The research, led by Georgia Kontogianni and Panagiotis Kouzis as co-first authors, with Georgios Kararigas and Themis Alissafi as co-senior authors, brings together teams from the Biomedical Research Foundation Academy of Athens, the National and Kapodistrian University of Athens, the University of Tirana, the University of Tours, the University of Crete, FORTH and the University of Nicosia. The group focused on regulatory T cells, or Tregs, the specialized CD4-positive lymphocyte population that acts as the immune system’s built-in restraint. Tregs patrol the periphery and suppress excessive activation of self-reactive lymphocytes, maintaining what immunologists call peripheral tolerance. Previous work had shown that Tregs are directly affected by checkpoint inhibitors, which target inhibitory signaling pathways that Tregs themselves rely upon, and that during immune-related adverse events these cells undergo extensive inflammatory and metabolic reprogramming, acquiring traits that echo the effector cells they are supposed to restrain. What remained unknown was whether this reprogramming follows the same trajectory in men and women.

The rationale for asking this question is grounded in decades of immunological observation. Women mount stronger humoral and cellular immune responses to infection and vaccination, are far more susceptible to many autoimmune diseases, including lupus, Hashimoto’s thyroiditis and multiple sclerosis, and show sex-dependent differences in Treg biology and inflammatory signaling. Estrogen, androgen and X-linked genetic factors all feed into immune gene regulation. If biological sex shapes the baseline behavior of Tregs, it seemed plausible that it would also dictate how those cells respond when checkpoint blockade or spontaneous autoimmunity pushes the immune system toward self-attack. Testing that idea at scale required a bioinformatics approach rather than a single-cohort experiment, because the relevant patient populations are scattered across clinical studies.

The team therefore performed a sex-based integrative transcriptomic analysis of bulk RNA sequencing datasets generated from peripheral blood Tregs. The datasets came from patients with advanced melanoma and additional solid tumors treated with anti-PD-1 immunotherapy, from individuals living with chronic autoimmune diseases, and from healthy controls. By separating the data into male and female cohorts, the researchers could run differential gene expression, pathway enrichment and network-level analyses independently for each sex, mapping the inflammatory, metabolic and tolerance-associated transcriptional programs that accompany each condition. This design allowed a direct comparison not only between disease and health, but between the sexes and between two distinct contexts of tolerance breakdown: drug-induced toxicity and spontaneous autoimmunity.

The results reveal a layered picture. During spontaneous autoimmune disease, female-derived Tregs exhibited markedly broader transcriptional remodeling than their male counterparts. The female cells showed extensive enrichment of biological processes spanning immune regulation, inflammatory signaling, metabolism and oxidative stress, suggesting that in women, loss of tolerance engages a wide swath of the Treg transcriptional machinery simultaneously. Male-derived Tregs, by contrast, displayed comparatively restricted remodeling, involving a narrower set of immune-related and metabolic pathways. In other words, the depth and breadth of the regulatory cell’s response to autoimmunity is not the same in both sexes, a finding that helps explain why autoimmune susceptibility and severity so often diverge between men and women.

Immune-related adverse events, however, told a different story. Checkpoint inhibitor-associated toxicity produced a distinct pattern of sex-dependent transcriptional organization in Tregs, one that did not simply mirror what the researchers observed in autoimmunity. Although the two conditions share many underlying biological processes, since both represent failures of peripheral tolerance, the degree, directionality and network-level organization of the transcriptional responses differed substantially between the sexes and between disease settings. The same cells, facing the same fundamental problem of lost tolerance, appear to solve or succumb to it along different molecular routes depending on both the sex of the patient and the inflammatory context in which tolerance was disrupted. This argues against a single, uniform program of Treg failure and in favor of disease-specific, sex-shaped trajectories of reprogramming.

The technical implication is significant for how immunologists interpret single-cell and bulk transcriptomic studies. If male and female Tregs organize their responses differently at the pathway and network level, then pooling the sexes, as many studies historically have done, may obscure biologically meaningful signals or generate misleading averages. The authors emphasize that biological sex does not impose one common regulatory program on Tregs; rather, it shapes disease-specific transcriptional trajectories according to the inflammatory environment. That distinction matters for biomarker development, because a transcriptional signature predictive of severe colitis in a male patient treated with anti-PD-1 therapy may not hold for a female patient, and vice versa. Incorporating sex as a biological variable from the earliest stages of study design, rather than as an afterthought in the analysis, emerges as a practical necessity for personalized immunology.

The clinical stakes are considerable. Immune-related adverse events remain a leading cause of treatment interruption and morbidity in oncology, and clinicians currently lack reliable tools to predict which patients will develop severe toxicities. If Treg reprogramming trajectories diverge by sex, sex-stratified biomarkers could improve risk stratification before and during checkpoint inhibitor therapy, potentially guiding earlier intervention with immunosuppression or closer monitoring. Beyond oncology, the findings offer a molecular framework for a long-standing epidemiological puzzle: why women dominate the incidence of many autoimmune diseases while men, once affected, sometimes follow different disease courses. The observation that female Tregs undergo broader remodeling during autoimmunity provides a transcriptional correlate of that epidemiology, and a starting point for mechanistic work into the hormonal and genetic drivers behind it.

For now, the study stands as a reminder that the immune system is not a one-size-fits-all machine. The researchers, supported in part by a European Research Council grant under Horizon 2020 and by the Hellenic Foundation for Research and Innovation, and conducted within the framework of the COST Action ENOTTA, have shown that the cells tasked with keeping immunity in check remodel themselves along paths dictated by both biology and context. As checkpoint inhibitors continue to expand across tumor types and treatment lines, understanding how sex shapes the fragile balance between tumor rejection and tissue destruction may prove essential to making these powerful therapies safer for everyone.

Subject of Research: Sex-dependent transcriptional reprogramming of regulatory T cells in immune-related adverse events and autoimmunity

Article Title: Biological sex defines regulatory T-cell reprogramming trajectories in immune-related adverse events and autoimmunity

Article References: Kontogianni, G., Kouzis, P., Gogas, H., Rumano, M., Mulleman, D., Verginis, P., Kararigas, G., & Alissafi, T. (2026). Biological sex defines regulatory T-cell reprogramming trajectories in immune-related adverse events and autoimmunity. Biology of Sex Differences. https://doi.org/10.1186/s13293-026-00980-z

Image Credits: AI Generated

DOI: 10.1186/s13293-026-00980-z

Keywords: regulatory T cells, immune checkpoint inhibitors, immune-related adverse events, autoimmunity, biological sex, peripheral tolerance, transcriptomics, anti-PD-1 therapy, sexual dimorphism, personalized immunology, Biological, defines

The study addressed a persistent problem in advanced melanoma: immune checkpoint inhibitors can produce durable responses, but their benefits are unevenly distributed. Anti-PD-1 antibodies work by releasing a molecular brake on T cells. Tumors often exploit the PD-1 pathway by engaging PD-L1 on immune cells, dampening the attack that would otherwise help eliminate malignant cells. Blocking PD-1 can restore T-cell activity, yet some melanomas remain immunologically “cold,” with few active immune cells entering the tumor or with biological features that make immune recognition difficult. Acral melanoma, which arises on the palms, soles and nail beds, and mucosal melanoma, which develops in tissues such as the nasal cavity or gastrointestinal and genital tracts, are especially important in Asian populations and have often shown less favorable responses to immunotherapy alone.

The investigators, led by researchers at Xijing Hospital of the Fourth Military Medical University and collaborating institutions, analyzed medical records from 211 people treated between June 2020 and June 2024. All had stage III or IV unresectable melanoma and received first-line treatment with an anti-PD-1 monoclonal antibody plus IFN-α1b, either with or without anlotinib. The majority—179 patients—received the doublet regimen, while 32 received the triplet. IFN-α1b is a form of interferon, a signaling protein that can influence tumor-cell behavior and stimulate components of the immune system. In principle, combining interferon signaling with checkpoint blockade could make the tumor environment more receptive to immune attack, while anlotinib may add a separate pressure on the tumor’s blood-vessel supply and growth pathways.

Anlotinib is an orally administered multitarget tyrosine kinase inhibitor that interferes with signaling proteins involved in angiogenesis, the process by which tumors recruit new blood vessels. A growing tumor requires oxygen and nutrients, and cancer-associated blood vessels can also contribute to an immune-suppressive microenvironment. Abnormal tumor vasculature may prevent immune cells from reaching malignant tissue efficiently, while pro-angiogenic signals can support populations of cells that restrain immune responses. Blocking selected vascular growth signals could therefore have effects beyond starving the tumor: it may help normalize the disorganized blood-vessel network and potentially improve immune-cell access. The study did not prove that this mechanism explained the clinical results, but it provides a biological rationale for testing anlotinib alongside immunotherapy and interferon rather than viewing the drugs as unrelated additions.

To reduce the influence of differences between the treatment groups, the researchers used propensity score overlap weighting. This statistical method estimates the likelihood that each patient would receive one regimen based on measured characteristics, then gives greater analytical weight to patients whose profiles overlap between the two groups. The approach can make comparisons more balanced for factors recorded in the medical files, such as disease features and baseline health status. After weighting, the triplet regimen was associated with a 58 percent lower estimated risk of death than the doublet regimen, reflected by a hazard ratio of 0.42 and a 95 percent confidence interval of 0.24 to 0.72. The hazard ratio summarizes differences in the rate at which events occur over time; it is not a direct statement that 58 percent of individual patients will live longer.

The researchers also reported a progression-free survival advantage. Median progression-free survival—the time before cancer growth, spread or death—was 9.4 months with the triplet and 5.2 months with the doublet. The corresponding hazard ratio was 0.52, with a 95 percent confidence interval of 0.34 to 0.80. These results suggest that the three-drug strategy was associated both with a longer period before the disease worsened and with longer overall survival. Yet the objective response rates tell a more nuanced story. After weighting, tumors shrank by the study’s predefined response criteria in 45.1 percent of patients receiving the triplet and 38.1 percent receiving the doublet. That numerical difference was not statistically significant, with a P value of 0.165. The survival signal may therefore reflect not only how often tumors initially shrink, but also how long disease control lasts or how treatment affects patients whose tumors do not meet the formal definition of response.

Exploratory analyses pointed to potentially greater benefit in patients whose tumors had PD-L1 expression below 1 percent and in those with mucosal melanoma. PD-L1 is a protein used by some cancers to weaken T-cell activity, and low expression is often associated with less predictable benefit from PD-1 blockade. A regimen that alters blood-vessel signaling or stimulates broader immune pathways could, in theory, help overcome some of the limitations of checkpoint therapy in these tumors. However, the subgroup findings were adjusted for false discovery rate and remain hypothesis-generating. Subgroups were small, particularly within the 32-person triplet cohort, making estimates less stable and increasing the possibility that apparent differences arose through chance. The authors specifically emphasized that these observations require confirmation rather than immediate adoption as proof of a biomarker-defined treatment strategy.

The trade-off was increased toxicity. Grade 3 or 4 adverse events, representing severe or medically significant treatment-related effects, occurred in 21.9 percent of patients receiving the triplet compared with 8.9 percent of those receiving the doublet. Although the study reported no treatment-related deaths, the difference indicates that adding anlotinib may create a clinically meaningful burden requiring monitoring and dose management. The source report did not provide a detailed breakdown of every severe adverse event in its abstract, so the overall rates should not be interpreted as a complete safety profile for individual patients. In clinical practice, the risks of an additional targeted drug would need to be weighed against expected disease severity, organ function, previous conditions and the availability of established melanoma treatments.

The study’s most important limitation is its design. Treatment was chosen in routine care rather than assigned randomly, and only measured baseline factors could be balanced through statistical weighting. Differences that were not recorded—or that influenced physicians’ decisions—could still have shaped the results. The uneven sample sizes also make the triplet group vulnerable to imprecise estimates, while the single-center-heavy Chinese clinical context may limit how readily the findings apply to patients elsewhere. The results nonetheless highlight a potentially important direction for melanoma research: combining immune checkpoint blockade with agents that modify interferon signaling and the tumor vasculature. Properly powered prospective randomized trials will be needed to determine whether the apparent survival improvement persists when the three-drug regimen is tested under controlled conditions, and whether its benefits justify the additional toxicity.

Subject of Research: First-line anti-PD-1 therapy combined with IFN-α1b, with or without anlotinib, for unresectable advanced melanoma

Article Title: Efficacy and safety of anti-PD-1 combined with IFN-α1b with or without anlotinib as the first-line therapy in patients with unresectable advanced melanoma: a real-world cohort study

Article References: Zhao, B., Tang, W., Xu, J. et al. “Efficacy and safety of anti-PD-1 combined with IFN-α1b with or without anlotinib as the first-line therapy in patients with unresectable advanced melanoma: a real-world cohort study.” Cancer Immunology, Immunotherapy (2026). Original research article

Image Credits: AI Generated

DOI: 10.1007/s00262-026-04534-5

Keywords: advanced melanoma, anti-PD-1 immunotherapy, IFN-α1b, anlotinib, combination therapy, mucosal melanoma, PD-L1, overall survival, progression-free survival

Alpha-fetoprotein-producing gastric cancer represents a rare yet notably aggressive subtype of gastric tumors, distinguished by elevated serum alpha-fetoprotein (AFP) levels. These cancers often exhibit rapid progression and dismal survival rates. Among these patients, those whose tumors overexpress human epidermal growth factor receptor 2 (HER2) fare even worse. The molecular intricacies underlying HER2-positive AFPGC create formidable therapeutic challenges, prompting oncologists to seek innovative combination regimens beyond standard chemotherapy.

This study analyzed 29 HER2-positive AFPGC patients selected from a pool of 259 individuals with confirmed elevated AFP levels, all treated between May 2017 and May 2022. By integrating comprehensive clinical data, treatment histories, and survival outcomes, researchers meticulously assessed the efficacy of various therapeutic strategies. Notably, 16 patients received a quadruple combination therapy consisting of doublet chemotherapy, trastuzumab—a monoclonal antibody targeting HER2—and an anti-PD-1 immune checkpoint inhibitor, a drug class revolutionizing cancer immunotherapy.

Immune checkpoint inhibitors disrupt the mechanisms tumors use to evade immune surveillance. Anti-PD-1 antibodies reinvigorate T-cell activity, thereby enhancing the body’s natural defenses. Trastuzumab targets HER2 receptors, impeding tumor growth and survival signals. By combining these agents with chemotherapy, which exerts cytotoxic effects on rapidly dividing cells, the researchers hypothesized a synergistic effect that could overcome the inherent resistance of HER2-positive AFPGC.

The clinical results affirm this hypothesis. Patients treated with the quadruplet combination achieved an objective response rate (ORR) of 80% and a disease control rate (DCR) of 93.3%, surpassing outcomes observed in the broader patient cohort. Across the entire study population, the median progression-free survival (mPFS) was 10.27 months, and median overall survival (mOS) was 20.50 months. Although mPFS and mOS in the quadruple therapy group measured slightly lower at 7.47 and 14.87 months respectively, this group contained patients experiencing prolonged survival, indicating potential durable responses.

Intriguingly, six patients on the quadruple regimen remained progression-free beyond the study’s follow-up, and eight were alive at the time of last contact. The wide range of progression-free and overall survival times, stretching as far as 41 months in some cases, underscores the heterogeneity of tumor biology and patient responses. These findings signal that even within a high-risk population like HER2-positive AFPGC, select individuals may derive substantial long-term benefits from this integrated treatment approach.

To elucidate the tumor microenvironment’s role in modulating these responses, the researchers employed multiple immunofluorescence staining techniques. This advanced cellular profiling revealed a pronounced deficit of critical immune effector populations—including CD3+ T cells, CD8+ cytotoxic T cells, CD56+ natural killer cells, and CD68+ macrophages—within both tumor parenchyma and stroma of HER2-positive AFPGC patients. Such immune cell paucity contrasts with patterns observed in HER2-negative and AFP-normal gastric cancers, hinting at unique immunosuppressive landscapes that may influence therapeutic success.

These immune profiling insights deliver powerful mechanistic clues. The diminished presence of immune infiltrates in HER2-positive AFPGC could partly explain its aggressive nature and resistance to monotherapies. Consequently, incorporating immune checkpoint blockade into treatment regimens appears particularly justified, as it can potentially restore antitumor immunity missing in these tumors. The synergy noted with trastuzumab and chemotherapy further supports combined targeting of proliferative and immune evasion pathways.

The study’s retrospective real-world design offers valuable evidence reflective of routine clinical practice, complementing controlled clinical trials. Despite limitations inherent to single-center analyses and relatively small sample sizes, the data underscore the transformative potential of precision immuno-oncology strategies tailored to molecular tumor characteristics such as HER2 status and AFP production.

Importantly, these findings advocate for routine AFP monitoring alongside HER2 evaluation in gastric cancer diagnostics. Identifying patients with this dual biomarker profile may enable clinicians to stratify those who stand to benefit most from quadruple combination regimens, optimizing resource allocation and personalized care. As therapeutic options expand, such biomarker-driven approaches will be instrumental in refining treatment algorithms.

Future prospective investigations with larger cohorts and multi-institutional collaboration are warranted to validate these observations and elucidate mechanisms underpinning varied patient responses. Additionally, explorations into combinatorial biomarkers capturing both tumor and immune parameters could further refine prognostic models and guide adaptive treatment adjustments.

In conclusion, this seminal analysis signifies a critical step toward overcoming the formidable challenge posed by HER2-positive AFPGC. By employing a multipronged therapeutic approach integrating chemotherapy, HER2-targeted monoclonal antibodies, and immune checkpoint inhibitors, clinicians can now envision improved prognoses for a subset of these otherwise poor-outcome patients. The marriage of molecular oncology and immunotherapy heralds a new era of hope and scientific progress in the fight against this refractory cancer subtype.

As novel immuno-modulatory agents gain approval and refine existing therapeutic modalities, precision medicine strategies tailored to unique tumor-immune landscapes will likely become standard care for complex gastric cancers. This study shines a spotlight on the imperative to innovate beyond one-size-fits-all paradigms, encouraging continued exploration of combinatorial regimens that pragmatically harness the immune system’s power to eradicate malignant cells.

Undoubtedly, the reported survival benefits and profound immunological insights provided by this Tianjin Medical University team will catalyze further research and clinical trials, paving the way for improved quality of life and extended survival among patients burdened by HER2-positive alpha-fetoprotein-producing gastric cancer worldwide.

Subject of Research: HER2-positive alpha-fetoprotein-producing gastric cancer and the efficacy of combination therapy involving anti-PD-1 antibodies, trastuzumab, and chemotherapy.

Article Title: The combination of anti-PD-1 antibodies, trastuzumab and chemotherapy may improve the outcome of some patients with HER2-positive alpha-fetoprotein-producing gastric cancer: a retrospective real-world analysis from a single center.

Article References:
Ge, S., Wang, F., Li, H. et al. The combination of anti-PD-1 antibodies, trastuzumab and chemotherapy may improve the outcome of some patients with HER2-positive alpha-fetoprotein-producing gastric cancer: a retrospective real-world analysis from a single center. BMC Cancer 25, 1549 (2025). https://doi.org/10.1186/s12885-025-14808-3

Image Credits: Scienmag.com

DOI: https://doi.org/10.1186/s12885-025-14808-3