Low Levels Of CD4 Research Articles

P-1274. Mpox: Characterization and Clinical Outcomes of Patients Attended in Health Service Providing Institutions in Colombia

Abstract Background The monkeypox virus, part of the Orthopoxvirus genus, triggered a worldwide outbreak in 2022. While this outbreak had widespread effects, there's limited information on mpox's specific impact in Colombia, particularly in terms of how it is managed, its burden, and its epidemiology. This research seeks to examine the medical context, clinical variations, and health consequences in individuals diagnosed with mpox, especially those with HIV in Colombian health institutions Methods This retrospective study was conducted in health institutions in Colombia based on clinical records from Jan 2022 to Dec 2023. . Participants in the study were diagnosed through molecular methods and their clinical evolution was tracked through medical records. CD4 groups, BMI groups, viral suppression status, and HIV status were used to analyse the results. Results One thousand four hundred thirteen (1,413, 97.2% male) individuals, including 2.6% identified as healthcare workers were included in this study. Concomitant sexually transmitted diseases were common, affecting 30.1%, mainly syphilis (80%) and Neisseria gonorrhoeae (16.4%). 54% of the population (764/1413 individuals, 99.3% male) were persons living with HIV (PWH), and almost one-third (31%, n=284) of participants had concomitant sexually transmitted diseases and HIV PWH also had a higher proportion of gastrointestinal symptoms and genital symptoms. An important difference between HIV-positive and negative patients is that the former had a higher proportion of syphilis (p-value 0.5909) and other STIs (p-value 0.0026). Although PWH were younger (34.3±7.7 vs. 37.7±7.5), the difference in mean age was not statistically significant, although the differences in proportion were (15 to 44 years group, p-value 0.0097). Conclusion The evidence presented shows that half of the population were people living with HIV, and the presence of mpox was not statistically significant in people with unsuppressed viral load or with low CD4 levels. The evidence provided by this study, whose results were standardized by the CRFs, and emphasizes the importance of interdisciplinary attention for patients suffering from mpox. Special emphasis should be placed on individuals support and follow-up, focusing on detecting concomitant STIs. Disclosures All Authors: No reported disclosures

Correlation of Dental and Periodontal Status With HIV Presence and Initial CD4 Counts: An Albanian Prospective Observational Study.

Background Different pathologies are encountered more often in human immunodeficiency virus (HIV)-infected patients, such as bacterial, fungal, viral infection, and neoplastic diseases. Recently, studies have shown that HIV-infected individuals have poorer oral health outcomes, worse dentition, and aggressive forms of periodontitis. This study aims to investigate the dental and periodontal status of HIV-infected patients, the correlation between CD4+ level and the CD4 percentage with dentition, and periodontal status. Methodology A prospective observational study was conducted in the University Dental Clinic and the Infective Service of University Hospital Center "Mother Teresa" in Tirana, Albania. All patients newly diagnosed with HIV infection (35 patients, 40.7%) and those without HIV infection (51 patients, 59.3%) who underwent oral examination from April through July 2024 were included. Patients were grouped according to HIV status into two groups. This study evaluated the basic demographic characteristics, laboratory measurements, especially CD4 counts, oral hygiene, and the presence of dental and periodontal lesions. The dentition status was assessed using the values of decay teeth (DT), filled teeth (FT), and missing teeth (MT), presented as DMFT. The periodontal status was evaluated through a periodontal probe measuring community periodontal index (CPI) and loss of attachment (LOA), as recommended by the World Health Organization's Oral Health Assessment Form 2013. The Pearson's correlation coefficient (r) was used to evaluate the correlation between the levels of CD4+ and DMFT, CD4+ and CPI, CD4+ and LOA, CD4+/lymphocyte percentage (CD4%) and DMFT, CD4% and CPI, and CD4% and LOA. P-values ≤0.05 were considered statistically significant. Results HIV-infected patients had a worse dentition status, with higher DT, higher MT, and higher DMFT index values (9.71 ± 6.72 vs. 5.96 ± 4.49, p = 0.003) compared to those without HIV. HIV-infected patients also had a worse periodontal status, with higher CPI (2.63 ± 1.06 vs. 0.94 ± 0.68) and LOA (2.57 ± 1.06 vs. 0.94 ± 0.68) compared to those without HIV. An important negative correlation was found between CD4+ and dentition and periodontal status. A lower level of CD4+ was correlated with a higher DMFT (r = -0.52, p = 0.01, CPI (r = -0.38, p = 0.024, and LOA (r = -0.37, p = 0.029). Conclusions HIV-infected patients manifest a worse dentition and periodontal status, with the worsening strongly correlated with the initial CD4+ levels. Periodontal disease may serve as a significant clinical indicator for the early diagnosis of HIV and its progression. Dental professionals should be vigilant in assessing periodontal health, especially in high-risk populations, as it may prompt timely testing and intervention for HIV infection.

Mycobacterium avium complex (MAC) infection in severely immunocompromised people living with HIV: Findings from a five-year cohort.

We sought to clarify the current incidence, risk factors and symptoms of disseminated Mycobacterium avium complex (dMAC) infection in admitted people living with HIV in a hospital in the Southeast of Spain. 5-years observational, retrospective and single-centre study. Demographic, clinical and analytical variables, along with microbiological, treatment and follow-up were collected. Five cases of dMAC infection in severely immunocompromised people living with HIV people living were found. dMAC was diagnosed in 22.7% of patients under 100 CD4. All patients presented with fever and clinical manifestations of pneumonia, lymphadenopathy, or gastrointestinal symptoms. Despite low CD4 levels and high viral loads in some cases, primary prophylaxis had not been previously administered. Until 2018, U.S. American guidelines recommended antimycobacterial prophylaxis for patients with low CD4 cell counts, a practice not adopted in Europe. Untreated dMAC infection is associated with high morbidity and mortality rates. dMAC infection represents a prevalent disease in severely immunosuppressed people living with HIV. dMAC requires a high index of suspicion in this population, in order to perform mycobacterial cultures from different samples.

Prognostic relevance of sarcopenia and tumor‐infiltrating CD8+ T cells in patients with hepatocellular carcinoma

AbstractAimThe relationship between sarcopenia, tumor‐infiltrating lymphocytes (TILs), and long‐term survival in patients with hepatocellular carcinoma (HCC) has not been investigated. We aimed to evaluate the prognostic relevance of sarcopenia and TILs in patients with HCC.MethodsWe included 351 patients with HCC following liver resection. Sarcopenia was defined based on the skeletal muscle index using computed tomography. Tumor‐infiltrating CD4+ and CD8+ T cells, perforin, and granzyme B were examined in liver resection specimens.ResultsSarcopenia patients had a significantly lower lymphocyte count (p = 0.003), prognostic nutritional index (p = 0.017), and CD4+ and CD8+ T cell counts (p = 0.008 and p = 0.006, respectively). The overall survival (OS) and recurrence‐free survival (RFS) rates of sarcopenia patients were significantly lower than non‐sarcopenia patients (both p < 0.001). Multivariate analysis revealed that sarcopenia and low CD8 levels were strong independent poor prognostic factors for OS and RFS (both p < 0.001). Regardless of sarcopenia, patients with high CD8 levels had significantly better OS and RFS rates and increased expression of perforin and granzyme B. Particularly, sarcopenia patients with high CD8 levels had much better OS and RFS than those with low CD8 levels and were even comparable to non‐sarcopenia patients with high CD8 levels.ConclusionsSarcopenia and low CD8 levels are strong independent poor prognostic factors in patients with HCC. Furthermore, sarcopenia patients with high CD8 levels had favorable survival and activated local immunity, suggesting that tumor‐infiltrating CD8+ T cells may play a functionally important role in sarcopenia patients.

Effect of multidisciplinary team-style continuity of care and nutritional nursing on lung cancer: randomized study.

Aim: The clinical efficacy of systemic chemotherapy is limited due to the nonspecific delivery of anticancer drugs and is associated with serious systemic adverse effects. Therefore, integrated treatment and comprehensive care are particularly important for postoperative chemotherapy patients with lung cancer.Materials & methods: This study aimed to ascertain the application effect of multidisciplinary team (MDT)-style continuity of care combined with whole-process nutritional nursing in postoperative chemotherapy patients with lung cancer. Nutritional indices, immune function, adverse emotions, self-efficacy, self-care ability, quality of life and toxic reactions during chemotherapy were recorded in postoperative chemotherapy patients with lung cancer receiving routine care (control group) and MDT-style continuity of care combined with whole-process nutritional care (intervention group).Results: After care, the intervention group performed higher BMI, PA, TP and ALB, CD3+, CD4+ and CD4+/CD8+, lower levels of CD8+, lower self-rating anxiety scale, self-rating depression scale and QLQ-C30 symptom domain scores and higher general self-efficacy scale, exercise of self-care agency scale and QLQ-C30 functional domain scores versus the control group (all p<0.05).Conclusion: MDT-style continuity of care combined with whole-process nutritional care can improve the nutritional status of postoperative chemotherapy patients with lung cancer, and in turn enhance their quality of life.

Rapamycin Prevents Expansion of Costimulation Blockade-resistant CD8+ Alloreactive Memory Cells following Depletional Induction in Renal Transplant Recipients.

Alemtuzumab induction with belatacept/rapamycin-based maintenance immunotherapy (ABR) prevents kidney allograft rejection and specifically limits early costimulation blockade-resistant rejection (CoBRR). To evaluate the mechanisms by which this regimen alters CoBRR, we characterized the phenotype and functional response of preexisting memory cells to allogeneic endothelial cells using intracellular cytokine staining and flow cytometry. IL-7-induced lymphocyte proliferation in the presence or absence of rapamycin was assessed to characterize the phenotype of proliferating cells. Lymphocytes from 40 recipients who underwent transplant using the ABR regimen were studied longitudinally. The rapid immunoresponses of preexisting alloreactive cells to allogeneic endothelial cells were predominantly CD8+TNF-α+/IFN-γ+ cells. These cells were effector memory (TEM) and terminally differentiated effector memory cells lacking CD28 expression, and most were CD57+PD1-. Neither rapamycin nor belatacept directly inhibited these cells. IL-7, a cytokine induced during lymphopenia postdepletion, provoked dramatic CD8+ TEM cell proliferation and a low level of CD8+CD57+PD1- cell expansion invitro. The IL-7 stimulation induced CD8+ cell mTOR phosphorylation, and rapamycin treatment markedly inhibited IL-7-induced TEM and CD57+PD1- cell expansion. This effect was evident in patients receiving the ABR in that the repopulation of CD8+CD57+PD1- TEM cells was substantially suppressed for at least 36 mo after transplant. These findings help define one mechanism by which a costimulation blockade/rapamycin-based therapy following alemtuzumab induction minimizes CoBRR, namely that in the presence of rapamycin, costimulation-resistant alloreactive cells are disproportionately ineffective at repopulating following post-transplant T cell depletion.

Replacement Therapy with Blood Products in People Living with HIV.

Cytopenias or coagulation deficiencies can occur in people living with HIV (PLWH). The severity of these disorders is influenced by the low levels of CD4+ lymphocytes, viral load, and the stage of viral infection. The aim of our retrospective observational study was to determine the frequency of cytopenias and coagulation deficiencies in PLWH as well as the need for replacement therapy with blood products. We sought to determine whether there is an association between severe anemia or thrombocytopenia (requiring replacement therapy) and CD4+T lymphocyte levels. All 29 patients were critically ill, with 27 out of 29 (93%) in advanced stages of HIV disease and 23 out of 29 (79%) having CD4+ lymphocyte counts below 200 cells/microL. Most patients were either late presenters (45%) or had been lost to follow-up (41%). In addition to HIV infection, various conditions that could alter hematologic parameters were associated, including co-infections with hepatitis viruses, tuberculosis at various sites, malignant diseases, sepsis, SARS-CoV-2 infection, or other opportunistic infections. No significant correlation was found between severe anemia or severe thrombocytopenia or coagulation deficiencies and the CD4+T lymphocyte count. Our data suggest that these hematological disorders in patients with advanced HIV infection are more likely to be associated comorbidities rather than the HIV infection per se.

Decreased Expression of CD247 and CD4 Immune Marker Predicts Poor Prognosis in Triple Negative Breast Cancer.

Triple negative breast cancer (TNBC) is an aggressive from of breast cancer and is associated with poor prognosis. Tumor microenvironment of breast cancer consists of a wide range of cell types, including tumor-infiltrating lymphocytes (TILs). Accumulating evidence indicate that TILs play a crucial role in cancer progression and resistance to standard chemotherapy. We used online computational tools to evaluate the prognostic significance of CD247 and CD4 in TNBC. TNBC patients with lower expression of CD247 and CD4 have much shorter relapse- free survival and overall survival than the patients with higher expression of these genes. CD247 and CD4 expression show a strong positive correlation with tumor-infiltrating dendritic cells, B-cells, CD4+, CD8+, and neutrophils. We've concluded that low levels of CD247 and CD4 may stop immune cells from entering the area around the tumor, which stops cancer cells from being killed and gives the patient a bad outlook. These findings suggest that CD247 and CD4 may be useful biomarkers or as a target to understand the progression of TNBC. Our findings also suggest that CD247 and CD4 targeted therapeutics should be explored in detail, and could be a potentially used as atreatment strategy for TNBC.

Association between immune system parameter and clinical characteristics among patients with solid cancer

BACKGROUND Lymphopenia has been reported to be a major predictor of chemotherapy-related toxicity. This study aimed to investigate the correlation between neutrophils, lymphocytes, CD4, and CD8 in solid cancer patients and cancer clinical characteristics. METHODS This was a cross-sectional study of patients who will undergo chemotherapy at the Hematology and Medical Oncology Clinic, Cipto Mangunkusumo Hospital, from June to September 2023. Clinical characteristics, CD4 and CD8 levels, and neutrophil and lymphocyte counts were assessed at the first visit. A comparative test was carried out on the patients’ average CD4, CD8, neutrophil, and lymphocyte counts. RESULTS Types of cancer were associated with CD4 levels. Patients with head and neck cancer had lower CD4 levels (411.3 [119.3–1,427.5] cells/mm³) compared with colorectal (514.7 [129.2–861.3] cells/mm³), breast and gynecological (567.5 [180.1–939 cells/mm³), and other cancers (681.4 [175.1–2,056.9] cells/mm³), with p = 0.009. Patients aged ≥40 years had higher CD8 levels than those aged &lt;40 years (376.4 [142.8–1,293.1] cells/mm³ versus 565.3 [185.9–1,944] cells/mm³, p = 0.01). Additionally, lymphocyte count was associated with cancer type, with the lowest number in head and neck cancer (1,380 [280–2,660] μl, p = 0.044). CONCLUSIONS CD4 levels and lymphocyte counts were associated with the cancer type, whereas CD8 levels were influenced by age.

CD276 Promotes an Inhibitory Tumor Microenvironment in Hepatocellular Carcinoma and is Associated with Poor Prognosis.

CD276 is an emerging immune checkpoint molecule that has been implicated in various cancers. However, its specific role in hepatocellular carcinoma (HCC) remains unclear. This study examined the impact of CD276 on patient prognosis and the tumor microenvironment (TME). The Cancer Genome Atlas (TCGA) database was utilized to evaluate CD276 expression in HCC and the association between CD276 and immune indicators was also analyzed. The signaling pathways correlated with CD276 expression were identified by gene set enrichment analysis (GSEA). Different algorithms were used to assess immune cell infiltration. The effect of CD276 knockdown on HCC cell phenotypes and its relationship with macrophage polarization was examined using the cell counting kit 8 (CCK-8) assay and co-culture system. CD276 was upregulated in HCC and associated with unfavorable clinical outcomes. Hgh CD276 expression was associated with enrichment of the G2/M checkpoint, E2F targets, and mitotic spindles. CD276 expression was correlated with the infiltration of immune cells, including high level of tumor-associated macrophages and low levels of CD8+ T cells. Knockdown of CD276 decreased HCC cell proliferation and increased apoptosis. CD276 silencing in HCC cells and co-culture with THP-1-derived macrophages had a regulatory effect on macrophage polarization and macrophage-mediated cell proliferation and migration. CD276 expression in HCC is associated with unfavorable clinical outcomes and may contribute to the development of an immunosuppressive microenvironment. Specifically, CD276 was associated with alterations in immune cell infiltration, immune marker expression, and macrophage polarization during HCC progression, suggesting its potential as a prognostic indicator and promising target for immunotherapeutic intervention in HCC.

Development and validation of a prognostic immunoinflammatory index for patients with gastric cancer

BACKGROUND Studies have demonstrated the influence of immunity and inflammation on the development of tumors. Although single biomarkers of immunity and inflammation have been shown to be clinically predictive, the use of biomarkers integrating both to predict prognosis in patients with gastric cancer remains to be investigated. AIM To investigate the prognostic and clinical significance of inflammatory biomarkers and lymphocytes in patients undergoing surgical treatment for gastric cancer. METHODS Univariate COX regression analysis was performed to identify potential prognostic factors for patients with gastric cancer undergoing surgical treatment. Least absolute shrinkage and selection operator-COX (LASSO-COX) regression analysis was performed to integrate these factors and formulate a new prognostic immunoinflammatory index (PII). The correlation between PII and clinical characteristics was statistically analyzed. Nomograms incorporating the PII score were devised and validated based on the time-dependent area under the curve and decision curve analysis. RESULTS Patients exhibiting elevated neutrophil-to-lymphocyte ratio, platelet-to-lymphocyte ratio, and systemic immune inflammatory index displayed inferior progression-free survival (PFS) and overall survival (OS). Conversely, low levels of CD3(+), CD3(+) CD8(+), CD4(+)CD8(+), and CD3(+)CD16(+)CD56(+) T lymphocytes were associated with improved PFS and OS, while high CD19(+) T lymphocyte levels were linked to worse PFS and OS. The PII score demonstrated associations with tumor characteristics (primary tumor site and tumor size), establishing itself as an independent prognostic factor for both PFS and OS. Time-dependent area under the curve and decision curve analysis affirmed the effectiveness of the PII-based nomogram as a robust prognostic predictive model. CONCLUSION PII may be a reliable predictor of prognosis in patients with gastric cancer undergoing surgical treatment, and it offers insights into cancer-related immune-inflammatory responses, with potential significance in clinical practice.

An explainable machine learning-based model to predict intensive care unit admission among patients with community-acquired pneumonia and connective tissue disease

BackgroundThere is no individualized prediction model for intensive care unit (ICU) admission on patients with community-acquired pneumonia (CAP) and connective tissue disease (CTD) so far. In this study, we aimed to establish a machine learning-based model for predicting the need for ICU admission among those patients.MethodsThis was a retrospective study on patients admitted into a University Hospital in China between November 2008 and November 2021. Patients were included if they were diagnosed with CAP and CTD during admission and hospitalization. Data related to demographics, CTD types, comorbidities, vital signs and laboratory results during the first 24 h of hospitalization were collected. The baseline variables were screened to identify potential predictors via three methods, including univariate analysis, least absolute shrinkage and selection operator (Lasso) regression and Boruta algorithm. Nine supervised machine learning algorithms were used to build prediction models. We evaluated the performances of differentiation, calibration, and clinical utility of all models to determine the optimal model. The Shapley Additive Explanations (SHAP) and Local Interpretable Model-Agnostic Explanations (LIME) techniques were performed to interpret the optimal model.ResultsThe included patients were randomly divided into the training set (1070 patients) and the testing set (459 patients) at a ratio of 70:30. The intersection results of three feature selection approaches yielded 16 predictors. The eXtreme gradient boosting (XGBoost) model achieved the highest area under the receiver operating characteristic curve (AUC) (0.941) and accuracy (0.913) among various models. The calibration curve and decision curve analysis (DCA) both suggested that the XGBoost model outperformed other models. The SHAP summary plots illustrated the top 6 features with the greatest importance, including higher N-terminal pro-B-type natriuretic peptide (NT-proBNP) and C-reactive protein (CRP), lower level of CD4 + T cell, lymphocyte and serum sodium, and positive serum (1,3)-β-D-glucan test (G test).ConclusionWe successfully developed, evaluated and explained a machine learning-based model for predicting ICU admission in patients with CAP and CTD. The XGBoost model could be clinical referenced after external validation and improvement.

Anti-HIV Activity and Immunomodulatory Properties of Fractionated Crude Extracts of Alternaria alternata.

Developing new anti-human immunodeficiency virus (HIV) drug candidates that target different sites in HIV-1 replication, with better resistance profiles and lower drug toxicity, is essential to eradicating HIV. This study investigated the potential of fractionated crude extracts of Alternaria alternata as immunomodulatory or anti-HIV drug candidates. Solid-phase extraction (SPE) was used to fractionate A. alternata PO4PR2 using three different columns: MAX (Mixed-mode, strong Anion-eXchange), MCX (Mixed-mode, strong Cation-eXchange), and HLB (Hydrophilic-Lipophilic Balance) with methanol gradient methods (5%, 45%, and 95%). An MTT (3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide) assay was used to assess the cell viability and cytotoxicity of the fractionated crude extract A. alternata PO4PR2 in the TZM-bl cell lines. This was followed by a luciferase-based antiviral assay to assess the antiviral activity of A. alternata PO4PR2. A time of addition (TOA) assay was performed to ascertain the mechanism of inhibition employed by the fractionated crude extract of A. alternata PO4PR2 in the HIV life cycle. The p24 titer was determined using an ELISA, while a luciferase-based antiviral assay was used to evaluate the HIV percentage inhibition for different HIV-1 replication cycles. The TOA assay was established using antiviral drugs that target different sites in the HIV replication cycle. These included maraviroc, azidothymidine, raltegravir, and amprenavir. The immunomodulatory effect of the fractionated crude extracts on CD4+ T cells was measured by a flow cytometric analysis, for which fluorochrome-labelled monoclonal antibodies were used as markers for activation (CD38 and HLA-DR) and exhaustion (PD-1). The MCX fraction demonstrated a more significant anti-HIV inhibition than that of the fractions generated in other columns, with an IC50 of 0.3619 µg/mL, an HIV inhibition of 77%, 5% HLB (IC50: 0.7232 µg/mL; HIV inhibition of 64%), and 5% MAX (IC50: 5.240 µg/mL; HIV inhibition of 67%). It was evident from the time of addition data that the crude extract and the 5% MCX fraction inhibited viral binding (68%), reverse transcription (75%), integration (98%), and proteolysis (77%). It was shown that A. alternata (the MCX fraction) have a significant inhibitory effect on reverse transcription (75% HIV inhibition) and integration (100% HIV inhibition). The 5% MCX (p = 0.0062), 5% HLB (p = 0.0269), and 5% MAX (p = 0.0117) fractionated A. alternata crude extracts had low levels of CD4+ T cell (CD38 + HLA-DR+) activation compared to those of the AZT treatment, while CD4+ T cell activation was insignificant. The 5% MAX and HLB A. alternata fractions may possess immunomodulatory compounds with less anti-HIV-1 activity. A. alternata could be a key source of innovative anti-HIV drugs with immunomodulatory characteristics.

Clinical and molecular mechanisms for predicting the efficacy of immunotherapy for hepatocellular carcinoma based on gut microbiome.

e16222 Background: Hepatocellular carcinoma (HCC) is an aggressive malignant tumor, ranking as the fifth most common cancer and the third leading cause of cancer-related deaths globally, posing a significant global health issue. Increasing evidence suggests that the gut microbiome and their metabolites play a crucial role in the development and therapeutic processes of cancer. Immune checkpoint inhibitors (ICIs) have recently become a treatment option for unresectable advanced HCC. To further guide the selection of immunotherapy for clinical HCC patients, it is necessary to further elucidate the prediction of immunotherapy efficacy for HCC based on the microbiome. Methods: The study enrolled 26 unresectable advanced HCC patients and collected baseline clinical laboratory data. We administered ICIs therapy to HCC patients every three weeks. After 6 months of treatment, patients were divided the responsive group (R group) and the non-responsive group (NR group) based on imaging evaluation according to the Response Evaluation Criteria in Solid Tumors (RECIST 1.1). Prior to the application of anti-tumor treatment, fecal samples were collected from the patients for 16S rRNA microbiome sequencing analysis. Immunofluorescence was used to detect the expression of PD-L1 in tumor tissue, and immunohistochemistry was used to assess changes in lymphocyte subsets CD3+T cells, CD4+T cells, CD8+T cells and Treg cells, as well as the expression of TNF-α, γ-IFN, and GZMB. Additionally, the peripheral blood lymphocyte subsets CD3+T cells, CD4+T cells and CD8+T cells were analyzed by flow cytometry. Results: In our study, R group showed higher taxa richness and more gene counts than those of NR group. Comparing with NR group, the relative abundance of Firmicutes were significantly increased, whereas Proteobacteria and Bacteroidota were significantly decreased in R group at phylum level (all P &lt; 0.05). Our study showed that R group had high level of lymphocyte subsets CD3+T cells and CD8+T cells and low level of CD4+T cells and Treg cells compared to NR group (all P &lt; 0.05). And the expressions of PD-L1, TNF-α,γ-IFN and GZMB were upregulated in R group(all P &lt; 0.05). Conclusions: These findings suggested that the characteristics of gut-microbial diversity and composition at the early treatment period of ICIs therapy in HCC may have distinct implications on drug efficacy and disease prognosis. ICIs therapy reshaped HCC tumor microenvironment in parallel with regulating the gut microbiome and lymphocyte subsets immunity.

Impacts of tumor microenvironment during neoadjuvant chemotherapy in patients with esophageal squamous cell carcinoma.

With the advent of immune checkpoint inhibitors (ICIs), a better understanding of tumor microenvironment (TME) is becoming crucial in managing esophageal squamous cell carcinoma (ESCC) patients. We investigated the survival impact of TME status and changes in patients with ESCC who underwent neoadjuvant chemotherapy (NAC) followed by surgery (n = 264). We examined immunohistochemical status (CD4+, CD8+, CD20+, Foxp3+, HLA class-1+, CD204+, and programmed death ligand-1 [PD-L1+]) on 264 pre-NAC and 204 paired post-NAC specimens. Patients were classified by their pre- and post-NAC immune cell status and their changes following NAC. Our findings showed that pre-NAC TME status was not significantly associated with survival outcomes. In contrast, post-NAC TME status, such as low level of T cells, CD4+ T cells, and high PD-L1 combined positive score (CPS), were significantly associated with poor overall survival (OS). Notably, TME changes through NAC exerted significant survival impacts; patients with consistently low levels of T cells, low levels of CD4+ T cells, or high levels of PD-L1 (CPS) had very poor OS (3-year OS: 35.5%, 40.2%, and 33.3%, respectively). Tumor microenvironment changes of consistently low T cells, low CD4+ T cells, and high PD-L1 were independent predictors of poor OS in multivariate Cox hazards analyses, while factors indicating post-NAC status (T cells, CD4+, and PD-L1 [CPS]) alone were not. Therefore, we suggest that the consistently low T/high PD-L1 group could benefit from additional therapies, such as ICIs, and the importance of stratification by the TME, which has recently been recognized.

Correlation of CD4+ levels and caspase-3 in condyloma acuminata with HIV reactive patients

Background: Human papillomavirus is an infection that causes malignancy because of persistency and modulation of apoptotic pathways, especially caspase-3. Factors that can increase persistency, recurrency, and malignancy of HPV infection include HIV infection with low CD4 levels. There is disagreement or deep molecular understanding of the induction and modulation of apoptosis in HIV-mediated CD4+ T cell depletion, especially in CA. However, it is necessary to see how CD4+ levels can influence caspase-3, so it may open up new avenues for supporting investigation to consider the presence of malignancy or therapeutic strategies regarding CD4+ can induce apoptosis.Purpose: This study aimed to determine whether there is a correlation between CD4+ levels and caspase-3 expression in condyloma acuminata with HIV reactive patients.Methods: This is an analytic observational study with a cross-sectional design. The research was conducted at Dr. Moewardi General Hospital from August to December 2023. Nineteen patients with condyloma acuminata and HIV reactive were included in this study with a consecutive sampling technique. The expression of caspase-3 was assessed using immunohistochemical staining, looking at the percentage of stained cell nuclei and cytoplasm and CD4+ levels with flow cytometry examination—data analysis using Pearson correlation.Results: Respondents in this study were primarily men, self-employed with heterosexual orientation and genito-genital-oral sexual intercourse. The statistical analysis showed no significant relationship (p: 0.300, r: -0.251) between CD4+ levels and caspase-3 expression.Conclusions: While not statistically significant, CD4+ level is reduced in correlation with increased caspase-3 expressions.

Human regulatory memory B cells defined by expression of TIM-1 and TIGIT are dysfunctional in multiple sclerosis.

Regulatory B cells (Bregs) play a pivotal role in suppressing immune responses, yet there is still a lack of cell surface markers that can rigorously identify them. In mouse models for multiple sclerosis (MS), TIM-1 or TIGIT expression on B cells is required for maintaining self-tolerance and regulating autoimmunity to the central nervous system. Here we investigated the activities of human memory B cells that differentially express TIM-1 and TIGIT to determine their potential regulatory function in healthy donors and patients with relapsing-remitting (RR) MS. FACS-sorted TIM-1+/-TIGIT+/- memory B (memB) cells co-cultured with allogenic CD4+ T cells were analyzed for proliferation and induction of inflammatory markers using flow cytometry and cytokine quantification, to determine Th1/Th17 cell differentiation. Transcriptional differences were assessed by SMARTSeq2 RNA sequencing analysis. TIM-1-TIGIT- double negative (DN) memB cells strongly induce T cell proliferation and pro-inflammatory cytokine expression. The TIM-1+ memB cells enabled low levels of CD4+ T cell activation and gave rise to T cells that co-express IL-10 with IFNγ and IL-17A or FoxP3. T cells cultured with the TIM-1+TIGIT+ double positive (DP) memB cells exhibited reduced proliferation and IFNγ, IL-17A, TNFα, and GM-CSF expression, and exhibited strong regulation in Breg suppression assays. The functional activity suggests the DP memB cells are a bonafide Breg population. However, MS DP memB cells were less inhibitory than HC DP memB cells. A retrospective longitudinal study of anti-CD20 treated patients found that post-treatment DP memB cell frequency and absolute number were associated with response to therapy. Transcriptomic analyses indicated that the dysfunctional MS-derived DP memB/Breg population exhibited increased expression of genes associated with T cell activation and survival (CD80, ZNF10, PIK3CA), and had distinct gene expression compared to the TIGIT+ or TIM-1+ memB cells. These findings demonstrate that TIM-1/TIGIT expressing memory B cell subsets have distinct functionalities. Co-expression of TIM-1 and TIGIT defines a regulatory memory B cell subset that is functionally impaired in MS.

Abstract 2468: Phase II study of cryoablation and post-progression immune checkpoint inhibition in metastatic melanoma

Abstract Introduction: Efforts to overcome resistance to immune checkpoint inhibition (ICI) are paramount, and novel combination strategies are in development. Image-guided percutaneous cryoablation is an established minimally invasive oncologic treatment that has demonstrated immune modulatory effects. We hypothesized that cryoablation may prime the tumor microenvironment (TME) through direct modulation of the tumor, thereby generating an anti-tumor response in ICI refractory tumors. Methods: In this non-randomized phase II single-center study, subjects with unresectable melanoma progressing on ICI underwent cryoablation of an enlarging metastasis, and ICI was continued for a minimum of two additional cycles. The primary endpoint was safety and feasibility, with objective response rate (ORR) and disease control rate (DCR) in non-ablated lesions a key secondary endpoint. To better understand the inflammatory state of TME as well as factors determining response to cryoablation, we performed immunohistochemistry (PD-L1, CD8+ TIL) as well as single-cell RNA-sequencing (ScRNAseq) on evaluable pre- and post-cryoablation biopsies obtained from metastatic sites of 15 patients. Results: From May 2018 through July 2020, 17 patients were treated. This regimen was feasible with all patients ongoing cryoablation with no unexpected safety signal. The ORR and DCR was 23.5% (95% CI 6.81-49.9) and 41.2% (95% CI 18.44-67.08), respectively, with four patients experiencing PR and three patients SD. Durable control, defined as tumor control ≥6 months, was achieved in 35%. Median progression-free survival (PFS) was 59 days (95% CI 42, 179) with a 6-month PFS rate of 23.5% (95% CI 7.3%, 44.9%). Median overall survival (OS) was 387 days (95% CI 106, 608) with a 1-year OS rate of 53% (95% CI (27.6%, 73%). Correlative analysis demonstrated pre-ablation tumors were uniformly associated with a “cold” TME, as all samples were PD-L1 negative and the majority had low levels of CD8+ TIL, defined as ˂5% of tumor cells associated with CD8+ TILs. ScRNAseq revealed no clear trends of cell type composition to predict best response to cryoablation, however an increase in B/plasma cells was seen in patients with disease control, suggesting these cells may play a role in mounting a durable anti-tumor response after cryoablation. Conclusion: The anti-tumor efficacy of combining cryoablation and post-progression ICI was superior to what would be expected with the use of post-progression ICI alone with an acceptable safety profile. Our data support further study of this novel, synergistic therapeutic approach. Citation Format: Meghan J. Mooradian, Florian J. Fintelmann, Thomas J. LaSalle, Judit Simon, Alexander Graur, Alona Muzikansky, Mari Mino-Kenudson, Sophia Shalhout, Howard L. Kaufman, Russell W. Jenkins, Donald Lawrence, Aleigha Lawless, Tatyana Sharova, Raul N. Uppot, Jacy Fang, Emily M. Blaum, Anna L. Gonye, Irena Gushterova, Genevieve M. Boland, Nir Hacohen, Moshe Sade-Feldman, Ryan J. Sullivan. Phase II study of cryoablation and post-progression immune checkpoint inhibition in metastatic melanoma [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2468.

Pneumocystis jirovecii pneumonia: a case report

Introduction and importancePneumocystis jirovecii (PJP) pneumonia is a serious life-threatening condition in immunocompromised individuals and is often associated with human immunodeficiency virus (HIV) + patients. We describe a case of PJP pneumonia which provided a diagnostic challenge in a patient who presented with no known risk factors leading to a delay in initiation of appropriate antibiotic therapy.Case presentationA 71-year-old previously healthy white/Caucasian male presented with subacute hypoxic respiratory failure due to multifocal pneumonia with diffuse bilateral ground glass opacities with consolidations despite prior treatment with antibiotics and steroids. He was admitted and started on intravenous broad-spectrum antibiotics but continued to deteriorate, eventually requiring intubation and transfer to the ICU. Bronchoscopy revealed PJP and treatment was initiated, but the patient developed refractory shock and multiorgan failure, and ultimately died. It was later discovered that he was HIV-1 positive.Clinical discussionPJP, as a potential cause of his presentation, was not considered given that our patient lacked any overt risk factors for PJP pneumonia. He continued to worsen despite broad spectrum antibiotic therapy and hence bronchoscopy was pursued. His clinical profile, in hindsight, was suspicious for PJP pneumonia and early PJP-directed antibiotic therapy may have prevented a fatal outcome, as in this case. There was an element of cognitive bias across multiple providers which may have contributed to the delay in treatment despite his rapid clinical decline while on conventional pneumonia treatment protocol. His diagnosis was later evident when his BAL-DFA grew PJP in addition to his low levels of CD4 and CD8 cells. He was found to be HIV-1 positive five days after his death; there was a delay in this diagnosis since all positive HIV tests from the hospital are reported as ‘pending’ until the presumptive positive sample goes to the Connecticut Department of Public Health State laboratory for the confirmatory test. PJP-targeted therapies were initiated later in our patient’s hospital course when the infection had progressed to refractory septic shock with multiorgan failure and eventual death.ConclusionPJP pneumonia is a fatal disease if not recognized early in the course of illness, and the patient usually undergoes multiple antibiotic regimens before they are diagnosed and receive appropriate clinical care. The gold standard of diagnostic testing for PJP is by obtaining bronchial washings through a flexible bronchoscopy and the turnaround time for such results may take a few days to result. A significant proportion of patients may not have any overt risk factors of immunosuppression and early empiric treatment for PJP may be clinically appropriate as the delay in diagnosis may be associated with significant morbidity and mortality risk.

Characteristics of splenic PD-1+ γδT cells in Plasmodium yoelii nigeriensis infection

Although the functions of programmed death-1 (PD-1) on αβ T cells have been extensively reported, a role for PD-1 in regulating γδT cell function is only beginning to emerge. Here, we investigated the phenotypic and functional characteristics of PD-1-expressing γδT cells, and the molecular mechanism was also explored in the Plasmodium yoelii nigeriensis (P. yoelii NSM)-infected mice. Flow cytometry and single-cell RNA sequencing (scRNA-seq) were performed. An inverse agonist of RORα, SR3335, was used to investigate the role of RORα in regulating PD-1+ γδT cells. The results indicated that γδT cells continuously upregulated PD-1 expression during the infection period. Higher levels of CD94, IL-10, CX3CR1, and CD107a; and lower levels of CD25, CD69, and CD127 were found in PD-1+ γδT cells from infected mice than in PD-1− γδT cells. Furthermore, GO enrichment analysis revealed that the marker genes in PD-1+ γδT cells were involved in autophagy and processes utilizing autophagic mechanisms. ScRNA-seq results showed that RORα was increased significantly in PD-1+ γδT cells. GSEA identified that RORα was mainly involved in the regulation of I-kappaB kinase/NF-κB signaling and the positive regulation of cytokine production. Consistent with this, PD-1-expressing γδT cells upregulated RORα following Plasmodium yoelii infection. Additionally, in vitro studies revealed that higher levels of p-p65 were found in PD-1+ γδT cells after treatment with a RORα selective synthetic inhibitor. Collectively, these data suggest that RORα-mediated attenuation of NF-κB signaling may be fundamental for PD-1-expressing γδT cells to modulate host immune responses in the spleen of Plasmodium yoelii nigeriensis–infected C57BL/6 mice, and it requires further investigation.