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Tanned leather is a product of high intrinsic value and a sign of luxury. Chromium-tanned leather shavings are abundantly produced wastes which contain chromium that could be revalorized. The present research proposes the use of alkaline hydrolysis, aided by an inorganic anion, to promote the hydrolysis of the Cr–O bond. De-chromed solids were employed to synthesize carbon material, which subsequently was applied for dye removal from aqueous solution, while the chromium-rich solution was used as a re-tanning agent. The results demonstrated that it is possible to develop a method of chromium-protein separation, with a minimum degree of hydrolyzation. When an activating agent was employed, the prepared carbon had a surface area of 585 m2/g, with trace levels of chromium (< 0.1%), present mainly as Cr2O3 particles. The materials displayed an adsorption capacity of up to 47.60 mg/g and 27.58 mg/g for methylene blue and Congo red respectively. The chromium-rich solution contained proteins and amino acids that aid in its employment as a re-tanning agent. Leather that was re-tanned with this solution had a similar tear strength and a superior and tensile strength (38%), superior elongation at break (10%), and improved elongation at break (47%) than traditional Cr33-tanned leather. The present study represents a major step in the development of cleaner production of leather. [ABSTRACT FROM AUTHOR]

Abstract Cytotoxic T lymphocytes (CTLs) play critical antitumor roles, encompassing diverse subsets including CD4+, NK, and γδ T cells beyond conventional CD8+ CTLs. However, definitive CTLs biomarkers remain elusive, as cytotoxicity-molecule expression does not necessarily confer cytotoxic capacity. CTLs differentiation involves transcriptional regulation by factors such as T-bet and Blimp-1, although epigenetic regulation of CTLs is less clear. CTLs promote tumor killing through cytotoxic granules and death receptor pathways, but may also stimulate tumorigenesis in some contexts. Given that CTLs cytotoxicity varies across tumors, enhancing this function is critical. This review summarizes current knowledge on CTLs subsets, biomarkers, differentiation mechanisms, cancer-related functions, and strategies for improving cytotoxicity. Key outstanding questions include refining the CTLs definition, characterizing subtype diversity, elucidating differentiation and senescence pathways, delineating CTL-microbe relationships, and enabling multi-omics profiling. A more comprehensive understanding of CTLs biology will facilitate optimization of their immunotherapy applications. Overall, this review synthesizes the heterogeneity, regulation, functional roles, and enhancement strategies of CTLs in antitumor immunity, highlighting gaps in our knowledge of subtype diversity, definitive biomarkers, epigenetic control, microbial interactions, and multi-omics characterization. Addressing these questions will refine our understanding of CTLs immunology to better leverage cytotoxic functions against cancer.

Cytotoxic T lymphocytes (CTLs) play critical antitumor roles, encompassing diverse subsets including CD4+, NK, and γδ T cells beyond conventional CD8+ CTLs. However, definitive CTLs biomarkers remain elusive, as cytotoxicity-molecule expression does not necessarily confer cytotoxic capacity. CTLs differentiation involves transcriptional regulation by factors such as T-bet and Blimp-1, although epigenetic regulation of CTLs is less clear. CTLs promote tumor killing through cytotoxic granules and death receptor pathways, but may also stimulate tumorigenesis in some contexts. Given that CTLs cytotoxicity varies across tumors, enhancing this function is critical. This review summarizes current knowledge on CTLs subsets, biomarkers, differentiation mechanisms, cancer-related functions, and strategies for improving cytotoxicity. Key outstanding questions include refining the CTLs definition, characterizing subtype diversity, elucidating differentiation and senescence pathways, delineating CTL-microbe relationships, and enabling multi-omics profiling. A more comprehensive understanding of CTLs biology will facilitate optimization of their immunotherapy applications. Overall, this review synthesizes the heterogeneity, regulation, functional roles, and enhancement strategies of CTLs in antitumor immunity, highlighting gaps in our knowledge of subtype diversity, definitive biomarkers, epigenetic control, microbial interactions, and multi-omics characterization. Addressing these questions will refine our understanding of CTLs immunology to better leverage cytotoxic functions against cancer. [ABSTRACT FROM AUTHOR]

PurposeCD8 cytotoxic T cells (CTLs) play a critical role in the clearance of virally infected cells. SARS-CoV-2-specific CD8 T cells and functional CTLs in natural infections and following COVID-19 vaccine in primary antibody deficiency (PAD) have not been reported. In this study, we evaluated T cell response following COVID-19 or COVID-19 mRNA vaccination in patients with PADs by assessing SARS-CoV-2 tetramer-positive CD8 T cells and functional CTLs.MethodsSARS-CoV-2-specific CD8 and functional CTLs were examined in a patient with X-linked agammaglobulinemia (XLA) and a patient with common variable immunodeficiency (CVID) following COVID-19 infection, and in 5 patients with CVID and 5 healthy controls 1 month following 2nd dose of COVID-19 mRNA vaccine (Pfizer-BioNTech). Cells were stained with SARS-CoV-2 spike protein-specific tetramers, and for functional CTLs (CD8+ CD107a+ granzyme B+ perforin+), with monoclonal antibodies and isotype controls and analyzed by flow cytometry.ResultsSARS-CoV-2-specific tetramer + CD8 T cells and functional CTLs in the patient with XLA following COVID-19 infection were higher, as compared to healthy control subject following COVID-19 infection. On the other hand, SARS-CoV2-tetramer + CD8 T cells and functional CTLs were lower in CVID patient following COVID19 infection as compared to healthy control following COVID-19 infection. SARS-CoV2-tetramer + CD8 T cells and functional CTLs were significantly lower in SARS-CoV2-naive CVID patients (n = 10) following vaccination when compared to SARS-CoV-2-naive healthy vaccinated controls (n = 10).ConclusionsCVID is associated with reduced SARS-CoV-2-specific CD8 T cells and functional CTLs in both natural SARS-CoV-2 infection and in response to SARS-CoV-2 mRNA vaccine, whereas natural infection in XLA is associated with a robust SARS-CoV-2-specific CD8 and functional CTL responses.

CD8+ cytotoxic T lymphocytes (CTLs) play a crucial role in anti-tumor immunity. In a previous study, we identified a subset of murine effector CTLs expressing the hepatocyte growth factor (HGF) receptor, c-Met (c-Met+ CTLs), that are endowed with enhanced cytolytic capacity. HGF directly inhibited the cytolytic function of c-Met+ CTLs, both in 2D in vitro assays and in vivo, leading to reduced T cell responses against metastatic melanoma. To further investigate the role of c-Met+ CTLs in a three-dimensional (3D) setting, we studied their function within B16 melanoma spheroids and examined the impact of cell–cell contact on the modulation of inhibitory checkpoint molecules' expression, such as KLRG1, PD-1, and CTLA-4. Additionally, we evaluated the cytolytic capacity of human CTL clones expressing c-Met (c-Met+) and compared it to c-Met− CTL clones. Our results indicated that, similar to their murine counterparts, c-Met+ human CTL clones exhibited increased cytolytic activity compared to c-Met− CTL clones, and this enhanced function was negatively regulated by the presence of HGF. Taken together, our findings highlight the potential of targeting the HGF/c-Met pathway to modulate CTL-mediated anti-tumor immunity. This research holds promise for developing strategies to enhance the effectiveness of CTL-based immunotherapies against cancer. [ABSTRACT FROM AUTHOR]

Quantifying the dynamical traits and harshness of SARS-CoV-2/HIV coinfection, a modified deterministic seven compartmental mathematical model is framed at cellular level encapsulating the interrelationships between epithelial cells (predominant target cells of COVID-19 infection), CD4+ T cells (target cells of HIV/AIDS infection), SARS-CoV-2 virions, HIV virions and HIV-specific CTL response. As a way to comprehend the kinetics of SARS-CoV-2/HIV coinfection and to provide prediction of model outcomes, the positivity, boundedness and existence conditions of the biologically feasible equilibrium points have been studied. The criteria of local stability and global stability of the coinfection system around the non-negative equilibrium points have been investigated and the analytical results are enhanced numerically. To measure the influence of the model parameters in SARS-CoV-2/HIV transmission dynamics, sensitivity analysis has been performed. Semi-relative sensitivity analysis demonstrates the maximum deviation of the concentration of infected epithelial cells due to changes in the model parameters and states that six parameters are most sensitive. The numerical simulation stipulates that the COVID-19 infection is likely to be died out from a HIV patient body by increasing the apoptosis of infected epithelial cells and that is possible through the proliferation of the CD4+ T cells count of an individual living with HIV via antiretroviral therapy (ART). Additionally, elevated concentration of CD4+ T cells in people living with HIV assists in heightening the CTLs response to combat COVID-19 and other opportunistic infections.

Chemotherapy, in combination with immune checkpoint blockade (ICB) targeting to programmed death‐1 (PD‐1) or its ligand PD‐L1, is one of the first‐line treatments for patients with advanced non–small‐cell lung cancer (NSCLC). However, a large proportion of patients, especially those with PD‐L1 negative tumors, do not benefit from this treatment. This may be due to the existence of multiple immunosuppressive mechanisms other than the PD‐1/PD‐L1 axis. Human leukocyte antigen‐G (HLA‐G) has been identified as an immune checkpoint protein (ICP) and a neoexpressed tumor‐associated antigen (TAA) in a large proportion of solid tumors. In this study, we evaluated the induction of HLA‐G as well as PD‐L1 using sublethal doses of chemotherapeutics including pemetrexed in different NSCLC cell lines. Except for gefitinib, most of the chemotherapeutic agents enhanced HLA‐G and PD‐L1 expression in a dose‐dependent manner, whereas pemetrexed and carboplatin treatments showed the most consistent upregulation of PD‐L1 and HLA‐G in each cell line. In addition to protein levels, a novel finding of this study is that pemetrexed enhanced the glycosylation of HLA‐G and PD‐L1. Pemetrexed potentiated the cytotoxicity of cytotoxic T lymphocytes (CTLs) to treat NSCLC. Both in vitro and in vivo experiments revealed that CTL‐mediated cytotoxicity was most pronounced when both anti‐PD‐L1 and anti‐HLA‐G ICBs were combined with pemetrexed treatment. In conclusion, anti‐HLA‐G could be an intervention strategy in addition to the anti‐PD‐1/PD‐L1 pathway for NSCLC. Moreover, dual targeting of PD‐L1 and HLA‐G combined with pemetrexed might have a better extent of CTL‐based immunotherapy. [ABSTRACT FROM AUTHOR]

In order to improve the computational accuracy of the AOA (angle of arrival) location, an AOA location method based on CTLS (constrained total least squares) and incorporating the effect of station errors is investigated. Its approximate closed-form solution is derived by first calculating the least squares solution, then substituting the least squares solution into the approximate closed-form solution and iterating to calculate the final result. Subsequently, since the result of the Fisher information matrix is minimized when the stations are uniformly distributed, the final accuracy of the AOA target location is improved by finding the optimal relative position deployment strategy for the sensor-carrying station in 3D space, under the assumption that the sensor carriers are uniformly distributed. Simulations show that, in the presence of station errors, the accuracy of AOA location is improved by approximately 10% using this paper's calculation method, compared to the common least squares location method. In the process of target location, the Fisher information matrix is used to calculate the lower bound of the location result for the CRLB (Cramer–Rao lower bound), which is optimal when the azimuth is uniformly distributed and all on the same horizontal plane, if its pitch angle is 35.56°. [ABSTRACT FROM AUTHOR]

CD8+ cytotoxic T lymphocytes (CTLs) play a crucial role in anti-tumor immunity. In a previous study, we identified a subset of murine effector CTLs expressing the hepatocyte growth factor (HGF) receptor, c-Met (c-Met+ CTLs), that are endowed with enhanced cytolytic capacity. HGF directly inhibited the cytolytic function of c-Met+ CTLs, both in 2D in vitro assays and in vivo, leading to reduced T cell responses against metastatic melanoma. To further investigate the role of c-Met+ CTLs in a three-dimensional (3D) setting, we studied their function within B16 melanoma spheroids and examined the impact of cell–cell contact on the modulation of inhibitory checkpoint molecules’ expression, such as KLRG1, PD-1, and CTLA-4. Additionally, we evaluated the cytolytic capacity of human CTL clones expressing c-Met (c-Met+) and compared it to c-Met− CTL clones. Our results indicated that, similar to their murine counterparts, c-Met+ human CTL clones exhibited increased cytolytic activity compared to c-Met− CTL clones, and this enhanced function was negatively regulated by the presence of HGF. Taken together, our findings highlight the potential of targeting the HGF/c-Met pathway to modulate CTL-mediated anti-tumor immunity. This research holds promise for developing strategies to enhance the effectiveness of CTL-based immunotherapies against cancer.

Background: The aim of this study is to create an index called IRGPRI (immune-related gene prognostic risk index) that can be utilized for predicting the prognosis and assessing the efficacy of immune checkpoint inhibitors (ICIs) therapy in patients with non-small-cell lung cancer (NSCLC)., Methods: Distinguishing gene expression patterns (DEGs) were detected in CD8 + cytotoxic T lymphocytes (CTLs) compared to other cellular types such as CD4 T cells, B cells, plasma cells, and CD8 Tex using the advanced technology of Single-cell RNA Sequencing (scRNA-seq). The construction of IRGPRI was accomplished by employing LASSO Cox regression analysis. We conducted a comparative analysis on clinical characteristics and molecular features, such as pathway enrichment and gene mutation, among the distinct subgroups of IRGPRI. Furthermore, we explored the correlation between immunological characteristics and IRGPRI subgroups to comprehensively assess the effectiveness of ICIs in NSCLC patients., Results: A total of 109 genes were identified by intersecting immune-related genes with DEGs obtained from single-cell RNA sequencing data (GSE131907), specifically comparing CTLs to other cell types. From these, we selected 7 prognosis-related genes, namely TRBC1 , HLA-DMA , CTSH , RAC1 , CTSL , ANXA2 , and CEBPB . These genes were used to construct the IRGPRI. The prognosis of patients diagnosed with NSCLC was found to be significantly better in the low-risk group compared to the high-risk group, as demonstrated by Kaplan-Meier (K-M) survival analysis. This observation was further confirmed through the utilization of data from the GEO cohort. The low-risk group demonstrated an increase in pathways linked with immune response, whereas the high-risk group exhibited a higher prevalence of pathways related to cancer. Furthermore, it was noted in the TCGA cohort that there existed a significant rise in the mutation frequency of every gene within the high-risk group as opposed to the low-risk group. Missense variation emerged as the most prevalent form of mutation. According to the analysis of immune cell infiltration and function, the comprehensive findings suggest that the group with a low risk is characterized by an increased presence of plasma cells, CTLs, T cells follicular helper, Tregs, and Dendritic cell resting. Additionally, they exhibit a higher score in terms of immune function for B cells, CD8 + T cells, checkpoint activity, T cell inhibition and stimulation. Moreover, this low-risk group demonstrates greater efficacy when treated with ICIs therapy compared to the high-risk group., Conclusions: Our research effectively developed and verified a unique IRGPRI, showcasing its association with immune-related characteristics. As a result, the potential of IRGPRI as a valuable biomarker for predicting prognosis and evaluating the effectiveness of ICIs treatment in cancer is evident., Competing Interests: The authors declare the following financial interests/personal relationships which may be considered as potential competing interests:Yikun Yang reports article publishing charges was provided by Shenzhen Key Medical Discipline Construction Fund. Shengjin Cui reports article publishing charges was provided by Basic Medical and Health Research Project of Baoan District. If there are other authors, they declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (© 2024 Published by Elsevier Ltd.)

Drug-resistant cytomegalovirus (CMV) infection after hematopoietic stem cell transplantation (HSCT) often leads to morbidity and mortality. Several studies have shown that CMV-cytotoxic T lymphocytes (CTLs) can overcome drug-resistant CMV infection, but still many questions remain unanswered. Here, we present a case of refractory CMV infection after allogeneic HSCT (allo-HSCT). Donor-derived CMV-CTLs failed to eliminate the virus in unique peripheral blood on the first application, when 70 mg methylprednisolone (MP) was taken per day. After a second attempt with a combination of 8 mg MP with leflunomide, a complete and persisting clearance of all involved sites, including peripheral blood, urinary system, leptomeninges, and retina, was achieved. To summarize, intravenous infusion of CTLs can eliminate CMV in the oculi and central nervous system (CNS), and a low dosage of 8 mg MP has no interaction with CMV-CTLs.

Rationale: Future optimization of computed tomography (CT) lung cancer screening (CTLS) algorithms will depend on clinical outcomes data. Objectives: To report the outcomes of positive and suspicious findings in a clinical CTLS program. Methods: We retrospectively reviewed results for patients from our institution undergoing lung cancer screening from January 2012 through December 2018, with follow-up through December 2019. All exams were retrospectively rescored using Lung-RADS v1.1 (LR). Metrics assessed included positive, probably benign, and suspicious exam rates, frequency/nature of care escalation, and lung cancer detection rates after a positive, probably benign, and suspicious exam result and overall. We calculated time required to resolve suspicious exams as malignant or benign. Results were broken down by subcategories, reason for positive/suspicious designation, and screening round. Results: During the study period 4,301 individuals underwent a total of 10,897 exams. The number of positive (13.9%), suspicious (5.5%), and significant incidental (6.4%) findings was significantly higher at baseline screening. Cancer detection and false-positive rates were 2.0% and 12.3% at baseline versus 1.3% and 5.1% across subsequent screening rounds, respectively. Baseline solid nodule(s) 6 to <8 mm were the only probably benign findings resulting in lung cancer detection within 12 months. New solid nodules 6 to <8 mm were the only LR category 4A (LR4A) findings falling within the LR predicted cancer detection range of 5-15% (12.8%). 38.5% of LR4A cancers were detected within 3 months. Conclusions: Modification of the definition and suggested workup of positive and suspicious lung cancer screening findings appears warranted. [ABSTRACT FROM AUTHOR]

CD8+ cytotoxic T lymphocytes (CTLs) and CD4+ helper T (Th) cells play a critical role in protective immune responses to tumor cells. Particularly, Th9 cells exert anti-tumor activity by producing IL-9. TNF receptor (TNFR)-associated factor 6 (TRAF6) is an adaptor protein that mediates the signals from both the TNFR superfamily and Toll-like receptors (TLRs). We have previously reported that T cell-specific TRAF6-deficent (TRAF6ΔT) mice spontaneously developed systemic inflammatory diseases. However, the physiological role of TRAF6 in T cells in controlling anti-tumor immune responses remains largely unclear. Here, we found that tumor formation of syngeneic colon cancer cells inoculated in TRAF6ΔT mice was accelerated compared to that in control mice. Although TRAF6-deficient naïve T cells showed enhanced differentiation of Th9 cells in vitro , these T cells produced lower amounts of IL-9 in response to a specific antigen. Moreover, CD4+ tumor-infiltrating lymphocytes (TILs) in tumor-bearing TRAF6ΔT mice expressed lower levels of IL-9 than those in WT mice. Importantly, administration of recombinant IL-9 (rIL-9) strongly suppressed tumor progression in TRAF6ΔT mice. Furthermore, expression levels of the T-box transcription factor Eomesodermin (Eomes) and its target molecules IFN-γ, granzyme B and perforin, as well as cytotoxic activity, were reduced in TRAF6-deficient CD8+ T cells in vitro. TRAF6-deficient T cells were found to express significantly increased levels of immune checkpoint molecules, CTLA-4 and PD-1 on the cell surface. These results demonstrate that the TRAF6 signaling pathway in T cells regulates anti-tumor immunity through the activation of tumor specific Th9 cells and CTLs in a tumor microenvironment. • Tumor growth inoculated in T cell-specific TRAF6-deficent mice is accelerated. • TRAF6 is required for optimal production of IL-9 by Th9 cells responding to antigen. • TRAF6 is required for cytotoxic activity of CTLs responding to antigen. • Eomes and its target molecules IFN-γ, granzyme B and perforin are regulated by TRAF6. • Immune checkpoint molecules CTLA-4, PD-1 are negatively regulated by TRAF6 in T cell. [ABSTRACT FROM AUTHOR]

TNF-related apoptosis inducing ligand (TRAIL) is expressed on cytotoxic T lymphocytes (CTLs) and TRAIL is linked to progression of diabetes. However, the impact of high glucose on TRAIL expression and its related killing function in CTLs still remains largely elusive. Here, we report that TRAIL is substantially up-regulated in CTLs in environments with high glucose (HG) both in vitro and in vivo. Non-mitochondrial reactive oxygen species, NFκB and PI3K/Akt are essential in HG-induced TRAIL upregulation in CTLs. TRAILhigh CTLs induce apoptosis of pancreatic beta cell line 1.4E7. Treatment with metformin and vitamin D reduces HG-enhanced expression of TRAIL in CTLs and coherently protects 1.4E7 cells from TRAIL-mediated apoptosis. Our work suggests that HG-induced TRAILhigh CTLs might contribute to the destruction of pancreatic beta cells in a hyperglycemia condition. [ABSTRACT FROM AUTHOR]

TNF-related apoptosis inducing ligand (TRAIL) is expressed on cytotoxic T lymphocytes (CTLs) and TRAIL is linked to progression of diabetes. However, the impact of high glucose on TRAIL expression and its related killing function in CTLs still remains largely elusive. Here, we report that TRAIL is substantially up-regulated in CTLs in environments with high glucose (HG) both in vitro and in vivo. Non-mitochondrial reactive oxygen species, NFκB and PI3K/Akt are essential in HG-induced TRAIL upregulation in CTLs. TRAILhigh CTLs induce apoptosis of pancreatic beta cell line 1.4E7. Treatment with metformin and vitamin D reduces HG-enhanced expression of TRAIL in CTLs and coherently protects 1.4E7 cells from TRAIL-mediated apoptosis. Our work suggests that HG-induced TRAILhigh CTLs might contribute to the destruction of pancreatic beta cells in a hyperglycemia condition.

In order to improve the computational accuracy of the AOA (angle of arrival) location, an AOA location method based on CTLS (constrained total least squares) and incorporating the effect of station errors is investigated. Its approximate closed-form solution is derived by first calculating the least squares solution, then substituting the least squares solution into the approximate closed-form solution and iterating to calculate the final result. Subsequently, since the result of the Fisher information matrix is minimized when the stations are uniformly distributed, the final accuracy of the AOA target location is improved by finding the optimal relative position deployment strategy for the sensor-carrying station in 3D space, under the assumption that the sensor carriers are uniformly distributed. Simulations show that, in the presence of station errors, the accuracy of AOA location is improved by approximately 10% using this paper’s calculation method, compared to the common least squares location method. In the process of target location, the Fisher information matrix is used to calculate the lower bound of the location result for the CRLB (Cramer–Rao lower bound), which is optimal when the azimuth is uniformly distributed and all on the same horizontal plane, if its pitch angle is 35.56°.

Background: Metabolic activities of tumor cells lead to a depletion of nutrients within the tumor microenvironment, which results in the dysfunction of infiltrating T cells. Here, we explored how glutamine (gln) metabolism, which is essential for biosynthesis and cellular function, can affect the functions of cytotoxic T lymphocytes (CTLs). Methods: Activated CTLs were co-cultured with hepatoma cells. Western blot was used to analyze changes of proteins and ELISA was used to analyze changes of effector. RNA-sequencing was used to detect differentially expressed genes in CTLs. The status of the endoplasmic reticulum (ER) was investigated using transmission electron microscopy experiments. Results: Co-culturing CTLs and hepatoma cells revealed that CTLL-2 cells in the co-culture group expressed high levels of PD-1 (Programmed cell death protein 1), TIM-3 (T cell immunoglobulin and mucin domain-containing protein-3), GRP78 (Glucose regulated protein 78), and P-PERK (phosphorylated protein kinase RNA-activated-like endoplasmic reticulum kinase) and secreted low levels of Granzyme B and perforin. Additionally, the substructure of the ER was severely damaged. When CTLs were treated with an inhibitor of ER stress, their functions were restored. Next, complete medium without Gln was used to culture cells, causing CTLs to display dysfunction and ER stress. WB results revealed decreased expression levels of GLS2 and SLC1A5 (Solute carrier family 1 member 5) in CTLs in the co-culture group. Subsequently, glutaminase (GLS) inhibitors were added to the cultures. As expected, CTLs treated with a GLS2 inhibitor had increased protein content of PD-1 and TIM-3, decreased secretion of Granzyme B and perforin, and an enhanced ER stress response. Conclusions: In summary, CTLs are functionally downregulated induced by hepatoma cells through the Gln-GLS2-ERS pathway.

Discus fish (Symphysodon aequifasciatus) exhibit a unique parental care behavior: adult discus produces secretion through their skin, on which the larvae live after birth. The immune components in the skin mucus of parental discus would change during different parental care. C-type lectins (CTLs) could identify and eliminate pathogenic microorganisms and play important roles in innate immunity. Studies on CTLs of discus fish especially during parental care, however, are scarce. Here, we identified 186 CTL genes that distributed in 27 linkage groups based on discus genome. Phylogenetic analysis showed that S. aequifasciatus CTL (SaCTL) members were grouped into 14 subfamilies. A total of 80 gene replication events occurred, of which 15 pairs were subjected to segmental duplication and 65 pairs underwent tandem duplication. Ka/Ks ranged from 0.11 (SaCTL25 / SaCTL158) to 0.68 (SaCTL36 / SaCTL69), all undergoing purifying selection. RNA-seq analysis revealed that SaCTL members, including duplicated genes, in the skin of parental discus show distinct expression patterns in different care stages and between male and female parents. The SaCTL11 was differentially expressed in most care stages and reached the maximum after eggs spawned, but the expression of its paired SaCTL14 was low in each stage. The SaCTL39 increased first and then decreased, reaching a peak in eggs spawned, while paired SaCTL48 first decreased and then increased, reaching a peak in hatched eggs. The SaCTL50 was differentially expressed only in female fish during care, but not in male fish. These results provide new insights into the evolution and potential functional differentiation of CTLs in discus fish during parental care. • A total of 186 CTLs were identified in discus fish genome.Sixty-five pairs of CTLs underwent tandem duplication. • Fifteen pairs of CTLs underwent segmental duplication. • Discuss CTLs exhibited a stage-specific and sex-dependent expression pattern during parental care. [ABSTRACT FROM AUTHOR]

Glioblastoma (GBM), a highly lethal brain cancer, is notorious for immunosuppression, but the mechanisms remain unclear. Here, we documented a temporospatial patterning of tumor-associated myeloid cells (TAMs) corresponding to vascular changes during GBM progression. As tumor vessels transitioned from the initial dense regular network to later scant and engorged vasculature, TAMs shifted away from perivascular regions and trafficked to vascular-poor areas. This process was heavily influenced by the immunocompetence state of the host. Utilizing a sensitive fluorescent UnaG reporter to track tumor hypoxia, coupled with single-cell transcriptomics, we revealed that hypoxic niches attracted and sequestered TAMs and cytotoxic T lymphocytes (CTLs), where they were reprogrammed toward an immunosuppressive state. Mechanistically, we identified chemokine CCL8 and cytokine IL-1β as two hypoxic-niche factors critical for TAM trafficking and co-evolution of hypoxic zones into pseudopalisading patterns. Therefore, perturbation of TAM patterning in hypoxic zones may improve tumor control. [Display omitted] • Host immune status influences tumor vasculature and hypoxic-zone formation in GBM • Spatial patterning of TAM and CTL parallels hypoxic-zone maturation to pseudopalisades • Sequestered TAM and CTL in hypoxic zones are reprogrammed toward immunosuppression • TAM/CTL organization involves CCL8 and IL-1β as niche factors in hypoxic zones Glioblastoma is notorious for immunosuppression, but the mechanisms are unclear. Sattiraju et al. report that hypoxic zones in GBM attract and sequester tumor-associated myeloid cells and cytotoxic T cells, where they are reprogrammed into an immunosuppressive state. This process is influenced by the immunocompetence state of the host and involves CCL8 and IL-1B as niche factors in hypoxic zones. [ABSTRACT FROM AUTHOR]

Background: Diospyros peregrina is dioecious plant native to India and belonging to the family of Ebenaceae, is largely utilized in treatment of various ailments. Little has been known about the antitumor activity of Diospyros peregrina with only 1 previous study on Ehrlich Ascites Carcinoma in mice. Therefore, it prompted us to extensively explore the immunomodulatory effect in various cancer forms. The focal point of this study revolves around breast cancer, which is the second most common cancer in the world. In view of the increasing demands for noninvasive treatments, natural plant-based agents open up promising applications in cancer immunotherapy METHODS: CD4 + lymphocytes were isolated from the peripheral blood mononuclear cells (PBMCs) of breast cancer patients and normal donor blood samples using magnetic-activated cell sorting (MACS) and cultured separately. Utilizing the plastic surface adherence property, the macrophages were isolated from CD4 negative lymphocytes of both breast cancer patients and normal donors. For the presentation of tumor antigens invitro, macrophages were pulsed with breast tumor associated antigen (BTAA) in presence or absence of Diospyros peregrina fruit preparation (DFP). Differentially pulsed and irradiated macrophages were co-cultured with autologous and allogenic lymphocytes. Supernatants hence collected from CD4 + lymphocytes were utilized for cytokine profiling using ELISA and proliferation was assessed by MTT assay. Cytotoxic T lymphocytes (CTLs) generated from CD4 negative lymphocytes culture (2 × 10 5 ) was incubated with MCF-7 (2 × 10 4 ) to check cytotoxicity using LDH release assay. CD4 + lymphocytes were treated in presence or absence of DFP, were analyzed using immunoblotting and RT-qPCR, to check DFP mediated T helper (Th) cell differentiation through investigation of signatory cytokines and transcription factors., Results: It was found that DFP elevated the proliferation of CD4 + T lymphocytes (Th) in response to BTAA. DFP also helped in presenting BTAA pulsed macrophages directing in the cytotoxic T-lymphocyte mediated immune response. Results indicated that DFP preferentially highlighted Th1 commitment with type-1 specific cytokines IFN-g and IL-12 and was indifferent in Th2 manifestation. DFP was not only involved in the upregulation of Tbet mounted type-1 mediated immune response and activation of STAT1 but also it downregulated STAT6 and GATA3, the functional activators and regulators of type-2 immune response. Moreover, it was observed that DFP inhibited the tumor-promoting environment modulated through Tregs by downregulating Foxp3 and STAT5. Further, it was detected that DFP directs Th1 bias and results in attainment of better suppression of breast tumor CONCLUSION: The results collectively pointed out that DFP favored cell-mediated immune response from BTAA antigen presentation on macrophages and also helping in the robust proliferation of an entire spectrum of T helper lymphocytes which furthermore strengthen the underlying immune responses, hence, fencing the body, of the progression of breast cancer., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Background: Metabolic activities of tumor cells lead to a depletion of nutrients within the tumor microenvironment, which results in the dysfunction of infiltrating T cells. Here, we explored how glutamine (gln) metabolism, which is essential for biosynthesis and cellular function, can affect the functions of cytotoxic T lymphocytes (CTLs)., Methods: Activated CTLs were co-cultured with hepatoma cells. Western blot was used to analyze changes of proteins and ELISA was used to analyze changes of effector. RNA-sequencing was used to detect differentially expressed genes in CTLs. The status of the endoplasmic reticulum (ER) was investigated using transmission electron microscopy experiments., Results: Co-culturing CTLs and hepatoma cells revealed that CTLL-2 cells in the co-culture group expressed high levels of PD-1 (Programmed cell death protein 1), TIM-3 (T cell immunoglobulin and mucin domain-containing protein-3), GRP78 (Glucose regulated protein 78), and P-PERK (phosphorylated protein kinase RNA-activated-like endoplasmic reticulum kinase) and secreted low levels of Granzyme B and perforin. Additionally, the substructure of the ER was severely damaged. When CTLs were treated with an inhibitor of ER stress, their functions were restored. Next, complete medium without Gln was used to culture cells, causing CTLs to display dysfunction and ER stress. WB results revealed decreased expression levels of GLS2 and SLC1A5 (Solute carrier family 1 member 5) in CTLs in the co-culture group. Subsequently, glutaminase (GLS) inhibitors were added to the cultures. As expected, CTLs treated with a GLS2 inhibitor had increased protein content of PD-1 and TIM-3, decreased secretion of Granzyme B and perforin, and an enhanced ER stress response., Conclusions: In summary, CTLs are functionally downregulated induced by hepatoma cells through the Gln-GLS2-ERS pathway., Competing Interests: The authors declare no conflicts of interest., (© 2022 The Author(s). Published by IMR Press.)