Increased IgM Levels Research Articles

In Schizophrenia, Deficits in Natural IgM Isotype Antibodies Including those Directed to Malondialdehyde and Azelaic Acid Strongly Predict Negative Symptoms, Neurocognitive Impairments, and the Deficit Syndrome.

Schizophrenia is characterized by an interrelated activation of the immune-inflammatory response system (IRS) and the compensatory immune-regulatory system (CIRS), which downregulates the IRS. Deficit schizophrenia is characterized by a deficit in IgM-mediated autoimmune responses to tryptophan catabolites. The presence and correlates of IgM isotype antibodies to oxidative-specific epitopes (OSEs), nitroso (NO), and nitro (NO2) adducts in schizophrenia remain unknown. This study measured IgM antibodies to malondialdehyde (MDA), azelaic acid, phosphatidylinositol, oleic acid, NO-tryptophan, NO-albumin, NO-cysteinyl, and NO2-tyrosine in a sample of 80 schizophrenia patients, divided into those with and those without deficit schizophrenia, and 38 healthy controls. Deficit schizophrenia was characterized by significantly lower IgM antibody levels to all OSEs as compared with non-deficit schizophrenia and controls. Lowered IgM antibodies to MDA coupled with increased IgM levels to NO-cysteinyl and NO2-tyrosine strongly predict deficit schizophrenia versus non-deficit schizophrenia with an area under the ROC curve of 0.913. A large part of the variance (21.2-42.2%) in the negative symptoms of schizophrenia and excitation is explained by IgM antibody titers to MDA (inversely) and NO-cysteinyl and/or NO2-tyrosine (both positively). Lower IgM antibodies to MDA are significantly associated with impairments in episodic memory including direct and delayed recall. These findings further indicate that deficit schizophrenia is a distinct phenotype of schizophrenia, which is characterized by lower natural IgM antibody levels to OSEs and relative increments in nitrosylation and nitration of proteins. It is concluded that deficits in natural IgM attenuate CIRS functions and that this impairment may drive negative symptoms and impairments in episodic memory and thus deficit schizophrenia.

Peculiarities of PED virus pathogenesis in neonatal non-immune piglets on Ukraine farms

The article presents the results of study of the immune and tissue features of pathogenesis of PED virus in neonatal piglets in Ukraine. Clinical and epizootological analysis was performed in three separate agricultural enterprises with adverse epidemiological situation with PED virus. Blood samples and fragments of the small intestine of PEDv infected piglets were used to study the characteristics of the pathogenesis of PED virus. It has been determined that the pathogenesis of super-acute infection by the PED virus in 1–7 day old non-immune piglets proceeds according to the classic type of infectious process with a relay transmission of the pathogen. Moreover, diarrhea syndrome has induced the development of irreversible and lethal pathophysiological lesions by replication the PED virus in enterocytes of infected piglets. Pathogenetic changes observed in PEDv infected piglets were combined with the following components: acute inflammatory-necrotic process in the tissue of the intestine, destruction and desquamation of the epithelium of the small intestine. The infection process on day 3–5 of manifestation of clinical signs of PED virus is accompanied with an increase in the number of erythrocytes, leukocytes and monocytes, hemoglobin, hematocrit level and aminotransferase activity. In addition, PED virus infection induced an increase in IgM levels and the number of phagocytizing leukocytes with increased aggressiveness and digestive power. The duration of the release of PED virus with feces is 26–37 days after cessation of watery diarrhea. The concentration of PEDv in the feces of convalescent piglets ranged from 120 to 1700 equivalent genome in 1 g of tissue. Parasite coenotic associations of the microbiocenosis of the digestive tract of PED virus infected piglets did not have time to form into stable epitopes. There was a constant quantitative prevalence of transient microbiota and a decrease in the share of probiotic microorganisms. The intestinal microflora consists of random transient microbiota of the environment with pathogenic properties. The results presented show evidence that molecular and cell-mediated immunological events in non-immune piglets have the initial stage of the specific progress of immune response against PEDv.

Hepatotoxicity and immunotoxicity of MC-LR on silver carp

Microcystins produced by some cyanobacteria can cause damages to the liver and kidneys of aquatic animals. In the natural water with cyanobacterial blooms, silver carp may suffer from the most serious affect of the bloom due to their filtering these cyanobacteria and ingesting them as food. In the present study, silver carp was exposed to microcystin-LR by using the method of intraperitoneal injection first to determine the acute toxicity of microcystin-LR on silver carp and then to determine the activity of inflammatory protein and content of inflammatory factors from the serum of silver carp following a subacute exposure of microcystin-LR at doses of 104.9 μg kg–1 (1/5 of LD50) or 262.1 μg kg–1 (1/2 of LD50). The results showed that MC-LR exposure increased fish liver index and promoted the activities of fish serum alanine aminotransferase (ALT) and aspartate aminotransferase (AST), indicating the hepatotoxicity of MC-LR on the fish. Moreover, MC-LR exposure also increased the number of leukocytes, complement C3 level, lysozyme activity (at the first 9 h of exposure), and the contents of cytokines TNF-α, IL-1β and IFN-γ in fish serum. In addition, a significant increase in IgM level was observed in the serum and head kidney of silver carp following MC-LR exposure. This result suggests that semi-lethal doses of MC-LR exposure is not only hepatotoxic but also immunotoxic to silver carp.

Effects of endocrine disruption by 4-nonylphenol ethoxylate on the grwoth preformance and immune responses of female and male immature Koi carp (Cyprinus carpio carpio)

Nonylphenol (NP) is an endocrine disrupting chemical which has been shown to be able to modulate endocrine system of various organisms by different mechanisms. The objective of this study was to investigate the potential effects of 4-NP and 17-β-estradiol (E2) on immune parameters (IgM level and lysozyme activity) of Koi carp (Cyprinus the carpio carpio) in an attempt to expand the knowledge of the immune-reproductive system interactions. Fish were injected with ascending doses (10, 50,100 μg/g-1 body weight) of 4-nonylphenol (4-NP) and (2 μg/g-1 body mass) of 17-β-estradiol (E2) by vehicle during 3a weeks experiment. After 21 days, the fishes (180) were anesthetized, blood samples collected from their caudal vein, and after dissection, theywere grouped into male and female according to their gonadal characters. The results of plasmatic immune parameters measurement showed that 4-NP induced significant increase in IgM levels and lysozyme activity at 50μg/g -1 dosagewhile the their value decreased at higher doses of 4-NP (100 μg/g-1) compared with the control treatment (p<0.05). In addition,the treatment with 2 μg/g-1E2 injection, induced a significant decrease in IgM and lysozyme activity levels. These results indicated that 4-NP and E2 could potentially disturb the balance of immune system in immature koi carp with potential consequences for their endocrine interactions.

Elevated vWF Antigen Serum Levels Are Associated With Poor Prognosis, and Decreased Circulating ADAMTS-13 Antigen Levels Are Associated With Increased IgM Levels and Features of WM but not Increased vWF Levels in Patients With Symptomatic WM

BackgroundWaldenström's macroglobulinemia (WM) is a rare malignancy characterized by bone marrow infiltration by lymphoplasmacytic cells and the presence of a monoclonal IgM paraprotein. The interactions of lymphoplasmacytic cells with other cells in their microenvironment, including mast cells and endothelial cells, support their survival and proliferation and can induce resistance to therapy. von Willebrand factor (vWF) plays a key role in primary hemostasis but is also a marker of endothelial “stimulation.” High levels of vWF have been associated with an adverse prognosis in patients with symptomatic WM and might reflect the interactions between lymphoplasmacytic cells and other cells of their microenvironment. Materials and MethodsConsidering vWF and ADAMTS-13 (a disintegrin and metalloproteinase with a thrombospondin type 1 motif, member 13) as markers of endothelial dysfunction and activation, we evaluated the prognostic importance of vWF and ADAMTS-13 antigen levels in the serum of patients with previously untreated symptomatic WM to validate vWF as a possible prognostic marker for progression-free and overall survival. We also validated the measurement of vWF in the serum instead of citrated plasma and investigated the possible correlations of ADAMTS-13 antigen levels with disease characteristics. The analysis included 42 patients with symptomatic WM and 19 matched healthy controls. ResultsThe serum levels of vWF antigen provided significant prognostic information, and patients with levels ≥ 200 IU/dL had a very poor prognosis compared with patients with lower levels. The ADAMTS-13 antigen levels were decreased in WM patients and correlated with the IgM levels, β2-microglobulin, and extent of bone marrow infiltration. ConclusionvWF levels measured in the serum could become an important prognostic marker in patients with WM and requires further investigation.

Immunological features of parvovirus infection in different pregnancy periods.

Parvovirus infection in pregnant women caused the development of immuno­patho­logical reactions.129 infected pregnant women and 16 women with physiological pregnancy in the 1st, 2nd and 3rd trimesters were examined. The functional state of neutrophils (Nph) and monocytes (Mc) of peripheral blood was investigated, parameters of cellular and humoral immunity and cytokine profile of serum of pregnant women were studied. In infected pregnant women in different periods of gestation as compared with pregnant women of control groups there were observed 1) an increase of index of phagocytosis Nph and Mc against the background of reducе of their absorption capacity and increasing NBT-activity with a decrease of reserve capacities of phagocytes; 2) an activation of the cellular immunity with an increase of helper mechanisms of the immune response; 3) a reliable increase of IgM level in serum in different periods of pregnancy; 4) a reliable increase of the content of IFN-γ, FNP-α, IL-2, IL-8 inserum against the background of a slight decrease in the level of anti-inflammatory IL-10. The largest deviations from the control indicators were recorded in the 1st trimester of pregnancy.

X-linked immunodeficiency with hyper-immunoglobulin M

X-linked immunodeficiency with hyper-immunoglobulin(Ig) M is a primary immunodeficiency disease, mainly due to the mutation of coding CD40 ligand(CD40L), which results in the dysfunction of immunoglobulin class switching that causes in a normal or increased IgM level, and a marked decrease in IgG, IgA and IgE level.The clinical manifestations are recurrent infection, neutropenia, and autoimmune disease or tumor. Key words: X-linked immunodeficiency with hyper-immunoglobulin M; CD40L gene; Mutation

Immunoglobulin and B-cell disturbances in patients with chronic idiopathic neutropenia

Chronic idiopathic neutropenia (CIN) is a granulocytic disorder associated with presence of activated, myelosuppressive T-lymphocytes. In the present study we have evaluated constituents of humoral immunity in CIN patients (n=48) compared to healthy controls (n=52). CIN patients displayed lower serum IgG levels due to a reduction in IgG1, IgG3, IgG4 but not IgG2, lower IgA and increased IgM levels compared to controls. The proportion of CD19+ cells did not differ between patients and controls; however the proportion of the naïve IgD+/CD27− B-cells was increased and the proportion of class-switched memory IgD−/CD27+ B-cells was decreased in the patients. The percentage of CD40+ B-cells did not differ between patients and controls and no aberrations in the CD40-meadiated signal transduction pathway or in CD40-gene polymorphisms were identified. These data provide further evidence that immune disturbances are associated with the pathophysiology of CIN and point out for the first time the implication of the B-cell system.

Impact of parental obesity on neonatal markers of inflammation and immune response.

Background/ObjectivesMaternal obesity may influence neonatal and childhood morbidities through increased inflammation and/or altered immune response. Less is known about paternal obesity. We hypothesized that excessive parental weight contributes to elevated inflammation and altered immunoglobulin (Ig) profiles in neonates.Subjects/MethodsIn the Upstate KIDS Study maternal pre-pregnancy body mass index (BMI) was obtained from vital records and paternal BMI from maternal report. Biomarkers were measured from newborn dried blood spots (DBS) among neonates whose parents provided consent. Inflammatory scores were calculated by assigning one point for each of 5 pro-inflammatory biomarkers above the median and one point for an anti-inflammatory cytokine below the median. Linear regression models and generalized estimating equations were used to estimate mean differences (β) and 95% confidence intervals (CI) in the inflammatory score and Ig levels by parental overweight/obesity status compared to normal weight.ResultsAmong 2974 pregnancies, 51% were complicated by excessive maternal weight (BMI>25), 73% by excessive paternal weight, and 28% by excessive gestational weight gain. Maternal BMI categories of overweight (BMI 25.0-29.9) and obese class II/III (BMI≥35) were associated with increased neonatal inflammation scores (β=0.12, 95% CI: 0.02, 0.21; p=0.02, and β=0.13, CI: −0.002, 0.26; p=0.05, respectively) but no increase was observed in the obese class I group (BMI 30-34.9). Mothers with class I and class II/III obesity had newborns with increased IgM levels (β=0.11, CI: 0.04, 0.17; p=0.001 and β=0.12, CI: 0.05, 0.19); p<0.001, respectively). Paternal groups of overweight, obese class I and obese class II/III had decreased neonatal IgM levels (β=−0.08, CI: −0.13,-0.03, p=0.001; β=−0.07, CI: −0.13, −0.01, p=0.029 and β=−0.11, CI:−0.19,-0.04, p=0.003, respectively).ConclusionsExcessive maternal weight was generally associated with increased inflammation and IgM supporting previous observations of maternal obesity and immune dysregulation in offspring. The role of paternal obesity requires further study.

Clinical spectrum and features of activated phosphoinositide 3-kinase δ syndrome: A large patient cohort study

BackgroundActivated phosphoinositide 3-kinase δ syndrome (APDS) is a recently described combined immunodeficiency resulting from gain-of-function mutations in PIK3CD, the gene encoding the catalytic subunit of phosphoinositide 3-kinase δ (PI3Kδ).ObjectiveWe sought to review the clinical, immunologic, histopathologic, and radiologic features of APDS in a large genetically defined international cohort.MethodsWe applied a clinical questionnaire and performed review of medical notes, radiology, histopathology, and laboratory investigations of 53 patients with APDS.ResultsRecurrent sinopulmonary infections (98%) and nonneoplastic lymphoproliferation (75%) were common, often from childhood. Other significant complications included herpesvirus infections (49%), autoinflammatory disease (34%), and lymphoma (13%). Unexpectedly, neurodevelopmental delay occurred in 19% of the cohort, suggesting a role for PI3Kδ in the central nervous system; consistent with this, PI3Kδ is broadly expressed in the developing murine central nervous system. Thoracic imaging revealed high rates of mosaic attenuation (90%) and bronchiectasis (60%). Increased IgM levels (78%), IgG deficiency (43%), and CD4 lymphopenia (84%) were significant immunologic features. No immunologic marker reliably predicted clinical severity, which ranged from asymptomatic to death in early childhood. The majority of patients received immunoglobulin replacement and antibiotic prophylaxis, and 5 patients underwent hematopoietic stem cell transplantation. Five patients died from complications of APDS.ConclusionAPDS is a combined immunodeficiency with multiple clinical manifestations, many with incomplete penetrance and others with variable expressivity. The severity of complications in some patients supports consideration of hematopoietic stem cell transplantation for severe childhood disease. Clinical trials of selective PI3Kδ inhibitors offer new prospects for APDS treatment.

Clinical and immunologic phenotype associated with activated phosphoinositide 3-kinase δ syndrome 2: A cohort study

Activated phosphoinositide 3-kinase δ syndrome (APDS) 2 (p110δ-activating mutations causing senescent Tcells, lymphadenopathy, and immunodeficiency [PASLI]-R1), a recently described primary immunodeficiency, results from autosomal dominant mutations in PIK3R1, the gene encoding the regulatory subunit (p85α, p55α, and p50α) of class IA phosphoinositide 3-kinases. We sought to review the clinical, immunologic, and histopathologic phenotypes of APDS2 in a genetically defined international patient cohort. The medical and biological records of 36 patients with genetically diagnosed APDS2 were collected and reviewed. Mutations within splice acceptor and donor sites of exon 11 of the PIK3R1 gene lead to APDS2. Recurrent upper respiratory tract infections (100%), pneumonitis (71%), and chronic lymphoproliferation (89%, including adenopathy [75%], splenomegaly [43%], and upper respiratory tract lymphoid hyperplasia [48%]) were the most common features. Growth retardation was frequently noticed (45%). Other complications were mild neurodevelopmental delay (31%); malignant diseases (28%), most of them being B-cell lymphomas; autoimmunity (17%); bronchiectasis (18%); and chronic diarrhea (24%). Decreased serum IgA and IgG levels (87%), increased IgM levels (58%), B-cell lymphopenia (88%) associated with an increased frequency of transitional Bcells (93%), and decreased numbers of naive CD4 and naive CD8 cells but increased numbers of CD8 effector/memory Tcells were predominant immunologic features. The majority of patients (89%) received immunoglobulin replacement; 3 patients were treated with rituximab, and 6 were treated with rapamycin initiated after diagnosis of APDS2. Five patients died from APDS2-related complications. APDS2 is a combined immunodeficiency with a variable clinical phenotype. Complications are frequent, suchassevere bacterial and viral infections, lymphoproliferation, and lymphoma similar to APDS1/PASLI-CD. Immunoglobulin replacement therapy, rapamycin, and, likely in the near future, selective phosphoinositide 3-kinase δ inhibitors are possible treatment options.

Systemic and mucosal immune response of rainbow trout to immunization with an attenuated Flavobacterium psychrophilum vaccine strain by different routes

Teleosts possess three immunoglobulin (Ig) heavy chain isotypes viz., IgM, IgT and IgD and all three isotypes are reported in rainbow trout. The expression of these Ig isotypes in response to different immunization routes was investigated and results provide a better understanding of the role these Igs in different tissues. Rainbow trout (Oncorhynchus mykiss) were immunized with an attenuated Flavobacterium psychrophilum strain, 259-93-B.17 grown under iron limiting conditions, by intraperitoneal, anal intubation and immersion routes. Serum, gill mucus, skin mucus and intestinal mucus samples were collected at 0, 3, 7, 14, 28, 42 and 56 days post immunization by sacrificing four fish from each treatment group and the unimmunized control group, and the IgM levels were estimated by an enzyme linked immunosorbent assay (ELISA). In addition, blood, gill, skin and intestinal tissue samples were collected for Ig gene expression studies. The secretory IgM, IgD and IgT gene expression levels in these tissues were estimated by reverse transcription quantitative real time PCR (RT-qPCR). Levels of IgM in serum, gill and skin mucus increased significantly by 28 days after immunization in the intraperitoneally immunized group, while no significant increase in IgM level was observed in fish groups immunized by other routes. Secretory IgD and IgT expression levels were significantly upregulated in gills of fish immunized by the immersion route. Similarly, secretory IgT and IgD were upregulated in intestines of fish immunized by anal intubation route. The results confirm mucosal association of IgT and suggest that IgD may also be specialized in mucosal immunity and contribute to immediate protection to the fish at mucosal surfaces.

Immunological & metabolic responses to a therapeutic course of Basti in obesity.

Background & objectives:Basti (medicated enema) is a popular Ayurvedic intervention recommended for obesity. However, there are no data to show whether any physiological or biochemical changes occur following this treatment. This study was conducted to identify the immunological and metabolic changes in obese individuals after a therapeutic course of Basti.Methods:Thirty two obese individuals (18 and 60 yr) with a body mass index (BMI) ≥30 kg/m2 who received a therapeutic course of 16 enemas (Basti) followed by a specific diet and lifestyle regimen for a period of 32 days as their treatment for obesity, were enrolled in the study. Clinical examination, measurement of immune and metabolic markers were done before (S1), immediately after (S2) and 90 days after the completion of therapy (S3).Results:A significant reduction (P<0.001) in weight, BMI, upper arm and abdominal circumference was seen at S3, along with a decrease in serum interferon (IFN)-γ (P<0.02), interleukin (IL)-6 (P<0.02) and ferritin (P<0.05) and increase in IgM levels (P<0.02). Peripheral blood lymphocytes (PBLs) stimulated with anti-CD3 monoclonal antibodies showed significant increase in reactive oxygen species (ROS) generation and calcium flux after Basti. All organ function tests revealed no changes.Interpretation & conclusions:Our study documents that a therapeutic course of Basti modulates immune responses by regulating pro-inflammatory cytokines, immunoglobulins and functional properties of T-cells. These changes are associated with a reduction in the body weight which is maintained even after three months of treatment. The study also documents the safety of Basti procedure.

Circulating Adamts-13 Is Reduced in Patients with Waldenstrom’s Macroglobulinemia and Is Associated with Increased IgM Levels and Features of the Disease but Not with the Increased Levels of Von Willebrand Factor

ADAMTS-13 is a protease, member of the ADAMTS family (A Disintegrin and Metalloproteinase with ThromboSpondin type 1 repeats-13), which cleaves the cell bound large ultrapolymeric von Willebrand factor (vWF) strings. Circulating ADAMTS-13 is primarily synthesized and released from hepatic stellate and endothelial cells. Acquired functional deficiency of ADAMTS-13, usually due to inhibitory IgG autoantibodies, results in excessive platelet aggregation and disseminated vWF/platelet-rich thrombus formation. A possible association of low levels of ADAMTS-13 Ag with arterial thrombosis and endothelial dysfunction has also been reported. Hivert and colleagues (Blood 2012;120:3214-21) and our group have shown that increased levels of vWF, the only known substrate of ADAMTS-13, are associated with poorer prognosis in patients with symptomatic Waldenstrom’s Macroglobulinemia (WM). Thus, our aim was to investigate the possible association of ADAMTS-13 antigen (Ag) levels with features of WM and possible biologic implications of the ADAMTS-13 / vWF interaction in WM patients’ prognosis.Our study included 42 patients with symptomatic WM who were treated and followed in the Department of Clinical Therapeutics of the University of Athens (Greece), from 1999 to 2012, 22 patients with asymptomatic WM and 19 healthy controls of matched gender and age. For the purpose of this study we used stored serum, which had been collected before initiation of any therapy. ADAMTS-13 antigen levels were measured using a commercially available kit (R&D Systems, Minneapolis, MN, USA), which has a detection limit for ADAMTS-13 13 (ng/mL) with intra- and inter-Assay Precisions of <3.7% and <5.7%, respectively.The median age of patients with symptomatic WM was 65 years (range: 37-83 years) and 54% were males. Anemia (Hb <11.5 g/dL) was present in 78% of patients, low platelet counts (<100x109/L) in 17%, beta2-microglobulin >3 mg/dl in 56%, while 7.5% had serum LDH ≥250 U/L and 58% had serum albumin <3.5 g/dL. Median serum IgM was 3340 mg/L (range 246-9563 mg/dL). According to IPSSWM, 22% had low, 43% intermediate and 35% high risk WM, respectively. Main reasons to initiate therapy included cytopenias in 42% of patients, B-symptoms in 15%, hyperviscosity in 12%, neuropathy in 10% and other reasons in 21%. Primary therapy based on rituximab was given in 93% of the patients and 54% achieved at least 50% reduction of IgM.The median levels of ADAMTS-13 in patients with symptomatic WM were 848 ng/ml (range 471-1622 ng/ml), while those of patients with asymptomatic WM were 875 ng/ml (range 280.5-1466 ng/nl) and of healthy controls were 1170 ng/ml (range 770-1598 ng/ml). Thus, there was a statistically significant difference in the levels of ADAMTS-13 between patients with either symptomatic or asymptomatic WM and normal individuals (p<0.001 and p=0.002 respectively), while there was no difference between those with symptomatic and asymptomatic WM (p=0.934). We then evaluated the association of ADAMTS-13 antigen levels with features of symptomatic WM. There was a negative correlation between low levels of ADAMTS-13 with high levels of IgM (r=-0.544, p<0.001), beta2-microglobulin (r=-0.376, p=0.01) and with the infiltration of the BM by lymphoplasmatic cells (r=-0.338, p=0.03), while there was a positive correlation with serum albumin levels (r=0.373, p=0.016). Accordingly, in patients who required therapy because of hyperviscosity syndrome, serum ADAMTS-13 was lower (p=0.04). There was no association of the levels of ADAMTS-13 and vWF Ag either in patients with symptomatic or asymptomatic WM (p>0.5 for both), and thus, a direct link between increased vWF Ag levels, which are found in patients with WM, and compensatory ADAMTS-13 Ag levels cannot be supported by our data. The levels of ADAMTS-13 Ag had no prognostic association with overall or progression free survival in patients with symptomatic WM.In conclusion, this is the first evaluation of ADAMTS-13 Ag levels in the serum of patients with WM, in which we found that the levels of ADAMTS-13 Ag are lower compared to healthy controls and these reduced levels inversely correlate with IgM levels, beta2-microglobulin and the degree of bone marrow infiltration. Levels of ADAMTS-13 do not increase in compensation to increased levels of vWF Ag, a marker of endothelial activation and this fact may have biological implications yet to be investigated. DisclosuresNo relevant conflicts of interest to declare.

Dietary modulation of immune response and related gene expression profiles in mirror carp (Cyprinus carpio) using selected exotic feed ingredients

An investigation was conducted to evaluate the effect of feeding selected exotic ingredients on immune responses and expression of immune related genes in mirror carp (Cyprinus carpio). Fish were fed diets for a total of 56days. Fishmeal served as the main protein source in the control diet and two experimental diets consisted of fishmeal fixed at 34% provision of protein and the remaining 66% protein was provided either by earthworm meal (EW diet) or a combination of whey protein concentrate (8%) and casein (58%) (WPC diet). At the start of the trial fish were injected intraperitoneally with Aeromonas hydrophila bacterin and samples of carp blood were taken for gene expression, haematological and serological analysis. Compared to fish fed fishmeal, a significant increase in mRNA expression of the pro-inflammatory cytokines IL-1β (24h post injection) and TNFα (at 12h and 48h post injection) was observed in fish fed EW. Moreover a similar trend was observed for complement 3 (C3) gene, where fish fed EW showed significant elevations in mRNA expression values at both 12 and 48h post injection compared to control fed fish. In contrast, fish fed WPC showed a significant decrease in C3 and TNF-α mRNA expression compared to fish fed fishmeal (48 h post injection). Fish fed EW and WPC showed a significant increase in peripheral leukocyte levels compared to fish fed fishmeal 14d post injection. Fish fed fishmeal presented significantly higher circulatory IgM levels at 7d post injection compared to fish fed EW and WPC. In contrast, fish fed EW and WPC showed a significant increase in IgM levels at 28d post injection. The present study shows an up-regulation of all immune related genes in fish fed EW, compared to fish fed fish meal, which is indicative of an acute inflammatory response. In contrast it appears feeding WPC has an immunosuppressive effect towards TNFα and C3 expression in carp blood. This may be offset by the fact that feeding WPC to carp appears to aid in the increased production of circulatory IgM and leukocyte levels compared to fishmeal fed fish.

Generation of mice with conditional ablation of the Cd40lg gene: new insights on the role of CD40L

CD40 ligand (CD40L) acts as an immune modulator in activated T cells, and mutations in the extracellular domain are associated to X-linked hyper IgM syndrome. A role for platelet CD40L in mediating thrombotic and inflammatory processes in atherosclerosis has also been reported. Using the Cre/loxP recombination technology we generated four knockout lines of mice with deletion of the Cd40lg gene restricted to the hematopoietic system. Mouse lines with expression of Cre recombinase driven by the Tie2, Vav1, or CD4 promoters showed in vivo ablation of CD40L in leukocytes and platelets. In contrast, in mice with Cre expression driven by the megakaryocyte lineage-restricted Pf4 promoter, abolition of CD40L expression was observed in megakaryocytes cultured in vitro, but not in circulating platelets. Characterization of these animals revealed reduced in vivo thrombogenesis and defective activation of washed CD40L-deficient platelets, suggesting that membrane-bound CD40L is involved in the control of haemostasis acting as a platelet co-activator. In addition, we report the practically absence of CD40L in mouse and human endothelial cells, as well as the detection of an exon 3-deleted CD40L transcript in both platelets and leukocytes of mouse and human origin. Finally, compared with their corresponding littermate floxed controls, Cre+ mice carrying CD40-deficient leukocytes did not exhibit increased IgM levels, and reduction of IgA and IgG levels was statistically significant only in Tie2-Cre+ mice, suggesting that expression of CD40L in an earlier developmental step may be determinant in the regulation of the class switch recombination process.

Abstract 80: Loss of Id3 in B Cells Leads to Increased Adipose Tissue T15-IgM and Protects Against Diet-Induced Glucose Intolerance

Background B1 B cells [[Unable to Display Character: &amp;#8211;]] the major producers of anti-inflammatory natural T15-IgM antibodies that promote tissue homeostasis [[Unable to Display Character: &amp;#8211;]] have been identified in adipose tissue, but their role in adipose tissue response to high-fat diet (HFD)-induced obesity is unclear. Our group and others have shown that the helix-loop-helix protein Id3 is a key regulator of B cell function in disease; mice null for Id3 have increased IgM levels and are protected from HFD-induced obesity. In this study, we test the hypothesis that B cell-specific Id3 deletion protects against the consequences of HFD-induced obesity through increased local production of natural IgM antibodies. Methods and Results C57Bl/6 floxed Id3 (Id3fl/fl) and CD19cre/+ mice were crossed to create an Id3fl/fl CD19cre/+ mouse. While these mice had similar body and adipose depot mass as littermate controls after 12 weeks of HFD, they had significantly improved glucose clearance (2hr blood glucose: 314 ± 33 vs. 420 ± 38 mg/dl; n=7-11; p&lt;0.05). At baseline, these mice were found to have significantly more B1b B cell than controls in the peritoneal fluid (3.2E5 ± 0.4E5 vs. 0.9E5 ± 0.4E5 cells; n=5-8; p&lt;0.01), epididymal adipose tissue (4072 ± 923 vs. 1149 ± 652 cells/g fat; n=5-8; p&lt;0.05), and omental adipose tissue. There were no differences in B1a or B2 cells in any compartment, and the mice had similar T cell and macrophage numbers in epididymal adipose tissue. ELISA analysis on whole epididymal adipose tissue revealed significantly higher levels of both total IgM (0.64 ± 0.06 vs. 0.24 ± 0.01 ng/ug protein; n=3-7; p&lt;0.001) and T15-IgM (7.6 ± 1.5 vs. 2.8 ± 0.4 U/ng protein; n=3-7; p&lt;0.05) in mice null for Id3 in B cells compared to controls. No differences were observed in the serum. In addition, omental fat from Id3fl/fl CD19cre/+ mice cultured ex vivo secreted significantly more T15-IgM than omental fat from littermate controls (40.0 ± 5.9 vs. 17.5 ± 4.2 U/mg fat; n=3-4; p&lt;0.01). Conclusions Loss of Id3 expression in B cells leads to elevated B1b B cells and increased local T15-IgM production in omental adipose tissue, along with improved glucose tolerance in mice fed a HFD, suggesting that T15-IgM may protect against the metabolic dysfunction caused by obesity.

Lymphoplasmacytic non-Hodgkin lymphoma/Waldenström's macroglobulinemia with CD5+, CD23+, and CD10-

Lymphoplasmacytic non-Hodgkin lymphoma/Waldenström's macroglobulinemia with CD5+, CD23+, and CD10-

HUMORAL IMMUNE RESPONSE TO ACETAMIPRID EXPOSURE AND ITS MODULATION BY SUPPLEMENTATION WITH VITAMIN C IN RATS

Acetamiprid (AP) is a fairly new neonicotinoid insecticide. Its indiscriminate use both in agriculture and domestic areas against wide range of pests such as aphids and whiteflies is causing toxicity to man and animals. In recent years the effects of insecticides on immune response have received more attention. The present study was designed to evaluate the toxic effect of repeated oral (by gavage) administration of AP over six weeks on humoral immune response. Forty eight adult male Sprague Dawley rats were divided into four groups (12 animals each). Animals of the 1st group were untreated and served as control , rats of the 2nd group were orally given 1/ 10 LD50 (83.18 mg/ kg b. wt.) , animals of the 3rd group were administered orally vitamin C alone at a dose level of 200mg / kg b.wt, every other day for six weeks , rats of the 4th group were given AP similar to those of the 2nd group then administered orally with vitamin C similar to those of the 3rd group 30 minutes after each AP administration. At day one after the end of experiment , eight rats were picked up randomly from each group , blood samples were collected from each animal under slight ether anaethesia through heart puncture into plain vaccutainer tubes and sera were separated. Data obtained revealed that oral AP administration into rats induced a non significant increase in IgM level ; significant decrease and increase in IgG and IL – 6 levels respectively as compared to the control values. Supplementation with vitamin C potentiated significantly the effect of AP on IgM , WBCs count , monocytes and lymphocytes percentage , and reduced significantly its effects on IgG and IL-6. These data indicate that AP is capable of inducing changes in humoral immunity that could be selectively modulated by supplementation with vitamin C.

A NUP98-HOXD13 leukemic fusion gene leads to impaired class switch recombination and antibody production

Myelodysplastic syndrome is a clonal process characterized by ineffective hematopoiesis and progression to acute leukemia. Although many myelodysplastic syndrome and leukemic patients have compromised immunity, the role of underlying mutations in regulating immune function is poorly understood. Recent studies show that NUP98-HOXD13 (NHD13) fusion gene results in myelodysplastic syndrome and impairs lymphocyte differentiation in transgenic mice. In our studies, we sought to elucidate the mechanism by which NHD13 affects B-lymphocyte development and function. Based on our preliminary findings that transgenic mice had increased levels of IgM and reduced IgG1 and IgE, we hypothesized that the fusion gene might impair class switch recombination (CSR). Mice were immunologically challenged with dinitrophenol. NHD13 mice showed a marked reduction in B-lymphocyte differentiation in their bone marrow and spleen following dinitrophenol stimulation and had reduced production of dinitrophenol-specific antibodies. Spleen follicles from these mice were small and hypocellular, indicating failure of clonal expansion. When isolated NHD13 B lymphocytes were stimulated in vitro using Escherichia coli lipopolysaccharide or lipopolysaccharide + interleukin-4, they failed to undergo sufficient CSR and proliferation. Taken together, our findings show that expression of NUP98-HOXD13 impairs CSR and reduces the antibody-mediated immune response, in addition to its role in leukemia. Further delineation of the NUP98-HOXD13 transgene may reveal novel pathways involved in CSR.