Expression Levels Of CD28 Research Articles

Circulating Immune Landscape Profiling in Psoriasis Vulgaris and Psoriatic Arthritis by Mass Cytometry.

Psoriasis, a systemic disorder mediated by the immune system, can appear on the skin, joints, or both. Individuals with cutaneous psoriasis (PsC) have an elevated risk of developing psoriatic arthritis (PsA) during their lifetime. Despite this known association, the cellular and molecular mechanisms underlying this progression remain unclear. We performed high-dimensional, in-depth immunophenotyping of peripheral blood mononuclear cells (PBMCs) in patients with PsA and psoriasis vulgaris (PsV) by mass cytometry. Blood samples were collected before and after therapy for a longitudinal study. Then three sets of comparisons were made here: active PsA vs. active PsV, untreated PsV vs. treated PsV, and untreated PsA vs. treated PsA. Marked differences were observed in multiple lymphocyte subsets of PsA related to PsV, with expansion of CD4+ T cells, CD16- NK cells, and B cells. Notably, two critical markers, CD28 and CD127, specifically differentiated PsA from PsV. The expression levels of CD28 and CD127 on both Naïve T cells (TN) and central memory CD4+ T cells (TCM) were considerably higher in PsA than PsV. Meanwhile, after treatment, patients with PsV had higher levels of CD28hi CD127hi CD4+ TCM cells, CD28hi CD127hi CD4+ TN cells, and CD16- NK cells. In the circulation of PsA patients, the TN and CD4+ TCM are characterized with more abundant CD28 and CD127, which effectively distinguished PsA from PsV. This may indicate that individuals undergoing PsV could be stratified at high risk of developing PsA based on the circulating levels of CD28 and CD127 on specific cell subsets.

Research and Analysis of Molecules such as CD28, CD45RA, CD45RO, CD38, HLA-DR, and CD57 on T Cells in Multiple Myeloma.

For many years it has been postulated that the immune system controls the progress of multiple myeloma (MM). However, the phenotypes of T cells in MM remain to be elucidated. In this study, we compared the phenotypes of T cells, which were obtained from the peripheral blood, in MM patients with those in healthy donors (HD). The expression of CCR7, CD57, CD28, HLA-DR, CD38, CD45RA, and CD45RO were assessed on T cells from MM patients and HDs using multicolor flow cytometry (MFC). For this study, 17 newly diagnosed MM patients were selected, and 20 healthy people were selected as a control group. MFC was used to detect the markers on T cells. We detected significant increases in the expression levels of HLA-DR, CD38, and CD57on CD8+ T cells, significant decreases in the expression levels of CD28 and CD45RA on CD8+ T cells, and a decrease of CD4+ effec-tor T cells in MM patients, compared to the HD group. Our study shows that the accumulation of peripheral CD8+CD57+T cells, CD8+CD38high T cells, and CD8+HLA-DR+CD38high T cells is reflective of an ongoing antitumor T cell response and a progressive immune dysfunction in MM. During chemotherapy, the recovery of immune function can be monitored by detecting the proportion of activated molecules of T lymphocytes.

Increased expression of co-stimulatory molecules and enhancement of the IgG response in rats orally administered with a polyherbal formulation

BackgroundLink Samahan® (LS) is a standardized modern formulation of a polyherbal preparation used in the indigenous system of medicine in Sri Lanka. ObjectiveEvaluation of the immunostimulatory activity of LS and the molecular mechanisms that modulate the humoral immune response. Material and methodsImmunostimulatory activity of LS was tested in rats following oral administration on days 1-5 and 15-19 and immunization with bovine serum albumin (BSA) on day 1 and 15. Anti-BSA IgM and IgG response in rats treated with LS, water and sugar (as controls) were compared on days 0-35, using ELISA. The expression of co-stimulatory molecules on lymphocytes was assessed on days 0-8 and days 14-22 using RT-qPCR. ResultsIgM and IgG levels of LS-treated rats were increased significantly by day 7 and 21 respectively compared to controls (p < 0.05). IgG response of LS-treated group reached a higher magnitude compared to its IgM response. Gene expression of CD28 and CD40L on T cells (4.9-5.1 fold) and CD80, CD86 and CD40 on APCs (2.4-3.1 fold) were induced significantly by day 2 compared to their expression on day 0 (p < 0.05). The expression levels of CD28 and CD40L on day 2-4 and 16-18 were similar while the expression of CD80, CD86 and CD40 on day 16-18 was higher (3.7-5.1 folds) compared to their levels on day 2-4 (2.4-3.2). ConclusionsThese findings support an adjuvant effect of LS contributing to its immunostimulatory activity and increased expression of co-stimulatory molecules that contribute to boosting immune response.

Abstract A33: Impact of tumor-derived exosomes on CD28 expression in T cells

Abstract Osteosarcoma is a rare disease with a disproportionate impact, as it is seen mostly in children and adolescents. Despite aggressive treatment, more than 50% of patients succumb to their disease within 10 years of diagnosis, making safe and effective treatments a huge unmet need. Immunotherapy, including immune checkpoint blockade, is drastically changing cancer treatment by increasing the antitumor immune response. Mechanisms of resistance to immune checkpoint blockade are incompletely understood; therefore, there is currently no available method to specifically identify patients who will respond favorably to immunotherapy. Exosomes are extracellular vesicles that can carry proteins, lipids, mRNA, microRNA, and DNA and play a role in extracellular communication. Exosomes can be found in circulation, making them an attractive tool for diagnostic testing. One potential mechanism of immunotherapy resistance could be destabilization of the CD28 coreceptor in T cells by microRNAs secreted in tumor-derived exosomes. Utilizing next-generation RNA-sequencing, we previously identified 2 microRNAs in dogs and 2 microRNAs in humans that were inversely correlated with CD28 expression in 44 human and 39 canine osteosarcoma tissues. Current work entails isolating exosomes from canine and human osteosarcoma tumor cell lines. Using a species mismatched approach, we can identify canine microRNAs in human T cells that were treated with canine exosomes, and vice versa, to verify delivery of microRNAs to T cells via tumor-derived exosomes. We are also using the CRISPR-Cas system to genetically engineer a human osteosarcoma cell line (HOS) that expresses the exosome marker CD63 linked to GFP so we can track exosome uptake by T cells by flow cytometry and determine the effect on CD28 expression levels. Identifying mechanisms of resistance will lead to improved patient selection and identification of therapeutic targets to combat this resistance. Citation Format: Ashley J. Schulte, Lauren J. Mills, Kelly M. Makielski, Taylor A. DePauw, Jaime F. Modiano. Impact of tumor-derived exosomes on CD28 expression in T cells [abstract]. In: Proceedings of the AACR Special Conference on Advances in Liquid Biopsies; Jan 13-16, 2020; Miami, FL. Philadelphia (PA): AACR; Clin Cancer Res 2020;26(11_Suppl):Abstract nr A33.

Abstract P-247: PRELIMINARY STUDY OF T CELLS FUNCTIONS AFTER RESTORATION OF SPONTANEOUS CIRCULATION FROM CARDIAC ARREST IN SD RATS

Aims & Objectives: Establishing a rat model of ROSC from cardiac arrest induced by asphyxia to observe the numbers and differentiation of peripheral blood T cells and its subsets and the expression of surface molecules Methods Rats were induced cardiac arrest by asphyxia or only sham-operation. The proportion of T cells and CD4+, CD8+ subsets and the expression of surface molecules CTLA-4, PD-1, CD28 on T cells were tested by flow cytometry. TNF-α, IL-6, IL-10, IL-4, IFN-γ, IL-17A in serum were measured by ELISA. Results Both IL-6 and IL-10 showed highest levels within 3 hours after CA and decreased during the next 3 days. CD3+ lymphocyte in CA groups was significantly lower than sham-operated groups at 24 and 72 h post-ROSC. The expression levels of CD28, PD-1, and CTLA-4 on T cells were significantly increased in CA groups at 24 h post-ROSC. Moreover, the levels of IFN-γ were markedly decreased, while IL-4 was significantly increased in the CA groups compared with the sham operated groups at 24 and 72 h post-ROSC Conclusions After cardiac arrest, a systemic inflammatory response syndrome develops, T cell functions partially changed, which manifested in the decreased percentage of T cells, the up regulated co-inhibitory molecules, and the shift from Th1 to Th2

The Effect of a Competitive Futsal Match on T Lymphocyte Surface Receptor Signaling and Functions.

In this study, the lymphocyte activation status (surface expression of CD95, CD28, CD25, and CTLA-4), lymphocyte number, lymphocyte subpopulations, lymphocyte necrosis and/or apoptosis, and lymphocyte release of reactive oxygen species (ROS) were investigated in blood samples from 16 futsal athletes before and immediately following a competitive match. Lymphocytes were isolated from the blood samples, and the cellular parameters were assessed by flow cytometry. The futsal match induced lymphocytosis and lymphocyte apoptosis, as indicated by phosphatidylserine externalization, CD95 expression, and DNA fragmentation. Additionally, the competitive match induced the necrotic death of lymphocytes. No differences in the percentage of CD4+ and CD8+ T cells or in the T-helper/suppressor profile between before and immediately after the match were observed. Additionally, after the futsal match, the CD95 and CD28 expression levels were decreased, and the lymphocytes spontaneously released higher levels of ROS. Regardless of the origin, the situation-specific knowledge of lymphocyte behavior obtained herein may facilitate the design of strategies to control the processes that result in infection and tissue injury and that subsequently decrease athletic performance.

The significance of the co-stimulatory molecules expression atternation in the peripharal T lymphocytes of patients with chronic hepatitis B infection

Objective To investigate the significance of changes in expression of co-stimulatory molecules on T lymphocytes in patients with chronic hepatitis B (CHB) infection. Methods In a case-control study, a total of 82 CHB cases including 50 male cases and 32 female cases (the mean age was 42.32±3.74) were enrolled in the First Affiliated Hospital to Soochow University from October 2012 to November 2013, together with 30 cases health control (15 male cases and 15 female cases, and the mean age was 42.32±3.74). Patients were divided into three groups： 40 cases of non-treated, 30 cases of effective-treated and 12 cases of ineffective-treated with anti-viral drugs and immune-related therapy. The expression levels of CD28, CTLA-4, PD-1, Tim-3 on T cells subset from peripheral blood were determined by Flow Cytometry. Serum load of HBV DNA was detected by Real-time PCR and the serology markers such as HBeAg and ALT were detected by conventional methods The Kruskal-Wallis test was used to analysis groups comparison. Independent samples t-test and Mann-Whitney U test were used to two sample comparison. Spearman′s rank correlation test was used to analyze the correlation. Results The expression levels of CD28, CTLA-4 and PD-1 on CD4 in health control group was the highest compared to ineffective-treated group ［404.65 (331.65-536.09) vs 277.15 (249.90-344.25) (H=29.81, P<0.001); 32.89 (29.69-39.69) vs 19.26 (11.90-20.56) (H=43.13, P<0.001), respectively］. The expression level of PD-1 on CD8 in pre-treated group was the highest ,while expression level in health control group was the lowest ［15.47 (12.50-17.78) vs 3.05 (1.41-3.97) (H=56.60, P<0.001)］. Similarly, the expression level of Tim-3 on CD4 in ineffective-treated group was the highest, while Tim expression on CD4 in health control group was the lowest ［199.62 (55.61-239.45) vs 70.62 (53.88-112.32) (H=41.03, P<0.001)］. The expression level of Tim-3 on CD8 in pre-treated group was the highest, while it was the lowest in health control group ［82.50 (78.69-84.58) vs 3.07(1.56-6.87) (H=74.84, P<0.001)］. Compared to the group with low viral load, the expression level of PD-1 on CD8 both in the pre-treated group with high viral load and the post-treated group was significantly increased ［17.87 (13.38-20.94) (U=25.00, P<0.001); 16.95 (5.39-18.27) (U=63.50, P<0.001), respectively］. Importantly, in the post-treated group, the PD-1 expression on CD4 T (r=0.689, P<0.001) and on CD8 T (r=0.751, P<0.001) was also positively correlated with ALT. Conclusions The abnormal expression of these co-stimulatory molecules may be present in the antiviral treatment process of CHB. It may provide new clues for the reasonable treatment of CHB.（Chin J Lab Med，2014,37：105-109） Key words: Hepatitis B, chronic; T-lymphocytes; Antigens, CD28; CTLA-4 antige; Membrane proteins; Flow cytometry

Expression levels of CD28, CTLA-4, PD-1 and Tim-3 as novel indicators of T-cell immune function in patients with chronic hepatitis B virus infection

Chronic hepatitis B (CHB) is one of the most common types of infectious diseases worldwide. The interaction between hepatitis B virus (HBV) and the host immune response is vital for the clinical outcome of HBV infection. Costimulatory signals are key factors for the host immune response and play a critical role in innate immunity, particularly antiviral immunity. The aim of the present study was to investigate the correlation between the expression levels of costimulatory molecules and the different states of CHB infection, including the expression levels prior to and following treatment with antiviral agents. The expression levels of CD28, CTLA-4, PD-1, Tim-3 and T-cell subsets were determined by flow cytometry. The load of HBV DNA in the serum was detected by quantitative polymerase chain reaction and the serology markers, including HBeAg and alanine aminotransferase (ALT), were measured by conventional methods. Compared to the healthy control group, the expression levels of CD28 and CTLA-4 on CD4 T cells prior to and following treatment with antiviral agents (the pre- and post-treatment groups, respectively) were significantly decreased, while the expression levels of Tim-3 on CD4 and CD8 T cells were significantly increased. In addition, the expression levels of PD-1 on CD4 and CD8 T cells in the pre-treatment group were significantly increased compared to those in the post-treatment and healthy control groups. Moreover, the multivariate analysis revealed that the levels of ALT and HBV-DNA in the serum were significantly positively correlated with PD-1 expression levels. In conclusion, the expression levels of these costimulatory molecules reflect the immune dysfunction of T cells in patients with CHB and, combined with T-cell subset analysis may be used as a novel evaluation system of immune function in patients with HBV infection.

The “time-window” effect of early allergen exposure on a rat asthma model

Background The hygiene hypothesis has been proposed to explain the pathogenesis of asthma. Allergen exposure was shown to inhibit asthma in an animal model. But the optimal timing of allergen exposure remains unclear. This study aims to explore the time effcct of allergen exposure and the possible mechanisms. Methods Neonate Wistar rats were randomly divided into asthma group, control group and day 1, day 3, day 7, and day 14 groups. The day 1, day 3, day 7 and day 14 groups were injected with ovalbumin (OVA) subcutaneously on days 1, 3, 7 and 14 after birth, respectively. Six weeks later, all groups, except the control group, were sensitized and stimulated with OVA to make the asthma model. We observed the pulmonary pathologic changes, detected the regulatory T cells, and CD28 expression level in thymus and spleen by flow cytometry. Results The asthmatic inflammation in the day 1, day 3 and day 7 groups, but not the day 14 group, was alleviated. The asthma group and day 14 group had lower proportions of regulatory T cells in the thymus compared with the control group, day 1, day 3, and day 7 groups. There was no significant difference in the CD28 expression levels on the regulatory and conventional T cells among groups. But the control group and the day 1, day 3, and day 7 groups had relatively higher proportions of CD28 positive regulatory T cells in the thymus than the day 14 group and the asthma group. Conclusions There is a “time-window” for early allergen exposure. The impairment of regulatory T cells may promote the development of asthma. Allergen exposure in the “time-window” can make the thymus produce normal quantity of regulatory cells. The CD28 signal on regulatory T cells may participate in the production of regulatory T cells.

High Levels of Granzyme B Expression in Invasive Cervical Carcinoma Correlates to Poor Response to Treatment

The present study assessed, in cervical carcinoma, expression levels of seven immune response genes and sought correlation to response to treatment. The expression levels of CD28, CTLA4, ICOS, ICOSL, CD80 and CD86 and granzyme B genes were assessed by real-time RT-PCR in pre-treatment tumor fragments. During the six-month follow-up after treatment, 8 patients presented tumor and 10 survived free of tumor. The only gene whose expression levels were higher in patients with poor outcome (p = 0.03) was granzyme B. Further evaluation, in adequately powered prospective studies is warranted to confirm the data and to translate this observation to the clinical setting.