scholarly journals P02.02 Generating neo- and self-antigen screening libraries for class II HLA presentation

2020 ◽  
Vol 8 (Suppl 2) ◽  
pp. A20.2-A20
Author(s):  
V Pinamonti ◽  
N Felix ◽  
JM Lindner
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Tumor Antigens ◽  
Cell Receptor ◽  
Class Ii ◽  
Patient Specific ◽  
Spectrometric Analysis ◽  
Cross Presentation ◽  
Class Ii Mhc ◽  
Antigen Presenting

BackgroundThe identification of neo-antigens presented by tumor cells is an essential tool for cancer prevention, diagnosis, and therapy. Current approaches frequently involve mass spectrometric analysis, but these workflows do not concomitantly identify the cognate T-cell receptor. Likewise, TCR functional screens are often limited to a subset of predicted neo-epitopes.Materials and MethodsHere, we present a new method for the generation of an un-biased antigen-presenting library. Due to the genomic instability of tumors, patient-specific libraries will be cloned using random primers, ensuring the cloning of tumor-specific transcribed regions. This approach will not only address class I presentation of intracellular tumor antigens, but is also designed to simultaneously screen for cross-presentation on class II MHC complexes by professional antigen-presenting cells, an increasingly important component of anti-tumor immune responses. To guarantee presentation of genetically encoded antigens on class II MHC complexes, a signal motif for chaperone-mediated autophagy (CMA) is introduced in front of the cDNA sequence. Furthermore, antigens will be processed by the intracellular machinery, avoiding potential restrictions on spliced peptides.ConclusionsOnce established, these libraries can be exploited in high-throughput screens to functionally identify neo-antigens together with their corresponding T-cell receptor.Disclosure InformationV. Pinamonti: Other; Significant; Janssen. N. Felix: Other; Significant; Janssen. J.M. Lindner: Other; Significant; Janssen.

The Density of the Class II MHC T Cell Receptor Ligand Influences IFN-γ/IL-4 Ratios in Immune Responsesin Vivo

1998 ◽  
Vol 183 (1) ◽  
pp. 70-79 ◽  
Author(s):  
Lisa DiMolfetto ◽  
Heather A. Neal ◽  
Adrian Wu ◽  
Christina Reilly ◽  
David Lo
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Class Ii ◽  
Receptor Ligand ◽  
Class Ii Mhc ◽  
Ifn Γ

Binary and ternary complexes between T-cell receptor, class II MHC and superantigen in vitro

Nature ◽  
1994 ◽  
Vol 369 (6478) ◽  
pp. 324-327 ◽  
Author(s):  
Alpna Seth ◽  
Lawrence J. Stern ◽  
Tom H. M. Ottenhoff ◽  
Isaac Engel ◽  
Michael J. Owen ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Class Ii ◽  
Ternary Complexes ◽  
Binary And Ternary Complexes ◽  
Class Ii Mhc

scholarly journals The surface phenotype of dendritic cells purified from mouse thymus and spleen: investigation of the CD8 expression by a subpopulation of dendritic cells.

1992 ◽  
Vol 176 (1) ◽  
pp. 47-58 ◽  
Author(s):  
D Vremec ◽  
M Zorbas ◽  
R Scollay ◽  
D J Saunders ◽  
C F Ardavin ◽  
...  
Keyword(s):  
T Cells ◽  
Dendritic Cells ◽  
T Cell ◽  
Magnetic Beads ◽  
Cell Receptor ◽  
Class Ii ◽  
Immunofluorescent Staining ◽  
Alpha Chain ◽  
Class Ii Mhc ◽  
Antigen Presenting

A new procedure for rapid isolation of dendritic cells (DC) was devised, involving collagenase digestion of tissues, dissociation of lymphoid-DC complexes, selection of light-density cells, then depletion of lymphocytes and other non-DC by treatment with a mixture of lineage-specific monoclonal antibodies (mAbs) and removal with anti-immunoglobulin-coupled magnetic beads. This enriched population (approximately 80% DC) was further purified when required by fluorescence-activated cell sorting for cells expressing high levels of class II major histocompatibility complex (MHC). The isolated DC were characterized by immunofluorescent staining using a panel of 30 mAbs. Thymic DC were surface positive for a number of markers characteristic of T cells, but they were distinct from T-lineage cells in expressing high levels of class II MHC, in lacking expression of the T cell receptor (TCR)-CD3 complex, and having TCR beta and gamma genes in germline state. Splenic DC shared many markers with thymic DC, but were negative for most T cell markers, with the exception of CD8. A substantial proportion of DC from both thymus and spleen expressed CD8 at high levels, comparable with that on T cells. This appeared to be authentic CD8, and was produced by the DC themselves, since they contained CD8 alpha mRNA. Thymic DC presented both the CD8 alpha and beta chains on the cell surface (Ly-2+3+), although the alpha chain was in excess; the splenic DC expressed only the CD8 alpha chain (Ly-2+3-). It is suggested that the expression of CD8 could endow certain antigen-presenting DC with a veto function.

Affinity and kinetics of the interactions between an αβ T-cell receptor and its superantigen and class II-MHC/peptide ligands

1997 ◽  
Vol 34 (6) ◽  
pp. 493-503 ◽  
Author(s):  
Sanjay S. Khandekar ◽  
Pamela P. Brauer ◽  
Jerome W. Naylor ◽  
Hsiu-Ching Chang ◽  
Petra Kern ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Class Ii ◽  
Peptide Ligands ◽  
Class Ii Mhc ◽  
Kinetics Of

scholarly journals A T Cell Receptor–specific Blockade of Positive Selection

1999 ◽  
Vol 189 (1) ◽  
pp. 13-24 ◽  
Author(s):  
Kristin K. Baldwin ◽  
Philip A. Reay ◽  
Lawren Wu ◽  
Andrew Farr ◽  
Mark M. Davis
Keyword(s):  
T Cell ◽  
Positive Selection ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Peptide Sequence ◽  
Class Ii Mhc ◽  
Peptide Complexes ◽  
Antigen Presenting ◽  
Medullary Epithelial Cells ◽  
Selection Of

To investigate the influence of endogenous peptides on the developmental processes that occur during thymocyte selection, we have used monoclonal antibodies that preferentially recognize the major histocompatibility complex (MHC) molecule I-Ek when it is bound to the moth cytochrome c peptide (88–103). One of these antibodies (G35) specifically blocks the positive selection of transgenic thymocytes expressing a T cell receptor that is reactive to this peptide– MHC complex. Furthermore, G35 does not block the differentiation of transgenic T cells bearing receptors for a different I-Ek–peptide complex. This antibody recognizes a subset of endogenous I-Ek–peptide complexes found on a significant fraction of thymic antigen-presenting cells, including cortical and medullary epithelial cells. The sensitivity of G35 to minor alterations in peptide sequence suggests that the thymic peptide–MHC complexes that mediate the positive selection of a particular class II MHC–restricted thymocyte are structurally related to the complexes that can activate it in the periphery.

Structure and specificity of a class II MHC alloreactive gamma delta T cell receptor heterodimer

Science ◽  
1989 ◽  
Vol 245 (4919) ◽  
pp. 746-749 ◽  
Author(s):  
L. Matis ◽  
A. Fry ◽  
R. Cron ◽  
M. Cotterman ◽  
R. Dick ◽  
...  
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Class Ii ◽  
Gamma Delta ◽  
Class Ii Mhc ◽  
Gamma Delta T Cell

scholarly journals Positive and Negative Selection in Transgenic Mice Expressing a T-Cell Receptor Specific for Influenza Nucleoprotein and Endogenous Superantigen

1993 ◽  
Vol 3 (3) ◽  
pp. 159-174 ◽  
Author(s):  
Clio Mamalaki ◽  
James Elliott ◽  
Trisha Norton ◽  
Nicholas Yannoutsos ◽  
Alain R. Townsend ◽  
...  
Keyword(s):  
T Cells ◽  
T Cell ◽  
Transgenic Mice ◽  
T Cell Receptor ◽  
Cell Receptor ◽  
Class Ii ◽  
Cytotoxic T Cell ◽  
Dependent Manner ◽  
Class Ii Mhc ◽  
Peripheral T Cells

A transgenic mouse was generated expressing on most (>80%) of thymocytes and peripheral T cells a T-cell receptor isolated from a cytotoxic T-cell clone (F5). This clone is CD8+and recognizes αα366-374 of the nucleoprotein (NP 366-374) of influenza virus (A/NT/60/68), in the context of Class ,MHC Db(Townsend et al., 1986). The receptor utilizes the Vβ11 and Vα4 gene segments for the β chain and α chain, respectively (Palmer et al., 1989). The usage of Vβ11 makes this TcR reactive to Class II IE molecules and an endogenous ligand recently identified as a product of the endogenous mammary tumour viruses (Mtv) 8, 9, and 11 (Dyson et al., 1991). Here we report the development of F5 transgenic T cells and their function in mice of the appropriate MHC (C57BL/10 H-2b, IE-) or in mice expressing Class II MHC IE (e.g., CBA/Ca H-2kand BALB/c H-2d) and the endogenous Mtv ligands. Positive selection of CD8+T cells expressing the Vβ11 is seen in C57BL/10 transgenic mice (H-2b). Peripheral T cells from these mice are capable of killing target cells in an antigen-dependent manner after a period ofin vitroculture with IL-2. In the presence of Class II MHC IE molecules and the endogenous Mtv ligand, most of the single-positive cells carrying the transgenic T-cell receptor are absent in the thymus. Unexpectedly, CD8+peripheral T-cells in these (H-2kor H-2d) F5 mice are predominantly Vβ11 positive and also have the capacity to kill targets in an antigen-dependent manner. This is true even following backcrossing of the F5 TcR transgene to H-2dscid/scid mice, in which functional rearrangement of endogenous TcR alpha- and beta-chain genes is impaired.

Probing Molecular Interactions within Class II MHC Aq/Glycopeptide/T-Cell Receptor Complexes Associated with Collagen-Induced Arthritis

2007 ◽  
Vol 50 (23) ◽  
pp. 5627-5643 ◽  
Author(s):  
Ida E. Andersson ◽  
Balik Dzhambazov ◽  
Rikard Holmdahl ◽  
Anna Linusson ◽  
Jan Kihlberg
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Molecular Interactions ◽  
Cell Receptor ◽  
Class Ii ◽  
Collagen Induced Arthritis ◽  
Receptor Complexes ◽  
Class Ii Mhc

scholarly journals Molecular Requirements for T Cell Recognition by a Major Histocompatibility Complex Class II–restricted T Cell Receptor: The Involvement of the Fourth Hypervariable Loop of the Vα Domain

1999 ◽  
Vol 189 (3) ◽  
pp. 509-520 ◽  
Author(s):  
Jayant Thatte ◽  
Ayub Qadri ◽  
Caius Radu ◽  
E. Sally Ward
Keyword(s):  
T Cell ◽  
T Cell Receptor ◽  
Mhc Class Ii ◽  
T Cell Activation ◽  
Cell Activation ◽  
Cell Receptor ◽  
Cell Interaction ◽  
Class Ii ◽  
Wild Type ◽  
Antigen Presenting

The role of two central residues (K68, E69) of the fourth hypervariable loop of the Vα domain (HV4α) in antigen recognition by an MHC class II–restricted T cell receptor (TCR) has been analyzed. The TCR recognizes the NH2-terminal peptide of myelin basic protein (Ac1-11, acetylated at NH2 terminus) associated with the class II MHC molecule I-Au. Lysine 68 (K68) and glutamic acid 69 (E69) of HV4α have been mutated both individually and simultaneously to alanine (K68A, E69A). The responsiveness of transfectants bearing wild-type and mutated TCRs to Ac1-11–I-Au complexes has been analyzed in the presence and absence of expression of the coreceptor CD4. The data demonstrate that in the absence of CD4 expression, K68 plays a central role in antigen responsiveness. In contrast, the effect of mutating E69 to alanine is less marked. CD4 coexpression can partially compensate for the loss of activity of the K68A mutant transfectants, resulting in responses that, relative to those of the wild-type transfectants, are highly sensitive to anti-CD4 antibody blockade. The observations support models of T cell activation in which both the affinity of the TCR for cognate ligand and the involvement of coreceptors determine the outcome of the T cell–antigen-presenting cell interaction.

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