Boosting immune response with the invariant chain segments via association with non-peptide binding region of major histocompatibility complex class II molecules
© Chen et al.; licensee BioMed Central Ltd. 2012
Received: 29 March 2012
Accepted: 17 September 2012
Published: 27 September 2012
Based on binding of invariant chain (Ii) to major histocompatibility complex (MHC) class II molecules to form complexes, Ii-segment hybrids, Ii-key structure linking an epitope, or Ii class II-associated invariant chain peptide (CLIP) replaced with an epitope were used to increase immune response. It is currently unknown whether the Ii-segment cytosolic and transmembrane domains bind to the MHC non-peptide binding region (PBR) and consequently influence immune response. To investigate the potential role of Ii-segments in the immune response via MHC II/peptide complexes, a few hybrids containing Ii-segments and a multiepitope (F306) from Newcastle disease virus fusion protein (F) were constructed, and their binding effects on MHC II molecules and specific antibody production were compared using confocal microscopy, immunoprecipitation, western blotting and animal experiments.
One of the Ii-segment/F306 hybrids, containing ND (Asn–Asp) outside the F306 in the Ii-key structure (Ii-key/F306/ND), neither co-localized with MHC II molecules on plasma membrane nor bound to MHC II molecules to form complexes. However, stimulation of mice with the structure produced 4-fold higher antibody titers compared with F306 alone. The two other Ii-segment/F306 hybrids, in which the transmembrane and cytosolic domains of Ii were linked to this structure (Cyt/TM/Ii-key/F306/ND), partially co-localized on plasma membrane with MHC class II molecules and weakly bound MHC II molecules to form complexes. They induced mice to produce approximately 9-fold higher antibody titers compared with F306 alone. Furthermore, an Ii/F306 hybrid (F306 substituting CLIP) co-localized well with MHC II molecules on the membrane to form complexes, although it increased antibody titer about 3-fold relative to F306 alone.
These results suggest that Ii-segments improve specific immune response by binding to the non-PBR on MHC class II molecules and enabling membrane co-localization with MHC II molecules, resulting in the formation of relatively stable MHC II/peptide complexes on the plasma membrane, and signal transduction.
Major histocompatibility complex (MHC) class II molecules play an important role in antigen presentation, with the binding antigen peptide as a key step in initiating the specific immune response. In this process, the invariant chain (Ii) acts as a chaperone for the correct folding and the functional stability of MHC II molecules. Ii is described initially as a nonpolymorphic type II integral membrane protein, binding MHC II α and β chains to form αβ/Ii complexes. These αβ/Ii complexes have been identified on B cell and dendritic cell surfaces[3–5]. In epithelial cells, the endocytosis of αβ/Ii complexes is highly dependent on the Ii di-leucine motif[6, 7]. Ii limits HLA-DR egress from the endoplasmic reticulum and prevents loading of self-peptide[2, 9, 10]. In immature lysosomes, Ii is proteolyzed, and its class II-associated invariant chain peptide (CLIP) is replaced by an antigenic peptide. In B cells, the major pathway of Ii-associated MHC class II molecules involves direct access to the endolysosomal compartments for peptide loading.
The mouse Ii isoform Ii31 consists of the cytosolic and transmembrane domains and luminal domain that contains CLIP and the trimerization region. The cytosolic domain (Cyt) contains an endosome-targeting signal that is essential for Ii targeting to the endosomal compartment, via the plasma membrane alone or the MHC class II complex[15, 16]. The transmembrane domain (TM) plays a key role in the formation of Ii trimers and in the degradation of Ii, thereby influencing MHC class II molecular functions, including complex formation and antigen presentation. The CLIP binds MHC class II peptide binding region (PBR) and interacts with class II molecules[10, 20]. The trimerization region is involved in the generation of this endosomal localization signal.
A method to increase antigen-specific stimulation of T-helper cells entails the use of the Ii hybrids, in which a four-amino-acid sequence (LRMK) is linked to T-helper epitopes[21–23], or the Ii peptide (CLIP) region is replaced with the various epitopes[24, 25]. Animal models illustrate the efficiency of Ii hybrid methodology in using melanoma peptides[21, 26], subvirion influenza A (H5N1) HA, human papilloma virus 16 E7(8–22), Listeria Th and hepatitis C virus epitopes. The mechanistic hypothesis states that the Ii -key binds initially to an allosteric site just outside the MHC class II binding groove at the cell surface[26, 27]. This induces a conformational change in the trough, facilitating antigenic epitope charging[22, 28], and a concomitant increase in the potency of antigen presentation compared with the unmodified class II epitope[29, 30]. As vector, Ii-key and Ii can enhance the interferon (IFN)γ and interleukin (IL)-4 or IL-2 responses in enzyme-linked immunosorbent spot assay[21, 24], epitope-specific CD4+ T cell activation, or specific antibody production. The Ii-hybrids can also function in desensitizing allergy and inducing antigen-specific tolerance and ameliorating arthritis. All these findings indicate potential clinical use of such allosteric site-directed, Ii-segment drugs.
Construction and identification of the Ii-segment epitope hybrids
Primers, cloned Ii-segments andreconstructed vectors in this study
Primer sequences (5´-´)
Ii-segments in hybrids bound to non-PBR of MHC class II molecules
Some Ii segments co-localized with MHC class II molecules on the plasma membrane
All the Ii-segments increased immune response
We demonstrated by immunoprecipitation and western blotting that Ii segment cytosolic and transmembrane domains bound to MHC class II molecules, and by confocal microscopy that they co-localized on the plasma membrane with MHC class II molecules in transfected cells. Although the amino acid sequences lying outside the N and C terminal segments of CLIP, Ii-key and DN, neither bound to nor co-localized with MHC class II molecules, the both segments made the epitope in the hybrids concomitantly more stable when associated with a PBR .
Together with the cytosolic and transmembrane domains, these amino acid sequences associate with the non-PBR of MHC class II molecules to form complexes, hybrid/MHC II, which might be better for presenting epitopes to immune cells, improving specific antibody production.
Ii-key and DN facilitate epitope loading in MHC II PBR
The conformation of the MHC class II molecules plays an important role in peptide association[35–41]. Binding of peptide to MHC class II molecules involves several conformational changes, including a transient “peptide-receptive” conformation[41–43]. The efficient generation of long-lived peptide/class II complexes involves two stages: initial conditioning of MHC class II molecules in an acidic environment, forming a floppy MHC that increases the ability of class II molecules to enter a compact conformation, upon binding to specific peptides; whereas the mature peptide-loaded MHC class II molecules appear as compact heterodimers. Ii-key and DN lie just outside N and C termini of the CLIP region, respectively. Ii-key facilitates epitope loading of MHC class II molecules at the cell surface[26, 27]. DN, two relatively conserved residues and hypothetically similar to Ii-key, help CLIP bind to the MHC class II PBR, enabling effective and stable epitope (F306) loading of the PBR. Furthermore, the MHC class II molecules exist in at least two different conformations with respect to their peptide-binding ability; one more receptive to binding than the other. The peptide may configure a class II MHC structure[35, 36], resulting in a compact conformation facilitating charging of epitopes. Ii-key and DN possibly enhance the loading stability of Ii-key/F306 or Ii-key/F306/DN with MHC class II PBR, to induce a conformational change in the trough and facilitate effective epitope charging and generation of long-lived peptide/class II complexes, affecting T cell secretion of IFN and increased humoral immune response. However, the affinity between Ii-key and DN, and MHC class II molecule is not sufficient to form complexes in cotransfected cells or immunoprecipitation (Figure2).
Cytosolic and transmembrane domain binding to non-PBR improves epitope loading in MHC class II PBR
The Ii and the Ii-segment/F306 hybrids bind MHC class II α or β chains as complexes on the plasma membrane (Figures 2 and3), based on their trimerization region. The Cyt/TM/Ii-key/F306(CLIP)/DN lacks the trimerization region, although it also binds MHC class II molecules to form visible complexes (Figures 2 and3). The transmembrane domain has a role in the formation of αβ/Ii trimers: along with Ii-key and DN, it is the third factor for binding to MHC class II molecules and maintaining stability of epitope/MHC II complexes. However, this domain also co-localizes to the membrane with MHC class II molecules. The MHC class II molecules require localization on the membrane rafts for signal transduction. A preferential localization of peptide-bound MHC class II molecules on the membrane results in optimal antigen presentation[47–49]. In addition, the cytosolic domain contains an endosome-targeting signal for immune regulation[15, 16]. The binding of the non-PBR via Ii functional segments, co-localization with MHC II on the membrane, and signal transduction are sufficient for stable complex formation, antigen presentation and initial immune response.
Ii-segments are a potential immune carrier
The Ii/F306 hybrid bound MHC class II molecules strongly to form complexes on the plasma membrane (Figure3). It stimulated an intermediate immune response, which was lower than that of other hybrids containing cytosolic and transmembrane domains (Figure3). Under normal conditions, the MHCII/Ii complex is directed to endosomes[50, 51] and then to immature lysosomes, where the Ii is proteolyzed, and the CLIP is replaced by an antigenic peptide. The Ii/F306 hybrid contains a trimerization region at its C terminus, which enables the Ii hybrid/MHC II molecule to form stable complexes. However, tight binding between MHC class II molecules and Ii/F306 hybrid might prevent Ii/F306 hybrid release to bind other MHC class II molecules and activate other immune cells. In other words, the trimerization region disrupts contact with receptors at other cell surfaces, which is necessary for initiation of the immune response when MHC class II molecules present antigenic peptide. In contrast to Ii/F306 hybrid, the other Ii-segment/epitope hybrids such as Cyt/TM/Ii-key/F306/DN bind MHC class II molecules on the plasma membrane weakly, which is sufficient to form relatively stable complexes to induce an immune response, but also for its disassembly to bind and activate more immune cells. In brief, these Ii-segments may be used as a carrier to promote specific immune responses.
Cloning and construction of the hybrids
We cloned various Ii functional segments from mouse Ii cDNA using PCR and constructed the Ii-segment/F306 hybrids (Figure1) with a series of primers (Table1). We also cloned mouse H2-Aa and H2-Ab genes with the primers (Table1). An Ii/F306 hybrid, in which the CLIP region was replaced by F306, was constructed by overlap extension PCR. The constructed Ii-segments or hybrids were then inserted into eukaryotic vector pmCherry-C1 or pEGFP-C1 (Table1, Nos. 4–12), and the mouse H2-Aa and H2-Ab genes were inserted into pEGFP-N1 (Table1, Nos. 1 and 2) to enable identification by confocal microscopy. These Ii-segment/F306 hybrids were also inserted into prokaryotic expression vectors pGEX-4 T-1 (Table1, Nos. 13–18) for immunization antigen. Additionally, F306 was inserted into pET-32a (No. 17) for expression of the coating antigen used in the ELISA. Mouse H2-Ab genes were inserted into PCMV-Myc (Table1, No. 3) for the expression of eukaryotic protein by immunoprecipitation and western blotting. All the constructed hybrids were identified by sequencing.
Cell culture, transfection and confocal microscopy
COS7 cells were obtained from Biology Science College, University of Science and Technology of China. The cells were grown at 37 °C in the presence of 5% CO2 in Dulbecco’s modified Eagle’s medium with 10% fetal calf serum (FCS) (GIBCO, USA). Transfection of COS7 cells was done using Lipofectamine 2000 (Invitrogen) reagent following the manufacturer’s instructions. Briefly, the hybrid vectors and media (amount depending on the size of the well) were mixed together, and Lipofectamine 2000 (twice the amount of the hybrid vector) was mixed with the medium in a separate tube. The medium containing the hybrid vectors and the medium containing Lipofectamine were mixed together, allowed to sit for 15–20 min at room temperature, and added slowly to the well. After 5 h, the medium was replaced with fresh media containing penicillin and streptomycin, and 10% fetal bovine serum. After 24 h, images of the COS7 cells were acquired with a Zeiss confocal laser scanning microscope (CLSM) using a × 60 oil objective [excitation at 488 nm for red fluorescent protein and emission at 515 nm for green fluorescent protein (GFP)].
Immunoprecipitation and western blotting
Immunoprecipitation included cotransfection of the COS7 cells seeded in 25-cm2 plates with fusion genes H2-Ab/Myc and Ii/GFP, Ii/F306/GFP, Ii-key/F306/DN/GFP, Cyt/TM/Ii-key/F306/DN/GFP or Cyt/TM/Ii-key/F306/GFP, respectively. At 36 h post-transfection, the cells were harvested and lysed in 1 mL immunoprecipitation lysis buffer (50 mM Tris–HCl, 150 mM NaCl, 1% Nonidet P40, and 0.5% sodium deoxycholate, 1 Protease Inhibitor Cocktail Tablets) at 4°C for 1 h. The cells were then centrifuged at 12 000 g at 4°C for 1 h, and 20 μL Protein A/G Plus-Agarose beads (GE Healthcare, USA) were added to the supernatants and incubated at 4°C for 2 h. After centrifugation at 12 000 g for 20 s at 4°C, 2 μL antibody to Myc (Zhongshan Golden Bridge Biotechnology, Beijing, China) was added to the supernatants and incubated for 2 h at 4 °C. The immune complexes were isolated using 50 μL Protein A/G Plus-Agarose beads at 4°C overnight. Centrifugation involved suspending residue in 1 mL immunoprecipitation lysis buffer, buffer 2 (50 mM Tris–HCl, 500 mM NaCl, 0.1% Nonidet P40, 0.05% sodium deoxycholate) and buffer 3 (50 mM Tris–HCl, 0.1% Nonidet P40, 0.05% sodium deoxycholate) for 20 min and adding Protein A/G Plus-Agarose beads under the above conditions, and repeating three times. Subsequently, washed immunoprecipitates were separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and transferred onto the polyvinylidene fluoride membrane (Millipore, Schwalbach, Germany). The blots were blocked with 10% (v/v) FCS for 1 h and then probed for 1 h with a murine antibody to GFP (Zhongshan Golden Bridge Biotechnology), followed by washing and incubation for 2 h with horseradish peroxidase (HRP)-conjugated secondary Abs (goat anti-mouse IgG, Zhongshan Golden Bridge Biotechnology) and an ECL detection system (Pierce Roclford).
Expression and purification of antigens
A homologous series of F306 or Ii-segments/F306-epitope hybrids cloned into pGEX-4 T-1 and pET-32a was transfected into E. coli expression strain Rosetta. Antigen expression was induced by 1 mmol/L IPTG. All proteins were extracted in denaturing conditions according to the Qiagen protocol and purified by immobilized-metal affinity chromatography with Ni-NTA agarose beads following the manufacturer’s instructions (Amersham Biosciences, Little Chalfont, UK), and found to be >98% pure by analytical HPLC. They were dissolved in sterile distilled water (5 mg/mL) and stored at −70 °C.
Mice and immunization
Balb/c female mice (10 weeks old) were obtained from the Animal Centre of Anhui Medicine University and bred under specific pathogen-free conditions at the facility. All experimental procedures were performed following the Anhui Medicine University animal care guidelines under an approved protocol. Thirty five mice were divided into seven groups. Mice to be immunized were anesthetized and injected intraperitoneally with 50 μg of each protein antigen. The animals received the protein doses at week 0, with complete Freund’s adjuvant as a 1:1 (v/v) emulsion in 100 μL. The second immunization occurred at weeks 2 in incomplete Freund’s adjuvant, and the third immunization took place at week 3 without adjuvant were carried out. One control group of mice was injected as above without any antigen. The sera were prepared at week 4 from blood collected from mice via the tail vein, and were stored at −20°C until used for estimation of the antibody titers.
Detection of antibody with ELISA
Ninety-six-well EIA/RIA plates (COSTAR, USA) were coated with 5 μg/mL His-F306 peptide and then blocked with 0.05% Tween-20 in PBS (PBST) containing 1% bovine serum albumin. The sera were added to the top row of each plate, and serial 1:2 dilutions in PBST were then placed in subsequent rows. The plates were incubated for 45 min at room temperature and washed with PBST. A goat anti-mouse IgG HRP conjugate (Zhongshan Golden Bridge Biotechnology) diluted 1:5000 was used as a secondary antibody and incubated for 45 min, followed by addition of OPD peroxidase (Sigma, USA) used as a substrate. After 15 min of incubation at room temperature, the absorbance was measured at 405 nm.
Statistical differences were calculated by one-way analysis of variance with post-test. Significance was defined as P < 0.01. All functional assays, e.g., specific antibody titers in ELISA, were performed in quadruplicate.
The research was financially supported by a grant from the National Natural Science Foundation of China, Beijing, under award number 31172306.
- Strubin M, Berte C, Mach B: Alternative splicing and alternative initiation of translation explain the four forms of the Ia antigen-associated invariant chain. EMBO J. 1986, 5: 3483-3488.PubMedPubMed CentralGoogle Scholar
- Roche PA, Cresswell P: Invariant chain association with HLA-DR molecules inhibits immunogenic peptide binding. Nature. 1990, 345: 615-618. 10.1038/345615a0.PubMedView ArticleGoogle Scholar
- Lindner R: Transient surface delivery of invariant chain-MHC II complexes via endosomes: a quantitative study. Traffic. 2002, 3: 133-146. 10.1034/j.1600-0854.2002.030206.x.PubMedView ArticleGoogle Scholar
- Cella M, Sallusto F, Lanzavecchia A: Origin, maturation and antigen presenting function of dendritic cells. Curr Opin Immunol. 1997, 9: 10-16. 10.1016/S0952-7915(97)80153-7.PubMedView ArticleGoogle Scholar
- Saudrais C, Spehner D, de la Salle H, Bohbot A, Cazenave JP, Goud B, Hanau D, Salamero J: Intracellular pathway for the generation of functional MHC class II peptide complexes in immature human dendritic cells. J Immunol. 1998, 160: 2597-2607.PubMedGoogle Scholar
- Dugast M, Toussaint H, Dousset C, Benaroch P: AP2 clathrin adaptor complex, but not AP1, controls the access of the major histocompatibility complex (MHC) class II to endosomes. J Biol Chem. 2005, 280: 19656-19664. 10.1074/jbc.M501357200.PubMedView ArticleGoogle Scholar
- McCormick PJ, Martina JA, Bonifacino JS: Involvement of clathrin and AP-2 in the trafficking of MHC class II molecules to antigen-processing compartments. Proc Natl Acad Sci USA. 2005, 102: 7910-7915. 10.1073/pnas.0502206102.PubMedPubMed CentralView ArticleGoogle Scholar
- Schutze MP, Peterson PA, Jackson MR: An N-terminal double-arginine motif maintains type II membrane proteins in the endoplasmic reticulum. EMBO J. 1994, 13: 1696-1705.PubMedPubMed CentralGoogle Scholar
- Stumptner P, Benaroch P: Interaction of MHC class II molecules with the invariant chain: role of the invariant chain (81–90) region. EMBO J. 1997, 16: 5807-5818. 10.1038/sj.emboj.7590555.PubMedPubMed CentralView ArticleGoogle Scholar
- Romagnoli P, Germain RN: The CLIP region of invariant chain plays a critical role in regulating major histocompatibility complex class II folding, transport, and peptide occupancy. J Exp Med. 1994, 180: 1107-1113. 10.1084/jem.180.3.1107.PubMedView ArticleGoogle Scholar
- Hsing LC, Rudensky AY: The lysosomal cysteine proteases in MHC class II antigen presentation. Immunol Rev. 2005, 207: 229-241. 10.1111/j.0105-2896.2005.00310.x.PubMedView ArticleGoogle Scholar
- Busch R, Rinderknecht CH, Roh S, Lee AW, Harding JJ, Burster T, Hornell TM, Mellins ED: Achieving stability through editing and chaperoning: regulation of MHC class II peptide binding and expression. Immunol Rev. 2005, 207: 242-260. 10.1111/j.0105-2896.2005.00306.x.PubMedView ArticleGoogle Scholar
- Benaroch P, Yilla M, Raposo G, Ito K, Miwa K, Geuze HJ, Ploegh HL: How MHC class II molecules reach the endocytic pathway. EMBO J. 1995, 14: 37-49.PubMedPubMed CentralGoogle Scholar
- Koch N, Lauer W, Habicht J, Dobberstein B: Primary structure of thegene for the murineIa antigen-associated invariant chains(Ii). An alternatively spliced exon encodes a cysteine-rich domain highly homologous to a repetitive sequence of thyroglobulin. EMBO J. 1987, 6: 1677-1683.PubMedPubMed CentralGoogle Scholar
- Rudensky AY, Maric M, Eastman S, Shoemaker L, DeRoos PC, Blum JS: Intracellular assembly and transport of endogenous peptide-MHC class II complexes. Immunity. 1994, 1: 585-594. 10.1016/1074-7613(94)90048-5.PubMedView ArticleGoogle Scholar
- Vogt AB, Stern LJ, Amshoff C, Dobberstein B, Hammerling BJ, Kropshofer H: Interference of distinct invariant chain regions with superantigen contact area and antigenic peptide binding groove of HLA-DR. J Immunol. 1995, 155: 4757-4765.PubMedGoogle Scholar
- Ashman JB, Miller JJ: A role for the transmembrane domain in the trimerization of the MHC class II-associated invariant chain. Immunol. 1999, 163: 2704-2712.Google Scholar
- Frauwirth K, Shastri N: Mutation of the invariant chain transmembrane region inhibits II degradation, prolongs association with MHC class II, and selectively disrupts antigen presentation. Cell Immunol. 2001, 209: 97-108. 10.1006/cimm.2001.1796.PubMedView ArticleGoogle Scholar
- Dixon AM, Stanley BJ, Matthews EE, Dawson JP, Engelman DM: Invariant chain transmembrane domain trimerization: a step in MHC class II assembly. Biochemistry. 2006, 45: 5228-5234. 10.1021/bi052112e.PubMedView ArticleGoogle Scholar
- Jasanoff A, Wagner G, Wiley DC: Structure of a trimeric domain of the MHC class II-associated chaperonin and targeting protein Ii. EMBO J. 1998, 17: 6812-6818. 10.1093/emboj/17.23.6812.PubMedPubMed CentralView ArticleGoogle Scholar
- Kallinteris NL, Wu S, Lu X, Humphreys RE, von Hofe E, Xu M: Enhanced CD4+ T-cell response in DR4-transgenic mice to a hybrid peptide linking the Ii-Key segment of the invariant chain to the melanoma gp100(48–58) MHC class II epitope. J Immunother. 2005, 28: 352-358. 10.1097/01.cji.0000170362.45456.00.PubMedView ArticleGoogle Scholar
- Zinckgraf JW, Sposato M, Zielinski V, Powell D, Treanor JJ, von Hofe E: Identification of HLA class II H5N1 hemagglutinin epitopes following subvirion influenza A (H5N1) vaccination. Vaccine. 2009, 27: 5393-5401. 10.1016/j.vaccine.2009.06.081.PubMedView ArticleGoogle Scholar
- Xu M, Lu X, Sposato M, Zinckgraf JW, Wu S, von Hofe E: Ii-Key/HPV16 E7 hybrid peptide immunotherapy for HPV16+ cancers. Vaccine. 2009, 27: 4641-4647. 10.1016/j.vaccine.2009.05.054.PubMedView ArticleGoogle Scholar
- Nagata T, Aoshi T, Suzuki M, Uchijima M, Kim YH, Yang Z, Koide Y: Induction of protective immunity to Listeria monocytogenes by immunization with plasmid DNA expressing a helper T-cell epitope that replaces the class II-associated invariant chain peptide of the invariant chain. Infect Immun. 2002, 70: 2676-2680. 10.1128/IAI.70.5.2676-2680.2002.PubMedPubMed CentralView ArticleGoogle Scholar
- Gao M, Wang HP, Wang YN, Zhou Y, Wang QL: HCV-NS3 Th1 minigene vaccine based on invariant chain CLIP genetic substitution enhances CD4(+) Th1 cell responses in vivo. Vaccine. 2006, 24: 5491-7. 10.1016/j.vaccine.2006.04.004.PubMedView ArticleGoogle Scholar
- Adams S, Humphreys RE: Invariant chain peptides enhancing or inhibiting the presentation of antigenic peptides by major histocompatibility complex class II molecules. Eur J Immun. 1995, 25: 1693-1702. 10.1002/eji.1830250632.View ArticleGoogle Scholar
- Xu M, Lis J, Gulfo JV: MHC class II allosteric site drugs: New immunotherapeutics for malignant, infectious and autoimmune disease. Scan J Immunol. 2001, 54: 39-44. 10.1046/j.1365-3083.2001.00964.x.View ArticleGoogle Scholar
- Sotiriadou NN, Kallinteris NL, Gritzapis AD, Voutsas IF, Papamichail M, von Hofe E, Humphreys RE, Pavlis T, Perez SA, Baxevanis CN: Ii-Key/HER-2/neu(776–790) hybrid peptides induce more effective immunological responses over the native peptide in lymphocyte cultures from patients with HER-2/neu + tumors. Cancer Immunol Immunother. 2007, 56: 601-613. 10.1007/s00262-006-0213-z.PubMedView ArticleGoogle Scholar
- Humphreys RE, Adams S, Koldzic G: Increasing the potentcy of MHC class II-presented epitopes by linking to Ii-Key peptide. Vaccine. 2000, 18: 2693-2697. 10.1016/S0264-410X(00)00067-0.PubMedView ArticleGoogle Scholar
- Gillogly ME, Kallinteris NL, Xu M, Gulfo JV, Humphreys RE, Murray JL: Ii-Key/HER-2/neu MHC class-II antigenic epitope vaccine peptide for breast cancer. Cancer Immunol Immunother. 2004, 53: 490-496. 10.1007/s00262-003-0463-y.PubMedView ArticleGoogle Scholar
- Rhyner C, Kündig T, Akdis CA, Crameri R: Targeting the MHC II presentation pathway in allergy vaccine development. Biochem Soc Trans. 2007, 35: 833-834. 10.1042/BST0350833.PubMedView ArticleGoogle Scholar
- Gjertsson I, Laurie KL, Devitt J, Howe SJ, Thrasher AJ, Holmdahl R, Gustafsson K: Tolerance induction using lentiviral gene delivery delays onset and severity of collagen II arthritis. Mol Ther. 2009, 17: 632-640. 10.1038/mt.2009.299.PubMedPubMed CentralView ArticleGoogle Scholar
- Dong L, Yu W, Xu F, Liu G: Cloning and identifying of mouse invariant chain gene and its identification in prokaryotic cells. J Anhui Agri Sci. 2007, 35: 7856-7857. ChinesGoogle Scholar
- Xu F, Wu S, Yu W: Establishment of P815 cell line stably expressing Newcastle disease virus-F gene (NDV-F). J Agri Biotech. 2009, 17: 567-570. ChinesGoogle Scholar
- Sadegh-Nasseri S, Germain RN: A role for peptide in determining MHC class II structure. Nature. 1991, 353: 167-170. 10.1038/353167a0.PubMedView ArticleGoogle Scholar
- Sadegh-Nasseri S, Stern LJ, Wiley DC, Germain RN: MHC class II function preserved by lowaffinity peptide interactions preceding stable binding. Nature. 1994, 370: 647-650. 10.1038/370647a0.PubMedView ArticleGoogle Scholar
- Sato AK, Zarutskie JA, Rushe MM, Lomakin A, Natarajan SK, Adegh-Nasseri SS: Determinants of the peptide-induced conformational change in the human class II major histocompatibility complex protein HLA-DR1. J Biol Chem. 2000, 275: 2165-2173. 10.1074/jbc.275.3.2165.PubMedView ArticleGoogle Scholar
- Carven GJ, Stern LJ: Probing the ligand-induced conformational change in HLA-DR1 by selective chemical modification and mass spectrometric mapping. Biochemistry. 2005, 44: 13625-13637. 10.1021/bi050972p.PubMedView ArticleGoogle Scholar
- Dornmair K, Rothenhausler B, McConnell HM: Structural intermediates in the reactions of antigenic peptides with MHC molecules. Cold Spring Harb Symp Quant Biol. 1989, 54: 409-416. 10.1101/SQB.1989.054.01.050.PubMedView ArticleGoogle Scholar
- Witt SN, McConnell HM: Formation and dissociation of short-lived class II MHC-peptide complexes. Biochemistry. 1994, 33: 1861-1868. 10.1021/bi00173a032.PubMedView ArticleGoogle Scholar
- Schmitt L, Boniface JJ, Davis MM, McConnell HM: Conformational isomers of a class II MHCpeptide complex in solution. J Mol Biol. 1999, 286: 207-218. 10.1006/jmbi.1998.2463.PubMedView ArticleGoogle Scholar
- Natarajan SK, Assadi M, Sadegh-Nasser S: Stable peptide binding to MHC class II molecule is rapid and is determined by a receptive conformation shaped by prior association with low affinity peptides. J Immunol. 1999, 162: 4030-4036.PubMedGoogle Scholar
- Rabinowitz JD, Vrljic M, Kasson PM, Liang MN, Busch R, Boniface JJ: Formation of a highly peptide-receptive state of class II MHC. Immunity. 1998, 9: 699-709. 10.1016/S1074-7613(00)80667-6.PubMedView ArticleGoogle Scholar
- Germain RN, Hendrix LR: MHC class II structure, occupancy and surface expression determined by post-endoplasmic reticulum antigen binding. Nature. 1991, 353: 134-139. 10.1038/353134a0.PubMedView ArticleGoogle Scholar
- Natarajan SK, Assadi M, Sadegh-Nasseri S: Stable peptide binding to MHC class II molecule is rapid and is determined by a receptive conformation shaped by prior association with low affinity peptides. J Immunol. 1999, 162: 4030-4036.PubMedGoogle Scholar
- Huby RD, Dearman RJ, Kimber I: Intracellular phosphotyrosine induction by major histocompatibility complex class II requires co-aggregation with membrane rafts. J Biol Chem. 1999, 274: 22591-22596. 10.1074/jbc.274.32.22591.PubMedView ArticleGoogle Scholar
- Karacsonyi C, Knorr R, Fulbier A, Lindner R: Association of major histocompatibility complex II with cholesterol- and sphingolipid-rich membranes precedes peptide loading. J Biol Chem. 2004, 279: 34818-34826. 10.1074/jbc.M404608200.PubMedView ArticleGoogle Scholar
- Anderson HA, Hiltbold EM, Roche PA: Concentration of MHC class II molecules in lipid rafts facilitates antigen presentation. Nat Immunol. 2000, 1: 156-162. 10.1038/77842.PubMedView ArticleGoogle Scholar
- Hiltbold EM, Poloso NJ, Roche PA: MHC class II-peptide complexes and APC lipid rafts accumulate at the immunological synapse. J Immunol. 2003, 170: 1329-1338.PubMedView ArticleGoogle Scholar
- Pond L, Watts C: Functional early endosomes are required for maturation of major histocompatibility complex class II molecules in human B lymphoblastoid cells. J Biol Chem. 1999, 274: 18049-18054. 10.1074/jbc.274.25.18049.PubMedView ArticleGoogle Scholar
- Brachet V, Pehau-Arnaudet G, Desaymard C, Raposo G, Amigorena S: Early endosomes are required for major histocompatiblity complex class II transport to peptide-loading compartments. Mol Biol Cell. 1999, 10: 2891-2904.PubMedPubMed CentralView ArticleGoogle Scholar
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