Increased IgG Levels Research Articles

Study of Novel Autoantibodies in Schizophrenia.

A recent genome-wide association (GWA) study confirmed 108 genetic loci that were strongly associated with schizophrenia. Fifteen schizophrenia-associated genes were selected for this study based on a number of selection criteria including their high expression in both brain tissues and B-lymphocyte cells. We aimed to investigate whether individuals with schizophrenia showed different levels of plasma IgG antibodies against protein-derived fragments encoded by these 15 genes. A total of 356 plasma samples were used to analyze circulating IgG antibodies against 18 target peptide antigens using an in-house enzyme-linked immunosorbent assay. Of 18 antigens tested, 6 (derived from DPYD, MAD1L1, ZNF804A, DRD2, TRANK1, and MMP16, respectively) showed increased IgG levels and 3 (derived from TSNARE1, TCF4, and VRK2, respectively) showed decreased IgG levels in patients with schizophrenia compared with control subjects. Receiver operating characteristic (ROC) curve analysis revealed that the anti-TRANK1 IgG assay had the area under the ROC curve of 0.68 (95% CI = 0.62–0.73), with the highest sensitivity of 20.7% against specificity of 95.2% among all 18 tests. There was no difference in positivity of anti-double strand DNA IgG between the patient group and the control group and no correlation between total IgG levels and each individual IgG level tested. Although risperidone treatment showed confounding effects on overall IgG levels in the circulation (combined P = .005), anti-TRANK1 IgG levels did not appear to be significantly affected (t = 1.358, P = .176). In conclusion, this study suggests that circulating anti-TRANK1 IgG is likely to serve as a biomarker for identification of a subgroup of schizophrenia.

Abstract TP341: Pericyte Derived Laminin Play a Beneficial Role in Intracerebral Hemorrhage

Introduction: Laminin, a major component of the basement membrane, exists in many isoforms. Although loss of laminin is found in intracerebral hemorrhage (ICH), the exact biological function of laminin in ICH remains largely unknown. This is primarily due to its intrinsic complexity (various isoforms) and the lack of genetic tools. In a previous study, we generated a transgenic line with laminin deficiency specifically in pericytes (PKO). These PKO mice are grossly normal in pure C57Bl6 background at young age, although mild blood brain barrier (BBB) breakdown is detected in aged mutants. In this study, we aim to investigate the role of pericyte-derived laminin in brain pathology and disease progression in ICH. Methods: Collagenase was injected into the striatum of young (2-month-old) control and PKO mice to induce hemorrhage. At 2, 5, and 14 days post injury (dpi), hematoma size, neuronal death, vascular permeability, inflammatory cell infiltration, and neurological deficits were examined. Results: Compared to the controls, the PKO mice showed larger hematoma and increased neuronal deaths at 2 and 5 dpi, but more severe neurological deficits at 14 dpi. Additionally, substantially increased IgG level was found in PKO brains at 14 dpi, indicating BBB integrity disruption. Consistent with this observation, significantly higher levels of CD45 + leukocytes and Ly6G + granulocytes were detected in PKO brains at 14 dpi. Similarly, elevated CD3 staining was found in PKO brains at 5 and 14 dpi. These data suggest that the recovery of vascular integrity after ICH is delayed in PKO mice. Moreover, PKO brains also showed increased number of CD11b + microglia/macrophages at all three time points examined, suggesting a possible role of pericyte-derived laminin in microglia/macrophage activation and/or migration. Conclusion: Our data support a neuroprotective role of pericyte-derived laminin in ICH. This beneficial effect is probably due to its biological activities in: (1) promoting neuronal survival; (2) stabilizing newly formed vessels and enhancing their integrity; (3) inhibiting inflammatory cell infiltration; and (4) regulating the activation and/or migration of microglia/macrophages.

ВОЗМОЖНОСТИ НЕМЕДИКАМЕНТОЗНОЙ КОРРЕКЦИИ ИММУНОЛОГИЧЕСКИХ НАРУШЕНИЙ В ОСТРОМ ПЕРИОДЕ ИШЕМИЧЕСКОГО ИНСУЛЬТА

In order to study the effectiveness of reflexotherapy in correction of immunological disorders in an acute period of ischemic stroke, 45 patients were clinical and immunological examined (on the 2nd day of patients’s stay in hospital and 15 days after the beginning of the course of early rehabilitation). In the main group of patients (30 people), whose basic complex of rehabilitation measures was optimized by the inclusion of reflexotherapy, there was a significant improvement in a number of indicators of cellular and humoral immunity: reduction of leukocytes in peripheral blood (p˂0.05), an increase in lymphocyte count (p˂0.05), relative and absolute values of T-lymphocytes (CD3+) content (p˂0.01 and p˂0.05, respectively), immunoregulatory cells of T-helpers (CD4+) (p˂0.05), decrease in the number of B-lymphocytes to normal values (p˂0.01), and an increase in IgG level (p˂0.05). In the control group (15 people), where standard treatment was conducted, there was no pronounced dynamics of the indices. Thus, a comprehensive clinical and immunological study of the effectiveness of non-drug correction of immunological disorders in the acute period of ischemic stroke demonstrated high efficiency of acupuncture, with relative simplicity and safety of its application.

Blood Biochemical and Immunological Responses to Garlic Oil Administration in Growing Rabbits Diet

The effect of dietary supplementation garlic oil on performance and blood parameters of New Zealand White rabbits has been studied. The garlic oil was added at graded levels of 0.0, 0.25, 0.5 and 0.75 g/kg for T1 (control), T2, T3 and T4, respectively. A total of thirty six male rabbits, 7 weeks of age and with an initial weigh of 950 g were used. There were four treatments, each with three replicates (n = 3) in randomly divided design. The parameters investigated were growth performance, carcass yield, glucose, plasma lipids profile, immunoglobulin’s G (IgG, IgA and IgM), antioxidant status and testosterone hormone. The results showed that feeding diet supplemented with garlic oil had insignificant effect on body weight, body weight gain, feed intake, feed conversion ratio, triglycerides, cholesterol, LDL and HDL. Rabbits fed the 0.5 g/kg garlic oil diet had significantly increased IgG level, hence improved immune responses and Testosterone hormone of rabbits. The colony forming units of coliform bacteria showed a significantly lower number compared with control. The present results indicate that supplemented of garlic oil at 0.5 g/kg of diet has a positive effect on HDL, immunoglobulin’s G, antioxidant status and testosterone hormone in addition to its antibacterial effect.

Vitamin C supplementation improved the efficacy of foot-and-mouth disease vaccine

ABSTRACT Vitamin C (VC) is an essential micronutrient and plays important roles in various biological processes including immune responses. Vaccination can stimulate immune responses effectively for prevention of virual infection. Adequate VC intake is required for the immune system to function efficiently. In this study, we evaluated the biological function of VC on immune response to inactivated foot-and-mouth disease (FMD) vaccine. The C57BL/6 mice were injected intraperitoneally with different dose of VC daily following immunization with inactivated FMD vaccine. The results showed high dose of VC supplementation significantly increased IgG level. Similarly, high dose of VC supplementation enhanced Th2 and Th17 cellular immune responses and also increased MHCII expression on dendritic cells. Taken together, these data indicate that high dose of VC supplementation can modulate Th2 and Th17 cellular immune responses to inactivated FMD vaccine and manifest the necessity for effective vaccination.

Pentalinonsterol, a Constituent of Pentalinon andrieuxii, Possesses Potent Immunomodulatory Activity and Primes T Cell Immune Responses.

The use of natural products as adjuvants has emerged as a promising approach for the development of effective vaccine formulations. Pentalinonsterol (PEN) is a recently isolated compound from the roots of Pentalinon andrieuxii and has been shown to possess antileishmanial activity against Leishmania spp. The objective of this study was to examine the immunomodulatory properties of PEN and evaluate its potential as an adjuvant. Macrophages and bone-marrow-derived dendritic cells (BMDCs) were stimulated with PEN and tested for gene expression, cytokine production, and their ability to activate T cells in vitro. PEN was also evaluated for its ability to generate antigen-specific Th1 and Th2 responses in vivo, following ovalbumin (OVA) immunization using PEN as an adjuvant. The results obtained demonstrate that PEN enhances the expression of NF-κB and AP1 transcription factors, promotes gene expression of Tnfα, Il6, Nos2, and Arg1, and upregulates MHCII, CD80, and CD86 in macrophages. PEN also enhanced IL-12 production in BMDCs and promoted BMDC-mediated production of IFN-γ by T cells. Further, mice immunized with OVA and PEN showed enhanced antigen-specific Th1 and Th2 cytokines in their splenocytes and lymph node cells, as well as increased levels of IgG1 and IgG2 in their sera. Taken together, this study demonstrates that PEN is a potent immunomodulatory compound and potentially can be used as an adjuvant for vaccine development against infectious diseases.

Circulating Antibodies to Linear Peptide Antigens Derived from ANXA1 and FOXP3 in Lung Cancer.

Our previous studies revealed that concentrations of circulating antibodies to annexin A1 (ANXA1) and forkhead-box P3 (FOXP3) increased significantly in patients with non-small cell lung cancer (NSCLC). This study was thus undertaken to replicate our initial findings with different sample sets. Antibodies were tested in 108 patients with NSCLC and 216 controls, who were divided into the discovery (49 vs. 108) and validation (60 vs. 108) group based on the time of enrolment. Analysis of the discovery group showed a significant increase in circulating anti-ANXA1 IgG levels in the patient group compared with the control group (p=0.005) but the validation group simply exhibited a trend toward an increase in IgG levels in NSCLC (p=0.238), generating a combined p-value of 0.009. The findings of this study support the notion that circulating IgG antibodies to ANXA1 could be used as a biomarker for early diagnosis of NSCLC but failed to replicate such findings for FOXP3.

A Phase 1 Human Immunodeficiency Virus Vaccine Trial for Cross-Profiling the Kinetics of Serum and Mucosal Antibody Responses to CN54gp140 Modulated by Two Homologous Prime-Boost Vaccine Regimens.

A key aspect to finding an efficacious human immunodeficiency virus (HIV) vaccine is the optimization of vaccine schedules that can mediate the efficient maturation of protective immune responses. In the present study, we investigated the effect of alternate booster regimens on the immune responses to a candidate HIV-1 clade C CN54gp140 envelope protein, which was coadministered with the TLR4-agonist glucopyranosyl lipid A-aqueous formulation. Twelve study participants received a common three-dose intramuscular priming series followed by a final booster at either 6 or 12 months. The two homologous prime-boost regimens were well tolerated and induced CN54gp140-specific responses that were observed in both the systemic and mucosal compartments. Levels of vaccine-induced IgG-subclass antibodies correlated significantly with FcγR engagement, and both vaccine regimens were associated with strikingly similar patterns in antibody titer and FcγR-binding profiles. In both groups, identical changes in the antigen (Ag)-specific IgG-subclass fingerprint, leading to a decrease in IgG1 and an increase in IgG4 levels, were modulated by booster injections. Here, the dissection of immune profiles further supports the notion that prime-boost strategies are essential for the induction of diverse Ag-specific HIV-1 responses. The results reported here clearly demonstrate that identical responses were effectively and safely induced by both vaccine regimens, indicating that an accelerated 6-month regimen could be employed for the rapid induction of immune responses against CN54gp140 with no apparent impact on the overall quality of the induced immune response. (This study has been registered at http://ClinicalTrials.gov under registration no. NCT01966900.)

Periploca forrestii saponin ameliorates CIA via suppressing proinflammatory cytokines and nuclear factor kappa-B pathways.

ObjectivePeriploca forrestii Schltr has been used as a Chinese folk medicine for the treatment of rheumatism, arthralgia and fractures. However, the anti-arthritic activity of Periploca forrestii saponin (PFS) and the active compound has still not been revealed. This study aimed to investigate the protective effects and mechanisms of PFS on collagen type II (CII) collagen-induced arthritis (CIA) mice. We sought to investigate whether PFS and Periplocin could regulate osteoclastogenesis, and if so, further investigation on its mechanism of action.MethodsArthritis was induced in female BALB/c mice by CIA method. PFS was administered at a dose of 50 mg/kg body weight once daily for five weeks. The effects of treatment in mice were assessed by histological and biochemical evaluation in sera and paws. Anti-osteoclastogenic action of PFS and Periplocin was identified using an osteoclast formation model induced by RANKL.ResultsPFS ameliorated paw erythema and swelling, inhibited bone erosion in ankle joint histopathological examination. PFS treatment resulted in decreased IgG2a, and increased IgG1 levels in the serum of CIA mice. Decreased TNF-α, and increased interleukin (IL)-4 and IL-22 levels were also found in PFS-treated mice. PFS inhibited the I-κBα phosphorylation, blocked nuclear factor (NF)-κB/p65 phosphorylation and abrogated AP-1/c-Fos activity. PFS downregulated toll-like receptor (TLR) 4, STAT3 and MMP-9 expression in CIA mice and RANKL-induced osteoclastogenesis. PFS and Periplocin inhibited RANKL-induced osteoclast formation in a dose dependent manner within nongrowth inhibitory concentration, and PFS decreased osteoclastogenesis-related marker expression, including cathepsin K and MMP-9.ConclusionThis study revealed that the protective mechanism of PFS on CIA was associated with regulatory effects on proinflammatory factors and further on the crosstalk between NF-κB and c-Fos/AP-1 in vivo and in vitro. Therefore, PFS is a promising therapeutic alternative for the treatment of RA, evidencing the need to conduct further studies that can identify their active components in treating and preventing RA.

IgG isotypic antibodies to crude Plasmodium falciparum blood-stage antigen associated with placental malaria infection in parturient Cameroonian women.

Few studies have reported an association between placental malaria (PM) infection and levels of isotypic antibodies against non-pregnancy associated antigens. To determine and evaluate IgG isotypic antibody levels to crude P. falciparum blood stage in women with and without PM infection. Levels of IgG (IgG1-IgG4) and IgM to crude P. falciparum blood stage antigen were measured by ELISA in 271 parturient women. Placental malaria infection was determined by placental blood microscopy and placental histology. Age, parity and intermittent preventive treatment during pregnancy with sulphadoxine-pyrimethamine (IPTp-SP) usage were considered during analysis. P. falciparum-specific IgG1 (96.5%) and IgG3 (96.7%) antibodies were predominant compared with IgG2 (64.6%) and IgG4 (49.1%). Active PM infection was associated with significant increased levels of IgG1, IgG4 and IgM while lower levels of these antibodies were associated with uptake of two or more IPTp-SP doses. PM infection was the only independent factor associated with IgG4 levels. Mean IgG1 + IgG3/IgG2 + IgG4 and IgG1 + IgG2 +IgG3/ IgG4 ratios were higher among the PM-uninfected group while IgG4/IgG2 ratio prevailed in the infected group. PM infection and IPTp-SP dosage influenced P. falciparum-specific isotypic antibody responses to blood stage antigens. An increase in IgG4 levels in response to PM infection is of particular interest.

Autoimmune lymphoproliferative syndrome caused by homozygous FAS mutations with normal or residual protein expression

Autoimmune lymphoproliferative syndrome caused by homozygous FAS mutations with normal or residual protein expression

<i>CTLA-4, ICOS, PD1</i> and <i>PTPN22</i> Gene Polymorphisms and Susceptibility to Autoimmune Hepatitis Type 1

Autoimmune hepatitis type 1 (AIH-1) is a progressive inflammatory liver disorder in which HLA Class II gene polymorphism prevails as the most important genetic risk. However, other gene polymorphisms have been associated with this disease. The single nucleotide polymorphisms of four candidate genes (CTLA-4 +49A/G, ICOS c.1564 T/C, PD1.3 G/A, PTPN22 1858C/T) were selected in this study. One-hundred and ninety (190) mestizo Venezuelan unrelated individuals grouped in AIH-1 patients (n=70) and healthy subjects (n=120) were evaluated. Our results showed significantly increased frequency of the PTPN22 1858 C/T (p= 0.0014; pc= 0.0042; OR= 8.7; 99%CI: 1.82-41.54) and ICOS c.1564 T/C (p= 0.070; pc=0.21; OR= 2.08; 95%CI: 1.09-3.93) genotypes in the patient population compared to control group. There was no significant association between CTLA-4 +49A/G and PD1.3 G/A genotypes in both groups. In addition, the PTPN22 1858C/T polymorphism was associated to cirrhosis, treatment relapse, increased IgG levels and co-existence of other autoimmune diseases. Furthermore, the ICOS c.1564 T/C polymorphism was related to higher levels of globulins, IgG and presence of ANA. Conclusion: this data suggest for the first time, that PTPN22 1858 C/T and ICOS c.1564 T/C gene polymorphisms are associated with the development of AIH-1.

Change of circulating antibodies against CD25-derived peptide antigen in hepatocellular carcinoma.

Several studies have shown altered levels of plasma anti-CD25 antibodies in patients with malignancy in lung, esophagus and breast. The present study was thus designed to test whether circulating anti-CD25 antibody levels were changed in hepatocellular carcinoma (HCC). An enzyme-linked immunosorbent assay (ELISA) was developed in-house to detect plasma IgG antibodies to CD25-derived linear peptide antigens in 122 patients with HCC and 121 control subjects. Student's t-test showed that plasma anti-CD25 IgG levels were significantly higher in HCC patients than control subjects (t = 4.96, P < 0.001), in which male patients mainly contributed to the increased IgG levels (t = 5.11, P < 0.001). Further analysis showed that plasma anti-CD25 IgG levels were dependent on the stages of HCC although there was no significant correlation between plasma anti-CD25 IgG levels and BCLC stages (r = 0.145, P = 0.110, N = 122); a significant increase in anti-CD25 IgG levels was observed in HCC patients with stages B (t = 4.43, P < 0.001) and C+D (t = 4.89, P < 0.001) as compared with control subjects. Receiver operating characteristic (ROC) curve analysis showed that the area under the ROC curve (AUC) was 0.66 (SE = 0.035, 95% CI 0.60-0.73). The sensitivity of anti-CD25 IgG assay was 12.3% against a specificity of 99.2%. The present study suggests that circulating anti-CD25 IgG antibodies may have prognostic rather than early diagnostic values for HCC.

Combining in vitro protein detection and in vivo antibody detection identifies potential vaccine targets against Staphylococcus aureus during osteomyelitis

Currently, little is known about the in vivo human immune response against Staphylococcus aureus during a biofilm-associated infection, such as osteomyelitis, and how this relates to protein production in biofilms in vitro. Therefore, we characterized IgG responses in 10 patients with chronic osteomyelitis against 50 proteins of S. aureus, analyzed the presence of these proteins in biofilms of the infecting isolates on polystyrene (PS) and human bone in vitro, and explored the relation between in vivo and in vitro data. IgG levels against 15 different proteins were significantly increased in patients compared to healthy controls. Using a novel competitive Luminex-based assay, eight of these proteins [alpha toxin, Staphylococcus aureus formyl peptide receptor-like 1 inhibitor (FlipR), glucosaminidase, iron-responsive surface determinants A and H, the putative ABC transporter SACOL0688, staphylococcal complement inhibitor (SCIN), and serine–aspartate repeat-containing protein E (SdrE)] were also detected in a majority of the infecting isolates during biofilm formation in vitro. However, 4 other proteins were detected in only a minority of isolates in vitro while, vice versa, 7 proteins were detected in multiple isolates in vitro but not associated with significantly increased IgG levels in patients. Detection of proteins was largely confirmed using a transcriptomic approach. Our data provide further insights into potential therapeutic targets, such as for vaccination, to reduce S. aureus virulence and biofilm formation. At the same time, our data suggest that either in vitro or immunological in vivo data alone should be interpreted cautiously and that combined studies are necessary to identify potential targets.

Effect of 16 Weeks Six a Side Recreational Football on Serum Immunoglobulin and White Blood Cells in Untrained Males

Objectives: Present study examined how 16 weeks of six a side supervised recreational football affected serum immunoglobulin and white blood cells in untrained males. Methods/Analysis: We selected 27 untrained males having mean age (18.87±.79) years. These participants were then randomly allocated to two groups; football (n = 12) and control group (n = 15). Sixteen weeks of supervised recreational football training was offered to football group. Football sessions were conducted two times a week with 30min duration. Analysis of Covariance was employed to find any difference in two groups after 16 weeks of football training. Findings: Significant increase was seen after 16 weeks in total white blood cell count (P = 0.001), eosinophil (P = 0.006) and basophil (P = 0.005), while no significant improvement was observed in Neutrophil (P = 0.797), lymphocytes (P = 0.820), IgA (P = .158), IgG (P = .077) and IgM (P= .655). Monocytes and eosinophil counts decreased significantly whereas basophil counts increased significantly post football training. Novelty/ Improvement: Sixteen weeks of supervised recreation football training was effective in increasing total WBC count in untrained males. Our training was vigorous in nature as indicated by high average heart rate during the training sessions. Further, recreation football could not affect serum immunoglobulin significantly but there was some substantive increase in IgG levels.

Impact of Immunoglobulin Therapy in Intestinal Transplant Recipients With Posttransplantation Hypogammaglobulinemia

BackgroundSevere hypogammaglobulinemia (HGG) (IgG <400 mg/dL) following intestinal transplantation is common. Although IgG replacement therapy is commonly used, clinical outcomes associated with increasing IgG levels to >400 mg/dL are not well described. MethodsKaplan–Meier analysis was performed to estimate survival, the log-rank test to compare survival distributions between groups, and the Fisher exact test to determine the association between HGG and rejection. ResultsA total of 23 intestinal transplant (IT) recipients with a median age of 2.3 years (range, 0.7–41 years) at the time of HGG diagnosis were included. The types of transplants were liver–small bowel (73.9%), liver–small bowel–kidney (8.7%), and small bowel only (17.4%). The 3-year survival after the diagnosis of HGG was 50.2% (95% confidence interval [CI] = 28.2%–68.7%). There was no difference in survival (P = .67) when patients were dichotomized based upon IgG level at last follow-up (IgG ≥400 mg/dL, n = 14; and IgG <400 mg/dL, n = 9). There was no also evidence of an association between survival and: total dose (P = .58), frequency (P = .11), and number of IgG doses administered (P = .8). There was no difference in survival between patients receiving (n = 12) or not receiving (n = 11) cytomegalovirus hyperimmunoglobulin (P = .10). ConclusionsImproved survival rates were not found in our IT recipients with severe HGG with immunoglobulin therapy to IgG levels of ≥400 mg/dL, even when cytomegalovirus hyperimmunoglobulin was administered.

Aspergillus fumigatus promotes T helper type 2 responses through thymic stromal lymphopoietin production by human corneal epithelial cells.

Fungal keratitis is a major cause of blindness. To understand the mechanism of both innate and adaptive immunity in corneal infection is of great significance in the treatment and prevention of fungal keratitis. Our previous study concerned innate immunity. Here, we explored the potential role of thymic stromal lymphopoietin (TSLP) in adaptive immunity of fungal keratitis. Human corneal epithelial cells (HCECs) were stimulated with Aspergillus fumigatus hyphae (10(6) pieces per millilitre) with or without TSLP siRNA, and peripheral blood mononuclear cells (PBMCs) were cultured with or without TSLP. HCECs and PBMCs were co-cultured in a transwell system for various periods. Then we collected PBMCs and detected the proliferation and activation as well as T helper type 2 (Th2) differentiation by flow cytometry and quantitative real-time reverse transcription polymerase chain reaction. IgG and IgA levels in supernatants of PBMCs were measured by means of ELISA. Thymic stromal lymphopoietin could induce a Th2 response in vitro, and the expression of TSLP was highly increased in HCECs stimulated with A. fumigatus hyphae. A. fumigatus-infected HCECs were capable of promoting human lymphocyte proliferation and activating human CD4(+) T cells, CD8(+) T cells and B cells by up-regulating the expression of activation marker CD69. Importantly, Th2 differentiation of CD4(+) T cells was induced during co-culture with A. fumigatus-infected HCECs in a transwell system. Interestingly, blockade of TSLP using siRNA prevented the proliferation and activation of lymphocytes as well as Th2 differentiation. We also detected an increased IgG level that was associated with TSLP. These findings suggested that HCEC-derived TSLP has a key role in adaptive immune responses of fungal keratitis via skewing Th2 differentiation and promoting humoral immunity.

Effect of Heat Stress on Humoral Immunity and its Amelioration by Amla Powder (Emblica officinalis) Supplementation in Buffaloes

The effect of amla powder supplementation on heat stress and immunoglobulin (Ig) levels of summer stressed buffaloes were investigated. The study was carried out with 24 apparently healthy Murrah buffaloes during pre-summer and summer seasons. The animals were divided into 3 groups of 8 animals each viz. Pre-Summer group (Group I); Summer Control group (Group II); Summer Treatment group (Group III). Group III animals were supplemented with amla powder @ 200 mg/Kg body wt./day for 30 days. In summer stressed buffaloes (Group II), there was a significant rise in erythrocytic lipid peroxidation level and superoxide dismutase activity along with a decrease in plasma IgM and increase in IgG levels. Supplementation of amla powder to summer stressed buffaloes was able to lower lipid peroxidation levels and superoxide dismutase activity coupled with the increase in IgM and decrease in IgG levels. It was concluded that amla powder supplementation can ameliorate the adverse effects of summer stress on humoral immunity in Murrah buffaloes.

IgE and IgG4 Epitope Mapping of Food Allergens with a Peptide Microarray Immunoassay.

Peptide microarrays are a powerful tool to identify linear epitopes of food allergens in a high-throughput manner. The main advantages of the microarray-based immunoassay are the possibility to assay thousands of targets simultaneously, the requirement of a low volume of serum, the more robust statistical analysis, and the possibility to test simultaneously several immunoglobulin subclasses. Among them, the last one has a special interest in the field of food allergy, because the development of tolerance to food allergens has been associated with a decrease in IgE and an increase in IgG4 levels against linear epitopes. However, the main limitation to the clinical use of microarray is the automated analysis of the data. Recent studies mapping the linear epitopes of food allergens with peptide microarray immunoassays have identified peptide biomarkers that can be used for early diagnosis of food allergies and to predict their severity or the self-development of tolerance. Using this approach, we have worked on epitope mapping of the two most important food allergens in the Spanish population, cow's milk and chicken eggs. The final aim of these studies is to define subsets of peptides that could be used as biomarkers to improve the diagnosis and prognosis of food allergies. This chapter describes the protocol to produce microarrays using a library of overlapping peptides corresponding to the primary sequences of food allergens and data acquisition and analysis of IgE- and IgG4-binding epitopes.

Quantitative and Functional Antibody Responses to the 13-Valent Conjugate and/or 23-Valent Purified Polysaccharide Vaccine in Aging HIV-Infected Adults.

The number of aging human immunodeficiency virus-infected (HIV+) individuals living in the United States has substantially grown over the past two decades. Advanced age and HIV infection both increase susceptibility to Streptococcus pneumoniae infection due to B cell dysfunction. The combined impact of these factors on pneumococcal vaccine responses remains unknown. We assessed serum immunoglobulin (Ig) G and IgM levels and opsonophagocytic killing assay (OPA) titers to pneumococcal serotypes 14 and 23F in HIV+ subjects and HIV-uninfected (HIV-) controls 50-65 years old. HIV+ individuals with CD4+ T cells/μl (CD4) >200 and ≥1 year of antiretroviral therapy (ART) received either a dose of the 13-valent pneumococcal conjugate vaccine followed by the 23-valent pneumococcal polysaccharide vaccine 8 weeks later (PCV/PPV) as currently recommended (n=15) or a single dose of PPV only (n=22). HIV- controls received PCV/PPV (n=14). HIV+ PCV/PPV and PPV groups exhibited similar increases in IgG levels and OPA titers for both serotypes after immunization. Postvaccination IgM levels for serotype 23F, but not 14, were significantly higher in HIV+ PCV/PPV compared to PPV groups. IgG and IgM levels for serotype 14 and OPA titers to serotype 23F were significantly reduced in HIV+ compared to HIV- PCV/PPV groups. Serotype-specific IgG levels correlated with OPA titers for all groups. Our data suggest that the recommended PCV/PPV regimen may not significantly improve quantitative or functional antibody responses compared to PPV only in aging HIV+ subjects. Continued efforts aimed at improving vaccine responses in this high risk population are warranted.