Maria-Danae Jessel, Owen McGreevy, Lauryn McReynolds, Lekh N Dahal, Timothy Gilbert, Hassan Z Malik, Christopher E Goldring, Laura E Randle
Cholangiocarcinoma is a malignancy of significant unmet clinical need with limited therapeutic options. Most patients are diagnosed at advanced or metastatic stages, where surgical resection with curative intent is no longer an option. Gemcitabine-cisplatin chemotherapy has been the standard of care for these patients, remaining unchanged for over a decade. Recently, the addition of the programmed death ligand 1 inhibitor, durvalumab, to this regimen demonstrated an objective response rate of 26.7% in a phase III trial, becoming the new standard of care for advanced cholangiocarcinoma. Although considered a success in cholangiocarcinoma treatment, the results indicate that only a small proportion of patients respond to treatment with immune checkpoint inhibitors. Emerging evidence suggests that many cholangiocarcinoma tumors exhibit an immunologically ‘cold’ tumor microenvironment, characterised by predominance of immunosuppressive immune populations and limited infiltration of cytotoxic T cells, which contributes to their resistance to immune checkpoint inhibitors. This review provides a comprehensive overview of the research studies that have employed immunomodulatory strategies in cholangiocarcinoma aimed at priming the tumor microenvironment for a more effective response to immune checkpoint inhibitors. This update will also evaluate the strengths and limitations of current pre-clinical models of cholangiocarcinoma, with emphasis on more advanced translational models. These complex models remain underutilised, hindering the development of novel therapeutic approaches. We suggest that these complex preclinical models may help translation of therapies into clinical practice.
{"title":"Targeting the tumor microenvironment in cholangiocarcinoma to improve immune checkpoint blockade: potential strategies and translational pre-clinical models","authors":"Maria-Danae Jessel, Owen McGreevy, Lauryn McReynolds, Lekh N Dahal, Timothy Gilbert, Hassan Z Malik, Christopher E Goldring, Laura E Randle","doi":"10.1002/cti2.70057","DOIUrl":"https://doi.org/10.1002/cti2.70057","url":null,"abstract":"<p>Cholangiocarcinoma is a malignancy of significant unmet clinical need with limited therapeutic options. Most patients are diagnosed at advanced or metastatic stages, where surgical resection with curative intent is no longer an option. Gemcitabine-cisplatin chemotherapy has been the standard of care for these patients, remaining unchanged for over a decade. Recently, the addition of the programmed death ligand 1 inhibitor, durvalumab, to this regimen demonstrated an objective response rate of 26.7% in a phase III trial, becoming the new standard of care for advanced cholangiocarcinoma. Although considered a success in cholangiocarcinoma treatment, the results indicate that only a small proportion of patients respond to treatment with immune checkpoint inhibitors. Emerging evidence suggests that many cholangiocarcinoma tumors exhibit an immunologically ‘cold’ tumor microenvironment, characterised by predominance of immunosuppressive immune populations and limited infiltration of cytotoxic T cells, which contributes to their resistance to immune checkpoint inhibitors. This review provides a comprehensive overview of the research studies that have employed immunomodulatory strategies in cholangiocarcinoma aimed at priming the tumor microenvironment for a more effective response to immune checkpoint inhibitors. This update will also evaluate the strengths and limitations of current pre-clinical models of cholangiocarcinoma, with emphasis on more advanced translational models. These complex models remain underutilised, hindering the development of novel therapeutic approaches. We suggest that these complex preclinical models may help translation of therapies into clinical practice.</p>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 10","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-24","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70057","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145367143","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Pei Dai, Oliver Skinner, Xufeng Lin, Aiden Telfser, Stephanie Ruiz-Diaz, Rohit G Saldanha, Katie Frith, Ming-Wei Lin, Kahn Preece, Paul E Gray, Alberto Pinzon-Charry, Anna Sullivan, Stephen Adelstein, Winnie WY Tong, Matthew JS Parker, Laila Girgis, Brynn Wainstein, Samar Ojami, Elissa K Deenick, Leonard D Goldstein, Michael J Rogers, Tri Giang Phan
Objective
To develop a rapid functional assay to validate variants of uncertain significance (VUS) in the MEFV gene.
Methods
Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of Clostridium difficile toxin A was directly visualised by immunofluorescence microscopy. A semi-automated algorithm was developed to count cells and ASC specks.
Results
The semi-automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity.
Conclusion
The semi-automated ASC speck assay may be a useful test to functionally validate VUS in the MEFV gene and screen for pyrin inflammasome overactivity.
{"title":"A semi-automated ASC speck assay to evaluate pyrin inflammasome activation","authors":"Pei Dai, Oliver Skinner, Xufeng Lin, Aiden Telfser, Stephanie Ruiz-Diaz, Rohit G Saldanha, Katie Frith, Ming-Wei Lin, Kahn Preece, Paul E Gray, Alberto Pinzon-Charry, Anna Sullivan, Stephen Adelstein, Winnie WY Tong, Matthew JS Parker, Laila Girgis, Brynn Wainstein, Samar Ojami, Elissa K Deenick, Leonard D Goldstein, Michael J Rogers, Tri Giang Phan","doi":"10.1002/cti2.70054","DOIUrl":"https://doi.org/10.1002/cti2.70054","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objective</h3>\u0000 \u0000 <p>To develop a rapid functional assay to validate variants of uncertain significance (VUS) in the <i>MEFV</i> gene.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Overactivity of the pyrin inflammasome pathway and ASC speck oligomerisation in response to stimulation with low concentrations of <i>Clostridium difficile</i> toxin A was directly visualised by immunofluorescence microscopy. A semi-automated algorithm was developed to count cells and ASC specks.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The semi-automated ASC speck assay is able to discriminate between healthy controls and patients with familial Mediterranean fever (FMF) and pyrin inflammasome overactivity with high sensitivity. It is also able to discriminate pyrin inflammasome overactivity from other autoinflammatory disease controls with high specificity.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The semi-automated ASC speck assay may be a useful test to functionally validate VUS in the <i>MEFV</i> gene and screen for pyrin inflammasome overactivity.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 10","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70054","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145366859","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Hannah Law, Raymond H. Y. Louie, Omid R. Faridani, Alexandra Carey Hoppé, Mollie Boyd, Mengfei Chen, Jerome Samir, Solange Obeid, Brad Milner, Fabio Luciani, Anthony D. Kelleher, Vanessa Venturi, C. Mee Ling Munier
Objectives
CD4+ T cells partitioned into different cell lineages based on transcription factor, cytokine and chemokine expression have defined functions that work cooperatively to ensure optimum operation of the immune system. This study investigated the lineages and interactions of key CD4+ T-cell subsets within human lymph nodes (LNs) post-vaccination to assess the potential for adaptability and flexibility in an immune response.
Methods
Ultrasound-guided fine needle biopsies/aspirates were used to isolate T follicular helper (Tfh) and Precursor (Pre)-Tfh cells from the vaccine-draining and contralateral axillary LNs of five individuals at 5 days after influenza vaccination, followed by single-cell RNA sequencing, gene expression and T-cell receptor analysis.
Results
Tfh and Pre-Tfh cells within five clusters with distinct gene expression profiles, designated: Resting, Activated migrating, B-cell-interacting Tfh, Proliferating and Cytotoxic, exhibited attributes of more than one T-cell lineage and expanded T-cell clones were present in more than one cluster, suggesting divergent differentiation into different fate lineages from a common precursor. Inferred pseudotime suggested a bifurcating trajectory rooted in the resting cluster and terminating at either the Proliferating or Cytotoxic clusters. This analysis predicted the transition of cells through activation states and the gain and loss of lineage attributes and effector functions. Enriched gene pathways along the pseudotime trajectories were consistent with these functional transitions and involvement in the immune response to vaccination.
Conclusion
These results reveal the flexible potential within the Tfh lineage that could be leveraged to drive more efficient vaccine responses and inform rational vaccine design.
{"title":"Human axillary lymph node T follicular helper (Tfh) and Precursor-Tfh cells exhibit functional flexibility following seasonal influenza vaccination","authors":"Hannah Law, Raymond H. Y. Louie, Omid R. Faridani, Alexandra Carey Hoppé, Mollie Boyd, Mengfei Chen, Jerome Samir, Solange Obeid, Brad Milner, Fabio Luciani, Anthony D. Kelleher, Vanessa Venturi, C. Mee Ling Munier","doi":"10.1002/cti2.70056","DOIUrl":"https://doi.org/10.1002/cti2.70056","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>CD4<sup>+</sup> T cells partitioned into different cell lineages based on transcription factor, cytokine and chemokine expression have defined functions that work cooperatively to ensure optimum operation of the immune system. This study investigated the lineages and interactions of key CD4<sup>+</sup> T-cell subsets within human lymph nodes (LNs) post-vaccination to assess the potential for adaptability and flexibility in an immune response.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Ultrasound-guided fine needle biopsies/aspirates were used to isolate T follicular helper (Tfh) and Precursor (Pre)-Tfh cells from the vaccine-draining and contralateral axillary LNs of five individuals at 5 days after influenza vaccination, followed by single-cell RNA sequencing, gene expression and T-cell receptor analysis.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Tfh and Pre-Tfh cells within five clusters with distinct gene expression profiles, designated: Resting, Activated migrating, B-cell-interacting Tfh, Proliferating and Cytotoxic, exhibited attributes of more than one T-cell lineage and expanded T-cell clones were present in more than one cluster, suggesting divergent differentiation into different fate lineages from a common precursor. Inferred pseudotime suggested a bifurcating trajectory rooted in the resting cluster and terminating at either the Proliferating or Cytotoxic clusters. This analysis predicted the transition of cells through activation states and the gain and loss of lineage attributes and effector functions. Enriched gene pathways along the pseudotime trajectories were consistent with these functional transitions and involvement in the immune response to vaccination.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>These results reveal the flexible potential within the Tfh lineage that could be leveraged to drive more efficient vaccine responses and inform rational vaccine design.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 10","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70056","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145366858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Eosinophilia, characterised by elevated eosinophil levels, is associated with various allergic and inflammatory conditions. This study aimed to elucidate the genetic determinants of eosinophil levels by examining polygenic risk scores (PRS) and identifying significant single nucleotide polymorphisms (SNPs).
Methods
We conducted a comprehensive genome-wide association study (GWAS) on eosinophil levels using a large cohort, comprising 221 851 controls (eosinophil levels ≤ 5%) and 11 200 cases (eosinophil levels > 5%) in Taiwan. The analysis included covariates such as age, sex and the first 10 principal components to account for population stratification. Polygenic risk scores were calculated, and their associations with eosinophil levels were evaluated.
Results
The control group exhibited a mean eosinophil level of 47.81 (SD, 21.08) and was composed of 55.32% females and 44.68% males. The case group had a mean eosinophil level of 43.27 (SD, 22.27) and consisted of 38.6% females and 61.4% males. The GWAS identified several SNPs with significant associations, including rs7646596, rs8191981, rs140105250, rs77143352, rs12535759, rs11327184, rs16917546, rs76738175, rs11568075, rs9557175 and rs2801127, with rs7646596 showing the most significant P-value. PRS analysis indicated that individuals with higher polygenic risk scores were significantly more likely to have elevated eosinophil levels.
Conclusion
This study identified several genetic loci significantly associated with eosinophil levels, highlighting the polygenic nature of eosinophilia. The robust associations, particularly with SNP rs7646596, offer valuable insights into the genetic underpinnings of eosinophilia and related conditions.
{"title":"Genome-wide association study and polygenic risk scores of eosinophilia in Taiwan","authors":"Hsing-Fang Lu, Su Boon-Yong, Chi-Ya Yang, Jiu-Yao Wang, Yen-Ting Chang, Fuu-Jen Tsai","doi":"10.1002/cti2.70050","DOIUrl":"https://doi.org/10.1002/cti2.70050","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Eosinophilia, characterised by elevated eosinophil levels, is associated with various allergic and inflammatory conditions. This study aimed to elucidate the genetic determinants of eosinophil levels by examining polygenic risk scores (PRS) and identifying significant single nucleotide polymorphisms (SNPs).</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We conducted a comprehensive genome-wide association study (GWAS) on eosinophil levels using a large cohort, comprising 221 851 controls (eosinophil levels ≤ 5%) and 11 200 cases (eosinophil levels > 5%) in Taiwan. The analysis included covariates such as age, sex and the first 10 principal components to account for population stratification. Polygenic risk scores were calculated, and their associations with eosinophil levels were evaluated.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The control group exhibited a mean eosinophil level of 47.81 (SD, 21.08) and was composed of 55.32% females and 44.68% males. The case group had a mean eosinophil level of 43.27 (SD, 22.27) and consisted of 38.6% females and 61.4% males. The GWAS identified several SNPs with significant associations, including rs7646596, rs8191981, rs140105250, rs77143352, rs12535759, rs11327184, rs16917546, rs76738175, rs11568075, rs9557175 and rs2801127, with rs7646596 showing the most significant <i>P</i>-value. PRS analysis indicated that individuals with higher polygenic risk scores were significantly more likely to have elevated eosinophil levels.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>This study identified several genetic loci significantly associated with eosinophil levels, highlighting the polygenic nature of eosinophilia. The robust associations, particularly with SNP rs7646596, offer valuable insights into the genetic underpinnings of eosinophilia and related conditions.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 10","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70050","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145317753","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
As an autoimmune disorder, myasthenia gravis (MG) manifests as an autoimmune attack on postsynaptic neuromuscular junction proteins by pathogenic autoantibodies. This immune attack disrupts neurotransmission, resulting in fatigable skeletal muscle weakness with diurnal fluctuation. However, functional cure biomarkers for patients remain limited.
Methods
Peripheral blood collection was performed at three time points in patients with MG: before treatment (Pre), 1 month after treatment (Post) and functional cure (long-term follow-up, LF). Single-cell RNA sequencing was performed. The clinical examination results were collected and summarised.
Results
In general, patients with MG exhibited dynamic changes in immune cell composition and inflammatory features. In particular, monocytes were enriched in the LF group, and further subgroup analysis revealed enrichment of CD14+S100A12+ monocytes and depletion of CD14+FOS+ monocytes in the LF group. Moreover, inflammation scores were significantly different in the Pre, Post and LF groups.
Conclusion
Our study provides a comprehensive cell landscape for patients with MG, identifies two dysregulated monocytes, elucidates the inflammation status and offers a new perspective on understanding the aetiology of functional cure and potential therapeutic strategies for patients with MG.
{"title":"Single-cell RNA sequencing reveals cell immune status and dysregulated monocytes in patients with myasthenia gravis","authors":"Yufan Guo, Yu Gu, Yuting Jin, Xintao Wu, Yuting Lou, Pu Miao, Ye Wang, Bijun Zhang, Xueting Lin, Chudi Zhang, Jianhua Feng","doi":"10.1002/cti2.70052","DOIUrl":"https://doi.org/10.1002/cti2.70052","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>As an autoimmune disorder, myasthenia gravis (MG) manifests as an autoimmune attack on postsynaptic neuromuscular junction proteins by pathogenic autoantibodies. This immune attack disrupts neurotransmission, resulting in fatigable skeletal muscle weakness with diurnal fluctuation. However, functional cure biomarkers for patients remain limited.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Peripheral blood collection was performed at three time points in patients with MG: before treatment (Pre), 1 month after treatment (Post) and functional cure (long-term follow-up, LF). Single-cell RNA sequencing was performed. The clinical examination results were collected and summarised.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>In general, patients with MG exhibited dynamic changes in immune cell composition and inflammatory features. In particular, monocytes were enriched in the LF group, and further subgroup analysis revealed enrichment of CD14<sup>+</sup>S100A12<sup>+</sup> monocytes and depletion of CD14<sup>+</sup>FOS<sup>+</sup> monocytes in the LF group. Moreover, inflammation scores were significantly different in the Pre, Post and LF groups.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Our study provides a comprehensive cell landscape for patients with MG, identifies two dysregulated monocytes, elucidates the inflammation status and offers a new perspective on understanding the aetiology of functional cure and potential therapeutic strategies for patients with MG.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 10","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-10-05","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70052","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145227997","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Samuel Liwei Leong, Janesha C Maddumage, Stephanie Gras, Emma J Grant
Objectives
CD8+ T cells are protective against influenza and there is an interest in designing a future CD8+ T-cell-mediated vaccine. However, a significant challenge is the extensive polymorphism of Human Leukocyte Antigen class I (HLA-I) molecules, the targets of CD8+ T cells. Despite this, HLA supertypes have been defined as a subset of HLA-I molecules sharing similar peptide motif preferences that may present overlapping peptide repertoires. Therefore, selecting immunogenic peptides presented by a range of HLA-I molecules for inclusion in a vaccine may partially overcome the challenge presented by HLA-I polymorphism.
Methods
In this study, we investigated the presentation and immunogenicity of a known HLA-B*44:03-restricted influenza-derived peptide NS1195-203 across the HLA-B44 supertype. Using TFold and AlphaFold2, we predicted the structures of the NS1195-203 bound by the HLA-B44 supertype molecules, including HLA-B*44:02, HLA-B*44:03, HLA-B*40:01, HLA-B*40:01 and HLA-B*45:01. Peripheral blood mononuclear cells (PBMCs) isolated from donors expressing one of these HLA-B44 supertype molecules were used to generate CD8+ T-cell lines against the NS1195-203 peptide and assess its immunogenicity via intracellular cytokine staining assay.
Results
The structures predicted with TFold and AlphaFold2 of the NS1195-203 peptide in complex with the HLA-B44 allomorphs were overall similar, with some notable differences at the peptide P9-Trp. A polyfunctional NS1195-203-specific CD8+ T-cell response was observed in HLA-B*44:03+ and HLA-B*44:02+ samples; however, minimal responses were observed in the three other HLA-B44+ supertype molecules.
Conclusion
Although HLA molecules from the same supertype may be able to present the same peptide, this will not always result in CD8+ T-cell responses. As such, HLA-I supertypes, defined based on peptide binding motif and presentation, do not include information on immunogenicity and are not currently able to be used on their own to select epitopes as vaccine candidates. However, new knowledge on HLA supertypes may help curate sets of peptides that are potential vaccine targets and applicable to a range of HLA allomorphs.
{"title":"Lack of immunogenicity for an influenza-derived peptide across the HLA-B44 supertype molecules","authors":"Samuel Liwei Leong, Janesha C Maddumage, Stephanie Gras, Emma J Grant","doi":"10.1002/cti2.70051","DOIUrl":"https://doi.org/10.1002/cti2.70051","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>CD8<sup>+</sup> T cells are protective against influenza and there is an interest in designing a future CD8<sup>+</sup> T-cell-mediated vaccine. However, a significant challenge is the extensive polymorphism of Human Leukocyte Antigen class I (HLA-I) molecules, the targets of CD8<sup>+</sup> T cells. Despite this, HLA supertypes have been defined as a subset of HLA-I molecules sharing similar peptide motif preferences that may present overlapping peptide repertoires. Therefore, selecting immunogenic peptides presented by a range of HLA-I molecules for inclusion in a vaccine may partially overcome the challenge presented by HLA-I polymorphism.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>In this study, we investigated the presentation and immunogenicity of a known HLA-B*44:03-restricted influenza-derived peptide NS1<sub>195-203</sub> across the HLA-B44 supertype. Using TFold and AlphaFold2, we predicted the structures of the NS1<sub>195-203</sub> bound by the HLA-B44 supertype molecules, including HLA-B*44:02, HLA-B*44:03, HLA-B*40:01, HLA-B*40:01 and HLA-B*45:01. Peripheral blood mononuclear cells (PBMCs) isolated from donors expressing one of these HLA-B44 supertype molecules were used to generate CD8<sup>+</sup> T-cell lines against the NS1<sub>195-203</sub> peptide and assess its immunogenicity via intracellular cytokine staining assay.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The structures predicted with TFold and AlphaFold2 of the NS1<sub>195-203</sub> peptide in complex with the HLA-B44 allomorphs were overall similar, with some notable differences at the peptide P9-Trp. A polyfunctional NS1<sub>195-203</sub>-specific CD8<sup>+</sup> T-cell response was observed in HLA-B*44:03<sup>+</sup> and HLA-B*44:02<sup>+</sup> samples; however, minimal responses were observed in the three other HLA-B44<sup>+</sup> supertype molecules.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Although HLA molecules from the same supertype may be able to present the same peptide, this will not always result in CD8<sup>+</sup> T-cell responses. As such, HLA-I supertypes, defined based on peptide binding motif and presentation, do not include information on immunogenicity and are not currently able to be used on their own to select epitopes as vaccine candidates. However, new knowledge on HLA supertypes may help curate sets of peptides that are potential vaccine targets and applicable to a range of HLA allomorphs.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 9","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-19","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70051","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145101845","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Priya A Debisarun, Rutger J Röring, Özlem Bulut, Thijs ten Doesschate, Thomas W van der Vaart, Vinod Kumar, Helga Lemmers, Heidi Dijkstra, Axel B Janssen, Karin Veerman, Rob ter Heine, Reinout van Crevel, Jaap ten Oever, Leo AB Joosten, Marc J Bonten, Cornelis H van Werkhoven, Janneke HHM van de Wijgert, Mihai G Netea
Objective
Chronic systemic inflammation can lead to metabolic, cardiovascular and neurodegenerative complications, but the factors influencing it are incompletely understood. In this study, we evaluated several factors, including Bacille Calmette–Guérin (BCG) and influenza vaccination, SARS-CoV-2 infection and sex, that may impact systemic inflammation as assessed by targeted inflammatory plasma proteome analysis in healthy individuals.
Methods
Participants were randomised to BCG or placebo vaccination at the start of the Dutch SARS-CoV-2 epidemic in March/April 2020. They reported their influenza vaccination status for the most recent influenza season. Twelve weeks after BCG or placebo vaccination, we assessed relative concentrations of 69 proteins in plasma of 357 individuals.
Results
Both BCG and quadrivalent influenza vaccination were associated with overall trends towards reduced systemic inflammation in both sexes, but with a more pronounced effect in men. However, the impact on specific immunological proteins varied between BCG and influenza vaccinations. SARS-CoV-2 infection in the 12 weeks between randomisation and plasma sampling was also associated with overall trends towards reduced systemic inflammation, reaching significance for CXCL10 and TNF concentrations. Notably, individuals who had received BCG vaccination prior to SARS-CoV-2 infection did not exhibit this protein profile. Furthermore, elevated CXCL11 and OPG concentrations at 12 weeks were associated with subsequent respiratory symptoms during the additional 9 months of follow-up.
Conclusions
Our study revealed distinctive alterations in the plasma inflammation proteome associated with BCG vaccination, influenza vaccination, SARS-CoV-2 infection and sex. These findings are exploratory and hypothesis-generating and warrant further investigation in well-controlled longitudinal cohort studies.
{"title":"Long-term effects of influenza and Bacille Calmette–Guérin vaccination on systemic inflammation","authors":"Priya A Debisarun, Rutger J Röring, Özlem Bulut, Thijs ten Doesschate, Thomas W van der Vaart, Vinod Kumar, Helga Lemmers, Heidi Dijkstra, Axel B Janssen, Karin Veerman, Rob ter Heine, Reinout van Crevel, Jaap ten Oever, Leo AB Joosten, Marc J Bonten, Cornelis H van Werkhoven, Janneke HHM van de Wijgert, Mihai G Netea","doi":"10.1002/cti2.70047","DOIUrl":"https://doi.org/10.1002/cti2.70047","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objective</h3>\u0000 \u0000 <p>Chronic systemic inflammation can lead to metabolic, cardiovascular and neurodegenerative complications, but the factors influencing it are incompletely understood. In this study, we evaluated several factors, including Bacille Calmette–Guérin (BCG) and influenza vaccination, SARS-CoV-2 infection and sex, that may impact systemic inflammation as assessed by targeted inflammatory plasma proteome analysis in healthy individuals.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Participants were randomised to BCG or placebo vaccination at the start of the Dutch SARS-CoV-2 epidemic in March/April 2020. They reported their influenza vaccination status for the most recent influenza season. Twelve weeks after BCG or placebo vaccination, we assessed relative concentrations of 69 proteins in plasma of 357 individuals.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Both BCG and quadrivalent influenza vaccination were associated with overall trends towards reduced systemic inflammation in both sexes, but with a more pronounced effect in men. However, the impact on specific immunological proteins varied between BCG and influenza vaccinations. SARS-CoV-2 infection in the 12 weeks between randomisation and plasma sampling was also associated with overall trends towards reduced systemic inflammation, reaching significance for CXCL10 and TNF concentrations. Notably, individuals who had received BCG vaccination prior to SARS-CoV-2 infection did not exhibit this protein profile. Furthermore, elevated CXCL11 and OPG concentrations at 12 weeks were associated with subsequent respiratory symptoms during the additional 9 months of follow-up.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusions</h3>\u0000 \u0000 <p>Our study revealed distinctive alterations in the plasma inflammation proteome associated with BCG vaccination, influenza vaccination, SARS-CoV-2 infection and sex. These findings are exploratory and hypothesis-generating and warrant further investigation in well-controlled longitudinal cohort studies.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 9","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-09-11","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70047","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"145037858","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Xiaomeng Zhang, Rachel Xu, Dmitry Zorin, Evan G Pappas, Jiawei Tang, Yuchen Bai, Vicky M Qin, Bianca von Scheidt, Ruihong Huang, Weronika Kulakowska, Charbel Darido, Phillip K Darcy, Michael H Kershaw, Clare Y Slaney
Objectives
Chimeric antigen receptor (CAR) T cell therapies have transformed the treatment of B cell malignancies and show promise in other diseases, including autoimmune disorders and cardiac injury. However, broader application, particularly in solid tumours, is limited by challenges such as antigen escape and tumour heterogeneity. This study aimed to develop an anti-FLAG CAR capable of engaging FLAG-tagged secondary reagents, providing a flexible and adaptable targeting strategy.
Methods
We engineered a humanised anti-FLAG CAR to engage FLAG-tagged secondary reagents. The initial construct exhibited tonic signalling, which was addressed through structural optimisation. Therapeutic efficacy was assessed in solid tumour mouse models expressing either FLAG or a FLAG-tagged secondary targeting reagent.
Results
The initial anti-FLAG CAR showed functional activity but exhibited tonic signalling and exhaustion, limiting its therapeutic utility. Structural optimisation significantly reduced exhaustion and improved T cell persistence and functionality. The optimised CAR T cells effectively inhibited tumour growth in models using either FLAG- engineered tumour cells or a FLAG-tagged secondary targeting reagent.
Conclusion
Our findings underscore the importance of CAR design in minimising exhaustion and enhancing therapeutic efficacy. This work supports a modular CAR T cell platform with the potential to overcome tumour antigen heterogeneity and immune evasion in solid cancers.
{"title":"Optimised modular anti-FLAG CAR T cells for solid tumor therapy","authors":"Xiaomeng Zhang, Rachel Xu, Dmitry Zorin, Evan G Pappas, Jiawei Tang, Yuchen Bai, Vicky M Qin, Bianca von Scheidt, Ruihong Huang, Weronika Kulakowska, Charbel Darido, Phillip K Darcy, Michael H Kershaw, Clare Y Slaney","doi":"10.1002/cti2.70046","DOIUrl":"https://doi.org/10.1002/cti2.70046","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Chimeric antigen receptor (CAR) T cell therapies have transformed the treatment of B cell malignancies and show promise in other diseases, including autoimmune disorders and cardiac injury. However, broader application, particularly in solid tumours, is limited by challenges such as antigen escape and tumour heterogeneity. This study aimed to develop an anti-FLAG CAR capable of engaging FLAG-tagged secondary reagents, providing a flexible and adaptable targeting strategy.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>We engineered a humanised anti-FLAG CAR to engage FLAG-tagged secondary reagents. The initial construct exhibited tonic signalling, which was addressed through structural optimisation. Therapeutic efficacy was assessed in solid tumour mouse models expressing either FLAG or a FLAG-tagged secondary targeting reagent.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>The initial anti-FLAG CAR showed functional activity but exhibited tonic signalling and exhaustion, limiting its therapeutic utility. Structural optimisation significantly reduced exhaustion and improved T cell persistence and functionality. The optimised CAR T cells effectively inhibited tumour growth in models using either FLAG- engineered tumour cells or a FLAG-tagged secondary targeting reagent.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>Our findings underscore the importance of CAR design in minimising exhaustion and enhancing therapeutic efficacy. This work supports a modular CAR T cell platform with the potential to overcome tumour antigen heterogeneity and immune evasion in solid cancers.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 8","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-21","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70046","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144885332","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Lewis J Williams, Connie SN Li-Wai-Suen, Alex L Garnham, Stefanie M Bader, Constantine S Tam, Ashley Whitechurch, Monica A Slavin, Marcel Doerflinger, Benjamin W Teh
Objectives
Chronic lymphocytic leukaemia (CLL) patients are at increased risk for infection, with the risk even higher for relapsed and refractory patients. Clinical assessment of infection risk is increasingly challenging in the era of immune-based therapies, such as Bruton's tyrosine kinase inhibitors. A pilot study was conducted to elucidate possible predictive immune markers.
Methods
Patients with relapsed and refractory CLL treated with ibrutinib were evaluated. Peripheral blood mononuclear cells (PBMCs) collected at defined intervals (baseline, 3- and 6 months following commencement of ibrutinib) were analysed, with or without phorbol myristate acetate (PMA)/ionomycin stimulation, using Luminex and RNA sequencing. Luminex and gene expression profiles were compared between patients that who did and did not develop infection to identify immune signatures associated with infection over a subsequent 3-month period.
Results
Twenty-eight patients were included in this pilot study. Forty-six per cent of patients developed an infection (13 patients, 17 events) over 9 months of patient monitoring. Most infections were clinically diagnosed (72.7%) with the remainder microbiologically diagnosed bacterial (18.1%) and viral (9.2%) infections. Cell populations did not correlate with subsequent infection. An inflammatory transcriptome profile at 3 months following ibrutinib was associated with a subsequent infection episode. Increased whole protein response to PMA stimulation at 3 and 6 months was associated with subsequent risk for infections. Increased whole protein response to PMA stimulation was associated with subsequent risk of infection 3 months after commencing ibrutinib.
Conclusion
The combination of protein and RNA analysis can provide further insight into the interactions of immunotherapies and immunity but should be validated further in large cohorts.
{"title":"Elucidating novel immune profiles for predicting infection in high-risk cohorts: a pilot study in patients with relapsed and refractory chronic lymphocytic leukaemia","authors":"Lewis J Williams, Connie SN Li-Wai-Suen, Alex L Garnham, Stefanie M Bader, Constantine S Tam, Ashley Whitechurch, Monica A Slavin, Marcel Doerflinger, Benjamin W Teh","doi":"10.1002/cti2.70049","DOIUrl":"https://doi.org/10.1002/cti2.70049","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>Chronic lymphocytic leukaemia (CLL) patients are at increased risk for infection, with the risk even higher for relapsed and refractory patients. Clinical assessment of infection risk is increasingly challenging in the era of immune-based therapies, such as Bruton's tyrosine kinase inhibitors. A pilot study was conducted to elucidate possible predictive immune markers.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Patients with relapsed and refractory CLL treated with ibrutinib were evaluated. Peripheral blood mononuclear cells (PBMCs) collected at defined intervals (baseline, 3- and 6 months following commencement of ibrutinib) were analysed, with or without phorbol myristate acetate (PMA)/ionomycin stimulation, using Luminex and RNA sequencing. Luminex and gene expression profiles were compared between patients that who did and did not develop infection to identify immune signatures associated with infection over a subsequent 3-month period.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Twenty-eight patients were included in this pilot study. Forty-six per cent of patients developed an infection (13 patients, 17 events) over 9 months of patient monitoring. Most infections were clinically diagnosed (72.7%) with the remainder microbiologically diagnosed bacterial (18.1%) and viral (9.2%) infections. Cell populations did not correlate with subsequent infection. An inflammatory transcriptome profile at 3 months following ibrutinib was associated with a subsequent infection episode. Increased whole protein response to PMA stimulation at 3 and 6 months was associated with subsequent risk for infections. Increased whole protein response to PMA stimulation was associated with subsequent risk of infection 3 months after commencing ibrutinib.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>The combination of protein and RNA analysis can provide further insight into the interactions of immunotherapies and immunity but should be validated further in large cohorts.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 8","pages":""},"PeriodicalIF":3.8,"publicationDate":"2025-08-03","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70049","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144767270","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
The imbalance of Th17/Treg cells represents a key pathogenic mechanism in immune thrombocytopenia (ITP); however, the underlying regulatory mechanisms remain poorly understood. Dysregulated succinylation has been implicated in disease onset and progression. Therefore, this study aimed to investigate the role of succinylation in modulating the Th17/Treg balance in ITP and to elucidate the associated molecular pathways.
Methods
Whole blood samples were collected from ITP patients and mouse models. The frequencies of Treg and Th17 cells were quantified using flow cytometry. Treg- and Th17-associated biomarkers were analysed via enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction and immunoblotting. The regulatory relationship between SIRT7 and STAT3 succinylation was evaluated through co-immunoprecipitation, immunofluorescence and immunoblotting assays.
Results
Patients with ITP exhibited elevated Th17/Treg ratios, accompanied by increased global succinylation levels and reduced SIRT7 expression. Overexpression of SIRT7 restored the Th17/Treg imbalance in vitro. Mechanistically, SIRT7 overexpression suppressed STAT3 succinylation at K573, thereby inhibiting STAT3 activity and downstream signalling. Conversely, enforced STAT3 expression counteracted the effects of SIRT7 overexpression on Th17/Treg dynamics. In vivo experiments demonstrated that SIRT7 knockout exacerbated thrombocytopenia and further disrupted Th17/Treg homeostasis in murine models.
Conclusion
SIRT7 mitigates ITP progression by maintaining Th17/Treg equilibrium through desuccinylation of STAT3. These findings highlight SIRT7 as a potential therapeutic target for ITP treatment, offering novel insights into the epigenetic regulation of immune dysregulation in autoimmune diseases.
{"title":"SIRT7 ameliorates Th17/Treg imbalance by desuccinylation of STAT3 to improve immune thrombocytopenia","authors":"Jiao Ge, Xiaoyan Zhang, Fajuan Tang, Yan Liu","doi":"10.1002/cti2.70048","DOIUrl":"https://doi.org/10.1002/cti2.70048","url":null,"abstract":"<div>\u0000 \u0000 \u0000 <section>\u0000 \u0000 <h3> Objectives</h3>\u0000 \u0000 <p>The imbalance of Th17/Treg cells represents a key pathogenic mechanism in immune thrombocytopenia (ITP); however, the underlying regulatory mechanisms remain poorly understood. Dysregulated succinylation has been implicated in disease onset and progression. Therefore, this study aimed to investigate the role of succinylation in modulating the Th17/Treg balance in ITP and to elucidate the associated molecular pathways.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Methods</h3>\u0000 \u0000 <p>Whole blood samples were collected from ITP patients and mouse models. The frequencies of Treg and Th17 cells were quantified using flow cytometry. Treg- and Th17-associated biomarkers were analysed via enzyme-linked immunosorbent assay, quantitative real-time polymerase chain reaction and immunoblotting. The regulatory relationship between SIRT7 and STAT3 succinylation was evaluated through co-immunoprecipitation, immunofluorescence and immunoblotting assays.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Results</h3>\u0000 \u0000 <p>Patients with ITP exhibited elevated Th17/Treg ratios, accompanied by increased global succinylation levels and reduced SIRT7 expression. Overexpression of SIRT7 restored the Th17/Treg imbalance <i>in vitro</i>. Mechanistically, SIRT7 overexpression suppressed STAT3 succinylation at K573, thereby inhibiting STAT3 activity and downstream signalling. Conversely, enforced STAT3 expression counteracted the effects of SIRT7 overexpression on Th17/Treg dynamics. <i>In vivo</i> experiments demonstrated that SIRT7 knockout exacerbated thrombocytopenia and further disrupted Th17/Treg homeostasis in murine models.</p>\u0000 </section>\u0000 \u0000 <section>\u0000 \u0000 <h3> Conclusion</h3>\u0000 \u0000 <p>SIRT7 mitigates ITP progression by maintaining Th17/Treg equilibrium through desuccinylation of STAT3. These findings highlight SIRT7 as a potential therapeutic target for ITP treatment, offering novel insights into the epigenetic regulation of immune dysregulation in autoimmune diseases.</p>\u0000 </section>\u0000 </div>","PeriodicalId":152,"journal":{"name":"Clinical & Translational Immunology","volume":"14 7","pages":""},"PeriodicalIF":4.6,"publicationDate":"2025-07-15","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://onlinelibrary.wiley.com/doi/epdf/10.1002/cti2.70048","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"144635425","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"OA","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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