Presentation of newly synthesized glycoproteins to CD4+ T lymphocytes. An analysis using influenza hemagglutinin transport mutants.

Kittlesen, David J.; Brown, Lawrence R.; Braciale, Vivian L.; Sambrook, Joseph P.; Gething, Mary‐Jane; Braciale, Thomas J.

doi:10.1084/jem.177.4.1021

Cited by 23 publications

(24 citation statements)

References 42 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…This finding agrees with our previous studies (12) for ER-localized tumor Ag, and with many other studies that have examined presentation of endogenous Ag by professional APC (30,(61)(62)(63); however, it differs from the results of Lich et al (47), who found that endosomal acidification was not required for presentation of a cytosolic Ag by Ii-positive B lymphoblastoid cells. Inconsistency with this latter study cannot be easily explained, other than the superficial conclusion that a small percentage of Ags are processed through an alternative mechanism.…”

Section: Discussionsupporting

confidence: 92%

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Dissanayake

Tuera

Ostrand‐Rosenberg

2005

The Journal of Immunology

View full text Add to dashboard Cite

Cell-based vaccines consisting of invariant chain-negative tumor cells transfected with syngeneic MHC class II (MHC II) and costimulatory molecule genes are prophylactic and therapeutic agents for the treatment of murine primary and metastatic cancers. Vaccine efficacy is due to direct presentation of endogenously synthesized, MHC II-restricted tumor peptides to CD4+ T cells. Because the vaccine cells lack invariant chain, we have hypothesized that, unlike professional APC, the peptide-binding groove of newly synthesized MHC II molecules may be accessible to peptides, allowing newly synthesized MHC II molecules to bind peptides that have been generated in the proteasome and transported into the endoplasmic reticulum via the TAP complex. To test this hypothesis, we have compared the Ag presentation activity of multiple clones of TAP-negative and TAP-positive tumor cells transfected with I-Ak genes and the model Ag hen egg white lysozyme targeted to the endoplasmic reticulum or cytoplasm. Absence of TAP does not diminish Ag presentation of three hen egg white lysozyme epitopes. Likewise, cells treated with proteasomal and autophagy inhibitors are as effective APC as untreated cells. In contrast, drugs that block endosome function significantly inhibit Ag presentation. Coculture experiments demonstrate that the vaccine cells do not release endogenously synthesized molecules that are subsequently endocytosed and processed in endosomal compartments. Collectively, these data indicate that vaccine cell presentation of MHC II-restricted endogenously synthesized epitopes occurs via a mechanism independent of the proteasome and TAP complex, and uses a pathway that overlaps with the classical endosomal pathway for presentation of exogenously synthesized molecules.

show abstract

Section: Discussionsupporting

confidence: 92%

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Dissanayake

Tuera

Ostrand‐Rosenberg

2005

The Journal of Immunology

View full text Add to dashboard Cite

show abstract

“…Other reports have shown that in viral infections, endogenous Ag can be presented to MHC II-restricted CD4 ϩ T cells (13,16,21). This possibility will be investigated for the presentation of nuclear Ag IE1.…”

Section: Discussionmentioning

confidence: 99%

Ex Vivo Stimulation and Expansion of both CD4⁺and CD8⁺T Cells from Peripheral Blood Mononuclear Cells of Human Cytomegalovirus-Seropositive Blood Donors by Using a Soluble Recombinant Chimeric Protein, IE1-pp65

Vaz-Santiago¹,

Lulé

Rohrlich

et al. 2001

J Virol

View full text Add to dashboard Cite

Human cytomegalovirus (HCMV) infection is common and usually well controlled. Immunocompromised patients such as those undergoing bone marrow transplantation and infected newborns are especially vulnerable to HCMV disease (5,20,23).The immune control of HCMV replication appears to be mainly mediated by cellular immune responses. CD4 ϩ and CD8 ϩ T lymphocytes have been proposed to play a major role in the control of viral replication and in protection from disease. The contribution of anti-IE1-and anti-pp65-specific Tcell precursors to the total anti-HCMV immunity is now well established (7,8,15,19,34). IE1 is the major protein produced in the immediate-early phase of the HCMV replication cycle, and the matrix protein pp65 has been shown to be internalized immediately after the viral input without de novo synthesis and then to be available for presentation to specific CD8 ϩ cytotoxic T lymphocytes (CTL) (2, 19). The establishment of a rapid T-cell response before the synthesis of new infectious virions could provide an efficient means to avoid spreading of the virus. This strongly supports the idea that both CD4 ϩ and CD8 ϩ T cells directed against IE1 and pp65 could be critical for the generation of effective vaccines against HCMV and in anti-HCMV cell therapy.The transfer of anti-HCMV effector T cells to allogeneic bone marrow recipients results in protection from HCMV diseases associated with transplantation. The procedure is based on the use of HCMV-infected autologous fibroblasts to stimulate anti-HCMV-specific T cells in vitro (33). The authors showed that persistence of cytotoxic CD8 ϩ T cells in recipients was facilitated by a simultaneous recovery of CD4 ϩ helper T cells after bone marrow transplantation (33). More recently, the use of Epstein-Barr virus (EBV)-transformed B cells transduced with a recombinant retrovirus expressing pp65 has been suggested to allow the concomitant expansion of both anti-EBV-and anti-HCMV-specific T cells (31). A similar strategy used autologous B lymphoblastoid cells stably transfected with cDNA coding for either pp65 or IE1 for the generation of specific CD8 ϩ T-cell clones (26). Our approach to circumvent the use of infectious virus in ex vivo expansion protocols for cellular immunotherapy is based on a procedure allowing the simultaneous triggering of the anti-IE1 and -pp65 responses by means of a recombinant chimeric protein, IE1-pp65.In this paper, we report the construction and purification of a recombinant IE1-pp65 protein from insect cells. We demonstrate that an important proportion of anti-HCMV CD4 ϩ T cells was directed against IE1-pp65 in HCMV-seropositive donors and that the protein induced activation of HLA-DR3-restricted anti-IE1 CD4 ϩ T-cell clones, as assessed through gamma interferon (IFN-␥) secretion and cytotoxicity. Moreover, soluble IE1-pp65 was able to stimulate and to expand anti-pp65 CD8 ϩ T cells from peripheral blood mononuclear

show abstract

“…HA has been well characterized as an immunogen (29), and many T cell epitopes, such as the immunodominant 306 -318 epitope that binds to many different HLA-DR alleles including HLA-DR4 (30) have been defined. Both Ii chain-positive and -negative cell lines present endogenous HA to CD4-positive T cells, whether infected with live virus (9) or transfected with constructs for wild type or ER-retained mutants of HA (31). Presentation of endogenous HA to CD4-positive T cells was inhibited by chloroquine, suggesting endosomal processing (31).…”

mentioning

confidence: 99%

“…Both Ii chain-positive and -negative cell lines present endogenous HA to CD4-positive T cells, whether infected with live virus (9) or transfected with constructs for wild type or ER-retained mutants of HA (31). Presentation of endogenous HA to CD4-positive T cells was inhibited by chloroquine, suggesting endosomal processing (31). Most important for our analysis, the folding pathway and intracellular transport of HA has been well characterized (32)(33)(34)(35) and antibodies specific for different folding intermediates and native conformations were available (34).…”

mentioning

confidence: 99%

Major Histocompatibility Complex Class II-dependent Unfolding, Transport, and Degradation of Endogenous Proteins

Aichinger

Karlsson²,

Jackson³

et al. 1997

Journal of Biological Chemistry

View full text Add to dashboard Cite

We have analyzed the ability of major histocompatibility (MHC) class II molecules to capture proteins in the biosynthetic pathway and whether this may be associated with MHC class II-dependent antigen processing. When coexpressed with HLA-DR 4 molecules in HeLa cells, influenza hemagglutinin was inhibited from folding and trimerization in the biosynthetic pathway, targeted to endosomal compartments, and rapidly degraded. Due to the interaction with MHC class II molecules, therefore, unfolded forms of hemagglutinin were bypassing the quality control mechanism of the secretory pathway. More important, however, the transport, endocytosis, and rapid degradation of unfolded hemagglutinin in the presence of MHC class II molecules suggest that proteins captured in the endoplasmic reticulum by class II molecules may become substrates for antigen processing and presentation to CD4-positive T cells. In insect cells we show that this phenomenon is not restricted to a few proteins such as hemagglutinin. A highly heterogeneous mixture of proteins from the endoplasmic reticulum including coexpressed hemagglutinin can form stable complexes with soluble HLA-DR ␣ and ␤ chains that were transported into the supernatant. This mechanism may gain biological significance in abnormal situations associated with accumulation of unfolded or malfolded proteins in the endoplasmic reticulum, for example during viral infections.

show abstract

Presentation of newly synthesized glycoproteins to CD4+ T lymphocytes. An analysis using influenza hemagglutinin transport mutants.

Cited by 23 publications

References 42 publications

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Ex Vivo Stimulation and Expansion of both CD4⁺and CD8⁺T Cells from Peripheral Blood Mononuclear Cells of Human Cytomegalovirus-Seropositive Blood Donors by Using a Soluble Recombinant Chimeric Protein, IE1-pp65

Major Histocompatibility Complex Class II-dependent Unfolding, Transport, and Degradation of Endogenous Proteins

Contact Info

Product

Resources

About

Presentation of newly synthesized glycoproteins to CD4+ T lymphocytes. An analysis using influenza hemagglutinin transport mutants.

Cited by 23 publications

References 42 publications

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Presentation of Endogenously Synthesized MHC Class II-Restricted Epitopes by MHC Class II Cancer Vaccines Is Independent of Transporter Associated with Ag Processing and the Proteasome

Ex Vivo Stimulation and Expansion of both CD4+and CD8+T Cells from Peripheral Blood Mononuclear Cells of Human Cytomegalovirus-Seropositive Blood Donors by Using a Soluble Recombinant Chimeric Protein, IE1-pp65

Major Histocompatibility Complex Class II-dependent Unfolding, Transport, and Degradation of Endogenous Proteins

Contact Info

Product

Resources

About

Ex Vivo Stimulation and Expansion of both CD4⁺and CD8⁺T Cells from Peripheral Blood Mononuclear Cells of Human Cytomegalovirus-Seropositive Blood Donors by Using a Soluble Recombinant Chimeric Protein, IE1-pp65