Restricted islet-cell reactive T cell repertoire of early pancreatic islet infiltrates in NOD mice

Baker, Felix J.; Lee, Mark; Chien, Yueh‐hsiu; Davis, Mark M.

doi:10.1073/pnas.142284899

Cited by 68 publications

(87 citation statements)

References 65 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…In contrast with established primer sets for the amplification of TCR CDR3 regions from single T cells (10,13,14,45), our primers amplify full-length variable regions. In the elegant study by Ozawa and colleagues (9), they were able to generate matching TCR a/b cDNAs from approximately 20% of processed single cells using the 5 0 -RACE technique.…”

Section: Discussionmentioning

confidence: 99%

Functional TCR Retrieval from Single Antigen-Specific Human T Cells Reveals Multiple Novel Epitopes

Simon

Omokoko

Breitkreuz

et al. 2014

Cancer Immunology Research

View full text Add to dashboard Cite

The determination of the epitope specificity of disease-associated T-cell responses is relevant for the development of biomarkers and targeted immunotherapies against cancer, autoimmune, and infectious diseases. The lack of known T-cell epitopes and corresponding T-cell receptors (TCR) for novel antigens hinders the efficient development and monitoring of new therapies. We developed an integrated approach for the systematic retrieval and functional characterization of TCRs from single antigen-reactive T cells that includes the identification of epitope specificity. This is accomplished through the rapid cloning of full-length TCR-a and TCR-b chains directly from single antigen-specific CD8 þ or CD4 þ T lymphocytes. The functional validation of cloned TCRs is conducted using in vitro-transcribed RNA transfer for expression of TCRs in T cells and HLA molecules in antigen-presenting cells. This method avoids the work and bias associated with repetitive cycles of in vitro T-cell stimulation, and enables fast characterization of antigen-specific T-cell responses. We applied this strategy to viral and tumor-associated antigens (TAA), resulting in the retrieval of 56 unique functional antigenspecific TCRs from human CD8 þ and CD4 þ T cells (13 specific for CMV-pp65, 16 specific for the well-known TAA NY-ESO-1, and 27 for the novel TAA TPTE), which are directed against 39 different epitopes. The proof-of-concept studies with TAAs NY-ESO-1 and TPTE revealed multiple novel TCR specificities. Our approach enables the rational development of immunotherapy strategies by providing antigen-specific TCRs and immunogenic epitopes.

show abstract

Section: Discussionmentioning

confidence: 99%

Functional TCR Retrieval from Single Antigen-Specific Human T Cells Reveals Multiple Novel Epitopes

Simon

Omokoko

Breitkreuz

et al. 2014

Cancer Immunology Research

View full text Add to dashboard Cite

show abstract

“…The lack of Asp57 not only gives the potential to interact with peptides containing acidic residues at P9 but also with T-cell receptors with a negatively charged CDR3b loop. 36 The relationship between DR8 and these alleles was further confirmed by analyzing the theoretical capacity of DR8 peptides to binding DQ8, I-A g7 and DQ2, using the MHC2Pred tool for prediction of promiscuous MHC class II binders (http://www.imtech.res.in/raghava/ mhc2pred/). Of the 216 DR8 sequences analyzed, 59 and 51%, respectively, were potential good binders to DQ8 and I-Ag7, using a very stringent threshold of 1.…”

Section: Hla-dr8-binding Motif L Muixí Et Almentioning

confidence: 97%

The peptide-binding motif of HLA-DR8 shares important structural features with other type 1 diabetes-associated alleles

et al. 2011

View full text Add to dashboard Cite

The objective of this study was to characterize the peptide-binding motif of the major histocompatibility complex (MHC) class II HLA-DR8 molecule included in the type 1 diabetes-associated haplotype DRB1*0801-DQA1*0401/DQB1*0402 (DR8-DQ4), and compare it with that of other diabetes-associated MHC class II alleles; DR8-bound peptides were eluted from an HLA-DR homozygous lymphoblastoid cell line. The repertoire was characterized by peptide sequencing using a LTQ ion trap mass spectrometer coupled to a multidimensional liquid chromatography system. After validation of the spectra identification, the definition of the HLA-DR8 peptide-binding motif was achieved from the analysis of 486 natural ligands, based on serial alignments of all possible HLA-DR-binding cores. The DR8 motif showed a strong similarity with the peptide-binding motifs of other MHC class II diabetes-associated alleles, HLA-DQ8 and H-2 I-A g7 . Similar to HLA-DQ8 and H-2 I-A g7 , HLA-DR8 preferentially binds peptides with an acidic residue at position P9 of the binding core, indicating that DR8 is the susceptibility component of the DR8-DQ4 haplotype. Indeed, some DR8 peptides were identical to peptides previously identified as DQ8-or I-A g7 ligands, and several diabetes-specific peptides associated with DQ8 or I-A g7 could theoretically bind to HLA-DR8. These data further strengthen the association of HLA-DR8 with type I diabetes.

show abstract

“…1A shows a dot plot of pancreatic lymphocytes from a 9-wk-old NOD mouse in which B cells (B220 ϩ ) and T cells (CD3 ϩ ) are found in roughly even proportions (7.9 and 8.7%, respectively, B:T ratio, 0.48). The unstained population in this assay is due to pancreatic tissue components, including islets that segregate with lymphocytes in density, size, and granularity (24). To evaluate the possible contribution of non-isletinvading lymphocytes, such as those from blood vessels in the pancreas, we used nonautoimmune C57BL/6 mice, in which there is no insulitis, as controls.…”

Section: Lymphocytes In the Islets Of Prediabetic Nod Mice Form Tertimentioning

confidence: 99%

“…In particular, little is known about B lymphocytes within these foci of autoreactivity. Studies on T cell receptors have shown oligoclonality within isolated islets, with a polyclonal repertoire across the pancreas as a whole (23,24), but their relationship to regional lymph nodes was not explored. Studies of cellular dynamics revealed that B and T lymphocytes invade the islets concurrently (25) and that CD4 ϩ T cell priming and proliferation occur in draining pancreatic lymph nodes before the onset of insulitis (26 -28), suggesting a collaborative relationship between these secondary lymphoid organs and TLS in the islets.…”

mentioning

confidence: 99%

Tertiary Lymphoid Structures in the Pancreas Promote Selection of B Lymphocytes in Autoimmune Diabetes

Kendall¹,

Yu²,

Woodward

et al. 2007

The Journal of Immunology

View full text Add to dashboard Cite

Autoimmune diabetes occurs when invading lymphocytes destroy insulin-producing β cells in pancreatic islets. The role of lymphocytic aggregates at this inflammatory site is not understood. We find that B and T lymphocytes attacking islets in NOD mice organize into lymphoid structures with germinal centers. Analysis of BCR L chain genes was used to investigate selection of B lymphocytes in these tertiary lymphoid structures and in draining pancreatic lymph nodes. The pancreatic repertoire as a whole was found to be highly diverse, with the profile of L chain genes isolated from whole pancreas differing from that observed in regional lymph nodes. A Vκ14 L chain predominated within the complex pancreatic repertoire of NOD mice. Skewing toward Vκ4 genes was observed in the pancreas when the repertoire of NOD mice was restricted using a fixed Ig H chain transgene. Nucleotide sequencing of expressed Vκs identified shared mutations in some sequences consistent with Ag-driven selection and clonal expansion at the site of inflammation. Isolated islets contained oligoclonal B lymphocytes enriched for the germinal center marker GL7 and for sequences containing multiple mutations within CDRs, suggesting local T-B interactions. Together, these findings identify a process that selects B lymphocyte specificities within the pancreas, with further evolution of the selected repertoire at the inflamed site. This interpretation is reinforced by Ag-binding studies showing a large population of insulin-binding B lymphocytes in the pancreas compared with draining lymph nodes.

show abstract

Restricted islet-cell reactive T cell repertoire of early pancreatic islet infiltrates in NOD mice

Cited by 68 publications

References 65 publications

Functional TCR Retrieval from Single Antigen-Specific Human T Cells Reveals Multiple Novel Epitopes

Functional TCR Retrieval from Single Antigen-Specific Human T Cells Reveals Multiple Novel Epitopes

The peptide-binding motif of HLA-DR8 shares important structural features with other type 1 diabetes-associated alleles

Tertiary Lymphoid Structures in the Pancreas Promote Selection of B Lymphocytes in Autoimmune Diabetes

Contact Info

Product

Resources

About