Marker Of Cell-mediated Immunity Research Articles

#4277 EGF, GDF-15 AND NEOPTERIN AS PREDICTORS OF CHRONIC KIDNEY DISEASE PROGRESSION IN CHILDREN BY MEANS OF ARTIFICIAL NEURAL NETWORK MODELS

Abstract Background and Aims Progression of chronic kidney disease (CKD) is a compound process, where activation of immunocompetent cells and subclinical inflammation play pivotal role. Enhanced atrophy of the tubular cells, and finally, gradual fibrosis of tubulointerstitial tissue, are responsible for irreversible character of the disease. Multiple molecules influence above-mentioned processes. Growth differentiation factor (GDF)15, a member of TGF-β cytokine superfamily, is a marker of inflammation and an integrative signal in both acute and chronic stress conditions. Elevated serum concentrations of GDF15 were associated with increased risk of development and progression of CKD in adults, as well as with mortality in this group of patients. Our previous investigation revealed increased serum GDF15 concentrations in children on chronic dialysis. Epidermal growth factor (EGF), a tubule-specific protein, promotes proliferation, differentiation and migration of epithelial cells, and therefore, modulates regeneration of injured renal tubules. Decreased concentrations of EGF in urine were observed in variety of kidney diseases, including diabetic nephropathy, lupus nephritis or CKD. Our previous analysis of EGF serum concentrations in CKD children confirmed their decreased values on chronic dialysis. Neopterin is a product of activated monocytes and macrophages and serves as a marker of cell-mediated immunity. Elevated serum concentrations of neopterin were observed in CKD adult patients, our investigation revealed its increased values in children on chronic dialysis. None of the above mentioned markers was analyzed in the population of CKD children treated conservatively. Therefore, the aim of study was to assess the serum concentrations of EGF, GDF-15 and neopterin in children with CKD on conservative treatment and verify the usefulness of these markers in predicting CKD progression by means of artificial intelligence tools. Method The study group consisted of 153 children with pre-dialysis CKD stages 1-5 (stage 1 – 27 patients, stage 2 – 26 patients, stage 3 – 51 patients, stage 4 - 28 patients, stage 5 – 21 patients). EGF, GDF-15 and neopterin serum concentrations were assessed by ELISA. The patient database was implemented into the artificial neural network. In detail, the recursively selected subsets of input variables constituted the input layer of an artificial neural network built of perceptrons (multi-layer perceptron). Anthropometric data, biochemical parameters, EGF, GDF15 and neopterin were included into the model, serum creatinine and eGFR, as direct classifiers of CKD stage, were excluded. Various models were tested, regarding their accuracy, AUROC and Matthews correlation coefficient (MCC) values. Results EGF serum concentrations decreased gradually, whereas GDF15 and neopterin values rose systematically with CKD progression, keeping statistically significant inter-stage differences. Moreover, the most precise ANN model, among the tested artificial neural networks, contained EGF, GDF15 and neopterin as input parameters and classified patients into either CKD 1-3 or CKD 4-5 groups. This model has put new patients into appropriate classes with excellent Accuracy of 96.77%, AUROC 0.9169 and Matthews correlation coefficient (MCC) of 0.9157. Conclusion The presented model of an artificial neural network, with serum concentrations of EGF, GDF15 and neopterin as input parameters, may serve as a useful predictor of CKD progression in the pediatric population. It suggests the essential role of inflammatory processes, defined by newly discovered markers, in the renal function decline towards advanced stages of CKD in children.

Adenosine deaminase activity in serum and pleural fluid of patients with tuberculosis and those without tuberculosis

Background Adenosine deaminase (ADA) has been considered as a marker for cell-mediated immunity. The purpose of this study is to evaluate the usefulness of ADA as a biomarker in diagnosis of pleural effusion in tuberculosis (TB). Patients and methods Forty patients from Abbasia Chest Hospital who were suspected to have exudative pleural effusion and prepared for pleural biopsy were enrolled in the study. ADA in pleural fluid and serum was measured for all of them, and then data were reported and analyzed. Results Of the 40 patients included in the study, most of the 20 patients with TB pleurisy were diagnosed mainly by thoracoscope (16/20), whereas the rest four cases were diagnosed by sonar-guided biopsy. The other 20 patients, diagnosed as non-TB pleural effusion (6/20), mesothelioma (8/20), pneumonia, and adenocarcinoma (6/20). There were significant statistical difference between both groups regarding age, total leukocyte count, erythrocyte sedimentation rate, and tuberculin skin test, whereas no significant association regarding sex, clinical presentation, and radiological finding within any groups. ADA in effusion, serum, and index had a significant statistical difference between the two groups, with cutoff point value in effusion and serum of ∼31.4 and 29.5 U/l, respectively, and 1.1 for the index, with sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of ∼100, 90, 90, 100, and 95%, respectively, for ADA in effusion and serum, whereas for the index of ∼80, 65, 69, 76, and 73%, respectively. Conclusion Serum and pleural fluid ADA level significantly increases in patients with TB pleural effusion than non-TB pleural effusion with higher sensitivity, specificity, positive predictive value, negative predictive value, and accuracy in prediction.

Role of biochemical markers in rheumatoid arthritis

Rheumatoid arthritis is the most common chronic, progressive systemic inflammatory arthritis of unknown cause. It affects approximately 1-2% of the population worldwide. It is a form of autoimmune disease principally synovial tissues. As the pathophysiology involves affecting cell mediated immune response, adenosine deaminase may have a role in rheumatoid arthritis. As it involves bones and joints, it is likely that it alters the levels of Serum Calcium, Phosphorus and Magnesium as well as Alkaline Phosphatase which may be helpful in diagnosis or management of the disease. 1. To evaluate the role of serum adenosine deaminase (ADA) as a marker of cell mediated immunity in Rheumatoid Arthritis. 2. To estimate serum alkaline phosphatase, calcium, phosphorus and magnesium as prognostic markers of Rheumatoid arthritisA cross-sectional, case control study with convenient sampling was undertaken after Institutional Ethical Clearance & informed consent. 30 patients of Rheumatoid Arthritis presenting to the orthopaedic outpatient department of Owaisi Hospital and Research Centre, Hyderabad were taken as cases along with an equal number of age and sex matched controls from January 2021 to June 2021. ADA levels were estimated by colorimetric assay on ErbaChem 7 semiautoanalyser. Serum Alkaline phosphatase, Calcium, Phosphorus and Magnesium were estimated on Cobas C311 autoanalyzer. There was a significant increase in levels of Adenosine Deaminase, Alkaline Phosphatase and Serum Phosphorus, while the levels of Calcium and Magnesium were significantly reduced in rheumatoid arthritisADA as a marker of cell mediated immunity reflects monocyte / macrophage activity and was increased in our study suggesting immunological and inflammatory process in the pathogenesis of Rheumatoid Arthritis (R.A). It can be used as a non invasive marker of chronic inflammation and may provide additional information about disease prognosis.Increased levels of Alkaline Phosphatase and Phosphorus associated with low Calcium and Magnesium suggest increased bone resorption in R. A and are useful as markers of disease activity.

Changes in Cell-Mediated Immunity (IFN-γ and Granzyme B) Following Influenza Vaccination.

Interferon gamma (IFN-γ) is considered a key moderator of cell-mediated immunity. However, little is known about its association with granzyme B, which plays an important role in the effector function of cytotoxic T lymphocytes (CTLs). In the present study, we collected blood samples from 32 healthy adults before and after vaccination with inactivated influenza vaccine in 2017/18 to measure the levels of IFN-γ and granzyme B, which play roles in cell-mediated immunity, and hemagglutination inhibition (HAI) antibody, which plays a role in humoral immunity. The levels of IFN-γ and granzyme B were significantly correlated both before and after vaccination. Furthermore, the post-vaccine fold increases in the IFN-γ and granzyme B levels were significantly correlated. The levels of IFN-γ and granzyme B decreased five months after vaccination in more than half of the subjects who exhibited an increase in IFN-γ and granzyme B at two weeks post-vaccination. This is the first study to investigate the correlation between IFN-γ and granzyme B levels following influenza vaccination. Our study suggests that both IFN-γ and granzyme B can be used as markers of cell-mediated immunity.

Study of Serum Adenosine Deaminase Level in Type 2 Diabetes Mellitus and its Correlation with Glycemic Control

Diabetes mellitus is a cluster of abnormal metabolic disorder having common features of hyperglycaemia with a state of increased free radical activity. Chronic hyperglycemic status favours auto-oxidation and the formation of advanced glycation end products. Adenosine deaminase (ADA) is considered as a good marker of cell mediated immunity. Increased ADA activity in diabetic individuals could be due to altered insulin related T-lymphocyte function. Hyperglycaemia is associated with increased level of (ADA), which is one of the factor which leads to increase oxidative stress level by generatingthe reactve oxygen species (ROS) leading to insulin resistance. In our study, ADA level was significantly high in controlled diabetes mellitus type 2 (group II with HbA1c < 7) and was much higher in uncontrolled diabetics (group III with HbA1c > 7) compared to healthy controls (group I). The present study was aimed to find the level of Serum (ADA) among the patients with type 2 diabetes mellitus through a case control study and correlation of Adenosine deaminase with glycemic control (HbA1c). Comparison of the parameters fasting plasma glucose (FPG), post prandial plasma glucose (PPPG) HbA1c, and ADA between the 3 groups were done using Student t test and was statistically significant. Pearson’s coefficient correlation was done between ADA and HbA1c and a positive correlaion was seen that was also statistically significant. This indicates that ADA ses with the extent of severity of type 2 diabetes. Positive correlation of ADA with HbA1c provides the information that ADA can be considered to reflect the glycemic status of the individual.

MDSC (myeloid-derived suppressor cells) is an important immunosuppressing factor and functionally related with VEGF and IL-17 in patients with gastrointestinal cancer

Abstract Background Immune checkpoint inhibitors have been reported to be ineffective in patients with high levels of MDSC (myeloid-derived suppressor cells) and MDSC has been proven to be stimulated by inflammation involving VEGF and IL-17. We have reported that MDSC, IL-17 and VEGF are closely associated with immunosuppression and poor prognosis. Methods Stimulation index (SI) obtained by PHA-blastogenic response of lymphocyte was used as a marker of cell-mediated immunity and was tested in 658 patients with unresectable diseases including 260 esophageal, 110 gastric and 288 colorectal cancers. These patients received chemotherapy and SI were analyzed in correlation with nutritional status, treatment outcome (RECIST) and prognosis. These patients were divided into 2 groups with a median level of SI and, chemotherapy outcome and overall survival were compared between these groups (Study 1). To characterize the mechanisms of immunosuppression, PBMC (peripheral blood cells) were separated from the patients and the production of IL-17 (ELISA) by PBMC. Circulating levels of MDSC (CD14-CD11b+CD33+/flow cytometry), serum levels of VEGF (ELISA) were measured and analyzed in 43 patients with gastric cancer and 64 with colorectal cancer (Study 2). Results (Study 1) The overall survival was significantly worse in patients with lower SI than in those with higher SI in each disease, and the responses (RECIST) to chemotherapy were significantly better in patients with higher SI than those with lower SI. SI were significantly inversely correlated with NLR (neutrophil/lymphocyte ratio: inflammatory parameter) and nutritional factors including prealbumin, retinol-binding protein and transferrin. (Study 2) SI were significantly and inversely correlated with the levels of MDSC, the production of IL-17, and the levels of VEGF. The levels of MDSC, VEGF, IL-17 were significantly correlated with each other and inversely with nutritional parameters. Conclusion The key mechanisms of immunosuppression in these patients are systemic inflammation involving MDSC, VEGF and IL-17. Immune checkpoint inhibitors might be more effective with a decreased MDSC by controlling VEGF and IL-17. Legal entity responsible for the study Masahiko Shibata. Funding Has not received any funding. Disclosure All authors have declared no conflicts of interest.

Clinical Significance of Increased Serum Neopterin in Chronic Kidney Failure as a Biomarker of Cell-mediated Immunity.

SummaryBackgroundNeopterin is a pyrazino-pyrimidine compound which is used as a marker of cell-mediated immunity in a variety of diseases. It is known that neopterin levels increase in diseases where interferon-gamma (IFN-g) stimulation is present, and also as a result of deficiencies in renal function, given that the primary means of elimination of neopterin is through the kidneys. In this study, we aimed to investigate the role of increased neopterin levels as a prognostic biomarker in patients with impaired renal function.MethodsA total of 90 individuals including 63 patients with chronic kidney failure (CKF) and 27 healthy volunteers were included in the study. Serum neopterin concentrations were measured using the enzyme-linked immunosorbent assay. A Mann-Whitney U test and a Pearson Correlation Test were used in the statistical analysis, with a p value of <0.05 being considered statistically significant.ResultsThe mean age was 52.21±0.16 years in the patient group and 56.55±0.32 years in the control group. In the CKF patients, serum neopterin levels increased to a significantly greater degree than in the control group (p<0.001), while no statistically significant correlation was identified between serum neopterin levels and age (p>0.05).ConclusionsA significant increase was found in the serum neopterin levels in the CKF patients, due to both the triggering of the disease and the reduction of neopterin elimination.

Substituting whole grains for refined grains in a 6-wk randomized trial has a modest effect on gut microbiota and immune and inflammatory markers of healthy adults1–3

Background: Observational studies suggest an inverse association between whole-grain (WG) consumption and inflammation. However, evidence from interventional studies is limited, and few studies have included measurements of cell-mediated immunity.Objective: We assessed the effects of diets rich in WGs compared with refined grains (RGs) on immune and inflammatory responses, gut microbiota, and microbial products in healthy adults while maintaining subject body weights.Design: After a 2-wk provided-food run-in period of consuming a Western-style diet, 49 men and 32 postmenopausal women [age range: 40–65 y, body mass index (in kg/m2) <35] were assigned to consume 1 of 2 provided-food weight-maintenance diets for 6 wk.Results: Compared with the RG group, the WG group had increased plasma total alkyresorcinols (a measure of WG intake) (P < 0.0001), stool weight (P < 0.0001), stool frequency (P = 0.02), and short-chain fatty acid (SCFA) producer Lachnospira [false-discovery rate (FDR)-corrected P = 0.25] but decreased pro-inflammatory Enterobacteriaceae (FDR-corrected P = 0.25). Changes in stool acetate (P = 0.02) and total SCFAs (P = 0.05) were higher in the WG group than in the RG group. A positive association was shown between Lachnospira and acetate (FDR-corrected P = 0.002) or butyrate (FDR-corrected P = 0.005). We also showed that there was a higher percentage of terminal effector memory T cells (P = 0.03) and LPS-stimulated ex vivo production of tumor necrosis factor-α (P = 0.04) in the WG group than in the RG group, which were positively associated with plasma alkylresorcinol concentrations.Conclusion: The short-term consumption of WGs in a weight-maintenance diet increases stool weight and frequency and has modest positive effects on gut microbiota, SCFAs, effector memory T cells, and the acute innate immune response and no effect on other markers of cell-mediated immunity or systemic and gut inflammation. This trial was registered at clinicaltrials.gov as NCT01902394.

Immunological changes in depression.

Background:The role of Immune system in the pathophysiology of depressive disorders is a field of active research, however Indian literature is sparse. The present study was planned to assess the immunological response in depression.Materials and Methods:The study comprised of 100 subjects. There were fifty cases of depression satisfying the ICD-10 criteria with no physical illness and HIV negative status and fifty age and sex matched healthy volunteers. Depression was assessed on HRSD and BDI scales. Assessment of three markers each of cellular immunity (NK cells, CD4, CD8 cells) and humoral immunity (Il-2, IL-6 and CRP) was carried out on both groups and depressed patients were reassessed on all parameters after 08 weeks of treatment with antidepressants (SSRIs or TCAs).Results:NK Cells were significantly higher in the depressed group and CD 8 Cells and CD 4 Cells were higher in the control group (P = 0.001). Depressed group before treatment v/s control group differed significantly in the cell mediated immune markers. IL-2 levels were higher in the control group. The markers of cell mediated immunity i.e., NK cells, CD4, CD8 had increased significantly after treatment (P =< 0.001). The humoral immunity markers (CRP and IL-2) decreased significantly after treatment (P =< 0.001). However IL -6 levels were raised significantly in the subjects after treatment (P =< 0.001).Conclusion:Dysregulation of immune response occurs in depressed patients with changes in both cell mediated and humoral immunity. Further, antidepressant treatment affects the immune status of depressed patients.

Multiple immunological mechanisms of cancer cachexia in patients with solid tumors.

667 Background: Cancer cachexia is a mixed and multiple factorial condition and commonly seen in patients with advanced cancer, and has long been reported that host-immune function is critically participated. Chronic inflammation plays a key role in the progress of malignant diseases and has been reported to relate with immune suppression and nutritional impairment seen in patients with advanced diseases. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers and 112 patients with gastrointestinal cancer. These cells were used for the detection of MDSC (myeloid-derived suppressor cells: CD11b+CD14-CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI) and for the production assay of cytokines including IFN-G, IL-6 and IL-10. Serum levels of soluble cytokine receptors including sIL-2R and sTNF-R1, and anti-inflammatory molecules such as IL-10 and IL-1 receptor antagonist (IL-1RA) were also measured. For the evaluation of nutritional status, serum concentrations of rapid turnover protein (RTP) including prealbumin, transferrin and retinol binding protein was measured. NLR (neutrophil/lymphocyte ratio) was calculated and used as a marker of chronic inflammation. Results: The circulating levels of MDSC was significantly increased in esophageal, gastric and colorectal carcinomas than in normal volunteer and significantly inversely correlated with serum concentration of RTP and with SI, and correlated with markers for inflammation. The levels of sIL-2R and sTNF-R1 were increased in patients with gastric and colorectal and inversely correlated with the levels of RTP and SI. In patients with cachexia, serum concentrations of IL-10 were increased and the production of IFN-G was decreased. Conclusions: It is suggested that suppression of cell-mediated immune function exists in patients with cancer cachexia and the immunological mechanisms of nutritional damages in these patients are even partially driven by Th2-dominant condition of CD4(+) cells and MDSC. These complicated mechanisms are also involved in inflammation-related immunological condition such as SIRS and CARS.

English

Rheumatoid arthritis is a chronic systemic autoimmune disorder causing inflammation of the synovial joints leading to joint destruction as well as a variety of extra articular features. In this present study, serum Adenosine deaminase and Alkaline phosphatase levels were studied in patients of rheumatoid arthritis. Thirty patients in the age group of 30-60 years of both sexes and equal number of age and sex matched healthy controls were included in this study. In rheumatoid arthritis patients, serum Adenosine deaminase levels were high, the mean serum Adenosine deaminase levels were 60 ± 9.55 and in controls the mean Adenosine deaminase levels were 21 ± 3.04. The p value was significant at p < 0.001. We conclude that serum Adenosine deaminase can be used as a marker for cell mediated immunity to monitor disease activity in rheumatoid arthritis. Serum Alkaline phosphatase was found to be raised in rheumatoid arthritis, the mean serum Alkaline phosphatase levels were 291.63 ± 35.84 in patients of rheumatoid arthritis when compared to healthy controls (196.73 ± 32.71).The p value was significant at p<0.001. The present study has focused on investigating the serum total Alkaline phosphatase levels in Rheumatoid Arthritis Patients and The findings have indicated a raised serum Alkaline phosphatase in patients of rheumatoid arthritis when compared to healthy controls thereby suggesting the role of serum Alkaline phosphatase as a marker of disease activity in rheumatoid arthritis.

Serum Adenosine Deaminase as Inflammatory Marker in Rheumatoid Arthritis.

Rheumatoid arthritis (RA) is a prototypical inflammatory joint disease. The degree of inflammation is reflected in the extent of joint damage, which further has influence on the quality of life of patients with RA, including risk of atherosclerosis. Hence, besides clinical indices, estimation of degree of inflammation using biochemical markers helps in effecting optimum treatment strategies. C-reactive protein (CRP) is established as an inflammatory marker in patients with RA. Adenosine deaminase (ADA), an enzyme of purine metabolism is considered as a marker of cell mediated immunity and has also been suggested as a marker of inflammatory process in RA. The present study attempts to study the efficacy of serum ADA activity as an inflammatory marker in RA. Forty six RA patients and forty six age and sex matched healthy controls were included in the study. ADA activity and high sensitivity C-reactive protein (hsCRP) levels in serum were measured in all the subjects. Statistical analyses were done using Medcalc statistical software version 12.2.2. ADA activity and hsCRP levels were increased in RA patients compared to controls (p<0.0001 and 0.0001 respectively). Significant positive correlation was obtained between hsCRP and ADA in patients (r=0.316, p=0.033). Receiver operating characteristic (ROC) curve analysis revealed statistically significant area under curve (AUC) for ADA that is comparable to that obtained for hsCRP (0.776, p<0.0001 for ADA, 0.726, p<0.0001 for hsCRP). Similar diagnostic utility was obtained with ROC generated cut-off value of 25.3 IU/L (82.6% sensitivity and 65.2% specificity) and with control mean value of 23.48 IU/L (86.96% sensitivity and 54.35% specificity) for ADA. Findings of the present study indicate the importance of ADA as a marker of inflammation. Considering the higher sensitivity obtained, we propose control mean (23.48 IU/L) as a cut-off for serum ADA activity as an inflammatory marker. Owing to the simplicity and also the cost effectiveness of ADA assay, ADA may be recommended as a marker of inflammation in patients with RA. However, further larger and well controlled studies are needed to establish its role as inflammatory marker.

Effect of prochloraz fungicide on neopterin and biopterin concentrations in blood plasma of common carp

Neopterin and biopterin are often used as markers of cell mediated immunity. The aim of this study was to evaluate the effect of prochloraz on neopterin and biopterin concentrations in blood plasma of common carp as possible indicators for monitoring perturbations of the immune system caused by xenobiotics like pesticides. Prochloraz is a widely used imidazole fungicide in horticulture and agriculture. A total of 60 juvenile common carp were divided into four groups of 15 fish and exposed to prochloraz at concentrations of 0, 50, 150 and 380 µg·l-1, respectively. The total length of exposure was 28 days. Neopterin and biopterin concentrations were determined by reverse phase high performance liquid chromatography with fluorescence detection. Our results indicated that concentrations of neopterin (25 ± 7.6 nmol·l-1) and biopterin (190 ± 29 nmol·l-1) in plasma of untreated common carp were comparable with those in mammals. Neopterin concentrations significantly (P &lt; 0.01) increased after exposure to prochloraz in comparison to non-exposed fish, while biopterin concentrations were not influenced by exposure to prochloraz. This study contains new data on neopterin and biopterin concentrations in fish plasma and investigates neopterin and biopterin in their non-traditional role as markers of cell mediated immunity of fish associated with pesticide exposure.

The impact of childhood maltreatment on biological systems: Implications for clinical interventions

Childhood maltreatment represents a significant risk factor for the development of a number of mental and physical health outcomes. Converging evidence suggests that early adversity induces significant and persistent biological changes in individuals ('biological embedding'). The present review focuses on the impact of childhood maltreatment on the hypothalamic-pituitary-adrenal axis and immune system function in both children and adults. Research suggests that childhood maltreatment is associated with hypothalamic-pituitary-adrenal axis dysregulation and diurnal cortisol profiles, as well as stress reactivity. Furthermore, childhood maltreatment is associated with disruptions in various immune system markers including pro- and anti-inflammatory substances, and markers of cell-mediated immunity. The potential of interventions to reduce these negative biological effects in maltreated children is also discussed.

Prognostic Utility of Serum Neopterin in Obstructive Jaundice Secondary to Malignant Lesions Treated by Percutaneous Transhepatic Biliary Drainage

PurposeTo perform biochemical profiles before and after percutaneous transhepatic biliary drainage (PTBD) and investigate the potential utility of measuring C-reactive protein (CRP), circulating cytokines, and neopterin, a marker of cell-mediated immunity, to predict outcomes of patients with obstructive jaundice. Materıals and MethodsIn a prospective study, 47 patients with obstructive jaundice secondary to malignant lesions were evaluated before, at the fifth hour after, and on the fifth day after PTBD for neopterin, nitrate, tumor necrosis factor (TNF)-α, interleukin (IL)-6, IL-10, CRP levels, and liver function. ResultsNeopterin levels on day 5 after PTBD were significantly higher than the levels before treatment and at the fifth hour. However, nitrate, cytokine, white blood cell, albumin, and creatinine levels were not significantly different. On the fifth day after PTBD, CRP levels were significantly higher and total bilirubin, direct bilirubin, alkaline phosphatase, aspartate transaminase, and alanine transaminase values were lower than the before-treatment values. Seven patients (15%) died within 30 days after drainage. On the fifth day after PTBD, neopterin, IL-6, IL-10, and creatinine levels were significantly higher and albumin levels were lower in the early mortality group. The performance characteristics of neopterin and creatinine were statistically significant in predicting mortality. ConclusionsNeopterin levels increased after PTBD, indicating cellular immune activation. The nonsignificant change in cytokine levels may be related to low enduring release in malignancy. The extremely elevated levels of neopterin and creatinine after PTBD might serve as harbingers of early death in patients with cholestasis secondary to malignant lesions.

Whole inactivated equine influenza vaccine: Efficacy against a representative clade 2 equine influenza virus, IFNgamma synthesis and duration of humoral immunity

Equine influenza (EI) is a serious respiratory disease of horses induced by the equine influenza virus (EIV). Surveillance, quarantine procedures and vaccination are widely used to prevent or to contain the disease. This study aimed to further characterise the immune response induced by a non-updated inactivated EI and tetanus vaccine, including protection against a representative EIV isolate of the Florida clade 2 sublineage. Seven ponies were vaccinated twice with Duvaxyn IE-T Plus at an interval of four weeks. Five ponies remained unvaccinated. All ponies were experimentally infected with the EIV strain A/eq/Richmond/1/07 two weeks after the second vaccination. Clinical signs of disease were recorded and virus shedding was measured after experimental infection. Antibody response and EIV-specific IFNgamma synthesis, a marker of cell-mediated immunity, were measured at different time points of the study. Vaccination resulted in significant protection against clinical signs of disease induced by A/eq/Richmond/1/07 and reduced virus shedding when challenged at the peak of immunity. Antigenic drift has been shown to reduce protection against EIV infection. Inclusion of a more recent and representative EIV vaccine strain, as recommended by the OIE expert surveillance panel on equine influenza vaccine, may maximise field protection. In addition, significant levels of EIV-specific IFNgamma synthesis by peripheral blood lymphocytes were detected in immunised ponies, which provided a first evidence of CMI stimulation after vaccination with a whole inactivated EIV. Duration of humoral response was also retrospectively investigated in 14 horses vaccinated under field condition and following the appropriate immunisation schedule, up to 599 days after first immunisation. This study revealed that most immunised horses maintained significant levels of cross-reactive SRH antibody for a prolonged period of time, but individual monitoring may be beneficial to identify poor vaccine responders.

Abstract 5416: Myeloid-derived suppressor cells (MDSC) are increased and correlated with immune suppression and chronic inflammation in patients with cancer

Abstract Chronic inflammation plays a key role in the progress of malignant diseases and has been reported to relate with immune suppression seen in patients with advanced diseases. MDSC: myeloid-derived suppressor cells found as a new type of immune suppressor cells are enhanced by inflammation and have been reported to be present in blood circulation, lymphnodes and tumor tissues in patients with cancer. The suppression of immune function via depleting arginine and producing immune-suppressing cytokines including IL-10 has also been reported. In order to investigate MDSC in patients, peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers, 53 patients with gastrointestinal cancer, 65 with breast cancer, 8 with thyroid cancer, and 5 with ovarian cancer. These cells were used for the detection of MDSC (CD11b+CD14−CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI) and for the production assay of cytokines including IL-6 and IL-10. MDSC (%PBMC) of whole patients (5.68 + 5.92%), those of gastrointestinal cancer patients (3.76 + 4.90%) and those of breast cancer patients (2.92 + 4.13%) were significantly higher than those of normal volunteers (p&amp;lt;0.005, p=0.05, p&amp;lt;0.05, respectively, vs 1.59 + 1.08%). MDSC were inversely correlated with the SI (r=−0.271, p&amp;lt;0.05) in patients with gastrointestinal cancer and positively correlated with IL-6 production in whole patients(r=0.287, p=0.05) and with IL-10 production in patients with breast cancer (r=0.363, p&amp;lt;0.05). They were also significantly correlated to neutrophil counts (r=0.287, p&amp;lt;0.05) and neutrophil/lymphocyte ratios (NLR, r=0.623, p=0.05), and inversely correlated to lymphocyte counts (r=−0.251, p&amp;lt;0.01). Thus MDSC were successfully detected in peripheral blood and were significantly higher in patients with cancer. There were strong relationships among the inhibition of cell-mediated immunity, production of type 2 cytokines and inflammatory markers such as NLR. Therefore, MDSC seemed to work as a suppressor of immune reaction and may be an important marker of chronic inflammation in patients with cancer. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 5416. doi:1538-7445.AM2012-5416

Study of Adenosine Deaminase Activity As A Biochemical Marker of Cell Mediated Immunity in Tuberculous Meningitis, Tuberculous Pleural Effusion and Tuberculous Ascites

The present study conducted with the aim that ADA estimation is a simple cheaper and quicker biochemical test and could provide additional supportive evidence for the diagnosis of tuberculous meningitis, tuberculous pleural effusion, tuberculous ascites in clinically suspected cases and will therefore help in early institution of therapy to these patients. The data was collected and interpreted at department of Physiology and Department of Medicine Gandhi Medical College, Bhopal.DOI: http://dx.doi.org/10.3329/jom.v13i1.9065JOM 2012; 13(1): 32-38

The protective effects of ozone therapy in a rat model of acetaminophen-induced liver injury

ObjectivesAcetaminophen (APAP) overdose may cause acute liver injury. Ozone therapy (OT) is shown to reduce inflammation and necrosis in several entities. Thus, we have designed this study to evaluate the efficacy of OT in a rat model of APAP-induced liver injury. MethodsTwenty-seven Sprague-Dawley rats were divided into three groups: sham, APAP and APAP+OT groups. In the APAP and the APAP+OT groups, liver injury was induced by oral administration of 1g/kg APAP. The APAP+OT group received a single dose ozone/oxygen mixture (0.7mg/kg) intraperitoneally 1h after APAP administration. All animals were killed at 24 hour after APAP administration. Blood samples and liver tissues were harvested to determine liver injury and oxidative stress parameters. Liver tissues and blood samples were obtained for biochemical and histopathological analyses. ResultsAPAP administration caused necrosis in the liver after 24h. The degrees of liver necrosis of the APAP group were higher than the other groups (in both p<0.05, respectively). In the APAP+OT group, liver antioxidant enzymes activities were significantly higher than the APAP group (p<0.05), but were lower than the sham group (p<0.05). In the sham group, serum neopterin, a marker of cell-mediated immunity, concentrations (4.8±1.2 nmol/L) were lower than the APAP (14.7±1.4nmol/L) and APAP+OT groups (7.5±2.4nmol/L) (in both p<0.05, respectively). ConclusionOur results showed that OT prevented liver necrosis in rats and reduced neopterin levels. These findings suggest that the use of OT as an adjuvant therapy which might improve the outcome in APAP induced liver injury.

Relationship of myeloid-derived suppressor cells (MDSC) and immune suppression, nutritional impairment, and inflammatory markers in patients with gastrointestinal cancer.

675 Background: A suppression of cell mediated immunity and malnutrition are commonly seen in patients with advanced cancer and it has been reported that chronic inflammation plays a key role in induction of these conditions. MDSC: myeloid-derived suppressor cells, found as a new type of immune suppressor cells that is closely related to chronic inflammation, have reported to be present in blood circulation in patients with cancer. MDSC have been reported to suppress immune function through several pathways. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 18 normal healthy volunteers, and 53 patients with gastrointestinal cancer including 5 patients with esophageal, 16 with gastric, 26 with colorectal, 3 with bile duct, 1 with pancreatic and 1 with hepatocellular carcinomas, and these cells were used for the detection of MDSC (CD11b+CD14-CD33+) by flow cytometry. PBMC was also used for the PHA-blastogenesis of lymphocytes which is a marker of cell mediated immunity (stimulation indices: SI). Results: MDSC(%PBMC) of whole patients with gastrointestinal cancer, those with gastric cancer and those with colorectal cancer were higher than those of healthy volunteer (3.76+4.90%, 5.18+1.86%, 4.21+1.43% vs 1.59+1.08%, p&lt;0.05, p&lt;0.05, p&lt;0.10, respectively). MDSC of patients with gastrointestinal cancer were also inversely correlated to the SI (r=−0.271, p&lt;0.05) and to the serum concentrations of total protein (r=−0.490, p&lt;0.005). It also tended to correlate to neutrophil counts (r=0.287, p&lt;0.10) and neutrophil/lymphocyte ratios (NLR, r=0.623, p=0.10), and inversely did to lymphocyte counts (r=−0.251, p&lt;0.10).Thus MDSC was successfully detected in peripheral blood and was significantly higher in patients with gastrointestinal cancer. It was related to the inhibition of cell-mediated immunity, hypoaproteinemia and inflammatory markers such as NLR. Conclusions: MDSC seemed to work in the mechanisms of suppression of immune reaction and malnutrition, typically seen in cancer cachexia, in patients with gastrointestinal cancer and may be important as a marker of advancement of malignant diseases.