IgG Profiles Research Articles

Etiological profile of cirrhosis of liver from North-East India with reference to their anti-hepatitis A virus seroprevalence

Background: Cirrhosis of liver is common in north-east India. Hepatitis A infection in adults with chronic liver disease can cause acute on chronic liver failure associated with high mortality and morbidity. There have been reports of an epidemiological shift in hepatitis A virus (HAV) seroprevalence from South-East Asia and India. This study evaluated the etiological profile and seroprevalence of anti-HAV IgG in cirrhosis of liver patients. Patients and Methods: 160 hospitalized adult cases of decompensated cirrhosis of liver and 200 healthy controls were assessed for etiology and their anti-HAV IgG status by commercially available kits. Results : Most common cause of cirrhosis of liver in our region is ethanol related. 95% of cases and 89% of controls were seropositive for anti-HAV IgG (P = 0,181, insignificant difference). All cases above the age of 40 years were seropositive. Seroprevalence between sexes (M 97% and F 83.3%) was statistically insignificant. Only age showed a high coefficient of correlation (r = 0.854, statistically significant, P Conclusion: Alcohol is the most common etiology of cirrhosis of liver in north-east India. Socio-cultural milieu in our part may play a role with alcohol contributing to a major but preventable heath burden. Anti-HAV vaccination in our setting is not indicated routinely to cirrhosis of liver patients as it will not be cost-effective. However, young cirrhotics should be screened for anti-HAV antibody and if negative, may be offered vaccination. Screening should target young chronic liver disease patients in view of reports of decreasing seroprevalence across Asia as compared to one or two decades back.

Ethnic sensitivity assessment – pharmacokinetic comparability between Japanese and non-Japanese healthy subjects on selected mAbs

Objective: Ethnic sensitivity studies (ESSs), where safety and pharmacokinetics (PK) are assessed in Japanese subjects, are routinely conducted according to Japanese regulatory requirement before the subsequent clinical studies. The necessity of ESSs is questionable in case of mAbs, where inherent IgG characteristics are considered ethnically insensitive. This report investigated PK profiles and immunogenicity (IG) following a single administration of mAbs in Japanese and non-Japanese healthy subjects.Research design and methods: PK and IG comparison between Japanese and non-Japanese healthy subjects was made on mAbs data available from public domain and unpublished internal reports. PK comparison was made based on statistical approach as well as assumed typical IgG profile using modeling and simulation.Results: When compared directly, most mAbs showed no difference between ethnic groups. When profiles of various mAbs were fit to an assumed typical IgG PK model, the majority of mAbs follow the expected behavior regardless of ethnicities. Deviations from this behavior did not appear to be due to inherent ethnic differences. When the incidence of IG was assessed, only Adalimumab showed apparent ethnic difference.Conclusions: The overall lack of observational difference may facilitate discussion of mAbs’ early clinical development in Japan, including the utility of dedicated ESSs.

Specific IgM, IgG and IgG1 directed against Toxoplasma gondii detected by flow cytometry and their potential as serologic tools to support clinical indirect fundoscopic presumed diagnosis of ocular disease

In the present study we evaluated the anti-Toxoplasma gondii immunoglobulin profiles of a group of 118 individuals living in an endemic area. The aim of the study was to select biomarkers to support the ophthalmological diagnosis of retinal/retinochoroidal scars presumably caused by T. gondii infection. Overall anti-T. gondii reactivity of the IgM, IgG, IgA, IgE and IgG subclasses was investigated by flow cytometry-based anti-fixed tachyzoite antibodies (FC-AFTA) in four groups of subjects, referred to as: i) TOXO(L) — seropositive patients with retinal/retinochoroidal scars presumably caused by T. gondii infection; these patients were further subdivided according to morphological aspects of their ocular scar lesions as A, B or C; ii) TOXO(NL) — seropositive patients without ocular scar lesions; iii) NEG(L) — T. gondii seronegative patients presenting retinal lesions; and iv) NEG(NL) — T. gondii seronegative without retinal lesions (negative controls). Our data demonstrated that anti-T. gondii IgG profiles were able to discriminate the mean reactivity of TOXO(L) from all other clinical groups. Analysis of anti-T. gondii immunoglobulin profiles revealed that IgM and IgG were good biomarkers capable of discriminating between individual reactivity in patients with retinal/retinochoroidal scars presumably caused by T. gondii infection [TOXO(L)] from those caused by other clinical conditions. Furthermore, anti-T. gondii IgG1 reactivity was able to discriminate TOXO(L) from all other clinical groups. In conclusion, the pre-selected IgM, IgG and IgG1 anti-T. gondii antibody subclasses were able to segregate both TOXO(L) and the other subgroups, including the scar lesion group types (A, B, C), from other clinical conditions. These results suggest the applicability of this technique in the clinical laboratory to detect putative biomarker for diagnosis of ocular lesions in T. gondii-infected patients. Studies in other areas implementing the methods described in the present study would be of value and enable evaluation of a system for classification of presumed ocular toxoplasmosis scar lesions. This classification would make comparative studies on ocular toxoplasmosis conducted in different regions around the world possible.

Changes in anti-thyroglobulin IgG glycosylation patterns in Hashimoto's thyroiditis patients.

Objective:Sera of Hashimoto's thyroiditis (HT) patients are known to exhibit elevated levels of anti-thyroglobulin IgG (TgAb IgG). Therefore, TgAb IgG represents a hallmark of this debilitating autoimmune disease. The aim of our study was to investigate the differential expression of specific glycosylation patterns of TgAb IgG from HT patients and healthy blood donors.Methods:HT patients (n = 32) were divided into two subgroups, medium level group (mHT, n = 15) and high level group (hHT, n = 17), according to the serum levels of TgAb detected by electrochemiluminescence immunoassay. TgAb IgG was purified by affinity chromatography from the sera of the HT group and control group (n = 15). MALDI-QIT-TOF-MS/MS spectrometry was performed to identify the glycosylation profiles of purified TgAb IgG. Lectin microarray technology was used to compare the abundance of different glycans found on TgAb IgG between HT patients and controls, and between the mHT and hHT groups.Results:The results by MALDI-QIT-TOF-MS/MS showed that the glycosylation profiles of TgAb IgG were similar between the mHT, hHT, and control groups. Furthermore, the lectin microarray showed that compared to the control group (all P < .001), there were higher levels present of (1) mannose (detected as lectin LCA, VFA, and MNA-M); (2) terminal sialic acid (detected as SNA-I and PSA); (3) core fucose (detected as LcH); and (4) Gal(β1–4)GlcNAc(β1–2)Man glycans (detected as PHA-L) on TgAb IgG from the HT group. A similar trend was observed between the hHT and mHT group, with elevated levels of mannose, terminal sialic acid, core fucose, and Gal(β1–4)GlcNAc(β1–2)Man glycans on TgAb IgG found in the hHT group compared with the mHT group (all P < .05).Conclusions:TgAb IgG of HT patients exhibits higher glycosylation levels than those observed for TgAb IgG of healthy controls. Our results provide new clues for exploring the role of TgAb in the pathogenesis of HT.

Anti-pertussis antibody kinetics following DTaP-IPV booster vaccination in Norwegian children 7–8 years of age

At the age of 7–8 years a booster of diphtheria, tetanus, acellular pertussis and polio vaccine is recommended for children in Norway. In this cross-sectional study we have analysed the antibody levels against pertussis vaccine antigens in sera from 498 children aged 6–12 years. The purposes of this study were to investigate the duration of the booster response against the pertussis vaccine antigens pertussis toxin (PT) and filamentous haemagglutinin (FHA); to determine the presence of high levels of pertussis antibodies in absence of recent vaccination; and to analyse how booster immunisation may interfere with the serological pertussis diagnostics. Prior to the booster the IgG antibody levels against PT revealed a geometric mean of 7.3IU/ml. After the booster the geometric mean peak anti-PT IgG response reached to 45.6IU/ml, followed by a steady decline in antibody levels over the next few years. The IgG anti-FHA levels followed the anti-PT IgG profiles. Three years after the booster the geometric mean IgG levels were only slightly above pre-booster levels. Prior to the booster 44% of the sera contained ≤5IU/ml of anti-PT IgG compared to18% 3 years after and 30% 4 years after the booster. When recently vaccinated children were excluded, 6.2% of the children had anti-PT IgG levels above 50IU/ml which may indicate pertussis infection within the last 2 years. This study indicates that the currently used acellular pertussis vaccines induce moderate immune responses to the pertussis antigens and that the antibodies wane within few years after the booster. This lack of sustained immune response may partly be responsible for the increased number of pertussis cases observed in this age group during the last years.

Effect of Dietary Inulin Source on Piglet Performance, Immunoglobulin Concentration, and Plasma Lipid Profile

Abstract The aim of the study was to evaluate the effect of an inulin source (aqueous or aqueous-alcoholic extract, dried chicory root, or dried Jerusalem artichoke tubers) on piglet performance, plasma lipid profile, and immunoglobulin concentration. The study was conducted on 534 piglets (44 litters) allocated to five nutritional groups: group I (control) - fed basal diet, groups II and III receiving basal diet supplemented with 2% of inulin (aqueous and aqueous-alcoholic extract respectively), and groups IV and V -4% additive of dried artichoke or dried chicory respectively. During the trial, piglets’ body weight, feed intake, diarrhoea incidence, and mortality were controlled. Blood samples were collected twice from six animals of each group. In blood plasma, indices of lipid profile and concentrations of IgA, IgG and IgM were measured. The addition of inulin, regardless of its form (extracts or dried plants), significantly improved the rearing indices. In piglets of groups III, IV and V a significant improvement of daily weight gains and feed efficiency was noted. Inulin showed hypolipidemic activity (lowered total cholesterol level) and stimulated piglet immune system manifested by elevated IgA and IgG concentrations. Irrespective of the inulin source, a lower mortality rate resulting from the improvement of animal health was noted in all experimental groups.

Tailor-made pentablock copolymer based formulation for sustained ocular delivery of protein therapeutics.

The objective of this research article is to report the synthesis and evaluation of novel pentablock copolymers for controlled delivery of macromolecules in the treatment of posterior segment diseases. Novel biodegradable PB copolymers were synthesized by sequential ring-opening polymerization. Various ratios and molecular weights of each block (polyglycolic acid, polyethylene glycol, polylactic acid, and polycaprolactone) were selected for synthesis and to optimize release profile of FITC-BSA, IgG, and bevacizumab from nanoparticles (NPs) and thermosensitive gel. NPs were characterized for particle size, polydispersity, entrapment efficiency, and drug loading. In vitro release study of proteins from NPs alone and composite formulation (NPs suspended in thermosensitive gel) was performed. Composite formulations demonstrated no or negligible burst release with continuous near zero-order release in contrast to NPs alone. Hydrodynamic diameter of protein therapeutics and hydrophobicity of PB copolymer exhibited significant effect on entrapment efficiency and in vitro release profile. CD spectroscopy confirmed retention of structural conformation of released protein. Biological activity of released bevacizumab was confirmed by in vitro cell proliferation and cell migration assays. It can be concluded that novel PB polymers can serve a platform for sustained delivery of therapeutic proteins.

Whole CMV proteome pattern recognition analysis after HSCT identifies unique epitope targets associated with the CMV status.

Cytomegalovirus (CMV) infection represents a vital complication after Hematopoietic Stem Cell Transplantation (HSCT). We screened the entire CMV proteome to visualize the humoral target epitope-focus profile in serum after HSCT. IgG profiling from four patient groups (donor and/or recipient +/− for CMV) was performed at 6, 12 and 24 months after HSCT using microarray slides containing 17174 of 15mer-peptides overlapping by 4 aa covering 214 proteins from CMV. Data were analyzed using maSigPro, PAM and the ‘exclusive recognition analysis (ERA)’ to identify unique CMV epitope responses for each patient group. The ‘exclusive recognition analysis’ of serum epitope patterns segregated best 12 months after HSCT for the D+/R+ group (versus D−/R−). Epitopes were derived from UL123 (IE1), UL99 (pp28), UL32 (pp150), this changed at 24 months to 2 strongly recognized peptides provided from UL123 and UL100. Strongly (IgG) recognized CMV targets elicited also robust cytokine production in T-cells from patients after HSCT defined by intracellular cytokine staining (IL-2, TNF, IFN and IL-17). High-content peptide microarrays allow epitope profiling of entire viral proteomes; this approach can be useful to map relevant targets for diagnostics and therapy in patients with well defined clinical endpoints. Peptide microarray analysis visualizes the breadth of B-cell immune reconstitution after HSCT and provides a useful tool to gauge immune reconstitution.

Total Plasma N-Glycome Changes during Pregnancy

During pregnancy, the mother faces a major immunological challenge. Most of the major plasma proteins have important immunological functions, and altered levels of these major proteins have been reported during pregnancy, potentially providing immunosuppression. A large number of the high abundance plasma proteins are post-translationally modified by N-glycans, and while it is now understood that these glycans may also affect the immunological functions, their pattern has not been studied in relation to pregnancy. Here, the N-glycosylation profile of 32 pregnant women was determined over the course of their pregnancy using a multiplexed CGE-LIF method. Moreover, for 6 women, the glycosylation profiles of the proteins IgG, IgA, and alpha1-antitrypsin were monitored. For total plasma, 16 glycan signals showed differential expression during pregnancy. In general the levels of largely sialylated bi-, tri-, and tetra-antennary glycans were increased during pregnancy, while biantennary glycans with no more than one sialic acid were decreased. Similarly altered glycosylation profiles were observed for the individual proteins IgG, with a decrease of digalactosylated biantennary glycans after delivery, and alpha1-antitrypsin, on which increased levels of triantennary glycans were observed during pregnancy. Overall, these results show altered glycosylation profiles both for total plasma glycoproteins and on individual proteins during pregnancy, which may contribute to immunosuppression and have other biological functions.

Serum immunoglobulin G level in patients with lupus nephritis and the effect of treatment with corticosteroids and mycophenolate mofetil

Reduced serum IgG level is associated with increased risk of infection. We investigated the circulating IgG level and its determining factors in active lupus nephritis patients treated with corticosteroids and mycophenolate mofetil (MMF). This was a retrospective study on the longitudinal IgG profile in Class III/IV ± V lupus nephritis patients treated with prednisolone and MMF. 46 patients were included. At baseline, 34 (73.9%) patients (Group I) had normal or elevated IgG (1444.0 ± 600.5 mg/dL) while 12 (26.1%) (Group II) had IgG levels (567.8 ± 160.9 mg/dL) below the lower limit of normal. IgG levels at baseline, three, six and 12 months after treatment were 1215.4 ± 649.7 mg/dL, 843.9 ± 347.6 mg/dL, 914.5 ± 362.4 mg/dL and 1034.6 ± 452.5 mg/dL respectively. Treatment with prednisolone and MMF led to a significant drop in IgG after two weeks, reaching a nadir at eight weeks, followed by gradual normalization. Similar changes in IgG were observed in Group I patients but there was non-significant change in Group II within the first 24 weeks. Eighteen (39.1%) patients had low IgG by six months, and only one patient had IgG <300 mg/dL, at both three and six months. IgG level was negatively associated with proteinuria at six months (r = -0.711, p = 0.010). Five of 18 patients with low IgG had infections within the first year, while IgG levels below the lower limit of normal did not increase infection risk (relative risk 1.863; 95% confidence interval 0.466 to 6.818, p = 0.280). Reduced IgG occurred in 26% of active lupus nephritis patients and the IgG levels are significantly influenced by the severity of proteinuria. Treatment with prednisolone and MMF does not result in clinically important suppression of IgG.

24-OR

Aim Flow cytometric crossmatch (FCXM) has increasingly become the preferred method of crossmatching due to its increased sensitivity and specificity compared to the standard complement dependent cytotoxicity (CDC)-based methods. Here we report a case of undiagnosed lymphoma identified during a routine deceased donor final FCXM. Methods FCXM was performed on a Becton Dickinson FACSCalibur flow cytometer using pronased donor peripheral blood mononuclear cells. Staining included PerCP-CD3 for T-cells, PE-CD19 for B-cells, and FITC-goat anti-human IgG for the detection of surface-bound antibody. Results Donor was a 56-year-old Caucasian male admitted for cardio-pulmonary arrest and was evaluated for donation after circulatory death (DCD). His past known medical history included developmental delay, mental retardation, hypothyroid and general seizure disorder. During the final crossmatch of 11 renal recipient candidates, while the gated lymphocyte population displayed a typical FSC/SSC profile, a subpopulation of dimly stained CD19+ B-cells (28% dimmer) was observed with every serum crossmatched. This abnormal B-cell population constituted >50% of the entire CD19+ population and also displayed an elevated baseline FITC-IgG staining (31% brighter), indicating a denser-than-normal surface IgG profile. The T-cell population appeared normal for CD3 and FITC-IgG staining. Further immunophenotyping showed that this B-cell population was normally bright for CD20 and CD45; negative for CD5, CD10 and CD23; indeterminate for CD11c and CD38; and light chain staining displayed a Kappa restriction. Taken together, the immunophenotypic profile suggested a diagnosis of some form of mature B-cell non-Hodgkin’s lymphoma. This donor case was subsequently turned off. Conclusions This case report illustrates the possibility of identifying undiagnosed hematological malignancies using the conventional histocompatibility crossmatch setup even with only limited lymphocyte markers.

Autoantibody Profiling in Multiple Sclerosis Using Arrays of Human Protein Fragments

Profiling the autoantibody repertoire with large antigen collections is emerging as a powerful tool for the identification of biomarkers for autoimmune diseases. Here, a systematic and undirected approach was taken to screen for profiles of IgG in human plasma from 90 individuals with multiple sclerosis related diagnoses. Reactivity pattern of 11,520 protein fragments (representing ∼38% of all human protein encoding genes) were generated on planar protein microarrays built within the Human Protein Atlas. For more than 2,000 antigens IgG reactivity was observed, among which 64% were found only in single individuals. We used reactivity distributions among multiple sclerosis subgroups to select 384 antigens, which were then re-evaluated on planar microarrays, corroborated with suspension bead arrays in a larger cohort (n = 376) and confirmed for specificity in inhibition assays. Among the heterogeneous pattern within and across multiple sclerosis subtypes, differences in recognition frequencies were found for 51 antigens, which were enriched for proteins of transcriptional regulation. In conclusion, using protein fragments and complementary high-throughput protein array platforms facilitated an alternative route to discovery and verification of potentially disease-associated autoimmunity signatures, that are now proposed as additional antigens for large-scale validation studies across multiple sclerosis biobanks.

Suplementação dietética de levedura de cerveja e de minerais orgânicos sobre o desempenho e resposta imune em frangos de corte desafiados com a vacina de coccidiose

O objetivo desta pesquisa foi avaliar o efeito da suplementação de levedura de cerveja e minerais orgânicos sobre o desempenho, morfometria intestinal e resposta imune humoral de frangos de corte, desafiados com a vacina de coccidiose. Foram utilizados 384 pintos de corte, distribuídos aleatoriamente em um delineamento inteiramente casualizado com quatro tratamentos (1. controle, 2. minerais orgânicos 3. levedo de cerveja, 4. associação de minerais orgânicos e levedo de cerveja) e 4 repetições de 24 aves cada. No alojamento, 12 aves de cada repetição foram identificadas com anilha e receberam a vacina contra coccidiose via ocular. Aos 7, 21, 35 e 42 dias, foi coletado sangue para análise de proteínas séricas totais e perfil eletroforético de IgA e IgG e um segmento do duodeno para análise morfométrica. A suplementação de levedo de cerveja ou de minerais orgânicos não afetou o desempenho de frangos de corte ou a produção de anticorpos. O desafio imunológico com a vacina de coccidiose não foi suficientemente eficaz em provocar uma resposta imune capaz de alterar a morfometria da mucosa duodenal e a produção de anticorpos.

Immunoproteomic identification of antigenic salivary biomarkers detected by Ixodes ricinus-exposed rabbit sera

Ixodes ricinus, the primary vector of tick-borne disease in Europe, is currently expanding its distribution area and its activity in many countries. Antibody responses to tick salivary antigens have been proposed as an alternative marker of exposure to tick bites. However, the identification of the I. ricinus corresponding antigens remains elusive. Using rabbits artificially exposed to I. ricinus and 2 other European tick species (Rhipicephalus sanguineus and Dermacentor reticulatus) as controls, a cross-comparison of IgG profiles was performed against protein salivary gland extracts (pSGE) from these 3 tick species using immunoblots. Immunoblot analysis highlighted a singularity in the immune patterns according to tick species exposure and pSGE antigen source. Two protein bands were detected against I. ricinus pSGE only in rabbits exposed to I. ricinus bites. An immunoproteomic approach based on a fluorescence detection method was developed to unambiguously identify corresponding antigenic spots on 2-D gels. Among the unique I. ricinus salivary antigenic proteins detected by sera from rabbits exposed to this tick species, I. ricinus calreticulin was identified. Although tick calreticulin was previously proposed as a potential antigenic marker following exposure to ticks (particularly in North American tick species), the present study suggested that Ixodes calreticulin does not appear to be cross-recognized by the 2 other tick genera tested. Additional experiments are needed to confirm the use of I. ricinus calreticulin salivary protein as a potential discriminant antigenic biomarker to Ixodes tick exposure.

A simple method for assessment of human anti-Neu5Gc antibodies applied to Kawasaki disease (P3263)

Abstract N-Glycolylneuraminic acid (Neu5Gc) is a dietary immunogenic sugar that metabolically incorporates into diverse glycoconjugates in humans. Anti-Neu5Gc antibodies are detected in all human sera, play a role in several inflammation-mediated pathologies and are potential diagnostics of such diseases. Currently, it is difficult to determine their levels in individual human samples due to diverse Neu5Gc-epitopes recognition. Here we describe a simple method for detection of human anti-Neu5Gc antibodies. We exploit the difference between two mouse models that differ only by Neu5Gc-presence (wild-type) or Neu5Gc-absence (Cmah-/- knockout). We characterize mouse serum from both strains by HPLC, lectin and mass-spectrometry. We then use Cmah-/- knockout sera to inhibit all non-Neu5Gc-reactivity followed by binding to wild-type sera to detect overall anti-Neu5Gc response in a single assay. We applied this methodology to characterize anti-Neu5Gc IgG and IgA in sera of Kawasaki disease (KD) patients at various stages compared to controls. KD is an acute childhood febrile disease characterized by inflammation of coronary arteries that untreated may lead to coronary artery aneurysms, thrombosis and myocardial infarction. This estimated response is comparable to the average of detailed anti-Neu5Gc IgG profile analyzed by a sialoglycan microarray. Both assays revealed an elevated response in acute KD patients with normal coronaries compared to patients with aneurysm or dilated coronaries.

Serum autoantibodies to myelin peptides distinguish acute disseminated encephalomyelitis from relapsing– remitting multiple sclerosis

Background and objective: Acute disseminated encephalomyelitis (ADEM) and relapsing–remitting multiple sclerosis (RRMS) share overlapping clinical, radiologic and laboratory features at onset. Because autoantibodies may contribute to the pathogenesis of both diseases, we sought to identify autoantibody biomarkers that are capable of distinguishing them. Methods: We used custom antigen arrays to profile anti-myelin-peptide autoantibodies in sera derived from individuals with pediatric ADEM (n = 15), pediatric multiple sclerosis (Ped MS; n = 11) and adult MS (n = 15). Using isotype-specific secondary antibodies, we profiled both IgG and IgM reactivities. We used Statistical Analysis of Microarrays software to confirm the differences in autoantibody reactivity profiles between ADEM and MS samples. We used Prediction Analysis of Microarrays software to generate and validate prediction algorithms, based on the autoantibody reactivity profiles. Results: ADEM was characterized by IgG autoantibodies targeting epitopes derived from myelin basic protein, proteolipid protein, myelin-associated oligodendrocyte basic glycoprotein, and alpha-B-crystallin. In contrast, MS was characterized by IgM autoantibodies targeting myelin basic protein, proteolipid protein, myelin-associated oligodendrocyte basic glycoprotein and oligodendrocyte-specific protein. We generated and validated prediction algorithms that distinguish ADEM serum (sensitivity 62–86%; specificity 56–79%) from MS serum (sensitivity 40–87%; specificity 62–86%) on the basis of combined IgG and IgM anti-myelin autoantibody reactivity to a small number of myelin peptides. Conclusions: Combined profiles of serum IgG and IgM autoantibodies identified myelin antigens that may be useful for distinguishing MS from ADEM. Further studies are required to establish clinical utility. Further biological assays are required to delineate the pathogenic potential of these antibodies.

Seroconversion Rates of Helicobacter pylori Infection in Korean Adults

Studies on seroconversion and its reversion rate in Korean adults with Helicobacter pylori infection are very rare. The purpose of this study was to evaluate the overall seroprevalence, seroconversion rate, and seroreversion rate of H.pylori infection in an adult population. We performed this retrospective cohort study on healthy adults who had visited our health screening center at Asan Medical Center more than twice between January 2000 and December 2010. We reviewed the anti- H.pylori Ab IgG profiles of the enrolled people and their family members and the results of esophagogastroduodenoscopies and a self-reported questionnaire. A total of 67,212 people were enrolled in this study. The mean follow-up duration was 4.6years, and each participant visited the center for a mean of 3.8 visits. The overall proportions of participants demonstrating persistent seropositivity, persistent seronegativity, seroconversion, and seroreversion were 53.1%, 32.5%, 4.3%, and 10.1%, respectively. The annual seroconversion rate was 2.79%. The annual crude and spontaneous seroreversion rates of the entire study population were 3.64% and 2.42%, respectively. According to multivariate logistic regression, old age (HR=1.015), smoking (HR=1.216), alcohol consumption more than four times per week (HR=1.263), marriage (HR=2.735), and living with H.pylori-infected family members (HR=1.525) were identified as statistically significant risk factors associated with seroconversion. The annual seroconversion rate was 2.79% in our study population. Marriage and living with H.pylori-infected family members were important risk factors affecting seroconversion in our adult population.

A Simple Method for Assessment of Human Anti-Neu5Gc Antibodies Applied to Kawasaki Disease

N-Glycolylneuraminic acid (Neu5Gc) is an immunogenic sugar of dietary origin that metabolically incorporates into diverse native glycoconjugates in humans. Anti-Neu5Gc antibodies are detected in all human sera, though with variable levels and epitope-recognition profiles. These antibodies likely play a role in several inflammation-mediated pathologies including cardiovascular diseases and cancer. In cancer, they have dualistic and opposing roles, either stimulating or repressing disease, as a function of their dose, and some of these antibodies serve as carcinoma biomarkers. Thus, anti-Neu5Gc antibodies may signify risk of inflammation-mediated diseases, and changes in their levels could potentially be used to monitor disease progression and/or response to therapy. Currently, it is difficult to determine levels of anti-Neu5Gc antibodies in individual human samples because these antibodies recognize multiple Neu5Gc-epitopes. Here we describe a simple and specific method for detection and overall estimation of human anti-Neu5Gc antibodies. We exploit the difference between two mouse models that differ only by Neu5Gc-presence (wild-type) or Neu5Gc-absence (Cmah−/− knockout). We characterize mouse serum from both strains by HPLC, lectin and mass-spectrometry analysis and show the target Neu5Gc-epitopes. We then use Cmah−/− knockout sera to inhibit all non-Neu5Gc-reactivity followed by binding to wild-type sera to detect overall anti-Neu5Gc response in a single assay. We applied this methodology to characterize and quantify anti-Neu5Gc IgG and IgA in sera of patients with Kawasaki disease (KD) at various stages compared to controls. KD is an acute childhood febrile disease characterized by inflammation of coronary arteries that untreated may lead to coronary artery aneurysms with risk of thrombosis and myocardial infarction. This estimated response is comparable to the average of detailed anti-Neu5Gc IgG profile analyzed by a sialoglycan microarray. Both assays revealed an elevated response in acute KD patients with normal coronaries compared to patients with aneurysm or dilated coronaries. Implications of these findings are discussed.

EFFECT OF CLIMATIC CONDITIONS ON BLOOD PLASMA IgG AND LEPTIN PROFILES IN BUFFALOES

SUMMARY This experiment was conducted during 2010 (August and September) in two buffalo farms located at Giza (F1) and Qena (F2) governorates. A total of 10 buffaloes (3 to 6 parities) was assigned to two experimental groups (n=5 per location). Blood samples were collected fortnightly at 09:00 h before feeding all the experimental animals to assess Leptin and IgG profiles as affected by Temperature Humidity Index (THI). To calculate the THI, meteorological data were collected at the two locations of the experiment. The results demonstrated that Leptin concentration was greater (P<0.05) in F1 (4.15±0.41 ng/ml) than F2 (1.86±0.41 ng/ml). There was a negative (P<0.05) correlation between Leptin and THI (r = -0.37). No significant (P<0.05) difference in IgG levels was observed among animals in the two farms. The plasma IgG concentration in buffaloes reared at F1 (19450±1480 mg/l) was greater than that of F2 (18439±1480 mg/l). There was no correlation between the levels of leptin hormone and total IgG. In conclusion, the increase of heat load during August to September presented as THI on lactating buffalo decreases their leptin hormone and total IgG level in blood plasma.

Comparison of Commercial Tests for Detecting Multiple Anti-Ganglioside Autoantibodies in Patients with Well-Characterized Immune-Mediated Peripheral Neuropathies

To assess the performance of commercial anti-ganglioside antibody assays, we determined anti-ganglioside antibody IgG and IgM isotype profiles of patients with acute and chronic well-characterized immune-mediated peripheral neuropathies by one immunodot assays (Zentec/Ingen: Dotzen Ganglio Profile Ab, Euroimmun/BioAdvance: Euroline ganglioprofile), two line-immuno assay (GA Generic Assays/Labodia: Anti-Gangli osid Dot, Euroimmun/BioAdvance: Euroline ganglioprofile), and one enzyme-linked immunosorbent assay (ELISA) (Bühlmann: GanglioCombi). Specific antibody profiles were compared with those obtained by our validated standard in-house immunodot assay (IDA). We selected 33 sera with high levels of IgG and IgM anti-ganglioside antibodies from 15 patients with Guillain-Barre syndrome (GBS) subtypes and variants, 12 patients with CANOMAD syndrome (chronic ataxic neuropathy with ophthalmoplegia, M-paraprotein, cold agglutinins, disialosyl antibodies), 5 patients with chronic motor peripheral neuropathies, and 1 patient with sensory neuropathy and a control group composed of 10 patients with non-autoimmune neuropathy. The 3 commercial IDAs employing hydrophobic membranes and the ELISA demonstrated different carbohydrate epitopes on 6 to 12 glycolipid antigens used for anti-ganglioside antibody detection. Comparison with the validated in-house IDA showed large variations in sensitivity between tests and a more diverse reactivity to gangliosides than expected. The test with the largest panel of glycolipids detecting 11 anti-ganglioside antibody reactivities (GM1, GM2, GM3, GM4, GD1a, GD1b, GD2, GD3, GT1a, GT1b, GQ1b, and sulfatide) revealed the best concordance with our in-house assay. However, even with this test, differences were observed in the immunoreactivity against some gangliosides and weakly stained bands were not easy to interpret. Our data suggest an urgent need for standardization of commercial anti-ganglioside assays and the introduction of international anti-ganglioside antibody reference standards.