Immunoglobulin Markers Research Articles

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

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

Raman spectroscopy of lymphocytes from patients with the Epstein–Barr virus infection

In this study, Raman spectroscopy is applied to trace lymphocytes activation following contact with the Epstein–Barr virus (EBV) of the herpesvirus family. The biomarker of cell activation is found to be the 520 cm−1 band, indicating formation of immunoglobulins. The blood samples are obtained from patients diagnosed with infectious mononucleosis and treated at the University Hospital in Kraków. The lymphocytes’ Raman spectra are collected using a mapping technique, exciting samples with a 514.5 nm line of Ar + laser. Measurements are performed on the 1st, 4th, 6th, 12th and 30th day of hospitalization, until the patient has recovered. The highest intensity of the immunoglobulin marker is observed on the 4th day of hospitalization, while the results of the blood count of patients show the greatest increase in the number of lymphocytes at the beginning of hospitalization. No activated lymphocytes were observed in the blood of healthy volunteers. Some information is provided by the evaluation of B-cell activation by estimating the activated areas in the cells, which are determined by the presence of the Ig marker. The 900 cm−1 band and band around 1450 cm−1 are also analyzed as markers of the presence of the latent membrane protein, LMP2A (and 2B), of the EBV viral protein. The anomalous degree of depolarization observed in B-cells in the course of EBV infection appears to be due to the influence of a virus protein, disrupting BCR signal transduction.

Concordance rate between oligoclonal bands and the Kappa index in patients with suspected multiple sclerosis (MS).

Oligoclonal bands (OCBs) and Kappa free light chains (FLCs) in the cerebrospinal fluid (CSF) are sensitive markers of intrathecal immunoglobulin (Ig)G synthesis in patients with multiple sclerosis. To evaluate the concordance rate between OCBCs and the Kappa index (KI) in patients with suspected multiple sclerosis (MS). Patients with suspected MS were referred to a specialized CSF laboratory as part of their diagnostic investigation. Paired CSF and serum samples were collected and submitted to detection of OCBs and determination of the KI. Positive and negative results were determined with both methods, and the percentage of agreement between them was established. In total, 171 serum and CSF samples from 171 patients were included in the analysis. The mean age of the patients was of 40 ± 14.2 years; 18.9% of them were male, and 81.1% were female. The OCBs and KI presented concordant results in 161 (94.2%) samples: in 74 (43.3%), both were positive, and in 87 (50.9%), both were negative. In 10 cases, the results were discrepant: KI positive/OCB negative in 8 and OCB positive/KI negative in 2 cases. The KI and OCBs presented high concordance level. Currently, the detection of OCBs in the CSF is the standard method for MS diagnosis, but it is time-consuming, and its visual interpretation can be difficult. The results suggest that the KI is a good alternative for the detection of intrathecal immunoproduction in cases of suspected MS.

Assessment of serum concentration and urinary excretion of tumor necrosis factor receptor 1 and 2 and their potential as markers of immunoglobulin A nephropathy activity.

Tumor necrosis factor receptor 1 (TNFR1) and 2 (TNFR2) can be cleaved from the cell surface and circulate alone or in combination with tumor necrosis factor alpha (TNF-α). These soluble receptors may play a key role in regulating the inflammatory response. The study aimed to evaluate the role of TNFRs in regulating the inflammatory response in immunoglobulin A nephropathy (IgAN). The study included 26 patients with newly diagnosed and biopsy-confirmed IgAN and 20 healthy controls. Study material included blood and fresh urine collected the morning before kidney biopsy and therapy. The serum concentrations of TNFR1 (STNFR1) and TNFR2 (STNFR2) and urinary excretion of TNFR1 (UTNFR1) and TNFR2 (UTNFR2) were determined with immunoassay. Subsequently, the data were evaluated statistically. The STNFR1 and STNFR2 levels were higher in IgAN patients than in healthy subjects (4747.87 pg/mL and 2817.62 pg/mL compared to 2755.68 pg/mL (95% CI: from -2948.41 to -1035.97; p = 0.001) and 1437.83 pg/mL (95% CI: from -1958.50 to -419.60; p = 0.001). The power of the test was 98.5% for STNFR1 and 96% for STNFR2. Urinary concentrations only increased for TNFR1 (3551.29 compared to 2338.95 pg/mg of creatinine (Cr) (95% CI: from -2247.03 to -177.66; p = 0.023). The STNFR1 marker was characterized by a sensitivity of 73.08% and a specificity of 90.00% (p < 0.001). Our results suggest that TNFR1 and TNFR2 are good markers of TNF-α pathway activation in IgAN patients.

Humoral Inflammatory Markers of Total Immunoglobulin E (IgE) Exposure on Palm Oil Plantation Pesticide Sprayers

Pesticides are often used in oil palm plantations to prevent disturbing animals and plants. Pesticide sprayers are vulnerable to pesticide exposure which can reduce cholinesterase levels is an indicator of pesticide poisoning. Long-term pesticide exposure and excessive doses cause the production of free radicals in the body, thereby increasing inflammatory markers, one of which is total IgE. This study aimed to describe cholinesterase and total IgE levels in oil palm farmers. This study used the quantitative-descriptive research method. The sampling was conducted at PT. X in Sanggau District of West Kalimantan Province. Examining cholinesterase levels used the Architec c-8000 device with the DGKC butyrylthiocholine 37°C method, while total IgE used the immunochemmiluminescent method with the Immulite 2000 device. The examination was carried out at the Prodia National Reference Lab in Jakarta. Based on the cholinesterase measurement results taken from 133 workers who underwent medical check-ups, 20 data were taken that were close to the threshold value less than normal to examine total IgE levels. The results obtained from the overview of cholinesterase levels showed that 20 pesticide sprayers (100%) had normal cholinesterase levels, and four (20%) had normal total IgE levels. Pesticide exposure will decrease cholinesterase levels, and increase IgE levels.

Amyloid beta-related angiitis in a patient with light chain systemic Amyloidosis: A rare co-morbidity. (P9-5.013)

<h3>Objective:</h3> To report a rare comorbidity of amyloid beta-related angiitis (ABRA) and systemic light chain amyloidosis (AL) <h3>Background:</h3> ABRA is a rare central nervous system (CNS) disorder with overlapping features of primary angiitis of the CNS and cerebral amyloid angiopathy. ABRA pathophysiology differs from extremely rare CNS involvement in AL. This is the first report of ABRA and AL comorbidity. <h3>Design/Methods:</h3> Case Report <h3>Results:</h3> A 73 year old Caucasian female with AL, prior autologous stem cell transplantation followed by relapse and daratumumab therapy resulting in complete response, presented with subacute development of headaches, global dysphasia and right sided homonymous hemianopia. Brain magnetic resonance imaging (MRI) demonstrated an increased T2/Fluid attenuated inversion recovery signal along the left parietal occipital cortex with mild restricted diffusion and microhemorrhages. Cerebrospinal fluid (CSF) studies showed: pleocytosis (30 nucleated cells/mm3) consisting of mature granulocytes, small mature lymphocytes and rare plasma cells, no monotypic B-cell, overtly aberrant T-cell, or expanded large granular lymphocyte population, no malignant cells; hyperproteinorachia (214 mg/dL); and no markers of intrathecal immunoglobulin G synthesis. Infectious CSF studies, CSF autoimmune encephalopathy panel and rheumatological work up were unremarkable. Steroid treatment (methylprednisolone, 1000 mg/day for 5 days intravenously, followed by a slow oral taper) led to a resolution in visual field deficit and a significant improvement in speech patterns. New microhemorrhages with new foci of restricted diffusion in the left posterior parietal/occipital region and posterior right centrum semiovale were shown on brain MRI three months later. Brain biopsy was performed with no neurological complications and pathology was consistent with ABRA. Liquid chromatography-tandem mass spectrometry of the brain tissue detected a peptide profile consistent with amyloid-beta precursor type amyloid deposition. The patient continued on oral steroid taper and is starting cyclophosphamide. <h3>Conclusions:</h3> Although ABRA is pathophysiologically distinct from and unrelated to AL, those two conditions may rarely co-exist. <b>Disclosure:</b> Dr. Javed has nothing to disclose. The institution of Dr. Jensen has received research support from ASCO / Conquer Cancer Foundation. The institution of Dr. Jensen has received research support from AHRQ. Dr. A Tuchman has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Shattuck Labs. Dr. A Tuchman has received personal compensation in the range of $500-$4,999 for serving as a Consultant for Janssen. An immediate family member of Dr. Allenzara has received personal compensation for serving as an employee of Atai . An immediate family member of Dr. Allenzara has stock in Atai company stock . An immediate family member of Dr. Ishizawar has received personal compensation for serving as an employee of EMD MilliporeSigma. Dr. Cho has nothing to disclose. Dr. Yap has nothing to disclose. The institution of Dr. Dujmovic Basuroski has received research support from Alexion Pharmaceuticals, Inc. The institution of Dr. Dujmovic Basuroski has received research support from Biogen MA Inc. The institution of Dr. Dujmovic Basuroski has received research support from EMD Serono Research and Development Institute, Inc . The institution of Dr. Dujmovic Basuroski has received research support from Celgene Corporation/Bristol-Myers Squibb. The institution of Dr. Dujmovic Basuroski has received research support from The Bodford Family Transverse Myelitis Center Fund.

GM Allotypes and COVID-19. A Pilot Study Performed on Sicilian Patients.

Several studies suggest that genetic variants that influence the onset, maintenance and resolution of the immune response might be fundamental in predicting the evolution of COVID-19. In the present paper, we analysed the distribution of GM allotypes (the genetic markers of immunoglobulin γ chains) in symptomatic and asymptomatic COVID-19 patients and in healthy controls, all born and residing in Sicily. Indeed, the role played by GM allotypes in immune responses and infection control is well known. Our findings show that the GM23 allotype is significantly reduced in healthy controls. Interestingly, in a previous study, Sicilians carrying the GM23 allotype were associated with the risk of developing a symptomatic Human Cytomegalovirus infection. However, a note of caution should be considered, due to the small sample size of patients and controls.

Glomerular C4d deposition in proliferative glomerular diseases.

The aim of this study was to evaluate the immunohistochemical expression of C4d in native renal biopsies of proliferative glomerular diseases, complement pathways in these diseases, and assess the relationship of C4d with histological and clinicopathological parameters, other complement proteins, and immunoglobulin markers. This cross-sectional study was conducted during the year 2018-19 involving 107 native renal biopsies with histologically diagnosed cases of proliferative glomerular diseases. C4d immunohistochemical evaluation of renal tissue sections was performed using polyclonal antihuman C4d as the primary antibody. Patients were classified as positive and negative groups based on their glomerular C4d deposition. The overall prevalence of C4d positivity was 80.4% in proliferative glomerular diseases ranging between 60.0% in C3 glomerulonephritis to 92.9% in membranoproliferative glomerulonephritis. Mixed capillary and mesangial deposition were noted in all cases of proliferative glomerulonephritis. Classical pathway was dominantly involved in all glomerular diseases except C3 glomerulonephritis and IgA nephropathy. Multivariate logistic regression analysis revealed that glomerular IgG staining (aOR: 5.86, 95% CI: 1.26-27.14) and IgM staining (aOR: 3.90, 95%CI: 1.07-14.18) were significantly associated with C4d positivity. C4d staining along with immunoglobulin markers such as IgG and IgM and complement proteins can be useful in delineating different complement activation pathways in glomerular diseases and understanding the disease pathogenesis.

Advances in Neonatal Infections.

Despite continued advances and developments in neonatal medicine, neonatal sepsis is the third leading cause of neonatal mortality and a major public health problem, especially in developing countries. Sepsis accounts for mortality for almost 50% of global children under 5 years of age.Over the past 50 years, there have been many advances in the diagnosis, prevention, and treatment of neonatal infections. The diagnostic advances include better culture techniques that permit more rapid confirmation of the diagnosis, advent of polymerase chain reaction (PCR) to rapidly diagnose viral infections, use of biologic markers indicating evidence of infection, and a better understanding of immunoglobulin markers of infection. From a therapeutic stand point, there have been a variety of antibiotics, antifungals, and antiviral agents, better approaches to prevent sepsis, specific immunotherapy, for example, respiratory syncytial virus (RSV); bundled approach to prevention of deep-line infection and better antibiotic stewardship, leading to earlier discontinuation of antibiotic therapy.Hand hygiene remains the benchmark and gold standard for late-onset sepsis prevention. The challenge has been that each decade, newer resistant bacteria dominate as the cause of sepsis and newer viruses emerge, for example, human immunodeficiency virus, zika virus, and novel coronavirus disease 2019.Future treatment options might include stem cell therapy, other antimicrobial protein and peptides, and targeting of pattern recognition receptors in an effort to prevent and/or treat sepsis in this vulnerable population. Also, the microbiome of premature infants has a smaller proportion of beneficial bacteria and higher numbers of pathogenic bacteria compared with term infants, likely owing to higher frequencies of cesarean sections, antibiotic use, exposure to the hospital environment, and feeding nonhuman milk products. Modifying the microbiome with more mother's milk and shorter duration of antibiotics in noninfected babies should be a goal. KEY POINTS: · Neonatal sepsis remains a leading cause of mortality.. · Challenges include bacterial resistance and newer viruses.. · Future treatments may include newer antibiotics/antivirals and stem cell therapy..

Recombinant nodavirus vaccine produced in bacteria and administered without purification elicits humoral immunity and protects European sea bass against infection

Viral necrosis virus (NNV) or nodavirus causes fish viral encephalopathy and retinopathy worldwide. In some cases, mortalities in aquaculture industry can reach up to 100%, some species being especially sensitive as is the case of European sea bass (Dicentrarchus labrax), one of the main cultured species in the Mediterranean, with the consequent economical loses. Development of new vaccines against NNV is in the spotlight though few researches have focused in European sea bass. In this study we have generated a recombinant NNV (rNNV) vaccine produced in Escherichia coli expressing the capsid protein and administered it to European sea bass juveniles by two different routes (intraperitoneal and oral). The last being considered non-stressful and desired for fish farming of small fish, which in fact are the most affected by NNV. Oral vaccine was composed of feed pellets containing the recombinant whole bacteria, and injected vaccine was composed of recombinant bacteria previously lysed. Our results revealed production of specific anti-NNV IgM following the two vaccination procedures, levels that were further increased in orally-vaccinated group after challenge with NNV. Genes related to interferon (IFN), T-cell and immunoglobulin markers were scarcely regulated in head-kidney (HK), gut or brain. Vaccination by either route elicited a relative survival response of 100% after NNV challenge. To our knowledge, this is the first report of a recombinant vaccine followed by no purification steps which resulted in a complete protection in European sea bass when challenged with NNV.

GP.03 Immune deficiencies/dysregulations underpinning childhood limbic encephalitis: a case series and literature review

Background: Limbic encephalitis (LE) is a rare autoimmune syndrome affecting limbic system structures and causing variety of manifestations including memory changes, temporal epilepsies, and psychiatric symptoms. It is a rare disease in children but with a well-recognizable combination of clinical, neuroimaging and/or histological signature. Beyond the association with anti-neuronal auto-antibodies, no clear immune system phenotype has been associated with limbic encephalitis. Our aim is to characterize the clinical and paraclinical features of non-paraneoplasic limbic encephalitis and to correlate them with potential underlying immune deficiencies. Methods: Retrospective case series of seven patients with limbic encephalitis recruited at the Montreal Children’s Hospital (MCH) with a focus on the immune- and neuro-phenotypes, including anti-neuronal antibodies, lymphocyte sub-typing, key markers of immunoglobulin and complement systems. Literature review showed 77 cases of non-paraneoplastic non-NMDA limbic encephalitis. Results: Symptoms included temporal epilepsy (n=5), psychiatric symptoms such as ADHD or autistic symptoms (n=2), and memory changes (n=3). One patient was positive for both voltage gated potassium channel antibodies (VGKC) and anti-thyroid peroxidase antibodies (TPO) and two were positive only for anti TPO antibodies. One patient showed low CD19, and immunoglobulins. Three patients showed chronic low CD56 cell count. Conclusions: The study is still ongoing, but at least 3 patients already display some traits of immune dysregulation.

Serum IgD Levels in Allergic Children with Gastrointestinal Manifestations

A large number of studies investigating various biomarkers for allergy have been published over the past decades. To this date, no single or specific biomarker for allergy has been identified. Therefore, we examined markers of inflammation and immunoglobulins in children with allergic symptoms, among which children with GI allergy, in an attempt to find new and indicative markers for GI allergy. In the present study, total serum levels of immunoglobulin (Ig)D, CM-lgE, Ig-free light chain (Ig-fLC) kappa and lambda, thymic stromal lymphopoietin (TSLP), thymus and activation-regulated chemokine (TARC/CCL17) and galectin-9 were analyzed on 117 atopic infants and children, divided into different allergic manifestations: asthma (n=7), atopic dermatitis (n=56), eosinophilic esophagitis (n=20) and gastrointestinal (GI) allergy (n=34). Elevated levels of serum IgD, without the presence of IgE, was found in children with GI allergy. Serum IgD did not correlate with other laboratory parameters of atopy such as serum cow’s milk-lgE, Ig-fLC, TSLP, TARC and galectin-9. In conclusion, serum IgD is elevated in children with GI allergy, in the absence of IgE, and can be considered as a new indicative marker for GI allergy, however, its exact role in the pathogenesis of atopy awaits further studies.

Delta hepatitis: Toward improved diagnostics.

Delta hepatitis: Toward improved diagnostics.

Comparison of MRD Levels and Gene Expression Patterns in MLL-R Versus Non-MLL Infant ALL

Abstract Introduction Acute lymphoblastic leukaemia (ALL) in infants has poor overall survival despite being characterized by very few genetic aberrations per case. The most common genetic change, present in over 75% of cases, is the rearrangement of the mixed lineage leukaemia (MLL/KMT2A) gene (MLL-R) that also occurs in AML and mixed phenotype acute leukaemia (MPAL). Because infant ALL is rare and distinctive, most Australian and New Zealand patients are enrolled on specific clinical trials. In earlier infant ALL trials, MRD was not used for risk stratification firstly because of the difficulty of finding sensitive markers for specific immunoglobulin and T-cell receptor (Ig/TCR) gene rearrangements and the secondly because, in infant ALL, Ig/TCR markers are often present in sub-clones that can be lost at relapse due to clonal selection. MRD testing is performed in the current Interfant 06 trial preferentially using markers based on the genomic breakpoint sequences of MLL gene rearrangements. The treatment of infant ALL remains very challenging with relatively poor survival rates attributable to both toxicity and relapse. The objectives of this study were therefore to analyse MRD data for Interfant 06 patients enrolled at ANZCHOG centres and to perform a pilot experiment evaluating gene expression for key genes in infant ALL samples on a microfluidics platform, as a basis for identifying potential targeted therapies. Methods MRD was measured in bone marrow DNA from Interfant 06 patients enrolled since 2006, using sensitive Real-time Quantitative PCR (PCR-MRD) patient-specific assays to detect either MLL gene rearrangements (in 17) and/or conventional immunoglobulin and T-cell receptor (Ig/TCR) markers (21). Gene expression levels for 90 genes important in childhood cancers were measured by Taqman-based microfluidic assays in duplicate using cDNA from 2 micrograms of total RNA from 10 infant ALL samples (5 MLL, 5 non-MLL) at diagnosis (6) or relapse (4). Results Patient-specific PCR-MRD tests were developed for 27 out of 28 Australian and New Zealand infant ALL patients. 17/18 MLL-R ALL patients had MLL-R assays and 21/28 patients had Ig/TCR MRD tests, with only 1 (non-MLL) patient having no MRD markers. There was a wide range of MRD responses to induction therapy (Figure 1). Bone marrow MRD at the end of induction was high (&gt;1x10-3) in 44% of MLL-R ALL infants compared to 25% of non-MLL ALL infants and 15% of older children enrolled on ANZCHOG ALL8. In 8/13 MLL-R patients who had both types of marker, MRD levels were higher when measured by their MLL-R marker than by their Ig/TCR marker. In a set of 90 genes selected for expression analysis, higher levels were found for 17 genes in 2 or more of the 10 infant ALL samples evaluated. These more highly expressed genes included potential or known drug targets BCL2, ERBB2, ERBB4, ILRA2, CSF1R and PARP1. Conclusions The quantitation of MRD based on MLL rearrangements in ALL is effective and can also be used to monitor response to therapy in infant ALL as well as MLL-R cases of AML and MPAL. The combined application of MLL-R and Ig/TCR markers allowed 97% of infant ALL patients to be MRD monitored with a sensitive marker. In most patients with both type of MRD marker, higher levels of MRD were detected in end of induction samples using the MLL versus Ig/TCR tests. One interpretation is that Ig/TCR genes are rearranged after the MLL rearrangement in ALL sub-clones that are both more mature and more chemo-sensitive. This finding also confirms the current consensus that disease-related MLL-R markers provide better risk assessment than Ig/TCR markers. Our Fluidigm analysis has shown that quantitative measurement of multiple gene expressions is feasible on small RNA samples and can be used to rapidly screen for specific expression of genes coding for drug targets in ALL patients. Support: NHMRC Australia APP1057746, Sporting Chance Cancer Foundation. Figure 1. Comparison of MRD response to induction therapy in MLL-R infant ALL compared with non-MLL infant ALL (Interfant 06) and older children (ANZCHOG ALL8). Figure 1. Comparison of MRD response to induction therapy in MLL-R infant ALL compared with non-MLL infant ALL (Interfant 06) and older children (ANZCHOG ALL8). Disclosures No relevant conflicts of interest to declare.

Genetic markers of immunoglobulin G and immunity to cytomegalovirus in patients with breast cancer

Human cytomegalovirus (CMV), a ubiquitous herpesvirus, has been implicated in the etiology of breast cancer. It is clear that not all people exposed to CMV are equally likely to develop this malignancy, implying the presence of host genetic factors that might modulate the cancer-spurring properties of the virus. CMV has evolved sophisticated strategies for evading host immunosurveillance. One strategy involves encoding decoy Fcγ receptors (FcγR) that thwart the Fcγ-mediated effector functions, such as antibody-dependent cellular cytotoxicity. In this investigation, using an enzyme-linked immunosorbent assay (ELISA), we aimed to determine whether the decoy FcγR encoded by the CMV gene RL13 binds differentially to anti-CMV antibodies expressing different immunoglobulin GM (γ marker) allotypes, genetic markers of immunoglobulin G (IgG). Results of our ELISA binding studies showed that the absorbance values for the binding of the viral FcγR to the GM 17-expressing IgG antibodies were significantly higher than for the GM 3-expressing antibodies (0.60 vs. 0.36; p=0.0019). These findings provide mechanistic insights into the modulating role played by the genetic variants of IgG in the generation of immunity to CMV in patients with breast cancer.

Influence of a montmorency cherry juice blend on indices of exercise-induced stress and upper respiratory tract symptoms following marathon running--a pilot investigation.

BackgroundProlonged exercise, such as marathon running, has been associated with an increase in respiratory mucosal inflammation. The aim of this pilot study was to examine the effects of Montmorency cherry juice on markers of stress, immunity and inflammation following a Marathon.MethodsTwenty recreational Marathon runners consumed either cherry juice (CJ) or placebo (PL) before and after a Marathon race. Markers of mucosal immunity secretory immunoglobulin A (sIgA), immunoglobulin G (IgG), salivary cortisol, inflammation (CRP) and self-reported incidence and severity of upper respiratory tract symptoms (URTS) were measured before and following the race.ResultsAll variables except secretory IgA and IgG concentrations in saliva showed a significant time effect (P <0.01). Serum CRP showed a significant interaction and treatment effect (P < 0.01). The CRP increase at 24 and 48 h post-Marathon was lower (P < 0.01) in the CJ group compared to PL group. Mucosal immunity and salivary cortisol showed no interaction effect or treatment effect. The incidence and severity of URTS was significantly greater than baseline at 24 h and 48 h following the race in the PL group and was also greater than the CJ group (P < 0.05). No URTS were reported in the CJ group whereas 50 % of runners in the PL group reported URTS at 24 h and 48 h post-Marathon.ConclusionsThis is the first study that provides encouraging evidence of the potential role of Montmorency cherries in reducing the development of URTS post-Marathon possibly caused by exercise-induced hyperventilation trauma, and/or other infectious and non-infectious factors.

Estimation of serum, salivary immunoglobulin G, immunoglobulin A levels and total protein, hemoglobin in smokeless tobacco chewers and oral submucous fibrosis patients.

Background:Oral submucous fibrosis (OSMF) is a debilitating, potentially cancerous oral condition. Although areca nut is the most important causative agent, it is also considered that the disease is immunologically mediated.Aim of the Study:To establish that autoimmunity and nutritional deficiency play a role in the etiopathogenesis of OSMF.Objectives of the Study:To show that serum immunoglobulin markers (immunoglobulin-G [IgG], immunoglobulin-A [IgA]) and nutritional parameters such as total serum protein (TSP), Hemoglobin (Hb) play a role in causing OSMF and also to correlate serum, salivary IgG, IgA levels in OSMF patients.Settings and Design:A case-control study was done with 50 patients (25 patients who were provisionally diagnosed as OSMF - Group I, and 25 patients who were chronic smokeless tobacco chewers and who did not have any intraoral lesion - Group II).Materials and Methods:Five milliliters of blood and saliva were collected from both the groups. Quantitative analysis of serum, and salivary IgG, IgA was done by turbidometric immunoassay. TSP and Hemoglobin (Hb) were estimated by spectrophotometry.Statistical Analysis:Results were analyzed by independent samples t-test and one-way analysis of variance (ANOVA).Results:All patients of OSMF showed significant (P < 0.01) increase in serum IgG, IgA, and salivary IgG levels as compared to smokeless tobacco chewers. The salivary IgA levels showed a significant decrease in OSMF patients (P < 0.05). TSP and Hb levels showed significant (P < 0.01) decrease in OSMF patients as compared to smokeless tobacco chewers.Conclusion:The elevation of immunoglobulin levels supports the concept of autoimmunity. The decrease in TSP and Hb suggests that nutritional deficiency plays a defined role in the occurrence as well as a further progression of OSMF.

First Report of the Myeloma Canada Research Network (MCRN)-001 Trial Utilizing Bortezomib-Based Induction, Enhanced Conditioning with IV Busulfan + Melphalan (BuMel) and Lenalidomide Maintenance: Feasibility of a National Canadian Study Based on Achievement of Minimal Residual Disease (MRD) Negativity

Conventional immunoglobulin (Ig) markers have been used to define multiple myeloma (MM) responses, but assessment of marrow for minimal residual disease (MRD) may provide better information on disease status/prognosis (Paiva B, et al. Blood 2008; 112: 4017). We therefore initiated a national multi-center ASCT trial with the goal of producing a high rate of MRD-negativity by using bortezomib (btz)-based induction, enhancing the conditioning regimen and utilizing post-ASCT maintenance.This phase 2 open label-trial was conducted in 10 Canadian centers. After btz-based induction (usually CyBorD) in the absence of disease progression, patients (pts) received BuMel conditioning (IV busulfan 3.2 mg/kg days -5 to -3 or days -6 to -4 + melphalan 140 mg/m2 day -2 or day -3), followed by ASCT on day 0. On day 100 post-ASCT, lenalidomide (len) 10 mg/day was commenced, escalated to 15 mg/day after 3 cycles if appropriate, and continued until disease progression.Bone marrow aspirate (BMA) samples were shipped centrally for MRD analysis by 15-color multiparameter flow cytometry (MFC) before any therapy, prior to ASCT, on day 100 post-ASCT, every 3 mos for the 1st year and every 6 mos thereafter until progression.Between 03/2013 - 07/2014, 99 newly diagnosed MM pts provided untreated BMA samples for MRD analysis. To date, 42 of a planned target of 78 pts have completed induction therapy have undergone ASCT and 39 are evaluable so far. 25 of the 99 (25%) who provided initial marrow samples did not meet criteria for enrollment: 3 (3%) had poor BMA samples; 3 (3%) did not have confirmed MM; 6 (6%) did not proceed with ASCT (1 due to progression); 1 (1%) had received dexamethasone prior to MRD analysis; 1 (1%) died during induction and 11 (11%) withdrew consent/opted for standard conditioning.Median age of the 39 evaluable pts is 53 (39-67); 64% are male. Median serum β2-microglobulin level is 3.64 mg/L (1.7-20) and albumin 37 g/L (2.8-48.1); 17 pts have ISS stage I; 9 have stage II; 9 have stage III MM and 4 have missing data. Ig subtype includes IgGκ in 16 (40%), IgGλ in 4 (10%), IgAκ in 5 (13%), IgAλ in 8 (21%), IgMλ in 1 (3%), κ in 1 (3%); non-secretory in 2 (5%) and no data in 2 pts (5%).Post-ASCT, only 4 SAEs have occurred: atrial fibrillation (2), acute kidney injury (1) and sepsis (1). There have been no ASCT-related deaths, and no pt has progressed at a median follow-up of 7.8 mos (range: 4.8-10.1).The best Ig response post-induction in the 31 pts with available restaging data is CR in 5 (16%), VGPR in 9 (29%), PR in 13 (42%), and SD in 1 (3%). 27 pts have reached day 100 post-ASCT and 8 pts have been formally evaluated. In these 8, the Ig response is CR in 2 (25%), VGPR in 5 (63%) and PR in 1 (12%). Table 1 summarizes MRD results to date.Table 1Comparison of conventional Ig response rates and achievement of MRD negativity# Conventional Ig Responses [# MRD negative]Time point of assessment#EvaluableTotal # MRD negativeCRVGPRPRMRAfter btz-based induction3165 [3]9 [1]13 [2]0Day 100 post-ASCT822 [2]5[0]1[0]0During len maintenance5105 [1]00Conclusions: 1) MFC performed on pre-therapy marrow samples to allow subsequent evaluation for MRD was successful in 97% of pts using a central lab; 2) IV BuMel was well-tolerated with few SAEs and no ASCT-related deaths; 3) MRD and conventional Ig responses may not correlate well; 4) Further F/U is required to determine the dynamics of MRD achievement and long term outcomes with this approach. DisclosuresReece:Novartis: Honoraria, Research Funding; BMS: Research Funding; Merck: Research Funding; Millennium: Honoraria, Research Funding; Janssen: Consultancy, Honoraria, Research Funding; Celgene: Consultancy, Honoraria, Research Funding; Otsuka: Honoraria, Research Funding; Amgen: Honoraria. Off Label Use: Lenalidomide maintenance after ASCT. Venner:Celgene: Honoraria, Research Funding; Janssen: Honoraria. White:Janssen: Consultancy, Honoraria; Celgene: Consultancy, Honoraria. Sebag:Novartis: Honoraria; Janssen: Honoraria; Celgene: Honoraria. Song:Celgene: Honoraria; Otsuka: Honoraria; janssen: Honoraria. Tay:Celgene: Honoraria; Janssen: Honoraria. Kukreti:Celgene: Honoraria. Trudel:Glaxo Smith Kline: Honoraria, Research Funding; Novartis: Honoraria; Celgene: Honoraria; Oncoethix: Research Funding. Tiedemann:Celgene: Honoraria; Janssen: Honoraria. Chen:Millennium: Research Funding; Janssen: Honoraria; Celgene: Honoraria, Research Funding.

Markers of gut mucosal inflammation and cow's milk specific immunoglobulins in non‐IgE cow's milk allergy

BackgroundAllergy to cow’s milk protein (CMP) may cause gastrointestinal (GI) symptoms in the absence of CMP specific IgE. The immunological mechanisms involved in such disease are not fully understood. Therefore we examined markers of gut mucosal inflammation and the immunoglobulin profiles in children with Gl symptoms suspected of cow’s milk protein allergy (CMPA).Patients and methodsWe prospectively recruited infants and young children (n = 57; median age 8.7 months) with gastrointestinal complaints suspected of CMPA. The diagnosis of CMPA was made using the double-blind, placebo-controlled food challenge. Serum and stool samples were collected during CMP-free diet and after both placebo and active challenges. We analyzed the stool samples for calprotectin, human β-defensin 2 and IgA. In serum, we analyzed the levels of β-lactoglobulin and α-casein specific IgA, and IgG antibodies (total IgG and subclasses IgG1 and IgG4). Control group included children with e.g. dermatological or pulmonary problems, consuming normal diets.ResultsFecal calprotectin levels were higher in the challenge positive group (n = 18) than in the negative (n = 37), with respective geometric means 55 μg/g [95% confidence interval 38–81] and 29 [24–36] μg/g (p = 0.0039), during cow’s milk free diet. There were no significant inter-group differences in the fecal β-defensin and IgA levels. The CMP specific IgG and IgA were not elevated in patients with CMPA, but the levels of β-lactoglobulin-IgG4 (p = 0.0118) and α-casein-IgG4 (p = 0.0044), and total α-casein-IgG (p = 0.0054) and -IgA (p = 0.0050) in all patient samples (regardless of CMPA diagnosis) were significantly lower compared to the control group using dairy products.ConclusionsDespite cow’s milk elimination in children intolerant to cow’s milk there might be ongoing low-grade inflammation in the gut mucosa. CMP specific IgG or IgA should not be used to diagnose non-IgE CMPA. The observed frequency of impaired CMP specific total IgA, IgG and IgG4 production in patients following cow’s milk free diet warrants further studies.

AB0764 Improved public health may explain a decrease in the incidence of seropositive rheumatoid arthritis in the early middle-aged finnish population in northern savo

BackgroundTrends in the incidence of RA may reflect changes in the host-environment relationship.ObjectivesTo assess the incidence of seropositive rheumatoid arthritis (RA) during 30 year period in Northern Savo Finland and...