Decrease In CD4 Research Articles

Remodeling of the immune and stromal cell compartment by PD-1 blockade in mismatch repair-deficient colorectal cancer.

Immune checkpoint inhibitor (ICI) therapy can induce complete responses in mismatch repair-deficient and microsatellite instability-high (d-MMR/MSI-H) colorectal cancers (CRCs). However, the underlying mechanism for pathological complete response (pCR) to immunotherapy has not been completely understood. We utilize single-cell RNA sequencing (scRNA-seq) to investigate the dynamics of immune and stromal cells in 19 patients with d-MMR/MSI-H CRC who received neoadjuvant PD-1 blockade. We found that in tumors with pCR, there is a concerted decrease in CD8+ Trm-mitotic, CD4+ Tregs, proinflammatory IL1B+ Mono and CCL2+ Fibroblast following treatment, while the proportions of CD8+ Tem, CD4+ Th, CD20+ B, and HLA-DRA+ Endothelial cells increase. Proinflammatory features in the tumor microenvironment mediate the persistence of residual tumors by modulating CD8+ Tcells and other response-associated immune cell populations. Our study provides valuable resources and biological insights into the mechanism of successful ICI therapy and potential targets for improving treatment efficacy.

Machine learning-based investigation of the relationship between immune status and left ventricular hypertrophy in patients with end-stage kidney disease.

Left ventricular hypertrophy (LVH) is the most frequent cardiac complication among end-stage kidney disease (ESKD) patients, which has been identified as predictive of adverse outcomes. Emerging evidence has suggested that immune system is implicated in the development of cardiac hypertrophy in multiple diseases. We applied machine learning models to exploring the relation between immune status and LVH in ESKD patients. A cohort of 506 eligible patients undergoing immune status assessment and standard echocardiography simultaneously in our center were retrospectively analyzed. The association between immune parameters and the occurrence of LVH were evaluated through univariate and multivariate logistic analysis. To develop a predictive model, we utilized four distinct modeling approaches: support vector machine (SVM), logistic regression (LR), multi-layer perceptron (MLP), and random forest (RF). In comparison to the non-LVH group, ESKD patients with LVH exhibited significantly impaired immune function, as indicated by lower cell counts of CD3+ T cells, CD4+ T cells, CD8+ T cells, and B cells. Additionally, multivariable Cox regression analysis revealed that a decrease in CD3+ T cell count was an independent risk factor for LVH, while a decrease in NK cell count was associated with the severity of LVH. The RF model demonstrated superior performance, with an average area under the curve (AUC) of 0.942. Our findings indicate a strong association between immune parameters and LVH in ESKD patients. Moreover, the RF model exhibits excellent predictive ability in identifying ESKD patients at risk of developing LVH. Based on these results, immunomodulation may represent a promising approach for preventing and treating this disease.

Dissecting the role of H-2D bclass I molecule in the development of brain atrophy during Theiler’s murine encephalomyelitis virus (TMEV) infection

Abstract Brain atrophy is a common feature of many neurological diseases as diverse as Alzheimer’s disease, cerebral palsy, Huntington’s disease, multiple sclerosis, epilepsy, encephalitis, neurosyphilis, neuroAIDS, and Covid-19 infection. We have developed a murine model of brain atrophy using the Theiler’s murine encephalomyelitis virus (TMEV) infection mouse model of multiple sclerosis. In this study, we investigated the contribution of the MHC class I molecule, H-2Db, in generating an immune response associated with the onset of brain atrophy using mice with a C57BL/6 background. The H-2D bClass I molecule not only contributed to significantly more ventricular enlargement in a T2-weighted MRI at 45 d.p.i., but this ventricular enlargement correlated to CD8 T cell infiltration. To further define the cellular and molecular mechanisms of MHC class I molecules in TMEV induced brain atrophy we investigated the cell specific contribution of brain resident antigen presenting cells (APCs) in ventricular atrophy. We developed novel bi-transgenic mouse lines with tamoxifen induced conditional ablation of the H-2Db class I molecule in Cx3CR1+ brain resident myeloid cells. Cx3CR1cre+/Db Lox P mice presented with significantly less ventricular atrophy at 45 d.p.i., compared to cre- littermates along with a decrease in CD8 T cell infiltration. These results strongly imply that antigen presentation by H-2Db on myeloid cells, at least in part is responsible for the development of brain atrophy and provided insight into the molecular mechanism of this poorly understood neuropathology. NS103212, NS122174

Targeting lL-38 triggers γδ T cell-dependent anti-tumor immunity

Abstract The IL-1 family receptor antagonist IL-38 suppresses auto-inflammatory reactions in murine disease models. Given the connection between auto-immunity and anti-tumor immunity, we wondered if targeting IL-38 would affect the latter. The transgenic PyMT mammary carcinoma model was employed to target IL-38 genetically or with a neutralizing antibody. IL-38-deficient (KO) PyMT mice had a markedly delayed tumor growth compared to WT mice, accompanied by increased NK cells, CD8+, NKT, and γδ T cell infiltrates. Accordingly, in human mammary carcinoma, IL-38 expression negatively correlated with infiltrating T cell subsets. This pattern was largely recapitulated with a IL-38-neutralizing antibody, where antibody-treated PyMT WT mice showed a strong decrease in tumor growth compared to IgG control treated animals, accompanied by an increase in CD8+ and γδ T cells. In a therapeutic model of chemoresistance, IL-38 antibody in combination with doxorubicin prevented tumor relapse, which was accompanied by increased in NK cells, CD8+, CD4+, and γδ T cells. To analyze the involvement of γδ T cells, we used γδ-TCR blocking antibody compared to IgG control in IL-38 KO mice. The delay in growth of IL-38 KO PyMT tumors compared to WT PyMT tumors was abolished upon γδ-TCR blockade. This was accompanied by a strong decrease in CD8+ cells upon effective γδ-TCR blockade, suggesting a cross talking between CD8+ and γδ T cells. Whole transcriptome sequencing analysis of PyMT tumors upon IL-38 neutralization and γδ-TCR blockage suggested NOTCH signaling to mediate the link between CD8+ and γδ T cells. Taken together, these results provide evidence for a therapeutic potential of anti-IL-38 antibodies to activate anti-tumor immunity in mammary carcinoma. Supported by a grant from Deutsche Krebshilfe (70114051)

Effect of epidural blockade on immune parameters in the perioperative period: randomized controlled study

BACKGROUND: The potential benefits of anaesthesia techniques in the parameters of cellular immunity and prevention of metastasis in various types of cancer are attracting increasing attention.
 OBJECTIVE: Our aim was to estimate the influence of the epidural blockade as a component of the anesthesia support on the immunity indexes in the patients operated for gastric cancer.
 MATERIALS AND METHODS: 89 patients operated on for gastric cancer, divided into two groups were investigated: in the 1st group (n=27) there was general anesthesia (GA) in combination with extended epidural blockade; in the 2nd group (n=24) GA with postoperative intravenous administration of analgesics. T-cell counts (CD3+, CD4+ CD8+, CD4+/CD8+) and cytokines (IL-4, IL-6, IL-10, INF-y and TNF-a) were determined.
 RESULTS: The CD3+ T-cell levels in both groups were significantly (р 0.05) reduced immediately after surgery. On the 2nd and 3rd day the decrease in CD3+ T-cells was more pronounced (р 0.05) in the 2nd group compared to the 1st group. The content of CD4+ T-cells decreased by the 1st day of the postoperative period (р 0,05) but then gradually increased, the CD4+ level in the 1st group was significantly (р 0.05) higher than in the 2nd group. The changes of CD8+ content between the studied groups were not statistically significant. The immunoregulatory index in both groups decreased by1st day (р 0.05). On the 9th day the index in the 1st group returned to the basic values, while in the 2nd group it remained significantly lower. On the 1st and 3rd day, the levels of IL-4 and IL-6 significantly (р 0.05) increased, while in the 2nd group the levels were higher than in the 1st group (р 0.05). On the 9th day IL-4 and IL-6 indices returned to their initial values. The changes of IL-10 level had a similar pattern, while in the 1st group the rate was significantly higher (р 0.05). At all stages of investigation, the level of IFN- was significantly (р 0.05) higher in the 2nd group, the level of TNF- was higher in the 1st group (р 0.05).
 CONCLUSION: The use of postoperative epidural analgesia during gastric resection for malignant neoplasm is associated with a less suppressive effect on the content of subpopulations of T-lymphocytes and cytokines in comparison with intravenous administration of analgesics and may contribute to the activation of antitumor immune response. Prospective long-term studies are needed to evaluate physician epidural analgesia for oncological outcomes after surgery for gastric cancer.

Abstract CT197: Profiling of tumor infiltrating T cells in malignant pleural/peritoneal mesothelioma (MPM) and ovarian cancer patients as part of a Phase 1 clinical trial of gavo-cel (TC-210)

Abstract Gavocabtagene autoleucel (gavo-cel; TC-210) is a T cell receptor fusion construct (TRuC) cell therapy directed against mesothelin. As part of the phase 1 dose-escalation clinical trial (NCT03907852), patients with advanced solid tumors expressing mesothelin were administered a single infusion of gavo-cel. Thirty-two patients with late-stage disease (23 MPM, 8 ovarian and 1 cholangiocarcinoma) received gavo-cel at doses ranging from 5 × 107 - 5 × 108 TRuC cells/m2. The disease control rate (defined as achieving a response by RECISTv1.1 criteria or having stable disease [SD] for at least 12 weeks post-infusion) was 77%. Dose level 3 (1 × 108 TRuC cells/m2 following lymphodepletion with cyclophosphamide and fludarabine) was selected as the Recommended Phase 2 Dose (RP2D) for the ongoing phase 2 portion of the trial. Spatial distribution of immune cells, particularly T cells, in the tumor and stromal regions of tumor biopsies has been shown to identify patients that may respond to immunotherapies. To understand the tumor immune contexture, we profiled the T cell compartment in biopsies from five patients collected prior to and eight weeks post treatment using multiplexed immunofluorescence. We observed that the MPM patient with a partial response (PR) had a greater number of T cells at baseline in tumor biopsies when compared with the other SD patients. Notably, this patient with a PR, had a 3.6-fold increase in CD3+CD8+ T cell infiltrates into the tumor bed. In contrast, 2 SD patients with MPM had a minimal increase of T cells in the tumor compartment at 8 weeks post infusion. The density of TILs 8 weeks post infusion was much lower in the SD patients compared to the PR subject, indicating a potentially insufficient number of infiltrating T cells. Interestingly, 2/2 SD patients with ovarian cancer had a decrease in CD3+CD8+ T cells infiltrating into the tumor, consistent with an immune exclusion phenotype. The PR patient with MPM had minimal increase in PD-L1 and CD155 (a ligand for the inhibitory receptor, TIGIT) from negligible levels at baseline, whereas both SD patients with MPM had marked increases in PD-L1 and CD155, indicating potential resistance mechanisms. In summary, this preliminary data suggests although CD8+ TILs are increased following treatment with gavo-cel, the tumor regions in patients with SD show evidence of immune surveillance escape via PD-L1 and CD155 upregulation, thus acting as putative mechanisms of resistance. However, as seen in the PR patient with MPM there appears to be a critical number of TILs associating with response. We are currently evaluating the tumor immune contexture in phase 2, focusing on differences across tumor types with regards to TIL content, T cell influx, and correlations with response. We will also explore additional markers such as PD-1 and TIGIT, to profile the exhaustion state of immune cells and their spatial relationship with infiltrating gavo-cel T cells. Citation Format: Munisha Smalley, Michael Ross, Samantha F. Fowler, Lauren McLaughlin, Erin Jeter, Kevin Zikaras, Philip D. Bardwell, Alfonso Quintás-Cardama, Robert Tighe. Profiling of tumor infiltrating T cells in malignant pleural/peritoneal mesothelioma (MPM) and ovarian cancer patients as part of a Phase 1 clinical trial of gavo-cel (TC-210) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 2 (Clinical Trials and Late-Breaking Research); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(8_Suppl):Abstract nr CT197.

Abstract 2138: Immune features of irAEs and aPD1 response in urothelial cancer patients of the RADIOHEAD study, as detected in blood by mass cytometry immune profiling

Abstract Checkpoint immunotherapies (CPI) have resulted in long lasting clinical responses; however, CPI treatment comes with the risk of immune-related adverse events (irAEs). Previous reports have established a correlation between irAE occurrence and better CPI patient outcomes, yet these correlations and their underlying biological mechanisms are poorly understood. To better understand the immunological mechanisms behind the correlation of irAE presentation and response, we performed immune profiling on 40 urothelial carcinoma (UC) patients receiving aPD-1 as part of the RADIOHEAD study (Lucas et al. SITC 2022, abstract #1256). A 44-marker mass cytometry panel was used to profile pretreatment and early on-treatment PBMCs to capture cell subsets and functional states across innate and adaptive immune cells. We performed analysis on gated immune cell populations and through unsupervised clustering. Group comparisons were made by adjusted Kruskal-Wallis test, whereas survival correlates were assessed through univariate Cox regression analysis. For clinical outcome analyses, we defined responders as patients who had overall survival greater than 12 months (n=26) and noted 8 patients developed irAEs. Several immune features early on-treatment were associated with overall survival. KLRG1+ naive and HLA-DR+ CD38+ central memory CD8 T cells were negatively correlated with survival, whereas Treg and TIM-3+ classical monocytes were positively associated (HR= 2.6, 3.2, 0.4, 0.05; p=0.0002, 0.00005, 0.01,0.009 respectively). Next, we classified patients into the following groups: responders with irAE (R-irAE, n=5), responders without irAE (n=21), non-responders with irAE (n=3) and non-responders without irAE (n=10). We found that several immune features were unique to R-irAE patients early on aPD-1 treatment. R-irAE patients had up to 30% more CD74+ cDCs, up to 4-fold more TIM-3+ classical monocytes, and had a greater fold decrease in CD16+ NK cells from baseline compared to other patients (p<0.005, 0.001,0.008 respectively). They also had up to 2-fold more Treg cells early on treatment, including a subset of activated Tregs marked by expression of CD38, CTLA-4, ICOS and TIGIT (p=0.05) which trended up to significance at pretreatment (p=0.075). In this study, we demonstrate that high-dimensional immune profiling by mass cytometry can detect novel blood-based biomarkers associated with clinical outcomes. Although CPI response and irAE incidence both require activation of a patient’s immune system, different mechanisms likely apply. Based on our findings, there are markers associated with responders broadly, as well as specific to R-irAE patients. We plan to validate these findings and identify additional markers and mechanisms of response and irAE with immune profiling on more RADIOHEAD indications. Citation Format: Connor P. Healy, Johanna M. Sweere, Samantha I. Liang, Natalia Sigal, Elizabeth Enrico, Li-Chun Cheng, Anastasia Lucas, Marshall A. Thompson, Diane M. Da Silva, Justin A. Jarrell, Ramji Srinivasan, Matthew H. Spitzer, Ngan Nguyen, Christine N. Spencer. Immune features of irAEs and aPD1 response in urothelial cancer patients of the RADIOHEAD study, as detected in blood by mass cytometry immune profiling [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 2138.

Abstract 1798: ALDH inhibition as modulator of ovarian tumor associated immune cells

Abstract Rationale: Ovarian cancer (OC) is an aggressive disease with the 3rd highest mortality to incidence ratio of all cancers. Aldehyde dehydrogenase-1A enzymes play a key role in retinoic acid (RA) synthesis. RA regulates cell proliferation, differentiation, and survival by activating nuclear receptor transcription factors (RAR, RXR). In cancer cells, increased ALDH expression correlates with chemoresistance and poor prognosis. Our goal is to effectively utilize ALDH inhibitors (ALDHi) as novel therapeutics and identify their mechanism in cancer and immune cells. Based on RNAseq data, NR4A1 has emerged as a possible ALDHi target. After binding RXRγ, NR4A1 serves as a ‘molecular switch’ for cancer cell survival. Here, we explore the potential use of ALDHi as immune modulators in OC. Methods: T cells: The effect of several ALDHi on T cells following anti-CD3/CD28 polyclonal stimulation was tested via flow cytometry. Macrophages: Human OC ascites samples were cultured and treated for 72h with ALDHi or DMSO control. Flow cytometry was performed to identify ascites resident cell populations. Western blot to detect NR4A1 was conducted. Dendritic cells (DCs): Murine bone-marrow derived DCs were exposed to ALDHi or DMSO and expanded in culture; cells were counted via hematocytometer to assess myeloid cell expansion and DC differentiation. DC vaccine: ALDHi and DMSO were used with a DC vaccine and sonic hedgehog inhibitor in a study of ID8 tumor-bearing mice to assess effects on tumor progression and survival. Results: We identified several novel ALDHi that promote T cell proliferation and CD8 expansion in vitro. Exposure of 5 patient ascites samples to ALDHi showed a significant decrease in CD14+ and CD163+ cells, indicating a loss of M2 immune-suppressive macrophages vs DMSO. NR4A1 expression was decreased in ALDHi treated cells vs DMSO. In contrast, no deleterious effect was observed on ascites T cells. DC expansion: There was a 10-fold increase in DC proliferation when cells were exposed to ALDHi vs DMSO in culture. DC vaccine: In vivo, addition of ALDHi to a DC vaccine did not provide a survival significance. Discussion: In vitro, ALDHi have differential, complementary effects on tumor and immune cells in OC. In tumor cells, ALDHi lower chemoresistance and trigger necroptosis. ALDHi support the expansion of polyclonal CD8 T cells and type-1 DCs and inhibit T-regulatory cells. In contrast, ALDHi reduce M2 macrophages. Despite beneficial effects in vitro, suggesting anti-tumor competence, there was a modest effect on survival in tumor bearing mice. This may be explained by poor bioavailability of current ALDHi. Further analyses of cryopreserved murine tissues (tumor, lymph nodes, spleen) using flow and CHIP cytometry to assess surface markers, Western blot to test levels of NR4A1, and IHC to assess intratumoral immune cell invasion will continue to test target engagement. Alternative in vivo treatment regimens with other compounds are ongoing. Citation Format: Julia Knight, Bingsi Gao, Mainpal Rana, Ibrahim Uygun, Ronald Buckanovich, Anda Vlad. ALDH inhibition as modulator of ovarian tumor associated immune cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 1798.

Abstract 6629: Genomic and immunophenotypic landscape of acquired resistance to PD-(L)1 blockade in non-small cell lung cancer

Abstract Background: Despite immune checkpoint inhibition (ICI) has extended survival in patients (pts) with non-small cell lung cancer (NSCLC), acquired resistance to ICI develops after an initial clinical benefit. However, the mechanisms underlying the development of acquired resistance to ICI in NSCLC are largely unknown. Methods: Tumor genomic profiling, immune cell subsets assessment from standard haematoxylin/eosin (H&E) stained slides using machine learning, multiplexed immunofluorescence (CD8, PD1, PD-L1, Foxp3, CK7), and HLA-I immunohistochemistry (IHC) were performed on matched pre and post ICI tumor biopsies from pts with NSCLC treated with ICI at the Dana-Farber Cancer Institute who developed acquired resistance to ICI. Results: Among 1823 pts with advanced NSCLC who received ICI, 80 developed acquired resistance to treatment and had matched pre- and post-ICI tissue samples for correlative analysis. Putative resistance mutations were identified in 27.3% of cases. The most common acquired loss-of-function mutations included STK11 (9.1%), B2M (6.5%), NF1/2 (5.2%), SMARCA4 (5.2%), APC (3.9%), KEAP1 (2.6%), and JAK1 (2.6%). In addition, acquired activating mutations were found in PI3KCA (3.9%) and SOS1 (1.3%). In examining copy number variations (CNVs), we identified acquired CNVs in 49.3% (38/77) of ICI-resistant samples, including heterozygous loss of B2M (23.4%), STK11 (16.8%), SMARCA4 (16.9%), KEAP1 (15.6%), PTEN (10.4%), CDKN2A/B (9.1%) and PD-L1/PD-L2 genes (9.1%), among others. Importantly, we also noted acquired homozygous deletions in CDKN2A/B (9.1%), PD-L1/PD-L2 (1.3%) and JAK2 (1.3%), and amplification in MDM2 (5.2%), KRAS (3.9%) and MYC (2.6%), at the time of resistance to ICI. Among 16 cases with pre- and post-ICI H&E stained slides, digital pathology analysis showed a significant decrease in the density of intratumoral lymphocytes in ICI-resistant tumor samples compared to pre-ICI samples (88 vs 36 cells/mm2, P=0.03). Consistently, among 6 pts with pre- and post-ICI multiplexed immunofluorescence we observed a significant decrease in CD8+ PD1+ T cells in ICI-resistant vs pre-ICI tumor samples (24 vs 10 cells/mm2, P=0.01). Among pts with tissue available for HLA-I IHC (N=9), we found a significant decrease in HLA-I expression by H-score at the time of acquired resistance (300 vs 260, P=0.03). In control cohorts of pts with pre- and post-chemotherapy (CT) (N=32) or targeted therapies (TT) (N=89) tumor genomic profiling no acquired mutations in STK11, KEAP1, B2M, JAK1, APC, SOS1 were detected. Similarly, intervening chemotherapy and targeted therapies were not associated with changes in lymphocytes density and HLA-I expression. Conclusion: Mechanisms of acquired resistance to ICI are heterogenous, including both genomic and immunophenotypic factors. New therapeutic strategies are required to delay and overcome ICI resistance in pts with NSCLC. Citation Format: Biagio Ricciuti, Giuseppe Lamberti, Sreekar Puchala, Navin Mahadevan, Joao Alessi, Xinan Wang, Yvonne Li, Federica Pecci, Alessandro Di Federico, Malini Gandhi, Adriana Barrichello, Victor Vaz, Andy Pangilinan, Danielle Haradon, Elinton Lee, Hersh Gupta, Mizuki Nishino, Andrew Cherniack, Scott Rodig, Lynette Sholl, Renato Umeton, MArk M. Awad. Genomic and immunophenotypic landscape of acquired resistance to PD-(L)1 blockade in non-small cell lung cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 6629.

Abstract 5874: HSF1 downregulation of CCL5 reduces CD8 T cell trafficking in breast cancer

Abstract Breast cancer is the leading cause of cancer related deaths in women. The presence of cytotoxic immune cells, specifically CD8+ T cells, in breast tumors is associated with better patient outcomes. Our goal is to understand mechanisms that regulate the infiltration of CD8+ T cells. This will in turn, improve the treatment and enhance patient overall survival. Utilizing a novel HSF1 activity gene signature, we observed that HSF1 activity was negatively associated with the presence of CD8+ T cells in breast cancer patients. Both CIBERSORT analysis of TCGA data as well as direct assessment of patient primary specimens demonstrated that patient tumors with high HSF1 activity had lower numbers of CD8+ T cells. To functionally test this relationship, HSF1 was knocked down in 4T1 breast cancer cells and grown orthotopically in BALB/c mice. Tumors with HSF1 knockdown showed lower tumor volumes and increased CD8+ T cell infiltration. We subjected the control and knockdown tumors to single- cell RNA sequencing analysis and found increased immune cell presence specifically CD8+ T cell presence in knockdown tumors. We further tested the effect of CD8+ T cells on the growth of HSF1 knockdown tumors by depleting CD8+ T cells in BALB/c mice. HSF1 knockdown tumors were significantly larger with CD8+ T cell depletion suggesting a functional role for HSF1 to inhibit CD8+ T cell infiltration and protect the tumor from immune-mediated killing. To investigate the mechanism by which HSF1 affects CD8+ T cells, we investigated whether HSF1 affects chemotactic cytokines. We found that loss of HSF1 significantly increased secretion of CCL5, a known chemoattractant for CD8+ T cells. Loss of HSF1 also increased mRNA levels of CCL5, suggesting a transcriptional effect on CCL5. To test the importance of CCL5 in the phenotype of HSF1 knockdown, a transwell migration assay was performed wherein T cells from BALB/c mice were placed in the upper chamber of a transwell chamber and the lower chamber contained conditioned medium from 4T1 cells with knockdown of HSF1 with or without knockdown of CCL5. The lower chamber was subjected to flow cytometry for CD3/CD8 to identify CD8+ T cells and we observed that loss of HSF1 increased migration of CD8+ T cells and this was ablated with additional loss of CCL5. These results suggest CCL5 is a significant mediator for the recruitment of CD8+ T cells when HSF1 is lost in cancer cells. We propose the model whereby high HSF1 activity in breast cancer cells suppresses expression and secretion of the chemo-attractant cytokine CCL5 that ultimately leads to a decrease in CD8+ T cells in breast tumors microenvironment. Citation Format: Curteisha LeeAnn Jacobs, Sakhi Shah, Wen-Cheng Lu, Richard Carpenter, Haimanti Ray, Xin Lu, Sha Cao. HSF1 downregulation of CCL5 reduces CD8 T cell trafficking in breast cancer. [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2023; Part 1 (Regular and Invited Abstracts); 2023 Apr 14-19; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2023;83(7_Suppl):Abstract nr 5874.

State of local immune in phegmon of the maxillofacial region and neck on the background local photodynamic therapy

Objective. To study the effect of local photodynamic therapy (PDT) on local immunity in patients with maxillofacial region and neck phlegmonas.Materials and methods. The study included 62 patients with phlegmon of the maxillofacial region and neck. Two groups of patients were formed. Patients of the 1st group were treated locally according to the traditional methods. Patients of the second group additionally underwent local PDT on the background of the complex of traditional therapeutic measures.Results. Under the influence of local PDT with the preparation «Fotolon» in patients with maxillofacial region phlegmonomas in the postoperative wound we observed activation of phagocytosis, an increase in the population of CD3 + T-lymphocytes, subpopulation CD4 + T-helper, immunoregulatory index (IRI) with a simultaneous decrease in the level of CD8 + T-lymphocytes, The decrease of CD25+ activated Tand B-lymphocytes carrying the receptor for interleukin 2, and CD95+ activated Tand B-lymphocytes carrying the Fas-receptor, the decrease of complement activity, circulating immune complexes (CIC), IgM, IgA levels with an increase of IgG.Conclusion. Local PDT activates local immunity in patients with maxillofacial and neck phlegmons. Exposure to PDT causes a more balanced immune system response by increasing the innate immune response in patients and a pronounced immunological reactivity in the region of the postoperative wound.

ВЛИЯНИЕ НИЗКОИНТЕНСИВНОГО ЛАЗЕРНОГО ИЗЛУЧЕНИЯ НА ИММУНОЛОГИЧЕСКИЕ ПОКАЗАТЕЛИ У ПАЦИЕНТОВ С ФЛЕГМОНАМИ ЧЕЛЮСТНО-ЛИЦЕВОЙ ОБЛАСТИ И ШЕИ

Background. The problem of treatment of phlegmon of the maxillofacial region and neck remains one of the most urgent problems of maxillofacial surgery. Aim. To determine the degree of influence of local Low Level Laser Therapy (LLLT) on the quantitative and functional characteristics of immunocytes and humoral factors in peripheral blood and lavage fluid in phlegmon of the maxillofacial region and neck. Material and methods. We have examined and treated 60 patients with phlegmon of the maxillofacial region and neck. Of these, 30 people made up group 1 – traditional treatment and 30 people were included into group 2 (traditional treatment supplemented with local LLLT). Also, 12 practically healthy individuals were examined to determine the regional norm of the selected indicators (control group). The total number of leukocytes and their subpopulations were determined in the blood and lavage fluid, immunophenotyping of lymphocytes was performed using monoclonal antibodies (CD3+, CD4+, CD8+, CD16+, CD19+, CD25+, CD95+), determination of the phagocytic index (PhI) and phagocytic number (PhN), the study of hemolytic complement activity (CH50), the presence of circulating immune complexes (CIC) and the level of immunoglobulins (Ig) of classes M, A, G. Results. Weekly local exposure of LLLT contributed to a decrease in leukocytes and neutrophils in the blood with an increase in their functional activity (PhN), an increase in the number of lymphocytes, levels of IgM, IgG, while reducing CH50 in blood serum. In the lavage fluid under the influence of LLLT, a decrease in leukocytes, neutrophils, activation of phagocytosis, an increase in lymphocytes due to the number of CD3+ and CD4+ T-lymphocytes, a decrease in CD8+ of cytostatic T-lymphocytes, CD25+ activated T- and B-lymphocytes, CD95+T- and B-lymphocytes, decrease in CH50, IgM level and increase in IgG were revealed. Conclusion. Comprehensive treatment of patients with phlegmon of the maxillofacial region supplemented with LLLT has a modulating effect on cellular and humoral immunity factors by enhancing the local immune response.

Altered T-Cell Profile in Sickle Cell Disease

Background: Impaired immune status due to altered T-cell response in sickle cell disease (SCD) might provide substantial insight into immune activity in SCD patients. Materials & methods: A total of30 healthy control, 20 SCD patients in a crisis state and 38 SCD patients in a steady state were evaluated for T-cell subsets. Results: A significant decrease in CD8+ (p=0.012) and CD8+45RA-197+ (p=0.015) T-cells were observed among SCD patients. Naive T-cells(45RA+197+;p<0.01) were elevated and effector (RA-197-) and central memory (RA-197+) T-cells were grossly reduced in the crisis state. Negative regression of naive T-cells with CD8+57+ affirmed immune inactivation. The predictor score reflected 100% sensitivity for predicting the crisis state (area under the curve=0.851; p<0.001). Conclusion: Monitoring naive T-cells with predictive scores can help assess the early shift from a steady state to a crisis state.

Age at SARS-CoV-2 infection and psychological and physical recovery among Chinese health care workers with severe COVID-19 at 28 months after discharge: A cohort study.

No prior study had reported the psychological and physical recovery of patients with COVID-19 2~3 years after discharge from the hospital. Moreover, it is not clear whether there is any difference in the health status of the patients with COVID-19 of different ages after discharge from the hospital. Embedding in the "Rehabilitation Care Project for Medical Staff Infected with COVID-19" in China, this study included 271 health care workers (HCWs) with severe COVID-19. Their status of health-related quality of life, persistent symptoms, functional fitness and immune function at 28 months after discharge were followed, and compared according to tertiles of age at SARS-CoV-2 infection (group of younger (≤ 33 years); medium (34-42 years); and older (≥43 years)). Multivariate linear regression and multivariable adjusted logistic regression models were applied in investigating the associations of age at SARS-CoV-2 infection and outcomes. At 28 months after discharge, 76% of the HCWs with severe COVID-19 had symptom of fatigue/weakness; 18.7% of the HCWs with severe COVID-19 did not fully recover their functional fitness; the decrease of CD3+ T cells, CD8+ T cells and the increase of natural killer cells accounted for 6.6, 6.6, and 5.5%, respectively. Compared with the HCWs with severe COVID-19 in younger group, HCWs with severe COVID-19 in older group had lower scores regarding physical functioning, role physical, bodily pain and role emotional; HCWs with severe COVID-19 in older group had higher risk of cough, joint pain, hearing loss and sleep disorder; HCWs with severe COVID-19 in older group scored lower on flexibility test. The variance of relative numbers of CD3+ T cells, CD8+ T cells and natural killer cells among HCWs with severe COVID-19 of different age groups were significant. This study demonstrated that older HCWs with severe COVID-19 recovered slower than those with younger age regarding health-related quality of life, persistent symptoms, functional fitness and immune function at 28 months after discharge. Effective exercise interventions regarding flexibility should be performed timely to speed their rehabilitation, especially among those with older age.

Mesenchymal stem cell suppresses the efficacy of CAR-T toward killing lymphoma cells by modulating the microenvironment through stanniocalcin-1.

Stem cells play critical roles both in the development of cancer and therapy resistance. Although mesenchymal stem cells (MSCs) can actively migrate to tumor sites, their impact on CAR-T immunotherapy has been little addressed. Using an in vitro cell co-culture model including lymphoma cells and macrophages, here we report that CAR-T cell-mediated cytotoxicity was significantly inhibited in the presence of MSCs. MSCs caused an increase of CD4+ T cells and Treg cells but a decrease of CD8+ T cells. In addition, MSCs stimulated the expression of indoleamine 2,3-dioxygenase (IDO) and programmed cell death-ligand 1 (PD-L1) which contributes to the immune-suppressive function of tumors. Moreover, MSCs suppressed key components of the NLRP3 inflammasome by modulating mitochondrial ROS release. Interestingly, all these suppressive events hindering CAR-T efficacy could be abrogated if the STC1 gene, which encodes the glycoprotein hormone staniocalcin-1, was knockdown in MSC. Using xenograft mice, we confirmed that CAR-T function could also be inhibited by MSC in vivo and STC1 played a critical role. These data revealed a novel function of MSC and staniocalcin-1 in suppressing CAR-T efficacy, which should be considered in cancer therapy and may also have potential applications in controlling the toxicity arising from the excessive immune response.

Modeling Blast Crisis Using Mutagenized Chronic Myeloid Leukemia-Derived Induced Pluripotent Stem Cells (iPSCs).

To model CML progression in vitro and generate a blast crisis (BC-CML) model in vitro in order to identify new targets. Three different CML-derived iPSC lines were mutagenized with the alkylating agent ENU on a daily basis for 60 days. Cells were analyzed at D12 of hematopoietic differentiation for their phenotype, clonogenicity, and transcriptomic profile. Single-cell RNA-Seq analysis has been performed at three different time points during hematopoietic differentiation in ENU-treated and untreated cells. One of the CML-iPSCs, compared to its non-mutagenized counterpart, generated myeloid blasts after hematopoietic differentiation, exhibiting monoblastic patterns and expression of cMPO, CD45, CD34, CD33, and CD13. Single-cell transcriptomics revealed a delay of differentiation in the mutated condition as compared to the control with increased levels of MSX1 (mesodermal marker) and a decrease in CD45 and CD41. Bulk transcriptomics analyzed along with the GSE4170 GEO dataset reveal a significant overlap between ENU-treated cells and primary BC cells. Among overexpressed genes, CD25 was identified, and its relevance was confirmed in a cohort of CML patients. iPSCs are a valuable tool to model CML progression and to identify new targets. Here, we show the relevance of CD25 identified in the iPSC model as a marker of CML progression.

Significance of tumor-associated neutrophils, lymphocytes, and neutrophil-to-lymphocyte ratio in non-invasive and invasive bladder urothelial carcinoma.

Tumor-infiltrating neutrophils and lymphocytes play essential roles in promoting or combating various neoplasms. This study aimed to investigate the association between tumor-infiltrating neutrophils and lymphocytes and the neutrophil-to-lymphocyte ratio in the progression of urothelial carcinoma. A total of 106 patients diagnosed with urothelial carcinoma were was. Pathological examination for tumor grade and stage and for tumor-infiltrating neutrophils, both CD4 and CD8+ T lymphocytes, as well as the neutrophil- to-lymphocyte ratio were evaluated. The presence of neutrophils and the neutrophil-to-lymphocyte ratio correlated with high-grade urothelial neoplasms. In both low- and high-grade tumors, the lymphocytes increased during progression from a non-invasive neoplasm to an early-invasive neoplasm. CD8+ T lymphocytes increased in low-grade non-muscle-invasive tumors compared to non-invasive tumors. Additionally, there was a significant decrease in CD8+ T lymphocytes during progression to muscle-invasive tumors. Our results suggest that tumor-infiltrating neutrophils and CD8+ T lymphocytes have a significant effect on tumor grade and progression.

An immune and epithelial-mesenchymal transition-related risk model and immunotherapy strategy for grade II and III gliomas.

Grade II and III gliomas are heterogeneous and aggressive diseases. More efficient prognosis models and treatment methods are needed. This study aims to construct a new risk model and propose a new strategy for grade II and III gliomas. The data were downloaded from The Cancer Genome Atlas (TCGA), the Gene Expression Omnibus (GEO), gene set enrichment analysis (GSEA), and the EMTome website for analysis. The Human Cell Landscape website and the Genomics of Drug Sensitivity in Cancer website were used for single-cell analysis and drug susceptibility analysis. Gene set enrichment analysis, gene function enrichment analysis, univariate and multivariate Cox regression analyses, Pearson's correlation analysis, log-rank test, Kaplan-Meier survival analysis, and ROC analysis were performed. We constructed an immune-related prognostic model associated with the isocitrate dehydrogenase 1 (IDH1) mutation status. By analyzing the immune microenvironment of patients with different risk scores, we found that high-risk patients were more likely to have an inflammatory immune microenvironment and a higher programmed death ligand-1 (PD-L1) expression level. Epithelial-mesenchymal transition (EMT)-related gene sets were significantly enriched in the high-risk group, and the epithelial-mesenchymal transition phenotype was associated with a decrease in CD8+ T cells and an increase in M2 macrophages. Transforming growth factor-β (TGF-β) signaling was the most important signaling in inducing epithelial-mesenchymal transition, and TGFB1/TGFBR1 was correlated with an increase in CD8+ T cytopenia and M2 macrophages. Survival analysis showed that simultaneous low expression of TGFBR1 and PD-L1 had better survival results. Through single-cell analysis, we found that TGFB1 is closely related to microglia and macrophages, especially M2 macrophages. Finally, we discussed the sensitivity of TGFB1 inhibitors in gliomas using cell line susceptibility data. These results demonstrated a potential immunotherapy strategy in combination with the TGFB1/TGFBR1 inhibitor and PD-1/PD-L1 inhibitor for grade II and III gliomas.

THE INFLUENCE OF THE WAR FACTOR ON AGE CHANGES IN THE SUBPOPULATION STRUCTURE OF LEUCOCYTES AS A COMPONENT OF INFLAMMAGING IN THE GROUPS OF MILITARY PERSONNEL AND CLEAN-UP WORKERS OF THE CHORNOBYL ACCIDENT.

to evaluate the subpopulation structure of peripheral blood leukocytes as a component of inflammaging in the servicemen of the Armed Forces of Ukraine and clean-up workers of the accident at the Chornobyl nuclear power plant. Three hundred ninety-seven males aged 30-82 (54.99 ± 8.85) years examined. Among them: clean-up workers of the accident at the Chornobyl nuclear power plant, aged 54-64 (59.95 ± 1.88) years, who were examined before the beginning and during the Russian military invasion of Ukraine; 24 civilians of the control group, aged 50-77 (58.87 ± 7.18) years and 15 persons of the age control group, aged 42-49 (46.00 ± 2.26) years, who examined before the war has started; the comparison group-W - 27 civilians (age: 51-82 (65.44 ± 9.13)) and the age control group-W - 14 civilians (age: 31-49 (40.64 ± 5.59)) who examined after the war has started; 73 servicemen of the Armed Forces of Ukraine aged 30-49 (42.20 ± 5.35) years and 54 servicemen aged 50-59 (54.07 ± 2.86) years. The subpopulation distribution of peripheral blood leukocytes was analyzed using flow cytometry. A decrease in the relative number of CD45+ 14- granulocytes, CD3+ HLA-DR+ T-lymphocytes, and CD3-57+ NK against the background of an increase in the number of CD45+ 14+ monocytes, CD3-HLA-DR+ B-lymphocytes found in the majority of people in the examination groups, the study of which conducted after the beginning of the war, CD4+ T-lymphocytes and CD3+ 57+ NKT. The relative number of CD45+ 14- granulocytes increased against the background of a decrease in CD45+ 14- lymphocytes was noted in clean-up workers who were examined before the war started.About 30% of civilians over 50 years of age who were examined before the war started had a decrease in CD19+ B- lymphocytes, CD4+ and CD8+ T-lymphocytes, CD3-57+ NK, CD3+ 57+ NKT, and the CD4+ / CD8+ ratio. Conducted correlation analysis revealed a weak to moderate inverse correlation between the relative number of CD19+ B-lymphocytes and age in civilians, regardless of when they were examined, and the servicemen of the Armed Forces of Ukraine.Inverse correlations of HLA-DR+ B-lymphocytes, CD45+ 14+ monocytes, CD4+ T-lymphocytes, CD4+ / CD8+ ratio, and CD3-57+ NK and CD3+ 57+ NKT with age were noted in civilians who were examined after the war has started. A direct correlation between CD3+ 57+ NKT and age was found in the servicemen of the Armed Forces of Ukraine. The immunological profile of both clean-up workers in the Chornobyl nuclear power plant and the servicemen of the Armed Forces of Ukraine or civilians was similar compared to the group with reference indicators. Anintergroup study revealed changes in the subpopulation distribution of peripheral blood leukocytes, which may be associated with both aging and changes in living conditions during military operations. Specific changes in cellular immunity can either tip the balance towards inflammaging or indicate already existing immunopathologicalreactions.

CD34+ progenitor cells as diagnostic and therapeutic targets in Alzheimer's disease.

CD34+ progenitor cells as diagnostic and therapeutic targets in Alzheimer's disease.