expressed HSV-1 proteins was achieved with antiserum for flow cytometry and fluorescence staining.