Cytokines as Immunological Markers for Follow up of Disease Activity During the Treatment of Pulmonary Tuberculosis

Background: Cytokines play a major role in protection against Mycobacterium tuberculosis infection and regulate the immune responses at a cellular level. Cytokine profile determines clinical outcome of the disease and responses to treatment as well. A T helper 1 (Th1) cytokine interferon gamma (IFN-U) is one of the most important cytokines which activate the macrophages to produce tumor necrosis factor-alpha (TNF-V). Excessive production of TNF-V have been implicated in immunopathogenesis of tuberculosis. A T helper 2 (Th2) response leads to release of IL-4, and IL-10 promoting an anti-inflammatory macrophage response. Interleukin-4(IL-4), has been implicated to down-regulate IFN-U, and thus has a harmful effect on TB patients. IL10 cytokine has thecapacity to inhibit Th1 activation and thus terminates cell mediated immune responses.Objective:The objective of the present study was to determine Th1 and Th2 cytokine profile in patients with tuberculosis to identify immunological marker for follow up of the disease activity, and to study the outcome of treatment.Methods: To examine this, blood samples were collected from newly diagnosed HIV negative pulmonary tuberculosis patients and from apparently healthy individuals as controls following an informed consent. Blood samples were as well collected at several intervals during the treatment with anti-tuberculosis drugs. Levels of IFN-U, TNF-V, IL-4 and IL-10 were measured pre and during treatment using commercial available enzyme-linked immune-sorbent assay (ELISA). Data were analyzed using SPSS 20. Receiver Operating Characteristic (ROC) Curve analysis has beencarried out to assess their discriminative power and to determine cut-off values. Analysis has been carried out further by calculating other measures of diagnostic test accuracy.Results: The median serum level of IL-4 was 20 and 35 pg/ml higher in new cases (untreated patients) and in patients under treatment with oral anti-tuberculosis, respectively, compared with that of controls (p=001). Levels of TNF- were significantly increased in patients before and afterthe treatment than those in control (p=0.001). New cases had the highest median level (10pg/ml) followed by those under treatment group (6pg/ml). Levels for IFN-U were not statistically different between patients and controls (p=0.351). Median levels of IL10 were similar in both controls and new cases groups (35pg/ml), but lower in patients under treatment group (20pg/ml). Increase in levels of IL-4 during treatment showed that Th2 immune responses still present and may indicate active disease and thus IL4 cytokine may be a possible marker for the disease activity.Conclusion: serum levels of TNF- in TB patients is useful in the evaluation of the disease activity during therapy, not replacing clinical parameters of disease activity in TB. Similar to TNF-, IL-4 can also be used as marker for TB severity. On the other hand IL-4test can be used to diagnose TBin highly exposed suspects where a positive result is more likely to indicate TB.Keywords: Tuberculosis; Cytokines; Immunological markers

Diagnostic value of Autoantibodies to GAD65 and IA-2 in Patients with Latent Autoimmune Diabetes in Adult (LADA)

Background: Latent autoimmune diabetes in adults (LADA) accounts for11 % of all cases of diabetes and often misdiagnosed as type 2diabetes. LADA resembles type 1diabetes and shares common physiological characteristics of type 1 but it does not affect children and has beenclassified distinctly as being separate from juvenile diabetes.  Autoantibodies against glutamic acid decarboxylase 65 (GADA) and tyrosine phosphatase (IA-2) are found frequently in patients withLADA. The presence of these autoantibodies in LADA predicts inevitable M cell failure and poor response to oral hypoglycemic therapy i.e., patients with LADA do not respond to oral hypoglycemic therapy.Objective: To determine an immunological marker to diagnose patients not responding to oral hypoglycemic therapy.Patients and methods: A facility-based cross sectional study was  conducted in Jabbir Abu Eliz Diabetes Center, located at Khartoum 2. Venous blood samples were obtained from the study patients. They were divided into three groups, group1 included 27 diabetic patients treated withinsulin, group2 included 15 diabetic patients of type 2 diabetes as controls, and group3 included 15 newly diagnosed patients older than 35 years at onset of diabetes. A standardized pre-tested administered questionnaire was used for data collection and the collected data were analyzed.Results: Males encountered in the study were 28 (49.1%). On patient recently diagnosed to have type 2 diabetes mellitus (T2DM) was positive for autoantibodies to GDA/IA-2. These autoantibodies were also positive in 15 patients with diabetes mellitus type 1 (T1DM)Conclusions: Autoimmune diagnostics is of particular importance in adults to discriminate between type 1 and type 2 diabetes and to assess the diagnosis of latent autoimmune diabetes in adults. The current study results revealed that autoantibodies to GAD/IA-2 are good marker for diagnosis of latent onset DM type 1. On the other hand, data indicate that the vast majority of cases of type 1 diabetes may be considered as immune-mediated, that multiple autoantibody to GAD/IA-2 analysisare of prognostic value to predict complications e.g., retinopathy. The current study recommends using of anti-GAD/IA-2 antibodies as marker for diagnosis of latent autoimmune diabetes in adults (LADA) who are not responding to oral hypoglycemic and may be at risk for getting complications. On the other hand, the study recommends using of anti-GAD/IA-2 antibodies for prognosis of the clinical progression of diabetes type 1 for prediction of insulin dependence

Association between Diabetes Mellitus type 1 and Celiac Disease: case-control study among Sudanese patients 2009-2011

Background: Gluten sensitive enteropathy (celiac disease (CD)) has a strong association with diabetes mellitus (type 1DM). Since, 2-3% of CD patients have selective IgA deficiency, the majority of the available tests may fail to show the auto-antibodies (the IgA endomysial antibody(EMA). To prevent such a false negativity, a new Enzyme Linked Immune Sorbent Assay (ELISA) test has been introduced to detect both IgG and IgA antibodies reactive with tissue transglutaminase (tTG), an autoantigen in CD patients.Objectives: This study has been conducted to detect celiac disease among Sudanese patients with type 1 autoimmune diabetes using anti-tissue transglutinamase antibodies as a diagnostic tool.Patients and Methods: Samples were collected from sixty nine randomly selected patients (38 males and 31 females) and their age ranged between 3-22 years with DM type 1 who were attending the outpatient clinics in Gabir Abu Eliz diabetic Center and Omdurman Pediatric EmergencyHospital. Blood samples were collected from 25 healthy individuals as controls. Levels of tTG specific IgA, tTG specific IgG and anti-endomysial antibodies of IgA class were measured in sera collected from both cases and from controls. All the results were analyzed using StatisticalPackages of Social Sciences (SPSS) version 17 and MicroSoft office excel.Results: Seven out of 69 patients with DM type 1 (10.1%) were identified as having CD using IgG anti-tTG and 5 (7.2%) of them were positive for IgA anti-tTG and IgA anti-endomysial antibodies. The mean of both anti-tTG IgA and IgG titers were higher in diabetic patients (M±SD = 12.30±41.0 and 7.2±13.1 respectively) when compared with the control group (M±SD =1.8±1.1 and 1.8±0.9 respectively), however, only anti-tTG IgG antibodies titer achieved statistical significance.Discussion and conclusion: The present study revealed that patients with DM type I have an increased tendency to develop CD. The increased association of CD and selective IgA deficiency is a potential source of false-negative IgA, therefore testing for IgG class autoantibodies isrecommended if celiac disease is suspected. Antibodies to tTG antigen fall once a gluten-free diet has begun, thus facilitating monitoring of dietary compliance. Thus, anti-tTG antibodies are highly sensitive marker for celiac disease with 95- 100 % sensitivty, and specificity of 90 to 97 %.Keywords: Diabetes mellitus type I; Celiac disease and diabetes mellitus type I association; IgG anti-tissue transglutaminase antibodies; IgA anti-endomysial antibodie

The use of impregnated curtains does not affect antibody responses against Plasmodium falciparum and complexity of infecting parasite populations in children from Burkina Faso

Abstract In Burkina Faso, where malaria is hyper-endemic and transmission intensity is very high, the majority of malaria-related morbidity and mortality occurs in children less than 5 years of age. A control measure such as the use of insecticide-treated curtains (ITC) significantly reduces transmission of malaria infection. Concerns remain whether reduced transmission intensity may lead to a delay in the development of immunity in younger children and even to a partial loss of already acquired immunity. In this study, the levels of P. falciparum-specific IgG subclasses, the number of infecting parasite clones determined by PCR-based genotyping of the msp2 gene and the parasite density were analysed in 154 asymptomatic children (3–6 years) living in 16 villages (8 with and 8 without ITC) in the vicinity of Ouagadougou, the capital of Burkina Faso. In addition, the parasite inhibitory effects of Ig fractions, prepared from selected children, in co-operation with normal human monocytes were studied. Blood samples from asymptomatic ITC-users showed a significant decrease in P. falciparum prevalence as well as in parasite density. However, no significant difference was observed in P. falciparum-specific antibodies or in parasite multiplicity of infection between the two groups. Furthermore, Ig fractions from children of both groups showed similar levels of inhibitory activity against autologous parasite growth both on their own and in co-operation with monocytes. © 2004 Elsevier B.V. All rights reserved. Keywords: Wild isolate of P. falciparum; Insecticide-treated curtains; Immunit

Comparative Quantification of Arterial Lipid by Intravascular Photoacoustic-Ultrasound Imaging and Near-Infrared Spectroscopy-Intravascular Ultrasound

Intravascular photoacoustic-ultrasound (IVPA-US) imaging and near-infrared spectroscopy-intravascular ultrasound (NIRS-IVUS) are two hybrid modalities that detect arterial lipid, with comparison necessary to understand the relative advantages of each. We performed in vivo and ex vivo IVPA-US imaging of the iliac arteries of Ossabaw swine with metabolic syndrome (MetS) and lean swine to investigate sensitivity for early-stage atherosclerosis. We repeated imaging ex vivo with NIRS-IVUS for comparison to IVPA-US and histology. Both modalities showed significantly greater lipid in MetS vs. lean swine, but only IVPA-US localized the lipid as perivascular. To investigate late-stage atherosclerosis, we performed ex vivo IVPA-US imaging of a human coronary artery with comparison to NIRS-IVUS and histology. Two advanced fibroatheromas were identified, with agreement between IVPA-measured lipid area and NIRS-derived lipid content. As confirmed histologically, IVPA-US has sensitivity to detect lipid content similar to NIRS-IVUS and provides additional depth resolution, enabling quantification and localization of lipid cores within plaques

Changes in the levels of cytokines, chemokines and malaria-specific antibodies in response to Plasmodium falciparum infection in children living in sympatry in Mali

<p>Abstract</p> <p>Background</p> <p>The Fulani are known to be less susceptible to <it>Plasmodium falciparum </it>malaria as reflected by lower parasitaemia and fewer clinical symptoms than other sympatric ethnic groups. So far most studies in these groups have been performed on adults, which is why little is known about these responses in children. This study was designed to provide more information on this gap.</p> <p>Methods</p> <p>Circulating inflammatory factors and antibody levels in children from the Fulani and Dogon ethnic groups were measured. The inflammatory cytokines; interleukin (IL)-1beta, IL-6, IL-8, IL-10, IL-12p70, tumor necrosis factor (TNF) and the chemokines; regulated on activation normal T cell expressed and secreted (RANTES), monokine-induced by IFN-gamma (MIG), monocyte chemotactic protein (MCP)-1 and IFN-gamma-inducible protein (IP)-10 were measured by cytometric bead arrays. The levels of interferon (IFN)-alpha, IFN-gamma and malaria-specific antibodies; immunoglobulin (Ig) G, IgM and IgG subclasses (IgG1-IgG4) were measured by ELISA.</p> <p>Results</p> <p>The results revealed that the Fulani children had higher levels of all tested cytokines compared to the Dogon, in particular IFN-gamma, a cytokine known to be involved in parasite clearance. Out of all the tested chemokines, only MCP-1 was increased in the Fulani compared to the Dogon. When dividing the children into infected and uninfected individuals, infected Dogon had significantly lower levels of RANTES compared to their uninfected peers, and significantly higher levels of MIG and IP-10 as well as MCP-1, although the latter did not reach statistical significance. In contrast, such patterns were not seen in the infected Fulani children and their chemokine levels remained unchanged upon infection compared to uninfected counterparts. Furthermore, the Fulani also had higher titres of malaria-specific IgG and IgM as well as IgG1-3 subclasses compared to the Dogon.</p> <p>Conclusions</p> <p>Taken together, this study demonstrates, in accordance with previous work, that Fulani children mount a stronger inflammatory and antibody response against <it>P. falciparum </it>parasites compared to the Dogon and that these differences are evident already at an early age. The inflammatory responses in the Fulani were not influenced by an active infection which could explain why less clinical symptoms are seen in this group.</p

Usefulness and limitations of transthoracic echocardiography in heart transplantation recipients

Transthoracic echocardiography is a primary non-invasive modality for investigation of heart transplant recipients. It is a versatile tool which provides comprehensive information about cardiac structure and function. Echocardiographic examinations can be easily performed at the bedside and serially repeated without any patient's discomfort. This review highlights the usefulness of Doppler echocardiography in the assessment of left ventricular and right ventricular systolic and diastolic function, of left ventricular mass, valvular heart disease, pulmonary arterial hypertension and pericardial effusion in heart transplant recipients. The main experiences performed by either standard Doppler echocardiography and new high-tech ultrasound technologies are summarised, pointing out advantages and limitations of the described techniques in diagnosing acute allograft rejection and cardiac graft vasculopathy. Despite the sustained efforts of echocardiographic technique in predicting the biopsy state, endocardial myocardial biopsies are still regarded as the gold standard for detection of acute allograft rejection. Conversely, stress echocardiography is able to identify accurately cardiac graft vasculopathy and has a recognised prognostic in this clinical setting. A normal stress-echo justifies postponement of invasive studies. Another use of transthoracic echocardiography is the monitorisation and the visualisation of the catheter during the performance of endomyocardial biopsy. Bedside stress echocardiography is even useful to select appropriately heart donors with brain death. The ultrasound monitoring is simple and effective for monitoring a safe performance of biopsy procedures

Antibody-Mediated Growth Inhibition of Plasmodium falciparum: Relationship to Age and Protection from Parasitemia in Kenyan Children and Adults

BACKGROUND: Antibodies that impair Plasmodium falciparum merozoite invasion and intraerythrocytic development are one of several mechanisms that mediate naturally acquired immunity to malaria. Attempts to correlate anti-malaria antibodies with risk of infection and morbidity have yielded inconsistent results. Growth inhibition assays (GIA) offer a convenient method to quantify functional antibody activity against blood stage malaria. METHODS: A treatment-time-to-infection study was conducted over 12-weeks in a malaria holoendemic area of Kenya. Plasma collected from healthy individuals (98 children and 99 adults) before artemether-lumefantrine treatment was tested by GIA in three separate laboratories. RESULTS: Median GIA levels varied with P. falciparum line (D10, 8.8%; 3D7, 34.9%; FVO, 51.4% inhibition). The magnitude of growth inhibition decreased with age in all P. falciparum lines tested with the highest median levels among children \u3c4 years compared to adults (e.g. 3D7, 45.4% vs. 30.0% respectively, p = 0.0003). Time-to-infection measured by weekly blood smears was significantly associated with level of GIA controlling for age. Upper quartile inhibition activity was associated with less risk of infection compared to individuals with lower levels (e.g. 3D7, hazard ratio = 1.535, 95% CI = 1.012-2.329; p = 0.0438). Various GIA methodologies had little effect on measured parasite growth inhibition. CONCLUSION: Plasma antibody-mediated growth inhibition of blood stage P. falciparum decreases with age in residents of a malaria holoendemic area. Growth inhibition assay may be a useful surrogate of protection against infection when outcome is controlled for age

Functional Memory B Cells and Long-Lived Plasma Cells Are Generated after a Single Plasmodium chabaudi Infection in Mice

Antibodies have long been shown to play a critical role in naturally acquired immunity to malaria, but it has been suggested that Plasmodium-specific antibodies in humans may not be long lived. The cellular mechanisms underlying B cell and antibody responses are difficult to study in human infections; therefore, we have investigated the kinetics, duration and characteristics of the Plasmodium-specific memory B cell response in an infection of P. chabaudi in mice. Memory B cells and plasma cells specific for the C-terminal region of Merozoite Surface Protein 1 were detectable for more than eight months following primary infection. Furthermore, a classical memory response comprised predominantly of the T-cell dependent isotypes IgG2c, IgG2b and IgG1 was elicited upon rechallenge with the homologous parasite, confirming the generation of functional memory B cells. Using cyclophosphamide treatment to discriminate between long-lived and short-lived plasma cells, we demonstrated long-lived cells secreting Plasmodium-specific IgG in both bone marrow and in spleens of infected mice. The presence of these long-lived cells was independent of the presence of chronic infection, as removal of parasites with anti-malarial drugs had no impact on their numbers. Thus, in this model of malaria, both functional Plasmodium-specific memory B cells and long-lived plasma cells can be generated, suggesting that defects in generating these cell populations may not be the reason for generating short-lived antibody responses