Lack of tumor recognition by hTERT peptide 540-548-specific CD8+ T cells from melanoma patients reveals inefficient antigen processing

AbstractCancer cells frequently display defects in their antigen-processing pathway and thereby evade CD8 T cell immunity. We described a novel category of cancer antigens, named TEIPP, that emerge on cancers with functional loss of the peptide pump TAP. TEIPPs are non-mutated neoantigens despite their ‘self’ origin by virtue of their absence on normal tissues. Here, we describe the development of a synthetic long peptide (SLP) vaccine for the most immunogenic TEIPP antigen identified thus far, derived from the TAP-independent LRPAP1 signal sequence. LRPAP121–30-specific CD8 T cells were present in blood of all tested healthy donors as well as patients with non-small cell lung adenocarcinoma. SLPs with natural flanking, however, failed to be cross-presented by monocyte-derived dendritic cells. Since the C-terminus of LRPAP121–30 is an unconventional and weakly binding serine (S), we investigated if replacement of this anchor would result in efficient cross-presentation. Exchange into a valine (V) resulted in higher HLA-A2 binding affinity and enhanced T cell stimulation. Importantly, CD8 T cells isolated using the V-variant were able to bind tetramers with the natural S-variant and respond to TAP-deficient cancer cells. A functional screen with an array of N-terminal and C-terminal extended SLPs pointed at the 24-mer V-SLP, elongated at the N-terminus, as most optimal vaccine candidate. This SLP was efficiently cross-presented and consistently induced a strong polyclonal LRPAP121–30-specific CD8 T cells from the endogenous T cell repertoire. Thus, we designed a TEIPP SLP vaccine from the LRPAP1 signal sequence ready for validation in clinical trials.

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Digital Immunophenotyping Predicts Disease Free and Overall Survival in Early Stage Melanoma Patients

Cells ◽

10.3390/cells10020422 ◽

2021 ◽

Vol 10 (2) ◽

pp. 422

Author(s):

Francesco De Logu ◽

Francesca Galli ◽

Romina Nassini ◽

Filippo Ugolini ◽

Sara Simi ◽

...

Keyword(s):

Overall Survival ◽

T Cells ◽

Cd8 T Cells ◽

Early Stage ◽

High Density ◽

Stage Ii ◽

Prognostic Impact ◽

Training Cohort ◽

Melanoma Patients ◽

Disease Free

Background: the prognostic significance of tumor infiltrating lymphocytes (TILs) in intermediate/thick primary cutaneous melanoma (PCM) remains controversial, partially because conventional evaluation is not reliable, due to inter-observer variability and diverse scoring methods. We aimed to assess the prognostic impact of the density and spatial distribution of immune cells in early stage intermediate/thick PCM. Materials and Methods: digital image acquisition and quantitative analysis of tissue immune biomarkers (CD3, CD4, CD8, CD68, PD-L1, CD163, FOX-P3, and PD-1) was carried out in a training cohort, which included patients with primary PCM ≥ 2 mm diagnosed, treated, and followed-up prospectively in three Italian centers. Results were validated in an independent Italian cohort. Results: in the training cohort, 100 Stage II–III melanoma patients were valuable. At multivariable analysis, a longer disease free survival (DFS) was statistically associated with higher levels of CD4+ intratumoral T-cells (aHR [100 cell/mm2 increase] 0.98, 95%CI 0.95–1.00, p = 0.041) and CD163+ inner peritumoral (aHR [high vs. low] 0.56, 95%CI 0.32–0.99, p = 0.047). A statistically significant longer DFS (aHR [high-high vs. low-low] 0.52, 95%CI 0.28–0.99, p = 0.047) and overall survival (OS) (aHR [high-high vs. low-low] 0.39, 95%CI 0.18–0.85, p = 0.018) was found in patients with a high density of both intratumoral CD8+ T-cells and CD68+ macrophages as compared to those with low density of both intratumoral CD8+ T-cells and CD68+ macrophages. Consistently, in the validation cohort, patients with high density of both intratumoral CD8+ and CD3+ T-cells were associated to a statistically better DFS (aHR[high-high vs. low-low] 0.24, 95%CI 0.10–0.56, p < 0.001) and those with high density of both intratumoral CD8+ and CD68+ were associated to a statistically longer OS (aHR[high-high vs. low-low] 0.28, 95%CI 0.09–0.86, p = 0.025). Conclusion: our findings suggest that a specific preexisting profile of T cells and macrophages distribution in melanomas may predict the risk of recurrence and death with potential implications for the stratification of stage II–III melanoma patients.

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CXC Chemokine Receptor 3 Expression by Activated CD8+ T cells Is Associated with Survival in Melanoma Patients with Stage III Disease

Cancer Research ◽

10.1158/0008-5472.can-04-2059 ◽

2004 ◽

Vol 64 (21) ◽

pp. 7697-7701 ◽

Cited By ~ 97

Author(s):

Irene M. Mullins ◽

Craig L. Slingluff ◽

Jae K. Lee ◽

Courtney F. Garbee ◽

Jianfen Shu ◽

...

Keyword(s):

T Cells ◽

Chemokine Receptor ◽

Cd8 T Cells ◽

Stage Iii ◽

Cxc Chemokine ◽

Melanoma Patients

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Chemical Castration of Melanoma Patients Does Not Increase the Frequency of Tumor-specific CD4 and CD8 T Cells After Peptide Vaccination

Journal of Immunotherapy ◽

10.1097/cji.0b013e31829419f3 ◽

2013 ◽

Vol 36 (4) ◽

pp. 276-286 ◽

Cited By ~ 2

Author(s):

Luis M. Vence ◽

Chiyu Wang ◽

Himabindu Pappu ◽

Ryan E. Anson ◽

Tejal A. Patel ◽

...

Keyword(s):

T Cells ◽

Cd8 T Cells ◽

Peptide Vaccination ◽

Chemical Castration ◽

Melanoma Patients

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Differential Responsiveness to IL-2, IL-7, and IL-15 Common Receptor γ Chain Cytokines by Antigen-specific Peripheral Blood Naive or Memory Cytotoxic CD8+ T Cells From Healthy Donors and Melanoma Patients

Journal of Immunotherapy ◽

10.1097/cji.0b013e3181998e03 ◽

2009 ◽

Vol 32 (3) ◽

pp. 252-261 ◽

Cited By ~ 9

Author(s):

Rachel Rosenthal ◽

Célia Groeper ◽

Laura Bracci ◽

Michel Adamina ◽

Chantal Feder-Mengus ◽

...

Keyword(s):

T Cells ◽

Cd8 T Cells ◽

Peripheral Blood ◽

Common Receptor ◽

Healthy Donors ◽

Melanoma Patients ◽

Differential Responsiveness

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Bim and PD-1 identify effector CD8 T cells responsive to anti-PD-1 therapy in metastatic melanoma patients

Journal for ImmunoTherapy of Cancer ◽

10.1186/2051-1426-3-s2-p298 ◽

2015 ◽

Vol 3 (S2) ◽

Author(s):

Roxana Dronca ◽

Xin Liu ◽

Kottschade Lisa ◽

Rob Mcwilliams ◽

Svetomir Markovic ◽

...

Keyword(s):

T Cells ◽

Metastatic Melanoma ◽

Cd8 T Cells ◽

Melanoma Patients

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NKG2D Polymorphism in Melanoma Patients from Southeastern Spain

Cancers ◽

10.3390/cancers11040438 ◽

2019 ◽

Vol 11 (4) ◽

pp. 438 ◽

Cited By ~ 1

Author(s):

Lourdes Gimeno ◽

Helios Martínez-Banaclocha ◽

M. Bernardo ◽

José Bolarin ◽

Luis Marín ◽

...

Keyword(s):

T Cells ◽

Natural Killer ◽

Cd8 T Cells ◽

Nk Cell ◽

Critical Role ◽

Healthy Controls ◽

Nucleotide Polymorphisms ◽

Southeastern Spain ◽

Melanoma Patients ◽

Risk Of Cancer

Background: Natural killer (NK) and CD8+ T cells are involved in the immune response against melanoma. C-Type lectin-like NK cell receptors are located in the Natural Killer Complex (NKC) region 12p13.2-p12.3 and play a critical role in regulating the activity of NK and CD8+ T cells. An association between polymorphisms in the NKC region, including the NKG2D gene and NKG2A promoter, and the risk of cancer has been previously described. The aim of this study was to analyze the association of polymorphisms in the NKC region with cutaneous melanoma in patients from southeastern Spain. Methods: Seven single-nucleotide polymorphisms (SNPs) in the NKG2D gene (NKC3,4,7,9,10,11,12), and one SNP in the NKG2A promoter (NKC17) were genotyped by a TaqMan 5′ Nuclease Assay in 233 melanoma patients and 200 matched healthy controls. Results: A linkage disequilibrium analysis of the SNPs performed in the NKC region revealed two blocks of haplotypes (Hb-1 and Hb-2) with 14 and seven different haplotype subtypes, respectively. The third most frequent haplotype from the block Hb-2—NK3 (CAT haplotype)—was significantly more frequent on melanoma patients than on healthy controls (p = 0.00009, Pc = 0.0006). No further associations were found when NKC SNPs were considered independently. Conclusions: Our results suggest an association between NKG2D polymorphisms and the risk of cutaneous malignant melanoma.

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The nonpolymorphic MHC Qa-1b mediates CD8+ T cell surveillance of antigen-processing defects

Journal of Experimental Medicine ◽

10.1084/jem.20091429 ◽

2009 ◽

Vol 207 (1) ◽

pp. 207-221 ◽

Cited By ~ 65

Author(s):

Cláudia C. Oliveira ◽

Peter A. van Veelen ◽

Bianca Querido ◽

Arnoud de Ru ◽

Marjolein Sluijter ◽

...

Keyword(s):

T Cells ◽

T Cell ◽

Cd8 T Cells ◽

Antigen Processing ◽

Germ Line ◽

Wide Spectrum ◽

Cell Subset ◽

Target Cells ◽

Peptide Epitopes ◽

Leader Peptides

The nonclassical major histocompatibility complex (MHC) Qa-1b accommodates monomorphic leader peptides and functions as a ligand for germ line receptors CD94/NKG2, which are expressed by natural killer cells and CD8+ T cells. We here describe that the conserved peptides are replaced by a novel peptide repertoire of surprising diversity as a result of impairments in the antigen-processing pathway. This novel peptide repertoire represents immunogenic neoantigens for CD8+ T cells, as we found that these Qa-1b–restricted T cells dominantly participated in the response to tumors with processing deficiencies. A surprisingly wide spectrum of target cells, irrespective of transformation status, MHC background, or type of processing deficiency, was recognized by this T cell subset, complying with the conserved nature of Qa-1b. Target cell recognition depended on T cell receptor and Qa-1b interaction, and immunization with identified peptide epitopes demonstrated in vivo priming of CD8+ T cells. Our data reveal that Qa-1b, and most likely its human homologue human leukocyte antigen-E, is important for the defense against processing-deficient cells by displacing the monomorphic leader peptides, which relieves the inhibition through CD94/NKG2A on lymphocytes, and by presenting a novel repertoire of immunogenic peptides, which recruits a subset of cytotoxic CD8+ T cells.

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