Introduction of soluble protein into the class I pathway of antigen processing and presentation

Moore, Mark; Carbone, Federico; Bevan, Michael J.

doi:10.1016/s0092-8674(88)91043-4

Cited by 1,030 publications

(735 citation statements)

References 27 publications

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“…Cells were loaded with OVA by osmotic shock as described previously [64]. In brief, *15 Â 10 7 cells were incubated in 1 mL hypertonic medium (0.5 M sucrose, 10% polyethylene glycol 1000, and 10 mM HEPES in RPMI 1640, pH 7.2) containing 10 mg/mL OVA (Calbiochem) for 10 min at 37 C. Pre-warmed hypotonic medium (13 mL; 40% H 2 O, 60% RPMI 1640) was added and the cells were incubated for an additional 2 min at 37 C. The cells were centrifuged, washed twice with cold PBS, and irradiated (1500 rad).…”

Section: Generation Of Ova-loaded B2m-/-cellsmentioning

confidence: 99%

“…The multi-copy OVA-HA expression vector pFvL210 was generated by cloning of a C-terminally HA-tagged version of the chicken OVA cDNA from pAc-neo-OVA1 [64] into the BamHI site of pTCG [65]. The plasmid was transformed into yeast strain UCC7164 (a derivative of S288C) [66] and transformed cells were grown in synthetic media lacking tryptophan to select for the plasmid.…”

Section: Yeastmentioning

confidence: 99%

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CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

et al. 2008

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Mouse splenic dendritic cell (DC) subsets possess distinct antigen-presentation abilities. CD8 + DC are specialized in cross-presentation of antigens to CD8 + T cells, whereas CD8 -DC are more efficient in antigen presentation to CD4 + T cells. In this study, we examined the capacity of CD8 + and CD8 -DC subsets to present fungal antigens in MHC class I and II molecules to CD8 + and CD4 + T cells, respectively. We used ovalbuminexpressing Saccharomyces cerevisiae (yeast-OVA) as a fungal model system. Both CD8 + and CD8 -DC subsets phagocytosed yeast in equal amounts and uptake was mediated via dectin-1. In addition, both DC subsets induced similar OVA-specific CD4 + T cell proliferation after incubation with yeast-OVA. However, the induction of OVA-specific CD8 + T cell activation was largely restricted to the CD8 -DC subset. Furthermore, only CD8 -DC produced cytokines such as IL-10 and TNF-a and increased IL-23p19 and IL-23p40 mRNA levels in response to yeast. Our results strongly suggest that DC subsets have different functions in the elicitation of adaptive immune responses in vivo. IntroductionFungal infections with pathogenic yeasts such as Candida albicans, Aspergillus fumigatus, Histoplasma capsulatum and Cryptococcus neoformans are a major problem in immunocompromised persons. These opportunistic pathogens usually do not pose problems in healthy individuals, but frequently become invasive in patients suffering from HIV, patients with malignancies or transplant recipients, resulting in life-threatening diseases [1][2][3].Both innate and adaptive immune responses are essential to induce protection against fungi. Dendritic cells (DC) play a central role in the adaptive immune response by capturing fungi and presenting their antigens in major histocompatibility complex (MHC) class I and MHC class II molecules to T cells [4, 5]. DC express a panel of microbial recognition receptors such as Toll-like receptors (TLR) and C-type lectin receptors that are essential for the elicitation of adaptive immune responses [6]. Of these receptors, TLR2 and the DCassociated C-type lectin (dectin-1) are specifically involved in recognition of fungal components [7][8][9]. Zymosan and fungal pathogens like C. albicans and A. fumigatus are sensed by TLR2 that signals via myeloid differentiation factor 88 (MyD88), resulting in NF-jB activation and the production of cytokines [10][11][12][13]. The importance of this signaling cascade can be inferred from the fact that MyD88-knockout mice show an increased susceptibility to fungal infections [14,15].Dectin-1 is expressed on myeloid cells like macrophages, neutrophils, monocytes, and even a subset of T cells [16]. Dectin-1 mediates fungal recognition and Revised 2/11/07 Accepted 11/12/07 [DOI 10.1002/eji.200737647] Key words: Antigen presentation Á Dendritic cell Á Fungal Á T cell Abbreviations: b2m: b2-microglobulin Á Dectin-1: DC-associated C-type lectin-1 Á CR3: complement receptor 3 Á MR: mannose receptor Á SIGNR1: ICAM-3-grabbing nonintegrin (DC-SIGN) homolog, SIGN-related...

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Section: Generation Of Ova-loaded B2m-/-cellsmentioning

confidence: 99%

Section: Yeastmentioning

confidence: 99%

CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

et al. 2008

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“…Peptides were internalized in the cytoplasm of homogenized spleen cells from BALB/c mice by osmotic lysis of pinosomes induced by hypertonic shock as described (20,24) using 300 pg/ml of peptide.…”

Section: Internalization Of Peptides In Cellsmentioning

confidence: 99%

“…Based on the observations that the tripalmitoil-S-glycerylcysteinyl-seryl-serine moiety used in the construct of Deres et al ( 13) behaves as a TD,, in another type of construct (18) and the ovalbumin tryptic fragment 229-276, which contains a predicted TDi, according to Sette's algorithm ( 19) and a proven TD, (20) but not the 242-276 (which only contains the TD,) is able to induce CTL in vivo (1 I), we speculated that synthetic colinear peptide constructs of the type TDh-TD, or TD,-TDh might be able to induce CTL in vivo. Ishioka et al (12) have suggested a similar proposition; however, to our knowledge, no experimental evidence has yet been published to validate these hypothesis.…”

Section: Introductionmentioning

confidence: 99%

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

Lasarte

Sarobe

Gullón

et al. 1992

Cellular Immunology

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Peptide constructs were engineered by colinear synthesis of two short synthetic peptide determinants: a determinant recognized by T helper cells (TD,) and a determinant recognized by T cytotoxic cells (TD,). Three types of constructs were synthesized: TD,-TDr, , TD,-TD, , and TD,,-KK-TD,, where KK are two lysine residues. In vivo immunization with free construct induced cytolytic lymphocytes (CTL) only in the case of TD,-TD,,. However, immunization with spleen cells to which these constructs had been internalized by hypertonic shock, induced CTL activity in all three cases. No CTL could be induced after immunization with free TD, in either protocol. These results indicate that cell internalization of the construct might be essential for CTL induction, and also, that "help" from the TDr, seems to be required. o 1992 Academic PKSS, IX.

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“…All the EL-4 derived transfectants including EL-4 transfected with OVA-hLa 51-58 (EL-4.OVA-hLa 51-58 ), EL-4 transfected with hLaA-OVA 257-264 (EL-4.hLaA-OVA 257-264 ), EL-4 transfected with hLa (EL-4.hLa) and EL-4 transfected by pAc-neo OVA (EG7, [34]) were maintained in DF-10 plus 0.3-0.5 mg/mL G418 (Geneticin, GIBCO, USA).…”

Section: Cell Culture and Reagentsmentioning

confidence: 99%

Direct antigen presentation by DC shapes the functional CD8⁺ T‐cell repertoire against the nuclear self‐antigen La‐SSB

et al. 2010

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Controversy still surrounds the importance of cross-presentation versus endogenous or direct presentation of MHC-I restricted Ag in CD8 1 T-cell (T CD81 ) immunity. It is even less clear what relative role these pathways play in shaping the T-cell repertoire specific for ubiquitous self-antigens, especially in cases where both Ag presentation pathways could potentially be involved. Here we provide evidence that a T CD81 repertoire specific for a determinant from the nuclear autoantigen La-SSB is largely shaped by direct presentation. In this system, mouse T CD81 reactive to a xenogeneic human La (hLa [51][52][53][54][55][56][57][58] ) K b peptide did not recognize directly presented peptide on either spleen cells from hLa-Tg mice or hLa transfected syngeneic cells. Interestingly, the same T CD81 were activated by in vivo challenge with allogeneic APC expressing either the Tg hLa or loaded with intact recombinant hLa protein, indicating functional cross-presentation of the hLa [51][52][53][54][55][56][57][58] . However, in irradiated bone marrow chimeric mice, DC expressing Tg hLa, but not WT DC that matured in hLa-Tg mice, constitutively presented the hLa [51][52][53][54][55][56][57][58] to T CD81 . These data demonstrate that although both the direct-and cross-presentation pathways are potentially operative in revealing hLa [51][52][53][54][55][56][57][58] to T CD81 , the T CD81 repertoire to this determinant is shaped quantitatively according to the efficiency of Ag presentation.Key words: Antigen processing/presentation . Autoimmunity . T-cell repertoire IntroductionAutoimmunity to nuclear antigens, in particular components of ribonucleoprotein complexes, is common in systemic autoimmune syndromes such as systemic lupus erythematosus and primary Sjögren's syndrome [1]. However, the mechanisms that shape the T-cell repertoire to these proteins and the role these auto-reactive T cells play in disease pathology are not well understood. Poor self-tolerance of some nuclear/cytoplasmic proteins and a propensity for them to become autoimmune targets could result from their sequestered localization [2], their low abundance or rapid turnover [3], and their exposure to antibodies during redistribution and concentration in apoptotic blebs [4]. Further, their association with RNA and the potential to stimulate TLR7 and 8 [5], coupled with defects in apoptosis and clearance of debris, renders some nuclear Ag as clinically relevant targets of autoimmunity [6,7]. Although much of the pathology in systemic autoimmunity is due to the deposition of complement-fixing immune complexes [8], CD81 T cells (T CD81 ) have increasingly been implicated in other autoimmune conditions [9,10]. However, the functional T CD81 repertoire specific to such nuclear autoantigens and the mode of MHC-I restricted antigen 330presentation (direct-versus cross-presentation) during the establishment of such T CD81 repertoire have not been investigated.The T CD81 repertoire is shaped by MHC-I restricted antigens presented on APC both in the thymus and...

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Introduction of soluble protein into the class I pathway of antigen processing and presentation

Cited by 1,030 publications

References 27 publications

CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

Direct antigen presentation by DC shapes the functional CD8⁺ T‐cell repertoire against the nuclear self‐antigen La‐SSB

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Introduction of soluble protein into the class I pathway of antigen processing and presentation

Cited by 1,030 publications

References 27 publications

CD8– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

CD8– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

Induction of cytotoxic T lymphocytes in mice against the principal neutralizing domain of HIV-1 by immunization with an engineered T-cytotoxic-T-helper synthetic peptide construct

Direct antigen presentation by DC shapes the functional CD8+ T‐cell repertoire against the nuclear self‐antigen La‐SSB

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CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

CD8^– dendritic cells preferentially cross‐present Saccharomyces cerevisiae antigens

Direct antigen presentation by DC shapes the functional CD8⁺ T‐cell repertoire against the nuclear self‐antigen La‐SSB