Clinical & Translational Immunology最新文献

Objectives

Circulating cytokines/chemokines are linked to checkpoint inhibitors (ICIs) in non-small cell lung cancer (NSCLC). The tumor-immune microenvironment (TIME) plays a significant role in modulating a broad range of cytokines and chemokines. This study aimed to explore the crosstalk between circulating cytokines/chemokines and TIME, related to ICIs outcomes.

Methods

We conducted a prospective cohort study of 81 participants with advanced or recurrent NSCLC who received ICIs. Pretreatment comprehensive 27 cytokines/chemokines analysis, immunohistochemistry (IHC) for CD8, Treg/FOXP3, and PD-L1(22C3) TPS, and RNA sequencing were conducted. Demographic characteristics were integrated in the analysis to define the crosstalk of significant TIME-related signatures. Circulating neutrophil-to-lymphocyte ratio (NLR) and IHC tumor-infiltrated neutrophil density were evaluated to correlate systemic and local inflammatory states with ICIs response.

Results

Pretreatment plasma IL-6 and IL-8 were the significant cytokines correlated with surrogate ICIs responses and ICIs progression-free survival (PFS). Despite several correlations between cytokines/chemokines and CD8⁺ TILs, Treg/FOXP3⁺ TILs, neither pretreatment IL-6 nor IL-8 was correlated with intra-tumoral or stromal Treg/FOXP3⁺ TILs, CD8⁺ TILs, and PD-L1. Four active neutrophil-related gene signatures overlapped and were significantly correlated with better ICIs outcomes and lower IL-6 levels. Among 69 genes in 4 neutrophil-related gene signatures, the overexpression of TREM1, SORL1, and HSD17B11 significantly contributed and inversely correlated with CIBERSORT memory B cells. Clinically, the stratification by low levels of IL-6/IL-8 and low NLR identified a subgroup with significantly improved survival outcomes, whereas IHC tumor-infiltrated neutrophil counts did not show a similar association.

Conclusion

Our study reveals a potential mechanistic axis linking circulating IL-6 and IL-8 to tumor-associated neutrophils, memory B cells, and therapeutic response. These findings underscore the crucial role of synergistic treatment in augmenting the efficacy of ICIs. The crosstalk between neutrophils and B cells in the orchestration of ICIs therapy for NSCLC was further elucidated through the roles of HSD17B11, SORL1, and TREM1.

Objectives

Anti-dsDNA antibodies are established biomarkers in systemic lupus erythematosus (SLE), yet the clinical utility of conventional assays remains limited by variable analytical performance. We evaluated a novel Fibre-Optic Surface Plasmon Resonance (FO-SPR) biosensor for real-time profiling of anti-dsDNA antibody–antigen interactions and investigated whether kinetic parameters are associated with disease severity, particularly lupus nephritis (LN).

Methods

Sera from 26 SLE patients (including 12 with LN), 14 disease controls and 16 healthy donors were analysed using FO-SPR biosensors coated with biotinylated dsDNA. The sensors generated real-time kinetic profiles from which maximum binding response (B_max), association rate and dissociation constant were computed. FO-SPR kinetic results were compared with a conventional chemiluminescent anti-dsDNA assay (CLIA) and correlated with disease activity and renal involvement.

Results

The FO-SPR platform enabled reliable detection of anti-dsDNA antibodies and clearly distinguished SLE patients with renal involvement: Eight of 12 LN patients showed B_max values above the 90th percentile of the non-SLE control group. In these patients, kinetic parameters revealed high-affinity anti-dsDNA profiles characterised by significantly higher association rates and maximal shifts and lower dissociation constants. The AUC-ROC for SPR-derived affinity parameters was 0.82 (P = 0.006), outperforming the CLIA assay to discriminate lupus patients with versus without nephritis. Furthermore, FO-SPR binding responses correlated with disease activity score in LN.

Conclusion

This study highlights the value of an innovative FO-SPR biosensor for real-time characterisation of anti-dsDNA antibody affinity. The kinetic profiles captured by this platform provide clinically relevant insights, particularly for distinguishing LN patients from other SLE phenotypes.

Objectives

Tuberculosis (TB) remains a global health challenge, with current antibiotic therapies being limited by long treatments, side effects and multidrug-resistant mycobacterial strains. In addition, Mycobacterium tuberculosis (Mtb), the main causative agent of TB, employs evasion mechanisms particularly within alveolar macrophages being the primary host cells. Conventional therapies fail to modulate macrophage function or effectively target host immunity, which is crucial in TB pathogenesis. Emerging evidence points to induced pluripotent stem cell-derived macrophages (iMacs) with enhanced bactericidal activity as a promising cell-based approach for TB treatment. Therefore, this study aimed to compare iMacs with blood monocyte-derived macrophages (MDMs) in response to Bacillus Calmette–Guérin (BCG), the live attenuated TB vaccine and heat-killed Mtb (HKMT).

Methods

iMacs and MDMs were challenged with BCG and HKMT to assess their functional responses. Key parameters evaluated included cell migration, phagocytosis kinetics, levels of autophagy- and apoptosis-related proteins, and cytokine production profiles following infection.

Results

iMacs displayed enhanced migration, faster phagocytosis and increased expression of autophagy- and apoptosis-related proteins compared with MDMs. Moreover, iMacs showed a stronger pro-inflammatory cytokine response and rapid return to baseline cytokine levels post-infection.

Conclusion

These findings support the potential of iMacs as an immunocompetent model for studying mycobacterial infections and as a tool for cell-based TB immunotherapies.

Objectives

Here, we characterised the diversity and persistence of immunological memory cells—particularly tissue-resident memory T (T_RM) and B (B_RM) cells—in the nasopharyngeal lymphoid tissues of healthy vaccinated (HV) individuals and those who experienced SARS-CoV-2 breakthrough infection (BR).

Methods

Nasopharynx (NP) samples were obtained using brushing from HV and BR subjects. Immune cell populations were analysed using transcriptomic profiling and flow cytometry.

Results

Transcriptomic profiling revealed that the NP of SARS-CoV-2-infected individuals exhibited distinctive signatures of lymphocyte-mediated immunity, underscoring its role as a key site for viral invasion and immune activation. Effector memory CD4+ and CD8+ T (T_EM) cells, along with non-germinal center (GC) B cells, predominated in the NP. Although overall frequencies of memory T cells were comparable between HV and BR groups, CD4+ T_EM and GC B cells were significantly enriched in the NP of BR individuals at least 1 year post infection. Notably, over 80% of CD4+ T_EM and 40% of CD8+ T_EM cells were T_RM, and more than 30% of memory B cells exhibited a B_RM phenotype. These populations of CD4+, CD8+ T_RM and B_RM persisted in the NP for over 2 years following SARS-CoV-2 infection or vaccination. In particular, CD4+ T_RM cells were significantly more abundant and durably maintained in the NP mucosa of BR individuals.

Conclusion

Our findings identify the nasopharynx as a key site of long-lived immunological memory, marked by persistent T_RM and B_RM cells after SARS-CoV-2 exposure.

Objectives

The early development of humoral immunity is important for long-term protection of the newborn. Here, we set out to discern these infant-produced antibodies from the vast background of maternal antibodies, which is challenging but essential to shed light on the infant-produced repertoire.

Methods

Using IgA1 and IgG1 antibody clonal repertoire analysis by mass spectrometry, we compared matched maternal serum, maternal milk, and infant serum samples, both at birth (T1) and at 7–11 weeks after delivery (T2) in four mother–infant dyads.

Results

We observed for both IgA1 and IgG1 unique infant-produced antibody repertoires at T2. For IgA1 at T2, no substantial clonal overlap was found between infant serum and breastmilk. The serum IgG1 clonal repertoires were highly alike at birth for mother and infant, but at T2, the contribution of the maternal clonal population in the infant had been drastically reduced, and a large portion of the T2 IgG1 repertoire originated from the infant.

Conclusions

Newborns produce their own antibody repertoires as early as a few months after birth. From this small study, no convincing evidence is found for transfer of milk antibodies into the infant circulation.

Objective

Examine the role of T cells in shaping and sustaining humoral immunity to SARS-CoV-2 and identify immune factors associated with durable antibody responses.

Methods

Unvaccinated adults (n = 67) with SARS-CoV-2 infection (Wuhan) of any severity were followed longitudinally for up to 14 months. Anti-Spike (S) binding and neutralising antibodies were measured. Bulk T cell IFNγ and IL-2 responses were assessed by Fluorospot to multiple viral proteins. S-specific T-cell subsets and functions were analysed in detail by flow cytometry in a subset of participants (n = 14), combining activation-induced markers (AIMs) and cytokines. Correlations between S-specific T-cell subsets, their polyfunctionality and antibody levels over time were defined.

Results

Over 14 months post infection, anti-S IgG and neutralising antibodies declined significantly but remained detectable in > 85% of participants. Bulk T-cell IFNγ and IL-2 responses persisted without significant reduction. However, S-specific CD4 T cells declined over time. A large proportion of these were peripheral helper T cells (Tph; CXCR5⁻PD-1⁺), which were the main producers of IL-21 and showed marked polyfunctionality during early convalescence, in contrast to circulating follicular helper T cells (cTfh). The early presence of polyfunctional, IL-21-producing Tph strongly correlated with S-specific IgG and neutralising antibodies early post-infection and predicted neutralising antibody maintenance a year later. Older age correlated with higher Tph and antibody levels.

Conclusion

Early S-specific T-cell responses—particularly polyfunctional, IL-21-producing Tph cells—play a key role in sustaining durable antibody levels post-infection, offering insights for future vaccine strategies and understanding long-term immunity.

Allogeneic haematopoietic stem cell transplantation (HSCT) is the only curative therapy for patients diagnosed with acute myeloid leukaemia (AML) and myelodysplastic syndrome (MDS). γδ+ T-cells, a rare subpopulation of T-cells with both innate and adaptive anticancer functions, have been understudied and the role of different γδ+ T-cell subsets after HSCT is unclear.

Objectives

We aimed to characterise the γδ+ T-cells reconstitution post-HSCT, investigating their association with graft-versus-host disease (GvHD) treatment and occurrence, and cytomegalovirus (CMV) reactivation.

Methods

Peripheral blood samples from AML and MDS patients and their respective donors were immunophenotyped by flow cytometry before and at several time points after HSCT. Additionally, next-generation sequencing of the TRG locus was performed to assess clonal dynamics and repertoire diversity.

Results

Early after HSCT, γδ+ T-cells showed a similar phenotype compared to pre-HSCT, and stable repertoires were observed up to 6 months post-HSCT. Patients with acute GVHD presented a higher frequency of CD8 in γδ+ T-cells, and γδ+ T-cell and subsets frequencies were not affected by anti-thymocyte globulin prophylaxis. Donor-derived Vδ2+ T-cell central memory phenotype was associated with a reduced risk of CMV reactivation in the recipient and with repertoire disturbance. Effector memory cells displaying a specific HLA-DR+ CD86+ phenotype were associated with CMV reactivation and a higher proportion of hyperexpanded clonotypes prior to transplantation.

Conclusion

Overall, Vδ1+ and Vδ2+ T-cell phenotypic profiles and γδ TCR repertoire remain stable post-HSCT. This stability is disrupted by CMV reactivation, which is potentially associated with γδ+ T-cells of donor origin and γδ TCR repertoire.

Objectives

Lung complications occur commonly post-haematopoietic stem cell transplant (HCT) in children and contribute significantly to mortality. The development of biomarkers predictive of post-HCT lung complications may improve outcomes for these children. Therefore, the primary objective of this study was to identify pre-transplant plasma biomarkers predictive of lung complications in children undergoing HCT.

Methods

Plasma samples from 117 pre-HCT patients were used to measure 78 soluble immune analytes, and extensive clinical metadata were retrospectively collected from the electronic medical record, including clinical characteristics of the post-HCT lung complications.

Results

Firstly, we showed that the plasma cytokine signature of children undergoing HCT differed significantly between children depending on the indication for HCT (malignant vs non-malignant). Secondly, we found that the plasma cytokine signature changed with the age of the patient. Thirdly, we showed that elevated pre-HCT plasma levels of CXCL9 and Chitinase 3-like 1, both induced by IFN-γ, were predictive of post-HCT lung complications in patients with a malignant indication for HCT [area under the curve (AUC): 0.70, P = 0.0007 and AUC: 0.68, P = 0.0016, respectively].

Conclusion

In conclusion, we have identified two potential biomarkers of post-HCT lung disease in paediatric patients and these require validation in future cohorts.

Objectives

Multiple sclerosis is an immune-mediated inflammatory disease of the central nervous system. Recently, kappa opioid receptor (KOR) agonists were found to promote remyelination and recovery in models of demyelination. However, the side effects of many full KOR agonists limit clinical use. Activation of KOR and mu opioid receptors (MOR) often has opposing effects. Therefore, it is plausible that activating KOR and antagonising MOR may confer a beneficial increase in therapeutic efficacy at lower doses.

Methods

This study investigated the hypothesis that combining the clinically available full KOR agonist nalfurafine (NalF) with a MOR antagonist/partial KOR agonist nalmefene (NalM) would provide greater than additive benefits in the murine model of immune-driven demyelination, experimental autoimmune encephalomyelitis (EAE).

Results

We found that NalM alone reduced EAE disease severity when administered therapeutically although NalM at the most effective doses did not enhance myelination or reduce lesions in the spinal cord. Moreover, a combination of NalF/NalM at suboptimal doses significantly reduced lesion size and promoted myelination in the spinal cord suggesting a better-than-additive effect on lesion resolution. This finding was supported by a bioinformatic analysis response that highlighted the myelination benefits of the suboptimal dose combination while the optimal dose combination of NalF/NalM stimulated increased immunomodulatory activity.

Conclusions

Findings from this study indicate that the remyelinating and immunomodulatory effects of the KOR agonist NalF can be significantly enhanced through interactions with the MOR antagonist NalM in a distinct dose-dependent manner.