Cell Lymphocyte Subsets Research Articles

Effects of lipopolysaccharide on T lymphocyte cell subsets and cytokine secretion in mesenteric lymph nodes of mice: Histological and molecular study

Lipololysaccharides (LPS) can disrupt the gut barrier. How dose LPS affects the immune performance of mesenteric lymph nodes? The results showed the hematological parameters significantly changed after LPS treatment. The length of intestinal villus was shortened and the depth of crypts was deepened, especially on the ileum. After LPS treatment 6 h, 12 h, the number of CD3+ T cells and CD4/CD8 in the mesenteric lymph nodes of ileum were reduced significantly; the levels of IFN-γ, TNF-ɑ and IL-2 were significantly decreased, and the levels of IL-6 and IL-10 were significantly increased in the ileum. The content of sIgA in the ileum was significantly decreased after LPS treatment 3 h, 6 h and was increased after LPS treatment 12 h. LPS through mesenteric lymph nodes, which induces the immune function reduced and the ileum injured obviously after treatment 6 h. Furthermore, the performance of intestinal immune performance was the lowest after LPS treatment 6 h.

Issue Highlights - May 2019.

Issue Highlights - May 2019.

Detection and Analysis of T Lymphocyte Subsets and B Lymphocytes in Patients with Acute Leukemia

To investigate the changes of T lymphocyte subsets, B lymphocyte and NK cells in peripheral blood of patients with acute leukemia at different periods and their significance. The peripheral T lymphocyte subsets and B lymphocyte of 95 patients with acute leukemia [(43 cases of acute lymphoblastic leukemia (ALL), 52 cases of acute myeloid leukemia (AML)] and 50 normal people were detected by flow cytometry respectively. The positive rate of CD3+, CD3+CD4+, CD3+CD8+, NK cells and CD4+/CD8+ in the patients with newly diagnosed acute leukemia were significantly lower than those in normal controls (P<0.05) , but increased obviously after complete remission. The positive rate of Treg cells in the patients with newly diagnosed leukemia group was significantly higher than that in normal controls (P<0.01) , but decreased obviously after complete remission. Positive rate of CD3-CD19+ cells in the patients with newly diagnosed acute myeloid leukemia was significantly lower , but higher in the patients with newly diagnosed acute lymphoblastic leukemia than that in normal controls with statistical significant difference (P<0.05) , and increased obviously in the patients with acute myeloid leukemia (P<0.05) after complete remission , but decreased in the patients with acute lymphoblastic leukemia (P<0.01) after complete remission or no-remission. Changes of T lymphocyte subsets, B lymphocytes and NK cells in the patients with newly diagnosed acute leukemia are significant, thus the detection of T lymphocyte cell subsets, B lymphocytes and NK cells can provide some evidences. for evaluation of the disease severity, curative efficiency and prognosis of patients with acute leukemia.

Elevated CD3lowdouble negative T lymphocyte is associated with pneumonia and its severity in pediatric patients

BackgroundPrevious studies have shown that the adaptive immunity function of T cells in disease states correlates with CD3 surface expression closely. During routine assessment of TBNK subsets in peripheral blood of pediatric patients by flow cytometry, we noticed that variable expression levels of CD3 on CD3+CD4−CD8− double-negative T (DNT) lymphocytes in different patients. The objective of this study was to assess the relationship of CD3 expression levels on DNT cells with disease severity.MethodsIn this prospective study, we investigated the frequencies of circulating CD4−CD8− DNT cell subsets with CD3low or CD3high phenotype by flow cytometry in 76 pediatric patients with pneumonia, 55 patients with severe pneumonia (SP), and 29 healthy controls (Con).ResultsThe numbers of circulating DNT cells were similar in all groups; however, the frequency of CD3low DNT cell subsets was significantly increased in patients with pneumonia (p < 0.001) and SP (p < 0.001). The elevated CD3low DNT cell frequency showed a positive correlation with the clinical severity of pneumonia. On sub-group analysis, the frequency of CD3low DNT cells was only elevated in children with pneumonia aged <5 years, while no association was observed with the causative pathogen of pneumonia.ConclusionsThese findings suggest that CD3 expression levels on DNT cell subsets of peripheral lymphocytes may be a valuable biomarker for evaluation of immune response in pediatric infectious disease. CD3low DNT cells were elevated in children with pneumonia aged <5 years, which indicates that it may be an important research target in pediatric infectious diseases.

PROGRESSIVE DECREASE IN T CELL QUANTITY AND FUNCTION IN A PATIENT WITH IL-12 PATHWAY DEFECT

Introduction T helper 1 (TH1) and IFN-g are key players in the cell-mediated immune response. Furthermore, IL-12 is an important cytokine that promotes TH1 cell differentiation and mediates its affects through signal transducer and activator of transcription 4 (STAT4). Mutations in the gene encoding the IL-12 receptor beta1 subunit have been identified in patients with disseminated Salmonella and nontuberculous mycobacterial (NTM) infections or disseminated Bacillus Calmette-Guerin (BCG) infection following vaccine administration. Case Description 5-month old female with history of recurrent salmonella enteritis and meningitis at 3 weeks and 3 months of age who presented to our Immunology clinic for an immune evaluation. Initial T cell lymphocyte subsets showed elevated CD3, CD4, CD8 and CD16/CD56 absolute values. Her mitogen culture proliferation assay was reported as low lymphocyte proliferation to ConA and PHA, however based on our assessment she had greater than tenfold response to stimulus and therefore normal function. The patient's lymphocyte subsets have been reassessed on a yearly basis and have showed progressive decline. This year, her CD4 and CD8 were 754 and 338 respectively. Mitogen proliferation culture re-assessment this year also showed significant decline in her T cell function and she was therefore started on Bactrim prophylaxis. Discussion Our patient showed progressive decline in her T cell subset values and decrease in overall function. Although classically MSMD (mendelian susceptibility to mycobacterial disease) patients showed increase susceptibility to disseminated BCG and Salmonella disease and is managed with interferon gamma and azithromycin prophylaxis, our patient now requires additional Bactrim prophylaxis. IL-12 Signaling Pathway

Characterization and Changes of Lymphocyte Subsets in Baricitinib-Treated Patients With Rheumatoid Arthritis: An Integrated Analysis.

ObjectiveBaricitinib is an orally administered inhibitor of JAK1 and JAK2 that has been shown to be effective in treating rheumatoid arthritis (RA). This study was undertaken to analyze changes in lymphocyte cell subsets during baricitinib treatment and to correlate these changes with clinical outcomes.MethodsAn integrated analysis was conducted by pooling data from 3 completed phase III trials comparing placebo with baricitinib treatment (RA‐BEAM, RA‐BUILD, and RA‐BEACON) and 1 ongoing long‐term extension study (RA‐BEYOND) in patients with active RA (n = 2,186).ResultsBaricitinib treatment was associated with an early transient increase in total lymphocyte count at week 4, which returned to baseline by week 12. Transient changes within normal reference ranges in T cells and subsets were observed with baricitinib treatment, up to week 104. B cells and relevant subpopulations increased after 4 weeks of baricitinib treatment, with no further increases noted through 104 weeks of treatment. Natural killer (NK) cells temporarily increased after 4 weeks of baricitinib treatment, before decreasing below baseline levels and then stabilizing over time. With baricitinib treatment, few correlations were observed between changes in lymphocyte subsets and clinical end points, and most correlations were also observed within the placebo group. A modest potential association between low NK cell numbers and treatment‐emergent infections was observed in the baricitinib 4 mg/day treatment group, but not for serious infections or herpes zoster.ConclusionOverall, these findings demonstrate that changes in lymphocyte subsets were largely within normal reference ranges across the baricitinib phase III RA clinical program and were not associated with increased risk of serious infections.

Antibiotic-induced dysbiosis effects on the murine gastrointestinal tract and their systemic repercussions

The gastrointestinal tract has become a focus of study recently. The crosstalk between microbiota, especially bacteria, and the intestinal mucosa has to be accurately balanced in order to maintain physiological homeostasis in the human body. This dynamic interaction results in different levels of short-chain fatty acids (SCFAs), IgA, and T cell lymphocyte subsets, which could lead the human body towards health or disease. The disruption of this microbiome characterises gut dysbiosis. Antibiotics are usually prescribed to fight against bacterial infection. They can also modulate the human microbiome, since it acts directly over organisational taxonomic units (OTUs) when taken orally. As a result, these pharmaceuticals enable gut dysbiosis and its systemic effects due to microbiome disturbance. Here, current data have been gathered from mice model experiments and epidemiological studies in an antibiotic-centred perspective. The presented data suggest the importance of translational studies in a murine model focusing on GIT homeostasis with bacterial groups since any changes to the GIT-microbiota have systemic repercussions in human health and disease.

Abstract 4053: Partial reversion of the tumor immunosuppressed environment by oncolytic adenoviruses armed with positive stimulators of the immune synapsis in breast cancer

Abstract Immunotherapy is currently one of the primary focuses of cancer research and has the potential to become the standard treatment for oncologic patients over the next decades. Promising clinical responses have been observed in patients treated with a variety of immune-targeted therapies such as checkpoint inhibitors, modified dendritic cells, and T cell or NK cell-based immunotherapies. However, the efficacy of these treatments is still very limited due to multiple factors related to tumor nature and microenvironment. Hence, combination therapies aimed at modulating the strong tumor immunosuppressive microenvironment and stimulating T-cell infiltration into the tumor lesions are urgently needed to overcome cancer intrinsic resistance mechanisms. Preclinical and clinical studies have shown the efficacy of oncolytic virotherapy to elicit antitumor immune responses and to increase infiltration of both innate and adaptive immune cells. These capabilities of the oncolytic viruses can be enhanced and exploited to improve response of patients to different immunotherapies. Here we tested the ability of two modified oncolytic adenoviruses termed Delta-24-RGDOX and Delta-24-GREAT, expressing OX40L and GITRL respectively, to modify the tumor microenvironment in different murine triple-negative breast cancer models. We hypothesized that intratumoral injection of these armed oncolytic adenoviruses will decrease immunosuppression within the tumor microenvironment. To this end, mice harboring fat pad tumors were treated with intratumoral doses of these armed oncolytic adenoviruses. Leukocytes from spleen and tumor tissues were analyzed for the relative presence of lymphocyte and myeloid cell subsets. In all mice treated with Delta-24-RGDOX or Delta-24-GITRL, CD8+ T-cell populations were significantly higher compared with those observed in the control mice. Significantly lower percentages of cell populations with immunosuppressive phenotypes, such as PD-1+ T cells (P&amp;lt;0.0001, Student t-test), CD4+CD25+ Foxp3+ regulatory T cells (P&amp;lt;0.0001, Student t-test) and MDSC (P&amp;lt;0.0001, Student t-test) were found in the mice treated with the armed oncolytic adenoviruses. Taken together, our results confirmed the strong immunomodulatory effect of enhanced oncolytic viruses, which can be easily combined with immune-modulating checkpoints, for the treatment of deadliest human cancers. Citation Format: Francisco W. Puerta Martinez, Yisel A. Rivera, Teresa Nguyen, Xuejun Fan, Rehnuma Shifat, Mohammad Belayat Hossain, Hong Jiang, Chandra Bartholomeusz, Lang F. Frederick, Juan Fueyo, Candelaria Gomez-Manzano. Partial reversion of the tumor immunosuppressed environment by oncolytic adenoviruses armed with positive stimulators of the immune synapsis in breast cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr 4053.

PO-372 IL-2 and IL-15 cytokine in vitro treatments induce NKG2D, CD158a and CD158b receptor expression on T, NKT- like and NK cell lymphocyte subsets from regional lymph nodes of melanoma patients

IntroductionRegional lymph nodes (LN)s are important immunological barriers in spreading of malignant tumours but also represent the most frequent early metastatic site in melanoma. Considering that cytokine therapy has shown substantial toxicity, there is a growing need for further in vitro testing of these agents to enlighten aspects of their regional application in malignancies. The aim of this study was to investigate the effect interleukin (IL)−2 and IL-15, cytokines with similar immune-enhancing effects, on the expression of activating (NKG2D) and inhibitory (CD158a,CD158b) receptors on CD8+ T, NKT-like and NK cell lymphocyte subsets from regional LNs of melanoma patients.Material and methodsMononuclear cells (MNC)s were purified from regional LNs of 35 melanoma patients (clinical stages II-IV) and in vitro cultured for 7 days in cell culture medium RPMI1640 (CM) alone, CM with 200 IU/ml rhIL-2 and CM with 25 ng/ml IL-15. Expression of NKG2D, CD158a and CD158b receptors on lymphocytes, CD8+ T, NKT-like and NK cells was estimated by flow cytometry. Statistical significance between the values obtained with cytokine compared to control CM treatments was evaluated by Wilcoxon signed rank test. NK cell cytotoxicity was evaluated by standard 51Cr release assay.Results and discussionsOur results show that IL-2 and IL-15 in vitro treatments significantly increase expression of activating NKG2D receptor on lymphocytes and their CD8+ T, NKT-like and NK cell subsets. Regarding investigated inhibitory receptors, the significant increase was obtained for CD158a only after IL-15 treatment on lymphocytes and their CD8+ T and NKT- like subsets. However, the expression of CD158b receptor significantly increased after IL-2 cytokine treatment on lymphocytes, CD8+ T and NK cells, and after IL-15 treatment on all investigated lymphocyte subsets.Both cytokines augmented NK cell antitumor cytotoxicity that positively correlated (p<0.01, Spearman signed rank test) with the expression of NKG2D activating receptor on NK cells from regional LNs of melanoma patients, irrespective of LN involvement. Conversely, the expression of inhibitory receptors did not correlate with NK cell cytotoxicity except for CD158b that after control CM treatment showed negative correlation.ConclusionAntitumor potential of immune cells from regional LNs induced with IL-2 and IL-15 cytokine treatments may indicate the importance of this immune cell population in the development of therapeutic strategies that involve local cytokine application.

IL-2 And IL-15 Induced NKG2D, CD158a and CD158b Expression on T, NKT- like and NK Cell Lymphocyte Subsets from Regional Lymph Nodes of Melanoma Patients.

Regional lymph nodes (LN)s represent important immunological barriers in spreading of malignant tumors. However, they are the most frequent early metastatic site in melanoma. Immunomodulatory agents including cytokines have been included in therapy of melanoma and have shown severe side effects and toxicity. In this sense, there is a growing need for bringing these agents to further in vitro testing that may enlighten aspects of their regional application. Therefore, the aim of this study was to investigate the effect of interleukin (IL)-2 and IL-15, the two cytokines with similar immune-enhancing effects, on the expression of activating NKG2D, inhibitory CD158a and CD158b receptors on CD8+ T, NKT-like and NK cell lymphocyte subsets from regional LNs of melanoma patients. In this study, we showed significant effects of IL-2 and IL-15 cytokine treatments on the expression of activating NKG2D and on inhibitory CD158a and CD158b receptors on lymphocytes, CD8+ T, NKT-like and NK cell lymphocyte subsets originating from regional LNs of melanoma patients. Furthermore, IL-2 and IL-15 by inducing the expression of NKG2D activating receptor on innate and on adaptive lymphocyte subsets and by augmenting NK cell antitumor cytotoxicity that correlated with the cytokine-induced NKG2D expression, increased antitumor potential of immune cells in regional LNs of melanoma patients irrespective of LN involvement. These findings indicate the importance of immune cell population from regional LNs of melanoma patients in the development of immune intervention strategies that may if applied locally increase antitumor potential to the level that controls tumor progressions.

EKSPRESI KORESEPTOR HUMAN IMMUNODEFICIENCY VIRUS CCR5 DAN CXCR4 PADA SUBSET SEL LIMFOSIT T SERTA MONOSIT

Chemokine receptors CCR5 and CXCR4 which lied on lymphocyte cell surface play important role in HIV infection and pathogenesis.The expression of these chemokine receptors will affect progressively the disease. The objectives of the study are to find the distributionof lymphocyte T cell subset and monocyte among the peripheral blood mononuclear cells and to know the determination of CCR5 andCXCR4 co receptors expression on T lymphocyte cells subset and monocyte. This study is a preliminary study to explore the distributionof co receptors CCR5 and CXCR4 expression in healthy people. The sample taken is peripheral blood mononuclear cells (PBMC) fromhealthy subjects. The identification of T lymphocyte cells subsets and monocyte, and the expression of CCR5 and CXCR4 co receptorswere determined using flowcytometry. The memory T cell (CD4+CD45RO) is found to be the largest proportion among T lymphocyte cell(66.2%), whereas the other T lymphocyte cell subset, regulatory T cell, which identified by CD25+ high expression was found between2.0-5.3% from the whole T lymphocyte cell. The proportion of CXCR4 co receptors was found higher compare to CCR5 co receptorson all T lymphocyte subsets and monocyte. Only small proportion of monocyte expresses both co receptors (2.85%), but most of the Tlymphocyte cell expressed both CCR5 and CXCR4. The expression of the CXCR4 on regulatory T cell (18.18%) is the lowest compared toother cells, but the fluorescence intensity of both co receptors was very high (CCR5 53.53 and CXCR4 49.33). The different distributionof CCR5 and CXCR4 co receptors among T lymphocyte cell subsets and monocyte will influence the vulnerability and the pathogenicityof HIV infection.

A Typical Immune T/B Subset Profile Characterizes Bicuspid Aortic Valve: In an Old Status?

Bicuspid valve disease is associated with the development of thoracic aortic aneurysm. The molecular mechanisms underlying this association still need to be clarified. Here, we evaluated the circulating levels of T and B lymphocyte subsets associated with the development of vascular diseases in patients with bicuspid aortic valve or tricuspid aortic valve with and without thoracic aortic aneurysm. We unveiled that the circulating levels of the MAIT, CD4+IL−17A+, and NKT T cell subsets were significantly reduced in bicuspid valve disease cases, when compared to tricuspid aortic valve cases in either the presence or the absence of thoracic aortic aneurysm. Among patients with tricuspid aortic valve, these cells were higher in those also affected by thoracic aortic aneurysm. Similar data were obtained by examining CD19+ B cells, naïve B cells (IgD+CD27−), memory unswitched B cells (IgD+CD27+), memory switched B cells (IgD−CD27+), and double-negative B cells (DN) (IgD−CD27−). These cells resulted to be lower in subjects with bicuspid valve disease with respect to patients with tricuspid aortic valve. In whole, our data indicate that patients with bicuspid valve disease show a quantitative reduction of T and B lymphocyte cell subsets. Future studies are encouraged to understand the molecular mechanisms underlying this observation and its pathophysiological significance.

Probiotic Enhanced Intestinal Immunity in Broilers against Subclinical Necrotic Enteritis

Along with banning of antibiotics, necrotic enteritis (NE), especially subclinical NE (SNE) whereby no clinical signs are present in chicks, has become one of the most threatening problems in poultry industry. Therefore, increasing attention has been focused on research and application of effective probiotic strains, as an alternative to antibiotics, to prevent SNE in broilers. In the present study, we evaluated the effects of Lactobacillus johnsonii BS15 on the prevention of SNE in broilers. Specifically, assessment determined the growth performance and indexes related to intestinal mucosal immunity in the ileum and cecal tonsil of broilers. A total of 300 1-day-old Cobb 500 chicks were randomly distributed into the following 5 groups: control group (fed with basal diet + de Man, Rogosa, and Sharpe liquid medium [normal diet]), SNE group (normal diet), BS15 group (basal diet + 1 × 106 colony-forming units BS15/g as fed [BS15 diet]), treatment group (normal diet [days 1–28] + BS15 diet [days 29–42]), and prevention group (BS15 diet [days 1–28] + normal diet [days 29–42]) throughout a 42-day experimental period. SNE infection was treated for all chicks in the SNE, BS15, treatment, and prevention groups. The present results demonstrated that BS15 supplementation of feeds in BS15 and prevention groups exerted a positive effect on preventing negative influences on growth performance; these negative influences included low body weight gain and increased feed conversion ratio caused by SNE. Although no changes were detected in all determined indexes in cecal tonsils, BS15-treated broilers were free from SNE-caused damage in villi in the ileum. BS15 inhibited SNE-caused decrease in immunoglobulins in the ileum. In the lamina propria of ileum, T cell subsets of lymphocytes influenced by SNE were also controlled by BS15. BS15 affected antioxidant abilities of the ileum and controlled SNE-induced mitochondrion-mediated apoptosis by positively changing contents and/or mRNA expression levels of apoptosis-related proteins. These findings indicate that BS15 supplementation may prevent SNE-affected growth decline mainly through enhancing intestinal immunity in broilers.

Abstract 1687: Systemic immunological changes during first line chemotherapy in patients with high-grade serous ovarian cancer

Abstract Ovarian cancer remains the most lethal gynecological malignancy and new therapeutic strategies are urgently needed. High-grade serous ovarian cancer (HGSC), in particular, is associated with a five-year survival of only 40%. A promising new approach for treating HGSC is immunotherapy, which has resulted in complete and durable responses, albeit in a minority of patients. One potential approach for improving these response rates is by combining chemo- and immunotherapy. Indeed, carboplatin/taxol chemotherapy was shown to augment immune responses in cervical cancer patients by depleting circulating myeloid suppressor cells. As patients with HGSC are treated in first line with similar carboplatin/taxol chemotherapy, we explored the effects of chemotherapy on systemic immunity in HGSC patients. Within this prospective observational study, 75 patients with suspected ovarian cancer were included. Blood was collected at three different time points during first line chemotherapy treatment, namely: prior to chemotherapy, between the third and fourth cycle of chemotherapy and 4-6 weeks after the sixth cycle of chemotherapy. All patients received 6 cycles of carboplatin/taxol chemotherapy and cytoreductive surgery (either as primary debulking, or after 3 cycles of neo-adjuvant chemotherapy). Peripheral blood mononuclear cells were isolated and analyzed for a total of 36 immune markers using 9 flow cytometry panels, in total analyzing 49 immune cell subsets. Results were compared to an age-matched cohort consisting of women surgically treated for a benign disease, and a cohort of healthy young volunteers. 75 patients have been included so far from which 22 were diagnosed with benign disease, 3 were diagnosed with another malignancy, and 50 were diagnosed with OC. Of the 50 OC patients, 18 were diagnosed with HGSC and from 9 HGSC patients, multiple time points were available for analysis of chemotherapy-dependent changes in the immune cell subsets. All patients developed leukopenia as a result of chemotherapeutic treatment. Age-related changes in lymphocyte and myeloid cell subsets were observed in all HGSC patients and patients with benign disease. Chemotherapy-dependent depletion of myeloid cells was observed in a subset of patients. Depletion of myeloid subsets was equally distributed among monocytes, macrophages and dendritic cells. No HGSC- or chemotherapy-dependent changes in T cell subsets or migration and activation markers were observed. Taken together, we observed no major systemic changes in immune cell subsets during carboplatin/taxol chemotherapy treatment, suggesting that combined chemo-immunotherapeutic strategies could be feasible during first line treatment of HGSC. Citation Format: Fenne L. Komdeur, Florine A. Eggink, Ninke Leffers, Hagma H. Workel, Kim L. Brunekreeft, Annechien Plat, Marco de Bruyn, Hans W. Nijman. Systemic immunological changes during first line chemotherapy in patients with high-grade serous ovarian cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 1687. doi:10.1158/1538-7445.AM2017-1687

Oral Administration of β-Glucosylceramide for the Treatment of Insulin Resistance and Nonalcoholic Steatohepatitis: Results of a Double-Blind, Placebo-Controlled Trial.

β-glucosylceramide (GC) is a naturally occurring glycosphingolipid that was shown to improve hepatic steatosis, steatohepatitis, and insulin resistance in animal models of nonalcoholic fatty liver disease. In this study, we evaluated the safety and efficacy of oral administration of GC in subjects with nonalcoholic steatohepatitis (NASH). Twenty-three patients with biopsy proven NASH were enrolled in a double-blind, placebo-controlled trial. Patients were orally administered daily with 7.5 mg of GC. Patients were followed for safety, liver enzymes, HbA1c, insulin sensitivity, lipid profile, hepatic fat content as measured by magnetic resonance imaging (MRI), and NASH score on liver biopsy. No treatment-related adverse events were observed during treatment. In a per protocol analysis of data, oral administration of GC decreased the hepatic fat content as measured by MRI in GC-treated compared with placebo. HbA1C decreased in patients treated with GC. GC treatment was associated with a milder decrease in the high-density lipoprotein serum levels. The beneficial effects were associated with a decrease in CD4 and NKT cell subsets of lymphocytes. Due to the small number of subjects enrolled, differences did reach statistical significance. Oral administration of GC is safe and biologically active in patients with NASH and insulin resistance.

919 - IL-2 and IL-15 cytokine treatments increase NKG2D receptor expression on T, NKT-like and NK cell lymphocyte subsets from regional lymph nodes of melanoma patients regardless of lymph node involvement

919 - IL-2 and IL-15 cytokine treatments increase NKG2D receptor expression on T, NKT-like and NK cell lymphocyte subsets from regional lymph nodes of melanoma patients regardless of lymph node involvement

Graded versus Intermittent Exercise Effects on Lymphocytes in Chronic Fatigue Syndrome.

There is increasing evidence of immune system dysfunction in chronic fatigue syndrome (CFS), but little is known of the regular exercise effects on immune cell parameters. This pilot study investigated the effects of graded and intermittent exercise on CD4 lymphocyte subset counts and activation compared with usual care. Twenty-four CFS patients (50.2 ± 10 yr) were randomized to graded exercise (GE), intermittent exercise (IE), or usual care (UC) groups; 18 sedentary non-CFS participants (50.6 ± 10 yr) were controls (CTL) for blood and immunological comparisons. Outcome measures were pre- and postintervention flow cytometric analyses of circulating lymphocyte subset cell counts; expression of CD3, CD4, CD25, and CD134; full blood counts; and V˙O2peak. Preintervention, CD3 cell counts, and expression of CD4, CD25, CD134, and CD4CD25CD134 were significantly lower in GE, IE, and UC compared with CTL (P < 0.05). Total lymphocyte concentration was significantly lower in GE and IE groups compared with CTL. There were significant postintervention increases in i) expression of CD4 and CD4CD25CD134 for GE and IE, but CD25 and CD134 for IE only; ii) circulating counts of CD3 and CD4 for GE, and CD3, CD4, CD8, CD3CD4CD8, CD3CD16CD56, CD19, and CD45 for IE; iii) neutrophil concentration for GE; and iv) V˙O2peak and elapsed test time for IE and GE, V˙Epeak for IE. Twelve weeks of GE and IE training significantly improved CD4 lymphocyte activation and aerobic capacity without exacerbating CFS symptoms. IE may be a more effective exercise modality with regard to enhanced CD4 activation in CFS patients.

Simultaneous detection of circulating immunological parameters and tumor biomarkers in early stage breast cancer patients during adjuvant chemotherapy

Chemotherapy-induced immune suppression has mainly been studied in patients with advanced cancer, but the influence of chemotherapy on the immune system in early stage cancer patients has so far not been studied systematically. The aim of the present study was to monitor the immune system during anthracycline- and taxane-based adjuvant chemotherapy in early stage breast cancer patients, to assess the impact of circulating tumor cells on selected immune parameters and to reveal putative angiogenic effects of circulating endothelial cells. Peripheral blood samples from 20 early stage breast cancer patients were analyzed using a flow cytometric multi-color of antibodies to enumerate lymphocyte and dendritic cell subsets, as well as endothelial and tumor cells. An enzyme-linked immunosorbent assay (ELISA) was used to measure the levels of various serological factors. During chemotherapy, all immunological parameters and angiogenesis surrogate biomarkers showed significant decreases. The numbers of circulating tumor cells showed significant inverse correlations with the numbers of T helper cells, a lymphocyte subset directly related to effective anti-tumor responses. Reduced T helper cell numbers may contribute to systemic immunosuppression and, as such, the activation of dormant tumor cells. From our results we conclude that adjuvant chemotherapy suppresses immune function in early stage breast cancer patients. In addition, we conclude that the presence of circulating tumor cells, defined as pan-cytokeratin(+), CD326(+), CD45(-) cells, may serve as an important indicator of a patient's immune status. Further investigations are needed to firmly define circulating tumor cells as a predictor for the success of breast cancer adjuvant chemotherapy.

Hyperexpansion of Functional Viral-Specific CD8+ T Cells in Lymphopenia-Associated MCMV Pneumonitis.

Cytomegalovirus (CMV) is a significant cause of morbidity and mortality in immunocompromised hosts, many of whom undergo significant periods of lymphopenia. However, the impact of lymphopenia and subsequent immune reconstitution on T cell responses and pulmonary pathology are poorly understood. Using a model of primary murine CMV infection in mice treated with cyclophosphamide (CY), the relationship of CD8+ T cell reconstitution to pneumonitis pathology was studied. Female BALB/c mice were infected with murine CMV (MCMV) with/without CY on day 1 post-infection. Lung pathology and viral specific T cell responses were assessed on days 7-28. T cell lymphocyte subsets, effector responses, and MCMV specificity were assessed at baseline and after in vitro stimulation of cells with immediate-early peptide pp89. CY treatment of MCMV-infected mice resulted in interstitial pneumonitis not seen with MCMV alone. Compared to MCMV alone, on day 14, MCMV/CY mice had greater number of CD8+ T cells, a fourfold increase in absolute number of pp89 tetramer-specific CD8+ cells, and an eightfold increase in MCMV specific T cell effector responses (IFN-γ; p<0.001). This expansion was preceded by transient lymphopenia, increased viral titers, and, most strikingly, a 10-fold increased proliferative capacity in MCMV/CY mice. In the setting of CY-associated lymphopenia, concurrent MCMV infection alters immune reconstitution toward a hyperexpanded MCMV-specific CD8+ effector T cell pool that correlates with significant lung immunopathology.

Decreased Interferon γ Production in CD3+ and CD3- CD56+ Lymphocyte Subsets in Metastatic Regional Lymph Nodes of Melanoma Patients.

As lymphogenic dissemination is very common in melanoma, regional lymph nodes (LN)s represent first immunological barriers to tumor invasion and play a complex role in antitumor immune defense. In this sense, their most prominent role is the presentation of tumor-derived antigens to naïve T cells and generation of cell-mediated adaptive immune response. Since tumor micro-environment affects immune cell function in this study we have evaluated the ability of T cells and NK cells in metastatic (involved) and non-metastatic regional LNs to produce interferon γ (IFNγ), a pleiotropic cytokine that regulates adaptive antitumor immune response. Our results show reduced IFNγ production in both T and NK lymphocyte subsets and decreased prevalence of T cells in metastatic regional LNs of melanoma patients. The decrease of IFNγ production in T cells was more pronounced with increased number of involved regional LNs indicating tumor-induced functional impairment of both T and NK cell lymphocyte subsets in involved regional LNs. Therefore, shown low IFNγ production in metastatic LNs may represent an obstacle in adaptive cell-mediated antitumor immune response and hence may enable tumor progression.