Total IgG Levels Research Articles

The Development of a Murine Model for Forcipomyia taiwana (Biting Midge) Allergy

Background Forcipomyia taiwana (biting midge) allergy is the most prevalent biting insect allergy in Taiwan. An animal model corresponding to the human immuno-pathologic features of midge allergy is needed for investigating the mechanisms and therapies. This study successfully developed a murine model of Forcipomyia taiwana allergy.MethodsBALB/c mice were sensitized intra-peritoneally with midge extract on days 0, 7, 14, 21 then intra-dermally on days 28, 31 and 35. Serum midge-specific IgE, IgG1, and IgG2a were measured every 14 days by indirect ELISA. The mice were challenged intradermally with midge extract at day 40 and then sacrificed. Proliferation and cytokine production of splenocytes after stimulation with midge extract were determined by MTT assay and ELISA, respectively. The cytokine mRNA expression in response to midge stimulation was analyzed by RT-PCR.ResultsSerum IgE, total IgG, and IgG1 antibody levels against midge extract were significantly higher in the midge-sensitized mice than in the control mice. After the two-step sensitization, all mice in the midge-sensitized group displayed immediate itch and plasma extravasation reactions in response to challenge with midge extract. Skin histology from midge-sensitized mice showed marked eosinophil and lymphocyte infiltrations similar to that observed in humans. Stimulation of murine splenocytes with midge extract elicited significant proliferation, IL-4, IL-10, IL-13 and IFN-γ protein production, and up-regulation of mRNA in a dose-dependent manner in the midge-sensitized group, but not in the control group.ConclusionsA murine model of midge bite allergy has been successfully developed using a two-step sensitization protocol. The sensitized mice have very similar clinical and immunologic reactions to challenge with midge proteins as the reactions of human to midge bites. This murine model may be a useful platform for future research and the development of treatment strategies for insect bite allergy.

Impact of colostrum and plasma immunoglobulin intake on hippocampus structure during early postnatal development in pigs

The first milk, colostrum, is an important source of nutrients and an exclusive source of immunoglobulins (Ig), essential for the growth and protection from infection of newborn pigs. Colostrum intake has also been shown to affect the vitality and behaviour of neonatal pigs. The objective of this study was to evaluate the effects of feeding colostrum and plasma immunoglobulin on brain development in neonatal pigs.Positive correlations were found between growth, levels of total protein and IgG in blood plasma and hippocampus development in sow-reared piglets during the first 3 postnatal days. In piglets fed an elemental diet (ED) for 24h, a reduced body weight, a lower plasma protein level and a decreased level of astrocyte specific protein in the hippocampus was observed, as compared to those that were sow-reared. The latter was coincident with a reduced microgliogenesis and an essentially diminished number of neurons in the CA1 area of the hippocampus after 72h. Supplementation of the ED with purified plasma Ig, improved the gliogenesis and supported the trophic and immune status of the hippocampus.The data obtained indicate that the development of the hippocampus structure is improved by colostrum or an Ig-supplemented elemental diet in order to stimulate brain protein synthesis and its development during the early postnatal period.

Myocardial performance in children with autoimmune hepatitis: Doppler tissue imaging study

Purpose The aim of this study was to assess subclinical impact of autoimmune hepatitis (AIH) on global myocardial performance in children using Doppler tissue imaging (DTI) and to correlate it with total serum IgG. Methods We included 30 children with AIH (mean age = 12.67 ± 2.9 years) as the study group and 20 age and sex matched healthy children (mean age = 11.93 ± 2.66 years) as the control group. Conventional two-dimensional echocardiography was performed to both groups and DTI were used to determine Right ventricular (RV) and left ventricular (LV) Tei indexes. Total serum IgG levels at initial diagnosis of AIH were retrieved from our hospital data-base and were correlated to the cardiac functions of AIH patients. Results Right ventricular (RV) and left ventricular (LV) Tei indexes were significantly higher in AIH group (mean ± SD: 0.46 ± 0.088 vs. 0.26 ± 0.01, P 0.0001 and 0.45 ± 0.086 vs. 0.31 ± 0.02, P 0.0001 , respectively). Mitral and tricuspid systolic (Sm) velocities were significantly lower in AIH children (mean ± SD: 8.226 ± 1.73 vs. 9.93 ± 0.94 cm/s, P = 0.0002 and 10.85 ± 1.11 vs. 12.48 ± 0.53 cm/s, P 0.0001 , respectively). Total IgG concentrations were significantly higher in AIH children [mean ± SD: 2841.3 ± 607.17 vs. 862.67 ± 70.35 mg/dl, P 0.0001 ], and they correlated positively with the LV Tei index ( r = 0.69, P 0.0001 ) and the RV Tei index ( r = 0.61, P 0.0003 ) and correlated negatively with the mitral systolic (Sm) velocity ( r = −0.76, P 0.0001 ) and tricuspid systolic (Sm) velocity ( r = −0.66, P 0.0001 ), on the other hand, fractional shortening (FS) did not correlate with serum IgG concentrations ( r = - 0.04 , P = 0.821 ). Conclusion The DTI technique appears to be more sensitive than conventional echocardiography in the early detection of myocardial dysfunction in AIH children.

Enhancement of Immune Responses by Co-delivery of CCL19/MIP-3beta Chemokine Plasmid With HCV Core DNA/Protein Immunization

Background:Using molecular adjuvants offers an attractive strategy to augment DNA vaccine-mediated immune responses. Several studies have revealed that an efficient HCV vaccine model should be able to induce both humoral and cell mediated immune responses targeting the conserved regions of the virus to circumvent the immune escape mutants. The beta chemokine Macrophage Inflammatory Protein 3-beta (MIP-3beta) is a key modulator of dendritic cells (DCs) and T-cells interaction, functions during immune response induction and is secreted specifically by cells in the lymphoid tissues.Objectives:In the present study, we questioned whether co-administration of MIP-3beta gene could enhance the immune responses to HCV core in DNA vaccination.Materials and Methods:Expression and biological activity of MIP-3beta expressing plasmid were evaluated by ELISA and transwell migration assays, respectively. HCV core DNA vaccine ± plasmid expressing MIP-3beta were electroporated subcutaneously to the front foot pads of BALB/c mice on days 0 and 14, and HCV core protein booster was applied to all core-DNA-vaccine received mice on the day 28. Both cell mediated immunity (proliferation, IFN-γ and IL-4 cytokine release, IFN-γ ELISpot and cytotoxic Granzyme B release assays) and humoral immune responses (total IgG and IgG2a/IgG1 subtyping) were evaluated ten days after final immunization.Results:Mice covaccinated with MIP-3beta elicited an enhanced Th1 biased systemic immune response as evidenced by higher IFN-γ/IL-4 and anti-core IgG2a/IgG1 ratio, lymphoproliferation, strong cytolytic GrzB release and enhanced population of IFN-γ producing immunocytes. Likewise, the humoral immune response assumed as the total anti-core IgG level was augmented by MIP-3beta co-delivery.Conclusions:These results exhibited the immuno potentiator effects of MIP-3beta plasmid when coadministrated with the HCV core DNA vaccine. Complimentary studies integrating MIP-3beta as a genetic adjuvant in HCV-core-DNA vaccination models are warranted.

면역보강제로서 부가적인 역할을 가지는 복합 살모넬라 타이피무리움 고스트 세포

고스트 백신/LT-B 그룹과 고스트 백신 복합 균주를 각각 구강으로 투여했을 때, 복합고스트백신 그룹은 LT-B와 유사한 면역 보강제로서 역할을 수행할 수 있는 것으로 나타났다. 따라서 독력이 다른 살모넬라 균주와 LT-B을 대체하는 복합 균주 군을 조성하여 intramuscle (IM)로 주사했다. 그 결과, 복합균주 군은 LT-B을 운반하는 pMMP300과 유사한 총 IgG 함량 변화를 보였다. IgG1과 IgG2a는 LT-B 그룹과 유사한 면역 응답을 보였다. 질 sIgA는 LT-B와 약간 차이가 나타났지만, 전반적으로 낮은 면역응답을 보인 반면에, fecal sIgA는 LT-B보다 높은 면역 응답을 보였다. 독력 살모넬라에 대한 보호효과는 대조구보다 30% 높은 수치를 나타내었고, LT-B와 유사한 결과를 보였다. 이상의 결과로 볼 때, 살모넬라 복합균주는 면역보강제 LT-B를 대체할 수 있는 역할을 수행하는 것으로 추정된다. Ghost cells have been recognized as eliciting humoral and cell-mediated immune responses and have also been predicted to play a role as an immune adjuvant. In this study, we used the intramuscular (IM) route to inject BALB/c mice with four vaccine groups constructed from Salmonella typhimurium ghost (STG) cells originating from different virulent strains and complex STG groups instead of heat-labile toxin (LT)-B, a type of adjuvant. Although the complex STG groups exhibited a response after a short delay, the groups showed final total IgG levels similar to those of the LT-B group, which encodes LT-B from pMMP300. The IgG1 response to the <TEX>${\chi}$</TEX>3339 group was the highest response at 6 weeks, whereas IgG2a responses to the <TEX>${\chi}$</TEX>3339 and JOL389 groups were higher at 6 and 8 weeks compared to those of the LT-B group. The response of vaginal sIgA to the LT-B group was generally higher than that of the other groups, whereas fecal sIgA to the LT-B group exhibited lower responses. Protection to virulent S. typhimurium in all groups was above 80%, which was similar to the LT-B group. Taken together, we suggest that STG complex groups can be used as an immune adjuvant instead of LT-B.

Circulating Immunoglobulins Are Not Associated with Intraplaque Mast Cell Number and Other Vulnerable Plaque Characteristics in Patients with Carotid Artery Stenosis

BackgroundRecently, we have shown that intraplaque mast cell numbers are associated with atherosclerotic plaque vulnerability and with future cardiovascular events, which renders inhibition of mast cell activation of interest for future therapeutic interventions. However, the endogenous triggers that activate mast cells during the progression and destabilization of atherosclerotic lesions remain unidentified. Mast cells can be activated by immunoglobulins and in the present study, we aimed to establish whether specific immunoglobulins in plasma of patients scheduled for carotid endarterectomy were related to (activated) intraplaque mast cell numbers and plasma tryptase levels. In addition, the levels were related to other vulnerable plaque characteristics and baseline clinical data.Methods and ResultsOxLDL-IgG, total IgG and total IgE levels were measured in 135 patients who underwent carotid endarterectomy. No associations were observed between the tested plasma immunoglobulin levels and total mast cell numbers in atherosclerotic plaques. Furthermore, no associations were found between IgG levels and the following plaque characteristics: lipid core size, degree of calcification, number of macrophages or smooth muscle cells, amount of collagen and number of microvessels. Interestingly, statin use was negatively associated with plasma IgE and oxLDL-IgG levels.ConclusionsIn patients suffering from carotid artery disease, total IgE, total IgG and oxLDL-IgG levels do not associate with plaque mast cell numbers or other vulnerable plaque histopathological characteristics. This study thus does not provide evidence that the immunoglobulins tested in our cohort play a role in intraplaque mast cell activation or grade of atherosclerosis.

Antitumor Cell-Complex Vaccines Employing Genetically Modified Tumor Cells and Fibroblasts

The present study evaluates the immune response mediated by vaccination with cell complexes composed of irradiated B16 tumor cells and mouse fibroblasts genetically modified to produce GM-CSF. The animals were vaccinated with free B16 cells or cell complexes. We employed two gene plasmid constructions: one high producer (pMok) and a low producer (p2F). Tumor transplant was performed by injection of B16 tumor cells. Plasma levels of total IgG and its subtypes were measured by ELISA. Tumor volumes were measured and survival curves were obtained. The study resulted in a cell complex vaccine able to stimulate the immune system to produce specific anti-tumor membrane proteins (TMP) IgG. In the groups vaccinated with cells transfected with the low producer plasmid, IgG production was higher when we used free B16 cell rather than cell complexes. Nonspecific autoimmune response caused by cell complex was not greater than that induced by the tumor cells alone. Groups vaccinated with B16 transfected with low producer plasmid reached a tumor growth delay of 92% (p ≤ 0.01). When vaccinated with cell complex, the best group was that transfected with high producer plasmid, reaching a tumor growth inhibition of 56% (p ≤ 0.05). Significant survival (40%) was only observed in the groups vaccinated with free transfected B16 cells.

Tunisian radish (Raphanus sativus) extract prevents cadmium-induced immunotoxic and biochemical alterations in rats

Cadmium (Cd), a known carcinogen and potent immunotoxicant in humans and animals, is dispersed throughout the environment as a result of pollution from a variety of sources. Tunisian radish (Raphanus sativus) extract (TRE) is a known anti-oxidant and free radical scavenger that has been shown to help alleviate immune system disorders, including some induced by environmental toxicants. The present study was undertaken to investigate potential protective effects of TRE against Cd-induced immunotoxicities (and general toxicities) in situ. Cadmium chloride (at 2.5 mg CdCl2/kg BW) and TRE (5, 10, or 15 mg/kg BW) were given (alone or in combination [actually, in sequence of Cd and then TRE]) to rats daily by oral gavage for 2 weeks. Results indicated that treatment with CdCl2 alone resulted in significant decreases in plasma levels of total protein, triglycerides, creatine kinase, creatinine, IgG and IgA, T-lymphocyte sub-types (CD4+, CD3+, CD56+, and CD8+), and in thymic and hepatic indices (relative weights). In contrast, CdCl2 treatment caused significant increases in serum LDH, AST, and ALT, in the formation/release of pro-inflammatory cytokines (IL-1 and TNFα), and in the relative weights of host spleen and kidneys. Rats treated with TRE alone had no discernable changes compared to the controls with regard to all test parameters. Combined treatment of CdCl2 and TRE—at any dose—resulted in a significant improvement of all test parameters compared to those seen with Cd alone. These results illustrated (and provided further support for a continuing belief in) the beneficial effects of TRE in reducing the harmful outcomes of commonly encountered toxicants (like Cd) on the immune system and on overall host health status.

LBSapSal-vaccinated dogs exhibit increased circulating T-lymphocyte subsets (CD4+ and CD8+) as well as a reduction of parasitism after challenge with Leishmania infantum plus salivary gland of Lutzomyia longipalpis

BackgroundThe development of a protective vaccine against canine visceral leishmaniasis (CVL) is an alternative approach for interrupting the domestic cycle of Leishmania infantum. Given the importance of sand fly salivary proteins as potent immunogens obligatorily co-deposited during transmission of Leishmania parasites, their inclusion in an anti-Leishmania vaccine has been investigated in the last few decades. In this context, we previously immunized dogs with a vaccine composed of L. braziliensis antigens plus saponin as the adjuvant and sand fly salivary gland extract (LBSapSal vaccine). This vaccine elicited an increase in both anti-saliva and anti-Leishmania IgG isotypes, higher counts of specific circulating CD8+ T cells, and high NO production.MethodsWe investigated the immunogenicity and protective effect of LBSapSal vaccination after intradermal challenge with 1 × 107 late-log-phase L. infantum promastigotes in the presence of sand fly saliva of Lutzomyia longipalpis. The dogs were followed for up to 885 days after challenge.ResultsThe LBSapSal vaccine presents extensive antigenic diversity with persistent humoral and cellular immune responses, indicating resistance against CVL is triggered by high levels of total IgG and its subtypes (IgG1 and IgG2); expansion of circulating CD5+, CD4+, and CD8+ T lymphocytes and is Leishmania-specific; and reduction of splenic parasite load.ConclusionsThese results encourage further study of vaccine strategies addressing Leishmania antigens in combination with proteins present in the saliva of the vector.

Humoral immune responses to HIV in the mucosal secretions and sera of HIV-infected women.

Although sera and all external secretions contain antibodies to human immunodeficiency virus (HIV), their levels, specificity, isotypes, and relevant effector functions display a great degree of variability. Antibodies that bind HIV antigens and neutralize the virus are predominantly associated with the IgG isotype in sera and in all external secretions, even where total levels of IgG are much lower than those of IgA. Rectal fluid that contains high IgA, but low IgG levels, displayed low neutralizing activity independent of antibodies. Therefore, external secretions should be evaluated before and after selective depletion of Ig. At the systemic level, HIV-specific IgA may interfere with the effector functions of IgG, as suggested by recent studies of individuals systemically immunized with an experimental HIV vaccine. Although HIV-specific IgG and IgA antibodies may exhibit their protective activities at mucosal surfaces through interference with viral entry and local neutralization at the systemic level, such antibodies may display discordant effector functions.

Evaluation of efficacy, biodistribution and safety of antibiotic-free plasmid encoding somatostatin genes delivered by attenuated Salmonella enterica serovar Choleraesuis

We describe here a balanced-lethal system using an Asd+ expression plasmid pVGS/2SS-asd encoding two copies of somatostatin (SS) genes carried by Δasd/Δcrp double mutant Salmonella enterica serovar Choleraesuis (named C501). The advantage of this novel system is the use of asd (aspartate-semialdehyde dehydrogenase) gene as selection marker to replace the antibiotic resistance markers, thus eliminating the industrial cultivation and environmental problems. We then evaluated the efficacy, biodistribution and safety of antibiotic-free plasmid delivered by strains C501. Mice orally immunized with C501 (pVGS/2SS-asd) had significantly higher levels of anti-SS total IgG and IgA antibodies than control mice and demonstrated a bias toward Th2-associated responses (IgG1/IgG2a ratio>1). Safety evaluation indicated that vaccinated mice displayed no abnormal clinical signs and histological changes. Biodistribution result revealed that the GS/2SS message was detected in several examined tissues with the exception of ovary and brain, but was rapidly cleared from the body (approximately 10 days). Furthermore, the risk of integration of plasmid pVGS/2SS-asd into the host cellular genome was considered to be negligible. These results may have important implications for the use of vaccine strain C501 (pVGS/2SS-asd) in domestic animals and prompt new perspectives on the safety of DNA vaccines delivered by attenuated bacteria.

The use of humoral responses as a marker of CMV burden in HIV patients on ART requires consideration of T-cell recovery and persistent B-cell activation.

Objectives. Elevated humoral responses to cytomegalovirus (CMV) associate with increased risk of cardiovascular disease (CVD) in HIV patients on antiretroviral therapy (ART). To better understand the persistence of CMV humoral responses in relation to CVD, we determined trends in CMV antibody levels over the first 10 years on ART. Design. We describe longitudinal analyses of plasma from 13 HIV patients commencing ART with <210 CD4 T-cells/µL and 27 controls. Antibodies reactive with CMV (fibroblast lysate, gB and IE-1 antigens), EBV-VCA, and HIVgp41 were quantitated. B-cell activation was assessed via total IgG and sBAFF. Inflammation was assessed via sTNF-RI and sCD14. Results. Amongst CMV seropositive HIV patients, levels of antibody reactive with CMV (P = 0.03) and EBV-VCA (P = 0.02) peaked after 1 year on ART. Levels of total IgG, sCD14, and sTNF-RI declined to approximate those in controls after 10 years, but sBAFF (P = 0.0002), EBV-VCA (P = 0.001), and CMV (P = 0.0004) antibodies remained elevated. A strong correlation between sBAFF and CMVgB antibody was seen at 10 years (R = 0.93, P = 0.0009) and verified in a second cohort. Conclusions. CMV antibody titres peak on ART and remain high. A correlation between CMV antibody and sBAFF suggests a role for HIV-induced B-cell pathology that may affect its use as a marker of CMV burden.

IgG and the sum of ohe subclasses: what is it good for?

Determination of IgG subclasses has a role in the diagnosis of IgG4 disease, and possibly IgG subclass deficiency. It is recommended that the total IgG level be measured as a quality check, with the requirement that the sum of the subclasses be within 15%. Although widely adopted, there seems little scientific justification for such an approach. We performed a prospective audit of 297 samples sent for IgG subclass measurement. Fifty of 297 samples failed the 15% rule; subsequently, eight passed on repeating the IgG, and another six on repeating the subclasses. EPG on the 36 remaining samples showed no abnormalities in 22, polyclonal hypergammaglobulinaemia in seven, paraprotein in two, and a questionable band in five; seven of 36 had isolated increase in IgG4. We next examined bias in these assays, and retrospectively examined the specific effects of isolated IgG4 elevation. We performed mathematical simulations of over 3000 combinations of IgG subclass results, each subjected to 10,000 virtual nephelometric assays, to determine how frequently the 15% rule would be violated by random chance alone, as reflected by the co-efficient of variation. Our findings have very significant implications for the use of the IgG check in routine diagnostic IgG subclass estimation, and these will be presented.

IgG4-related kidney disease

IgG4-related kidney disease (IgG4-RKD) is a comprehensive term for renal lesions associated with IgG4-related disease, which is a recently recognized clinical entity characterized by a dense lymphoplasmacytic infiltrate rich in IgG4-positive plasma cells with fibrosis affecting several organs. Tubulointerstitial nephritis with increased IgG4-positive plasma cells and fibrosis is the most dominant feature of IgG4-RKD and may cause acute or chronic renal dysfunction, although some glomerular lesions such as membranous nephropathy are sometimes evident. Radiologically, several characteristic abnormalities are often demonstrated, sometimes mimicking malignancies. IgG4-RKD predominantly affects middle-aged to elderly men, and most patients have accompanying IgG4-related extrarenal lesions such as sialadenitis, lymphadenopathy, or type 1 autoimmune pancreatitis. Serology usually demonstrates high levels of serum total IgG and IgG4, and high levels of serum IgE and hypocomplementemia are also frequent features. Corticosteroid therapy is usually quite effective, leading to amelioration of the renal dysfunction and radiological and serological abnormalities. However, as any delay in treatment may result in irreversible renal failure, early diagnosis and appropriate therapy are very important. Despite these distinctive clinicopathological features of IgG4-RKD, its pathogenesis remains poorly understood. Awareness of this condition and accumulation of more cases worldwide are necessary.

Autoimmune thyroiditis in children and adolescents with type 1 diabetes mellitus is associated with elevated IgG4 but not with low vitamin D.

To assess levels of vitamin D and of immunoglobulin G subclasses in children and adolescents with type 1 Diabetes Mellitus with or without autoimmune thyroiditis. Among 213 patients with type 1 diabetes, the cases with thyroid-specific autoantibodies formed Group 1 [n=19, M/F: 7/12, median age 13 years (10.1-14.7)]. Nineteen age-, gender-, and diabetes duration-matched cases with type 1 diabetes without any other systemic disease were designated as controls [Group 2, M/F: 7/12, median age 12.9 years (10.5-14.9)]. Levels of thyroid hormones, vitamin D, total IgG and IgG subclasses, as well as IgG subclasses/total IgG ratios were similar between the groups. Five cases (26%) in Group 1 had IgG4 levels > + 2 SDS, whereas there were no such cases in Group 2 (p=0.046). These five patients had similar clinical features but higher median IgG4 levels and IgG4/Total IgG ratios compared to the subjects with IgG4 levels < + 2 SDS in Group 1 and Group 2. There was no difference of vitamin D levels between the groups. Only a small percentage of patients with type 1 diabetes also having autoimmune thyroiditis had elevated serum IgG4 levels, revealing the heterogeneity of autoimmune thyroiditis and existence of IgG4 thyroiditis in the pediatric age group. Total IgG, the other IgG subclasses, and vitamin D levels did not differ in patients with autoimmune thyroiditis and type 1 diabetes compared to those suffering only from type 1 diabetes.

Immunological Characterization in Malaria Patients with and without the Sickle-Cell Trait

Background: Malaria is one of the main causes of mortality in tropical zone. Specific immune responses are induced by parasite, including the release of cytokines from peripheral blood mononuclear cells. Sickle cell trait confers a high degree of resistance to severe and complicated malaria. The present study aims to assess immunological response of P. falciparum infection by measuring of total IgG level and IL-6, IL-12 & IL-18 levels for P. falciparum among Saudi Arabian patients with and without sickle-cell trait. Patients and Methods: Thirty patients who had clinical suspicion of malaria and sickle-cell trait attending Jazan general hospital in KSA were included in the study. Malaria patients with sickle-cell trait will be matched with a control (thirty patients diagnosed to have malaria but without sickle-cell trait). Diagnosis of malaria was done by Immunochromatography strip and blood film. Diagnosis of Sickle-cell trait was done by hemoglobin electrophoresis assessment of total IgG titre and Interleukin 6, 12, 18 levels using ELIZA. Results: Cytokines and IgG in uncomplicated clinical malaria (n = 22) and severe malaria (n = 7) were IL6 (83.1 pg/mL) versus (75.2 pg/mL), IL12 (19.4 pg/mL) versus (16.3 pg/mL), IL18 (22.45 pg/mL) versus (24.2 pg/mL) and IgG (13.3 SD) versus (4.5 SD). Differences in the IL6, IL12 and IgG were statistically significant (p value >0.02, >0.004 & >0.002 respectively). Among malaria patients with sickle cell trait, Cytokines and IgG in asymptomatic (n = 19) and uncomplicated clinical malaria (n = 11) were IL6 (88.9 pg/mL) versus (79.2 pg/mL), IL12 (24 pg/mL) versus (22.9 pg/mL), IL18 (24.2 pg/mL) versus (31.2 pg/mL) and IgG (27 SD) versus (7.35 SD). Differences in IgG were statistically significant (p value ≥ 0.003). Serum IL6 levels were higher in patients with uncomplicated clinical malaria without sickle cell trait (median 83.1 ± 5.1 pg/mL) than in patients with uncomplicated clinical malaria with sickle cell trait (medians 79.2 ± 4.5 pg/mL). This difference was statistically significant (p value = 0.003). Serum IgG levels were higher in patients with uncomplicated clinical malaria without sickle cell trait (median 13.3 ± 10.84) than in patients with uncomplicated clinical malaria with sickle cell trait (medians 7.35 ± 2.95) while in asymptomatic malaria and severe malaria, medians (27 ± 11.13 versus 4.5 ± 2.75). This difference was statistically highly significant (p value = 0.0009). Conclusion: Malaria protection by HbAS involves the enhancement of not only innate but also of acquired immunity to the parasite. Cytokins (IL6, IL12) and IgG play an important role in protection against severe malaria. The presence of HbAS is associated with increased acquired immunity to malaria. Further work will be done to work out how this change in immunity occurs but treatment of anyone with malaria, whatever their sickle cell status, is essential.

Immune response induced by the TAT-conjugated influenza M2e in mice

Matrix 2 protein ectodomain (M2e) of influenza virus appears to be a promising vaccine candidate because its sequence is highly conserved among virus strains. However, M2e is too meager to induce a strong immune response by itself. Several approaches are being used to increase the antigenicity of M2e. In an effort to enhance the M2e-specific immune response, we generated a TAT-conjugated M2e recombinant protein. Seven-week-old BALB/c mice were divided into three groups and transcutaneously immunized with 100 μg TAT-8×M2e (TAT conjugated with eight copies of M2e) and 8×M2e (eight copies of M2e) proteins on days 1, 15 and 29. The control mice were injected with PBS on the same days. Antibody titers specific for M2e were measured using indirect ELISA. Mice immunized with the TAT-8×M2e and 8×M2e proteins developed almost the same levels of M2e-specific total IgG and IgG1 antibodies. However, a higher level of M2e-specific IgG2a was observed in mice immunized with TAT-8×M2e than in those immunized with 8×M2e. These results suggest that TAT has an adjuvant effect that induces a Th1-type immune response. Therefore, the TAT-M2e vaccine can be applied to animals as a new influenza vaccine for enhancement of Th1-type immune responses.

Pharmacokinetics of intravenous immunoglobulin in multifocal motor neuropathy

Multifocal motor neuropathy (MMN) is often responsive to treatment with intravenous immunoglobulin (IVIg), but the optimal dose and intervals of IVIg maintenance treatment have not been established. Increase in IgG concentration (ΔIgG) after IVIg infusion has recently been identified as determinant of outcome in Guillain-Barré syndrome. ΔIgG may therefore represent a potentially useful biomarker to optimise IVIg dosing in patients with MMN. The aims of this study were to determine variability of IVIg pharmacokinetics in patients with MMN in relation to treatment response, and to establish whether interindividual differences in IVIg pharmacokinetics were associated with genetic polymorphisms of the endothelial IgG receptor (FcRn) which determines IgG half-life. Twenty-three patients with MMN receiving their first IVIg treatment at a cumulative dose of 2.0 g/kg in 5 days were included. A good treatment response was defined as an increase in muscle strength of at least one Medical Research Council point in minimally two muscle groups. IgG concentrations in serum were determined at baseline, at day 1 and day 5 of the IVIg course, and 3 weeks after treatment. FcRn copy number variation and differences in repeat length of the variable number of tandem repeats in the FcRn gene were determined by quantitative PCR and Sanger sequencing. Seventeen patients (74%) had a good response to treatment. Total IgG and ΔIgG levels showed large variation between patients. Mean ΔIgG was higher in IVIg responders than in non-responders, with the largest difference on day 1 (11.1 g/L vs 4.5 g/L, p=0.06), but our study lacked power to show statistically significant differences. Genetic variation in the FcRn gene was not associated with ΔIgG levels or response to treatment. IVIg pharmacokinetics varies in patients with MMN and may be associated with clinical response.

Cathepsin L1 mimotopes with adjuvant Quil A induces a Th1/Th2 immune response and confers significant protection against Fasciola hepatica infection in goats

Thirty goats were randomly allocated in five groups of six animals each, for immunization with 1 × 10(14) phage particles of clones 11, 13, and 13 with Quil A adjuvant and wild-type M13KE phage at the beginning and 4 weeks later. The control group received phosphate-buffered saline. All groups were challenged with 200 metacercariae at week 6 and slaughtered 14 weeks later. The mean worm burdens after challenge were reduced by 46.91% and 79.53% in goats vaccinated with clones 13 and 13 with Quil A (P < 0.05), respectively; no effect was observed in animals immunized with clone 11 and M13KE phage. Animals receiving clones 11, 13, and 13 with Quil A showed a significant reduction in eggs output. Vaccinated animals produced parasite-specific total IgG antibody which were boosted after challenge with metacercariae of F. hepatica. Furthermore, levels of anti-phage total IgG increased rapidly within 2 weeks of the first vaccination and were always significantly higher in all vaccinated goats than in the infected control group. The fluke burden of goats immunized with clones 13 and 13 with Quil A was significantly correlated with IgG2 and total IgG. Goats vaccinated with phage clones produced significantly high titres of IgG1 and IgG2 antibodies indicating a mixed Th1/Th2 response. These data indicate that cathepsin L1 mimotopes has a potential as a vaccine candidate against Fasciola hepatica, whose efficacy will be evaluated in other host species, including those of veterinary importance.

Chronic Lymphocytic Leukemia Patients Have a Preserved Cytomegalovirus-Specific Antibody Response despite Progressive Hypogammaglobulinemia

Chronic lymphocytic leukemia (CLL) is characterized by progressive hypogammaglobulinemia predisposing affected patients to a variety of infectious diseases but paradoxically not to cytomegalovirus (CMV) disease. Moreover, we found reactivity of a panel of CLL recombinant antibodies (CLL-rAbs) encoded by a germ-line allele with a single CMV protein, pUL32, despite differing antibody binding motifs. To put these findings into perspective, we studied prospectively relative frequency of viremia, kinetics of total and virus-specific IgG over time, and UL32 genetic variation in a cohort of therapy-naive patients (n=200). CMV-DNA was detected in 3% (6/200) of patients. The decay of total IgG was uniform (mean, 0.03; SD, 0.03) and correlated with that of IgG subclasses 1-4 in the paired samples available (n=64; p<0.001). Total CMV-specific IgG kinetics were more variable (mean, 0,02; SD, 0,06) and mean decay values differed significantly from those of total IgG (p=0.034). Boosts of CMV-specific antibody levels were observed in 49% (22/45) of CMV-seropositive patients. In contrast, VZV- and EBV-specific IgG levels decayed in parallel with total IgG levels (p=0.003 and p=0.001, respectively). VZV-specific IgG even became undetectable in 18% (9/50) of patients whereas CMV-specific ones remained detectable in all seropositive patients. The observed CMV-specific IgG kinetics were predicated upon the highly divergent kinetics of IgG specific for individual antigens - glycoprotein B-specific IgG were boosted in 51% and pUL32-specific IgG in 32% of patients. In conclusion, CLL patients have a preserved CMV-specific antibody response despite progressive decay of total IgG and IgG subclasses. CMV-specific IgG levels are frequently boosted in contrast to that of other herpesviruses indicative of a higher rate of CMV reactivation and antigen-presentation. In contrast to the reactivity of multiple different CLL-rAbs with pUL32, boosts of humoral immunity are triggered apparently by other CMV antigens than pUL32, like glycoprotein B.