Percentage Of CD4 Research Articles

Role of serum cd4+, cd25+, and foxp3+ cells’ segmental and nonsegmental vitiligo

Background Vitiligo is an autoimmune skin disease presented with depigmented macules and patches. Vitiligo has an impact on the quality of life. The etiopathogenesis of vitiligo is multifactorial, including genetic, immune dysregulation, and oxidative stress mechanisms. Lately, several researches have underlined the pivotal function of regulatory T cells (Tregs) in the vitiligo pathogenesis with lack of data regarding their role in segmental vitiligo (SV). Objective To investigate the role of Tregs in SV and nonsegmental vitiligo (NSV) pathogenesis and its correlation with disease activity and severity. Patients and methods This case–control study included 20 cases with NSV and 10 cases with SV in addition to 10 healthy volunteers. Vitiligo Area Scoring Index and Vitiligo Disease Activity scores were estimated for vitiligo cases and all the included participants were assessed for the percentage of serum CD4+, CD25+, and FOXP3+ T cells using flow cytometry staining buffer. Results There was significant reduction of the peripheral Tregs in NSV cases in comparison with healthy participants and negative correlation with their percentage to disease activity. On the other hand, there was insignificant difference between the percentage of peripheral Tregs in SV cases and healthy participants. Also, there was insignificant correlation between peripheral Tregs and both severity and activity of the disease among SV cases. Conclusion NSV cases showed significant reduction of the peripheral Tregs and negative correlation with disease activity, indicating the importance of Tregs in the etiopathogenesis of NSV and hence future targeting therapy. On the other hand, in SV cases, there was insignificant reduction of peripheral Tregs and insignificant correlation between their percentage and both severity and activity indicating mosaic etiopathogenesis.

The peripheral CD4+ T cells predict efficacy in non-small cell lung cancer (NSCLC) patients with the anti-PD-1 treatment.

Programmed cell death protein 1 (PD-1) inhibitor therapy has become a routine treatment for advanced non-small cell lung cancer (NSCLC). However, only some NSCLC patients would benefit from anti-PD-1 therapy. We urgently need to identify biomarkers associated with clinical response to change treatment strategies promptly for patients who fail to benefit from anti-PD-1 treatment. This study was aimed to explore whether circulating CD4+ T cells and CD8+ T cells could be biomarkers for predicting anti-PD-1 efficacy. In this study, 118 NSCLC patients who received anti-PD-1 therapy were enrolled. The percentages of circulating CD4+ T cells and CD8+ T cells before and after anti-PD-1 treatment were determined by flow cytometry. The programmed cell death ligand 1 (PD-L1) expression of tumor tissues was detected by immunocytochemistry. The anti-PD-1 treatment efficacy was assessed by immune response evaluation criteria in solid tumors (iRECIST). The percentage of CD4+ T cells and CD4+/CD8+ ratio in the peripheral blood (PB) was significantly elevated after anti-PD-1 treatment. In contrast, the percentage of CD8+ T cells in the PB was significantly decreased after anti-PD-1 treatment. Furthermore, we found that the percentages of CD4+ T cells and CD4+/CD8+ ratios considerably increased, and the percentages of CD8+ T cells significantly reduced in the effective group. On the contrary, the patients in the ineffective group showed no significant differences in the biomarkers. Multivariate logistic revealed that the percentage of CD4+ T cells at baseline was an independent predictor of anti-PD-1 treatment. The area under the curve (AUC) of the CD4+ T cells percentage was 0.7834 with a cut-off value of 28.53% (sensitivity =82.5%, specificity =66.23%). The percentage of CD4+ T cells at baseline could predict anti-PD-1 efficacy in NSCLC patients.

Peripheral blood T-cell modulation by omalizumab in chronic urticaria patients.

Chronic spontaneous urticaria (CSU) is a highly prevalent and difficult to manage cutaneous disease characterized by the presence of recurrent urticaria, angioedema, or both, for a period of 6 weeks or longer. One of the biological treatments used for patients with CSU with an autoimmune background and bad control of the disease is omalizumab, an anti-IgE monoclonal antibody. The understanding of the mechanism of action of this biological drug in CSU along with the identification of potential biomarkers of clinical response can be helpful in the personalized management of the disease. The purpose of this study was to analyze the effect of omalizumab on peripheral blood lymphocyte subpopulations in patients with CSU in order to identify potential biomarkers of treatment response. We analyzed 71 patients with CSU [33 under omalizumab and 38 under non-immunomodulatory drugs (treated with antihistamines; NID)] and 50 healthy controls. An exhaustive immunophenotyping of whole blood T-cell subpopulations, including naïve, central memory, effector memory, effector cells, Th1, Th2, and Th17 was performed by multiparametric flow cytometry. Moreover, in CSU patients, we analyzed markers of inflammation (ESR, DD, CRP), atopy (prick test, IgE quantification), and autoimmunity (anti-thyroid antibodies and indirect basophil activation test).To evaluate the clinical activity, the Urticaria Activity Score 7 (UAS 7) test was used. In patients with CSU under treatment with omalizumab, there was a significant decrease in the percentage of naïve and an increase in the percentage of central memory CD4 T cells as well as a decrease in the percentage of naïve and increase in the percentage of effector CD8 T-cell subsets. Moreover, patients under treatment with omalizumab had higher percentages of Th1 and Th2 cells than patients under treatment with NID. The immune monitoring of T-cell subpopulations in patients with CSU starting omalizumab, may be a useful strategy to analyze treatment response in the clinical practice.

Identification and characterization of circulating and adipose tissue infiltrated CD20+T cells from subjects with obesity that undergo bariatric surgery

T cells play critical roles in adipose tissue (AT) inflammation. The role of CD20+T cell in AT dysfunction and their contributing to insulin resistance (IR) and type 2 diabetes progression, is not known. The aim was to characterize CD20+T cells in omental (OAT), subcutaneous (SAT) and peripheral blood (PB) from subjects with obesity (OB, n = 42), by flow cytometry. Eight subjects were evaluated before (T1) and 12 months post (T2) bariatric/metabolic surgery (BMS). PB from subjects without obesity (nOB, n = 12) was also collected. Higher percentage of CD20+T cells was observed in OAT, compared to PB or SAT, in OB-T1. CD20 expression by PB CD4+T cells was inversely correlated with adiposity markers, while follicular-like CD20+T cells were positively correlated with impaired glucose tolerance (increased HbA1c). Notably, among OB-T1, IR establishment was marked by a lower percentage and absolute number of PB CD20+T cells, compared nOB. Obesity was associated with higher percentage of activated CD20+T cells; however, OAT-infiltrated CD20+T cells from OB-T1 with diabetes displayed the lowest activation. CD20+T cells infiltrating OAT from OB-T1 displayed a phenotype towards IFN-γ-producing Th1 and Tc1 cells. After BMS, the percentage of PB CD4+CD20+T cells increased, with reduced Th1 and increased Th17 phenotype. Whereas in OAT the percentage of CD20+T cells with Th1/17 and Tc1/17 phenotypes increased. Interestingly, OAT from OB pre/post BMS maintained higher frequency of effector memory CD20+T cells. In conclusion, CD20+T cells may play a prominent role in obesity-related AT inflammation.

A predictive model for progression to clinical arthritis in at-risk individuals with arthralgia based on lymphocyte subsets and ACPA.

The presence of ACPA significantly increases the risk of developing RA. Dysregulation of lymphocyte subpopulations was previously described in RA. Our objective was to propose the predictive model for progression to clinical arthritis based on peripheral lymphocyte subsets and ACPA in individuals who are at risk of RA. Our study included 207 at-risk individuals defined by the presence of arthralgias and either additional ACPA positivity or meeting the EULAR definition for clinically suspect arthralgia. For the construction of predictive models, 153 individuals with symptom duration ≥12 months who have not yet progressed to arthritis were included. The lymphocyte subsets were evaluated using flow cytometry and anti-CCP using ELISA. Out of all individuals with arthralgia, 41 progressed to arthritis. A logistic regression model with baseline peripheral blood lymphocyte subpopulations and ACPA as predictors was constructed. The resulting predictive model showed that high anti-CCP IgG, higher percentage of CD4+ T cells, and lower percentage of T and NK cells increased the probability of arthritis development. Moreover, the proposed classification decision tree showed that individuals having both high anti-CCP IgG and low NK cells have the highest risk of developing arthritis. We propose a predictive model based on baseline levels of lymphocyte subpopulations and ACPA to identify individuals with arthralgia with the highest risk of progression to clinical arthritis. The final model includes T cells and NK cells, which are involved in the pathogenesis of RA. This preliminary model requires further validation in larger at-risk cohorts.

Association between peripheral blood immunological status and intrathecal inflammatory markers differentiate multiple sclerosis clinical phenotypes.

The difference in the clinical course, response to therapy, and distribution of CNS inflammation in primary-progressive (PPMS) and relapsing-remitting multiple sclerosis (RRMS) suggests differences in the underlying immunological characteristics of the disease. We aimed to investigate differences in immunological profiles in relation to intrathecal inflammation in different MS forms. The peripheral blood (PB) proportions of CD4 + and CD8 + T-cells and CD19 + B-cells were retrospectively compared with the markers of intrathecal immunoglobulin G (IgG) synthesis at diagnosis: IgG index, percentage of intrathecal IgG synthesis (IF IgG), the number of oligoclonal bands (OCB), depending on the blood-brain barrier (BBB) function, and antibody specific index to neurotrophic viruses (MRZH reaction). Thirty-six controls, 71 RRMS and 25 PPMS were enrolled. PPMS had higher percentage of CD4 + T-cells compared to RRMS (P = 0.043) and controls (P = 0.003). The percentage of CD8 + T-cells and CD19 + B-cells, and respective absolute cell counts did not differ according to the MS phenotype. In RRMS with the dysfunctional BBB, the IgG index (r = 0.642, P = 0.012) correlated significantly with the CD19 + B-cells while the CD4 + T-cells inversely correlated with IF IgG (r=-0.574, P = 0.039). Interestingly, in PPMS the number of OCB was positively associated with CD4+ (r = 0.603, P = 0.015) and negatively associated with CD8 + T-cells (r=-0.554, P = 0.033), while IF IgG negatively correlated with CD8 + T-cells (r=-0.689, P = 0.003), but only in the preserved BBB function. The PB CD4 + T-cells and B-cells were associated with the intrathecal inflammation in RRMS with BBB dysfunction while CD8 + T-cells were involved in PPMS with CNS-compartmentalized inflammation.

Decrease of Tregs cells and increase of exhausted Treg cells as the predictors of COVID19 severity

BackgroundT cells and regulatory T cells (Tregs) play a critical role in viral infectious immunity. Exhaustion of T cells during infection and decreased Tregs both contribute to the exacerbation of the disease. In the present study, we assessed T cells and regulatory T cells of COVID-19 patients and a control group according to the expression of the PD-1 molecule. MethodsForty-two COVID-19 patients and 40 controls were enrolled in the study. In COVID-19 patients, blood samples were collected on the first day of their hospitalization. Regulatory T cells (CD4+, CD25+, FOXP3+), CD4+PD-1+, and PD-1+ regulatory T cells were assessed by flow cytometry. ResultsThe percentage of CD4+PD-1 + T cells in COVID-19 patients was significantly higher compared to the control group (P < 0.0001). The percentage of PD-1+ regulatory T cells was significantly increased in the patient group compared to the control group (P < 0.0001). However, the Treg percentage was significantly decreased in the patient group compared to the control group (P < 0.0001). The frequency of CD4+PD-1 + T cells, Tregs, and PD-1+ Tregs had acceptable sensitivity and specificity for assisting in the diagnosis of severe/critical COVID-19. The declined Tregs and enhanced CD4+CD25+, CD4+PD-1+, and PD-1 + T cells were associated with disease severity. ConclusionThe decrease in Tregs and the increase in exhaustion of these cells and T cells play an important role in COVID-19 pathogenesis. These immune parameters could be used as meaningful indicators for assisting in the diagnosis of severe/critical COVID-19.

Use of parvovirus B19-like particles in self-illuminated photodynamic therapy for solid tumors

Bioluminescence resonance energy transfer photodynamic therapy, which uses light generated by bioluminescent proteins to activate photosensitizers and produce reactive oxygen species without the need for external irradiation, has shown promising results in cancer models. However, the characterization of delivery systems that can incorporate the components of this therapy for preferential delivery to the tumor remains necessary. In this work, we have characterized parvovirus B19-like particles (B19V-VLPs) as a platform for a photosensitizer and a bioluminescent protein. By chemical and biorthogonal conjugation, we conjugated rose Bengal photosensitizer and firefly luciferase to B19V-VLPs and a protein for added specificity. The results showed that B19V-VLPs can withstand decoration with all three components without affecting its structure or stability. The conjugated luciferase showed activity and was able to activate rose Bengal to produce singlet oxygen without the need for external light. The photodynamic reaction generated by the functionalized VLPs-B19 can decrease the viability of tumor cells in vitro and affect tumor growth and metastasis in the 4 T1 model. Treatment with functionalized VLPs-B19 also increased the percentage of CD4 and CD8 cell populations in the spleen and in inguinal lymph nodes compared to vehicle-treated mice. Our results support B19V-VLPs as a delivery platform for bioluminescent photodynamic therapy components to solid tumors.

Inclusion of Lacticaseibacillus paracasei NSMJ15 in broiler diets induces changes in jejunal immune cell population and cecal microbiota.

The objective was to investigate growth performance, antioxidant enzyme activity, intestinal morphology, immune cell distribution, short chain fatty acid (SCFA) profile, and microbiota in broiler chickens fed a diet containing Lacticaseibacillus paracasei NSMJ15. A total of 120 1-day-old Ross 308 male broilers were allocated to 2 dietary treatments in a randomized complete block design. A control group was fed a corn-soybean meal control diet, and an NSMJ15-supplemented group was fed a control diet supplemented with 1 g/kg L. paracasei NSMJ15 at the expense of cornstarch. Each dietary treatment had 6 replicates with 10 birds per cage. Growth performance was recorded on day 9. On day 10, one bird representing median body weight was selected to collect serum for antioxidant enzyme activity, jejunal tissue for immune cell isolation and morphometric analysis, and cecal digesta for 16S rRNA gene sequencing and SCFA analysis. Supplementation of L. paracasei NSMJ15 did not affect growth performance, serum antioxidant enzyme activity, and jejunal histomorphology compared to the control group. In the NSMJ15-supplemented group, the population of CD3+CD4+CD8- T cells increased (p = 0.010), while the population of CD3+CD8+TCRγδ+ T cells decreased (p = 0.022) compared to the control group. The L. paracasei NSMJ15 supplementation decreased (p = 0.022) acetate concentration in the cecal digesta compared to the control group. The 16S rRNA gene sequencing analysis showed that NSMJ15-supplemented group differentially expressed (p<0.05) 10 more amplicon sequence variants compared to control group without affecting alpha and beta diversity indices of the cecal microbiota. Genera Mediterraneibacter and Negativibacillus were positively (p<0.05) correlated with CD4+ T cells, while genera Gemmiger, Coprococcus, Sellimonas, Massilimicrobiota, and Blautia were negatively (p<0.05) correlated with SCFA concentration. The results of the present study suggest dietary L. paracasei NSMJ15 supplementation may increase percentage of CD4+ T cells and decrease acetate concentration in broiler chickens by increasing the differential expression of specific microbial genera.

Identification of pediatric activated T-cell hepatitis using clinical immune studies

Background and aimsThe majority of indeterminate pediatric acute liver failure (PALF) cases are secondary to immune dysregulation, labeled activated T-cell hepatitis (TCHep). We aimed to describe a cohort of children with acute severe hepatitis and PALF and define how clinical immune labs may help identify the TCHep group. MethodsRetrospective review of children with acute hepatitis and PALF between March 2020 and August 2022. Patients were classified as known diagnosis, indeterminate hepatitis (IND-Hep), or TCHep (defined by liver biopsy with predominant CD8 T-cell inflammation or development of aplastic anemia). Results124 patients were identified: 83 with known diagnoses, 16 with TCHep, and 25 with IND-Hep. Patients with TCHep had significantly increased median total bilirubin levels (7.5 mg/dL (IQR 6.8–8.9) vs 1.5 mg/dL (IQR 1.0–3.6), p < 0.0001), soluble interleukin-2 receptor levels (4512 IU/mL (IQR 4073–5771) vs 2997 IU/mL (IQR 1957–3237), p = 0.02), and percent of CD8+ T-cells expressing perforin (14.5 % (IQR 8.0–20.0) vs 1.0 % (IQR 0.8–1.0), p = 0.004) and granzyme (37.5 % (IQR 15.8–54.8) vs 4.0 % (IQR 2.5–5.5), p = 0.004) compared to IND-Hep patients. Clinical flow cytometry showed that TCHep patients had significantly increased percent CD8+ T cells (29.0 % (IQR 24.5–33.5) vs 23.6 % (IQR 19.8–25.8), p = 0.04) and HLA-DR+ (16.0 % (IQR 14.5–24.5) vs 2.7 (1.8–5.3), p < 0.001) compared to IND-Hep patients indicative of increase in CD8+ T cells that are activated. ConclusionsPeripheral blood clinical immune studies demonstrate increased markers of CD8 T-cell activation, proliferation, and cytotoxic function for TCHep patients. These readily available immune function labs can be used to help distinguish patients with TCHep from those with other causes. This provides a non-invasive tool for early detection of potential TCHep before progression to liver failure.

Association between Circulating T Cells and the Gut Microbiome in Healthy Individuals: Findings from a Pilot Study.

Though the microbiome's impact on immune system homeostasis is well documented, the effect of circulating T cells on the gut microbiome remains unexamined. We analyzed data from 50 healthy volunteers in a pilot trial of aspirin, using immunophenotyping and 16S rRNA sequencing to evaluate the effect of baseline T cells on microbiome changes over 6 weeks. We employed an unsupervised sparse canonical correlation analysis (sCCA) and used multivariable linear regression models to evaluate the association between selected T cell subsets and selected bacterial genera after adjusting for covariates. In the cross-sectional analysis, percentages of naïve CD4+ T cells were positively associated with a relative abundance of Intestinimonas, and the percentage of activated CD8+ T cells was inversely associated with Cellulosibacter. In the longitudinal analysis, the baseline percentages of naïve CD4+ T cells and activated CD4+ T cells were inversely associated with a 6-week change in the relative abundance of Clostridium_XlVb and Anaerovorax, respectively. The baseline percentage of terminal effector CD4+ T cells was positively associated with the change in Flavonifractor. Notably, the microbiome taxa associated with T cell subsets exclusively belonged to the Bacillota phylum. These findings can guide future experimental studies focusing on the role of T cells in impacting gut microbiome homeostasis.

Absolute CD4 count and percentage values among Libyan patients with HIV by single-platform flow cytometry.

Single-platform flow cytometry technology together with CD45-gating is becoming the method of choice for absolute CD4 T cell enumeration. Immunological assessment of HIV patients by monitoring CD4 can provide valuable information on antiviral treatment response and disease progression. A total of 97 HIV-positive individuals were recruited from 2 hospitals in Tripoli, Libya, and 14 healthy blood donors. The HIV-infected individuals were classified by CD4+ count into HIV-positive (>200 cells/µL) or AIDS (≤200 cells/µL) groups. CD4+ and CD8+ cell counts were determined and compared among the groups and with similar published data. The mean ± SD CD4+ cell counts were 1106 ± 442.8 cells/µL in healthy individuals, 460 ± 219.7 cells/µL in the HIV-positive group, and 78 ± 64.3 cells/µL in the AIDS group. The mean ± SD CD4+/CD8+ ratio was 1.6 ± 0.58, 0.4 ± 0.22, and 0.1 ± 0.1, respectively. CD4+ counts in Libyan healthy adults might be higher than those reported in several studies in other regions, whereas CD4+ counts in Libyan AIDS patients seem lower. Reference values for T lymphocyte counts in Libyan healthy individuals should be investigated more extensively, and the reasons why Libyan AIDS patients seem to have such lower CD4+ counts should be examined.

Rare subgroup of T Lymphocytes in Oral and Maxillofacial Cancer Patients

Abstract Malignant tumors of head and neck region represent a heterogenous group of pathologies. Immune phenotype of each tumor might represent valuable information for a putative personalized treatment in the future. We decided to revisit very interesting previous data from a study we conducted in 2003 where we encountered a rare population of CD4+CD8+ double-positive T cells in lymph nodes surgically removed in patients with head and neck malignancies. The study included 27 patients (22 males and 5 females) who underwent surgical procedures that associated lymphadenectomy during March-July 2003 in the Maxillo-Facial Surgery Clinic of „Dan Theodorescu” University Dental Hospital in Bucharest. We encountered a high percentage of CD4+CD8+ T lymphocytes in peripheral blood of non-Hodgkin Lymphoma and poorly differentiated carcinoma, as well as in lymph nodes of both Hodgkin and non-Hodgkin lymphomas. We would like to emphasize the importance of immune phenotyping, when possible, to discriminate between tumor subtypes, evidence that establishing a true identity of the cells involved might be useful for future studies and/or personalized therapeutic strategies.

Changing Characteristics of Treg/Th17 Cells in the Nasal Mucosa of a Mouse Model of Allergic Rhinitis and the Effect of Intervention with Anti-IL-17 Antibody.

The incidence rate of allergic rhinitis (AR) is on the rise, which seriously affects the quality of life, work efficiency, mental state of patients, and sleeping in AR sleep. This experiment aimed to investigate the changes in Treg/Th17 cells in the nasal mucosa of an AR mouse model and the intervention effect of an Anti-IL-17 antibody. A mouse model of AR was induced by intraperitoneal ovalbumin (OVA) injection for sensitization and stimulation with nasal drops. The times of rubbing, sneezing, and symptomatology scores were counted and analyzed. Pathological damage to the nasal mucosa was observed by Hematoxylin-Eosin (HE) staining. Peripheral blood CD4+CD25+CD127- Treg cell levels and the content of Th17 cells were measured by flow cytometry (FCM). ELISA kits were used to detect the levels of relevant cytokines (IL-10 and TGF-β1) secreted by Treg cells. The intervention effect of Anti-IL-17 antibody was observed by giving Anti-IL-17 antibody treatment. The times of rubbing, sneezing, and symptomatology scores were significantly higher in mice in the OVA group than in the Control group (p < 0.001). The percentage of CD4+CD25+CD127- Treg cells in CD4+CD25+ T cells (p < 0.05) and the levels of IL-10 (p < 0.001) and TGF-β1 (p < 0.001) were significantly decreased. After OVA induction, the continuity of the nasal mucosa of mice was interrupted, the percentage of Th17 cells, IL-17, and IL-4 levels were increased, and IFN-γ levels were significantly reduced (p < 0.001). And protein expression of RORγt was significantly upregulated (p < 0.001). In addition, all of these results were reversed by Anti-IL-17 antibody treatment, significantly improving AR-related symptoms (p < 0.05). Anti-IL-17 antibodies may regulate the body's immune response by promoting CD4+CD25+CD127- Treg cell differentiation, thereby ameliorating the symptoms associated with AR.

Predicting mortality within 1 year of ART initiation in children and adolescents living with HIV in sub-Saharan Africa: a retrospective observational cohort study

Differentiated service delivery (DSD) for children and adolescents living with HIV can improve targeted resource use. We derived a mortality prediction score to guide clinical decision making for children and adolescents living with HIV. Data for this retrospective observational cohort study were evaluated for all children and adolescents living with HIV and initiating antiretroviral therapy (ART); aged 0-19 years; and enrolled at Baylor clinics in Eswatini, Malawi, Lesotho, Tanzania, and Uganda between 2005 and 2020. Data for clinical prediction, including anthropometric values, physical examination, ART, WHO stage, and laboratory tests were captured at ART initiation. Backward stepwise variable selection and logistic regression were performed to develop predictive models for mortality within 1 year of ART initiation. Probabilities of mortality were generated, compared with true outcomes, internally validated, and evaluated against WHO advanced HIV criteria. The study population included 16 958 children and adolescents living with HIV and initiated on ART between May 18, 2005, and Dec 18, 2020. Predictive variables for the most accurate model included: age, CD4 percentage, white blood cell count, haemoglobin concentration, platelet count, and BMI Z score as continuous variables, and WHO clinical stage and oedema, abnormal muscle tone and respiratory distress on examination as categorical variables. The area under the curve (AUC) of the predictive model was 0·851 (95% CI 0·839-0·863) in the training set and 0·822 (0·800-0·845) in the test set, compared with 0·606 (0·595-0·617) for the WHO advanced HIV criteria (p<0·0001). This study evaluated a large, multinational population to derive a mortality prediction tool for children and adolescents living with HIV. The model more accurately predicted clinical outcomes than the WHO advanced HIV criteria and has the potential to improve DSD for children and adolescents living with HIV in high-burden settings. National Institute of Health Fogarty International Center.

Comparison of tumor immune microenvironments (TIMEs) between primary and metastatic sites (Mets) in triple-negative breast cancer (TNBC).

1097 Background: The TIME is critical in determining response to immune checkpoint inhibitors (ICIs), but TIME differences across primary and mets in TNBC are not well understood. Since ICIs are standard of care for patients with TNBC, we studied the TIMEs of primary TNBC and mets to investigate how immune composition may affect ICI efficacy. Secondary analysis stratified cohorts by race and disease sites to compare TIMEs in Black or African American (B/AA) with White patients, since TNBC has a high prevalence of B/AA women who are underrepresented in studies and have a worse prognosis. Methods: We retrospectively analyzed de-identified next-generation sequencing data from TNBC patients (n=1,044) in the Tempus Database. Tumors from primary breast (PB, n=553), liver (n=153), lymph node (LN, n=174), lung (n=111), and bone (n=53) sites were sequenced with the Tempus xT DNA (648-gene panel) and xR RNA assays. Histologies included invasive ductal carcinoma (82%), lobular carcinoma (2.2%), mixed ductal and lobular carcinoma (1.7%), and other (14%). Demographics, TMB, MSI, PD-L1, and proportions of B, T (CD4+, CD8+), NK cells, and macrophages were compared across sites. LN were used as a positive control. Chi-squared/Fisher’s exact or Kruskal-Wallis tests were used to assess statistical significance (two-sided, evaluated at the 0.05 alpha level). Results: The cohort (median age=57 years, IQR 46-66) comprised a diverse population (65% White, 23% B/AA, 2.8% Asian, and 9.3% other). Liver exhibited a lower percentage of B cells and higher percentage of macrophages compared to PB (p&lt;0.0001 for both), lung (p&lt;0.0001 for both), and bone (B cells p&lt;0.0001, macrophages p&lt;0.05). Liver also exhibited lower percentages of CD8+ T cells compared to PB (p&lt;0.05) and CD4+ and NK cells compared to lung (CD4+ cells p&lt;0.01, NK cells p&lt;0.0001). Bone had lower percentages of CD8+ cells compared to PB (p&lt;0.0001), lung (p&lt;0.001), and liver (p&lt;0.01). Compared to lung, bone had a lower percentage of CD4+ cells (p&lt;0.0001) and higher percentage of macrophages (p&lt;0.01). PD-L1 positivity, TMB, and MSI exhibited no differences across sites. Secondary analysis compared the TIME between B/AA and White cohorts in PB (B/AA n=85 vs White n=204: %B cells, 15 vs 11, p=0.017; %CD8 cells, 4.9 vs 7.3, p=0.025), liver (B/AA n=15 vs White n=48: %B cells, 8 vs 4, p=0.2; %CD4, 23 vs 14, p=0.2; %CD8, 7.2 vs 4.6, p=0.07), and lung (B/AA n=9 vs White n=33: %B cells, 22 vs 11, p=0.021; %CD8, 7.9 vs 5.2, p=0.5). Conclusions: Compared to PB and lung, liver and bone had immune cell infiltrates indicating a less immunogenic TIME in the overall TNBC population. Although limited by sample size, this is one of the first studies to assess the TIME across race and sites of disease. These findings are hypothesis generating and provide further rationale to better understand how the TIME across disease sites and race may alter the efficacy of ICIs in TNBC.

Novel electrical therapy to improve sleep disturbance in patients with autoimmune rheumatic diseases.

Sleep disturbance is common in autoimmune rheumatism diseases (ARD) and it plays an important role in activating disease and affects the quality of life. This study aims to evaluate the efficacy and acceptability of the novel electrical therapy on sleep disturbance in ARD patients and its effect on immunologic factors. A total of 51 ARD patients (26 treatment group and 25 control group) with sleep disturbance were enrolled in this study. Sleep parameters and immunological indicators (serum level of 12 cytokines and immune function) were collected. The novel electrical therapy was prescribed for 15-30 min 3-6 times a day. The Pittsburg Sleep Index (PSQI) was assessed before and after 3 months' treatment by Mi Energy equipment. Immune function and serum levels of cytokines of all participants at baseline and after treatment were tested with flow cytometry and flow immunofluorescence, respectively. Correlation analysis was used to analyze the relationship between sleep disturbance and immunologic factors. Multiple linear regression analysis was employed to investigate the risk of sleep disturbance in ARD. The global score of PSQI (Baseline: 12.81 ± 4.07, After novel electrical therapy: 4.88 ± 2.76) was effectively improved after 3 months of adjuvant therapy by electrical therapy. We also found that serum levels of IL-8 and IL-1β statistically significantly decreased after novel electrical therapy. This adjuvant therapy can also significantly decrease the percentage of CD4 + CD8 + T cell, effector memory CD8 + T cell, Memory CD8 + T cell, Th17 cell, and plasma cell and significantly can increase the percentage of naïve CD8 + T cell, Th2 cell, and Tfh2 cell. Nevertheless, all serum level of 12 cytokines and the percentage of immune cells did not correlate with the PSQI global score except the Tc17 cell. Furthermore, age is an independent risk factor influencing PSQI scores (OR = 1.15, p < 0.05) in patients with autoimmune diseases through multiple linear regression analysis. Novel electrical therapy can effectively improve sleep disturbance in patients with ARD. It can also change the serum level of some cytokines (IL-8 and IL-1β) and percentage of immune cells (CD4 + CD8 + T cell, effector memory CD8 + T cell, Memory CD8 + T cell, Th17 cell, naïve CD8 + T cell, Th2 cell, Tfh2 cell, and plasma cell).

Comparison of T cell maturation profiles in the 1st and 5th wave of COVID-19 in the Polish population.

The coronavirus pandemic has become the most critical global health threat of this century and the greatest challenge to the human population. The search for simple and quick diagnostic methods enabling the identification of patients infected with the SARS-CoV-2 virus may be a valuable method to track infection. The aim of the study was the clinical and immunological characterization of patients by assessing the degrees of maturity of T lymphocytes from the 1st and 5th waves of coronavirus disease 2019 (COVID-19) in comparison to a healthy control group (HC). We determined leukocyte and T lymphocyte subpopulations (recent thymic emigrant (RTE), naïve, effector, central memory and effector memory) in patients from the 1st COVID-19 wave (n = 23), the 5th COVID-19 wave (n = 38) and HC (n=20) using a panel of monoclonal antibodies using multiparameter flow cytometry. We observed a lower median proportion of lymphocytes and NK cells, and elevated percentage and number of neutrophils in patients from the 5th wave compared to the 1st. We found a reduced percentage of CD4+ effector memory cells in the 1st wave group compared to the 5th wave (14.1 vs 23.2, p < 0.05), and a higher percentage of RTE and naïve CD8+ cells in the 1st wave compared to the 5th wave (p < 0.05). The effector memory CD8+ cells were highest in the 5th wave compared to both 1st wave and HC patients (respectively, 35.1 vs 18.1 vs 19.3%, p < 0.05). The 5th wave group showed significantly more differences compared to HC. Our results showed a clear increase of effector cells with a simultaneous decrease in virgin T cells in the 5th COVID-19 infection. Monitoring lymphocyte subsets during infection allows assessment of the patient's immune status and of readiness of lymphocytes to respond to the immune response, and may be necessary to improve clinical outcomes.

Electroacupuncture treatment improves postoperative ileus by inhibiting the Th1 cell-mediated inflammatory response through the vagus nerve.

Electroacupuncture (EA) has been reported to improve intestinal motility in mice with postoperative ileus (POI). Previous studies, however, have yielded heterogeneous results regarding the effect of EA on POI. Herein, a POI mouse model was constructed by intestinal manipulation. To evaluate the effect of EA treatment on colonic transit, the levels of inflammatory markers (macrophage inflammatory protein (MIP)-1α, interleukin (IL)-1β, IL-6, monocyte chemotactic protein (MCP)-1 and intercellular adhesion molecule (ICAM)-1) were detected by enzyme-linked immunosorbent assay (ELISA); immune cell infiltration was detected by immunohistochemical staining of myeloperoxidase (MPO), ectodysplasin (ED)-1 and ED-2, and the percentage of CD4+ interferon (IFN)-γ+ Th1 cells and IFN-γ secretion levels were determined. Activated Th1 cells and pentoxifylline, a cell differentiation inhibitor, were used to assess the role of Th1 cells in EA treatment of POI. Neostigmine administration and unilateral vagotomy were performed to confirm whether the effects of EA treatment on Th1 cells were mediated by the vagus nerve (VN). The results revealed that EA treatment at ST36 improved POI, as indicated by a decreased level of inflammatory-related markers and immune cell infiltration and shortened colonic transit time. The activated Th1 cells abolished the effects of EA treatment on POI. The effects of EA treatment on POI were enhanced by stimulation of the VN along with a decreased level of Th1 cells, but these effects were abolished by vagotomy along with an increased percentage of Th1 cells; this result indicates that the VN mediates the role of Th1 cells in the effects of EA treatment of POI. Our findings showed that the effects of EA treatment of POI were mainly mediated by Th1 cells through the stimulation of the VN and inhibition of the inflammatory response.

#2899 Role of the Th17.1 cells in IgA nephropathy

Abstract Background and Aims T helper (Th) cells play a crucial role in glomerulonephritis, including IgA nephropathy (IgAN). Recently identified Th17.1 cells, which are elevated in sarcoidosis, lupus, and Takayasu arteritis, express chemokine receptors CCR6 and CXCR3. These receptors are implicated in inflammatory cell infiltration and fibrosis in glomerulonephritis models. Notably, Th17.1 cells express MDR-1 (multidrug resistance protein) and the drug efflux protein p-glycoprotein, associated with corticosteroid resistance. However, the exact role of Th17.1 cells in IgAN remains unclear. Intriguingly, Th17.1 cells produce INF-γ and IL-17, cytokines implicated in IgAN pathogenesis. Method In this retrospective cross-sectional study, we examined circulating Th17.1 cells in 18 confirmed IgAN patients. Antibodies such as anti-CXCR3-FITC and anti-CCR6-PCy7 (Biolegend, U.S.A.) were used to stain Th subpopulations. We also used combinations of the following antibodies: anti-CD45-FITC, anti-CD4-PE, anti-CD8-ECD, anti-CD3-PCy5, anti-CD56-PCy5, anti-CD3-PCy7, anti-CD19-ECD, anti-CD127-FITC, anti-CD25-PCy5, and anti-CD4-PCy7 (Coulter Immunotech, France) to evaluate T, B, NK, and Treg cells. Multicolor flow cytometry was performed using a Navios cytometer, and data were analyzed using Kaluza software (Beckman Coulter, U.S.A.). Th effector subpopulations of CD4+ lymphocytes were identified based on chemokine receptor expression: Th17.1 (CXCR3+CCR6+). Th17.1 cells were quantified as a percentage of CD4+ lymphocytes. Th1, Th2, and Treg cells were also evaluated. We explored correlations between Th17.1 cell counts, estimated glomerular filtration rate (eGFR, mL/min/1.73 m²), and proteinuria (24-hour collection). Among the patients, 5 were classified as refractory (progressive eGFR decline &amp;gt; 5 mL/min/1.73 m² and 24-hour proteinuria &amp;gt; 1g despite steroid and immunosuppressant therapy). Results The study included 18 patients. The mean (95% CI) Th17.1 cell count was 20% (16-23), and the 24-hour proteinuria was 2.1g (1.4-2.7). A significant direct correlation was observed between Th17.1 cells and proteinuria (g/L, adjusted for 24 hours) (r = 0.50, P = 0.03). Refractory patients exhibited higher Th17.1 levels compared to others (25% [95% CI: 17-33] vs. 18% [95% CI: 14-21], P = 0.02) (Fig. 1). Patients with proteinuria &amp;lt; 0.5g/24 h had lower Th17.1 cell counts compared to those with higher proteinuria (16% [95% CI: 12-20] vs. 21% [95% CI: 17-25], P = 0.06. No correlation was found between eGFR, proteinuria, and Th1, Th2, Th17, or Treg cells. Conclusion These findings suggest that Th17.1 cells may contribute to the pathogenesis of IgAN and could potentially indicate the response to steroids or immunosuppressants in treatment. While the direct involvement of Th17.1 cells in kidney tissue remains unexplored, further investigation is warranted due to its therapeutic and prognostic implications.