Immunotherapy advances最新文献

Cytokine-induced memory-like natural killer cells (CIMLNK) represent a novel form of adoptive cellular therapy that is easy to manufacture and readily available. These cells are generated after overnight stimulation of purified natural killer (NK) cells with interleukin-12 (IL-12), interleukin-15 (IL-15), and interleukin-18 (IL-18). While CIMLNK has demonstrated efficacy in patients with relapsed or refractory acute myeloid leukemia (AML), its potential application in B-cell acute lymphoblastic leukemia (B-ALL) remains unclear. Tafasitamab (TAFA), a monoclonal antibody (mAb) directed against CD19, a surface antigen expressed on B-ALL cells, has been developed to augment anti-tumor efficacy through antibody-dependent cellular cytotoxicity (ADCC), a mechanism predominantly mediated by NK cells. Consequently, we sought to assess the susceptibility of B-ALL to the combination of CIMLNK and TAFA using three B-ALL cell lines: NALM6, SUP-B15, and RS4;11. The addition of TAFA significantly augmented the cytotoxic activity, degranulation capacity, and IFN-γ production of CIMLNK. TAFA-induced ADCC was found to be dose-dependent and was abolished after CD16 blockade. Furthermore, TAFA-mediated effects against NALM6 and SUP-B15 were more pronounced in CIMLNK compared to unstimulated NK cells. In vivo, the combination of CIMLNK and TAFA led to a more pronounced survival benefit in leukemia-bearing mice. In summary, our findings suggest that this combination holds promise as a potential alternative treatment option for patients with relapsed refractory B-ALL.

Macrophages engulf apoptotic bodies and cellular debris as part of homeostasis, but they can also phagocytose live cells, such as aged red blood cells. Pharmacologic reprogramming with the SMAC mimetic LCL161 in combination with T-cell-derived cytokines can induce macrophages to phagocytose live cancer cells in mouse models. Here we extend these findings to encompass a wide range of monovalent and bivalent SMAC mimetic compounds, demonstrating that live cell phagocytosis is a class effect of these agents. We demonstrate robust phagocytosis of live pancreatic and breast cancer cells by primary human macrophages across a range of healthy donors. Unlike mouse macrophages, where a combination of SMAC mimetics with lymphotoxin enhanced phagocytosis, human macrophages were more efficiently polarized to phagocytose live cells by the combination of SMAC mimetics and IFNg. We profiled phagocytic macrophages by transcriptional and proteomic methodologies, uncovering a positive feedback loop of autocrine TNFa production.

Background: Cancer immunotherapy with transgenic T-cell receptor-engineered T cells (TCR-T) enables the targeting of intracellular tumor-specific antigens; in contrast, chimeric antigen receptor-modified T cells (CAR-T) mediate tumor cell killing via the recognition of surface antigens. In the case of acute myeloid leukemia, the lack of leukemia-specific surface antigens limits the efficacy of CAR-T cells; therefore, TCR-T cells may represent a more targeted immunotherapy approach. However, the tumor immunosuppressive environment eliminates the best-functioning, high-avidity TCR-T cells, thus creating a need for novel, enhanced TCR-T cells.

Methods: The piggyBac transposon vector used for gene modification of T cells expresses a T-cell receptor specific to the WT1 tumour antigen, an NFAT promoter-regulated CAR specific to GM-CSF receptor, a CD3xCD33 bispecific T-cell engager, and a truncated EGFR suicide gene system. The transgenic T cells were generated by electroporation using a single expression vector, and the efficiency of these engineered TCR-T cells was evaluated using models that utilized AML cell lines and primary AML cells.

Results: The NFAT-driven GM-CSF CAR significantly enhances the antileukemic activity of WT1-specific TCR-T cells, which importantly maintain specificity for their HLA/peptide antigenic complex. Next, by inserting the CD3xCD33 bispecific T-cell engager into the transposon vector, both TCR-T cells and recruited non-transfected bystander T cells can efficiently target the CD33 antigen, providing more robust antileukemic effects.

Conclusion: The presented strategy, utilizing a single piggyBac transposon vector, enables the complex redirection of T-cell specificity against acute myeloid leukemia by inserting TCR, CAR, BiTE constructs, along with a tEGFR gene suicide system.

Background: Cytotoxic T lymphocytes (CTL) are key effectors in the antitumor immune response. However, their function is commonly suppressed in tumors in the form of exhausted CTL. Understanding mechanisms of suppression and of therapeutics to overcome them is of substantial basic and translational importance yet hindered by limited access to large numbers of exhausted CTL in vitro.

Methods: Here we use three-dimensional tissue culture to generate primary human CTL with suppressed function. Using functional assays, a 21-antibody flow cytometry panel and determination of calcium signaling and CTL tumor cell couple maintenance, we have characterized their phenotype.

Results: We show that these cells closely resemble exhausted CTL from tumors. For a better understanding of in vitro human primary CTL as key tools in therapeutic development, before and after induction of suppression, we have determined the dependence of CTL function on methodology of generation, antigen dose, and affinity across two T-cell receptors and multiple tumor cell lines. As a further determination of their phenotype, we have investigated the morphology and subcellular F-actin distributions of CTL as key regulators of effector function. Primary human CTL formed cell couples with tumor target cells even in the absence of antigen. Yet, the gradual stabilization of such cell couples was associated with increasing CTL effector function. Induction of suppression substantially destabilized CTL tumor cell couples.

Conclusion: This comprehensive characterization of the phenotype of in vitro primary human CTL, including a suppressed state, should facilitate their use in basic research, the development of CTL-targeting therapeutics and the determination of their mechanism of action.

Background: Chimeric antigen receptor (CAR) T-cells have been utilized for the treatment of several malignancies, including Non-Hodgkin lymphomas. A myriad of product- and patient-specific factors determines the extent of patient response, and determining which are most impactful requires analysis of clinical data.

Methods: We used population-level ordinary differential equation models to fit clinical flow cytometry and tumour biopsy data from the TRANSCEND-NHL-001 (NCT02631044) study [1]. We analyzed the impact of lymphodepletion, CAR T-cell phenotypes, and other factors on CAR T-cell dynamics for 30 days after infusion.

Results: We quantified the relative contribution of antigen-dependent and independent sources of proliferation on CAR T-cell dynamics, finding that both make a large contribution and that antigen-independent proliferation was highly correlated with patient IL-15 and IL-7 blood concentrations. The proportion of CAR T-cells in naïve, memory, or effector cells was found to have a limited impact on CAR T-cell dynamics, compared with lymphodepletion and tumour burden.

Conclusions: This study shows how models can be used to link endogenous T-cells, CAR T-cells, and their phenotypes, and may be useful for determining whether a given patient may be responding poorly to treatment, by observing the dynamics of their endogenous T-cells. The framework we developed can be utilized for other CAR T constructs and indications, to test product alterations or biological hypotheses at the population level.

Background: Camrelizumab has become the first-line treatment for most patients with advanced tumors. Among advanced tumor patients undergoing camrelizumab, the majority develop immunogenicity, resulting in the production of anti-drug antibodies (ADA). The impact of ADA on the efficacy and safety of camrelizumab treatment is currently unknown.

Method: Hematologic samples from 31 tumor patients treated with camrelizumab were collected to serve as an experimental cohort for ADA levels detection. Concurrently, a separate validation cohort consisting of 16 patients was established. Follow-up data on patients' OS and PFS were collected and analyzed.

Results: High ADA levels (≥1200 ng/ml) after the three cycles camrelizumab treatment were linked to poorer patient outcomes, as shown by significant differences between PD and PR (P = 0016) and PR and SD (P = .0439). This trend was also present in the validation cohort (PD vs PR, P = .0413). More importantly, high ADA levels after the three cycles camrelizumab treatment were associated with a significant reduction in OS (P = .0128) and PFS (P = .0004), with the validation cohort reporting comparable findings (OS: P = .0009; PFS: P = .0007). Additionally, camrelizumab concentration was negatively correlated with ADA levels (experimental cohort: R ² = 0.3876; validation cohort: R ² = 0.3702). Patients had higher ADA levels after the early phase of camrelizumab treatment.

Conclusion: High ADA levels were associated with shorter OS and PFS in patients after three cycles of camrelizumab therapy. Furthermore, patients had higher ADA levels after the early phase of treatment, specifically in the first three cycles with camrelizumab. It found that the higher the ADA concentration, the lower the serum camrelizumab concentration.

Dendritic cells (DCs) are critical sentinels of the immune system, serving as indispensable bridges between innate and adaptive immune responses. DCs are a heterogeneous population, with subsets playing specialized roles in immune defense, tolerance, or disease development. Among these, the recently redefined DC3 subset has gained attention for its unique features and potential roles in health and disease. This review focuses on the phenotypic, functional, and developmental diversity of DC subsets-primarily DC3s-and their contributions to cancer. The tumor microenvironment (TME) in solid tumors is characterized by varying degrees of immune cell infiltration, including DCs. Within the TME, DCs play diverse roles, either promoting anti-tumor responses or facilitating immune evasion. Key subsets include conventional type 1 and type 2 DCs (cDC1s and cDC2s), as well as plasmacytoid DCs (pDCs). DC3s share certain features with cDC2s and monocytes but are distinct in their phenotype, function, and ontogeny. Functionally, DC3s can prime and activate T cells, skewing CD4⁺ T cells towards Th17 and stimulating CD8⁺ T cells with a tissue-resident memory phenotype. In cancer, their presence correlates with diverse outcomes depending on the TME: DC3 presence is linked to increased survival in patients with pancreatic ductal adenocarcinoma and oropharyngeal cancer while in non-small-cell lung cancer and melanoma it is associated with immunosuppression. The emerging understanding of DC3s highlights the complexity of DC biology and its relevance to diseases. The dynamic immunomodulatory functions of DC3s open new avenues for developing targeted therapies against cancer and immune-mediated disorders.

Background: Regulatory B cells (Bregs) are critical mediators of immune modulation and tumor progression. However, their prognostic relevance and mechanistic roles in clear cell renal cell carcinoma (ccRCC) remain insufficiently explored.

Methods: A comprehensive pancancer strategy was implemented to assess the prognostic role of Breg cells. Spatial transcriptomics, multiplex immunofluorescence (mIF), and immunohistochemistry were performed to investigate Breg localization and immunosuppressive functionality in ccRCC. A machine learning-derived Breg signature (CMLBregS) was established and validated for risk stratification and immune profiling.

Results: Elevated Breg signatures were prominently observed in ccRCC and were associated with advanced T stage, higher tumor grades, and decreased progression-free survival. Spatial transcriptomics and mIF revealed that CD20⁺CD23⁺IL10V Breg cells exert immunosuppressive effects, with or without of the presence of tertiary lymphoid structures. The CMLBregS, comprising 16 Breg-related genes, effectively stratified built a binary classification system. A high-CMLBregS score was linked to an immunosuppressive TME characterized by upregulated IL-10 and TGF-β production, suppression of lymphocyte activation, reduced T cell proliferation, and dampened innate immune responses. Patients with higher CMLBregS scores demonstrated significantly worse clinical outcomes across multiple cohorts. Among CMLBregS-related genes, IRF4 emerged as a key prognostic marker, strongly correlating with IL-10 and PDCD1 expression. Notably, patients with elevated CMLBregS scores exhibited poorer responses to immune checkpoint blockade therapy and more aggressive disease progression during immunotherapy.

Conclusion: This study underscores the pivotal role of Bregs in promoting immune suppression and poor prognosis in ccRCC. The CMLBregS model offers a robust prognostic tool, identifies patients less likely to benefit from immunotherapy, and highlights IRF4 as a potential alternative target. These findings provide a foundation for future strategies aimed at overcoming Breg-mediated immunosuppression in ccRCC.

Regulatory B cells (Bregs) play a crucial role in maintaining immune tolerance and preventing autoimmune diseases. However, in autoimmune conditions, the quantity and function of Bregs are often impaired, leading to pro-inflammatory microenvironment and immune dysregulation. This review provides an in-depth examination of how Bregs are affected in various autoimmune diseases, including multiple sclerosis, rheumatoid arthritis, systemic lupus erythematosus, Sjögren's disease, autoimmune diabetes, and other autoimmune conditions. By summarizing the alterations in Bregs phenotype and function in these specific diseases, we conclude that the Bregs response is complex and variable, showing inconsistent trend across different diseases or even within the same disease. Thus, understanding the heterogeneous nature of Bregs in the autoimmune pathogenesis facilitates novel therapeutic strategies to re-establish immune equilibrium.

Background: Cancer immunotherapy with engineered T cells has become a standard treatment for certain haematological cancers. However, clinical trial outcomes for solid tumours are significantly lagging. A primary challenge in solid tumours is the lack of essential metabolites in the tumour microenvironment, such as glucose, due to poor vascularization and competition with tumour cells.

Methods: To address this, we modified T cells to use fructose as an alternative energy source by introducing ectopic GLUT5 expression.

Results: We show that "GLUT5-armored" T cells, engineered with either chimeric antigen receptors (CARs) or an ectopic T-cell receptor (TCR), achieve enhanced anti-tumour activity in low-glucose environments in both in vitro and in vivo models.

Conclusion: This straightforward modification is compatible with current clinical approaches and may improve the efficacy of T-cell therapies for solid tumours.