IgG Ab Levels Research Articles

Effects of Hydroxyurea (HU) On Lymphocyte Subsets and the Immune Response to Pneumococcal, Measles, Mumps and Rubella Vaccination in the Pediatric Hydroxyurea Phase III Clinical Trial - BABY HUG - (ClinicalTrials.gov Identifier: NCT00006400)

AbstractAbstract 243▪▪This icon denotes a clinically relevant abstractSusceptibility to encapsulated bacteria, particularly Streptococcus pneumonia, is well known in sickle cell disease (SCD). Hydroxyurea (HU) is commonly used in adults and children with SCD, but, little is known about the effects of HU on immune function in SCD and recommendations for immunization are lacking. As HU reversibly inhibits ribonucleotide reductase, causing cell cycle arrest at the G1-S interface, we postulated that HU might delay the transition from naïve to memory T cells, resulting in a delay in immunologic maturation, with deleterious effects on vaccine responses; therefore, T cell subsets, including conversion of naïve to memory T cells and antibody (Ab) responses to pneumococcal (Pnu), measles, mumps and rubella (MMR) vaccinations were studied during the BABY HUG Trial of HU for infants and toddlers with SCD. Methods:Blood was collected for measurement of: T cell subsets at entry (9–18 mos of age), age 24 mo and exit after 2 yrs of treatment; Pnu Ab levels were measured (by EIA after preabsorption with C-polysaccharide) at entry, before and 2–8 wks after 23-valent polysaccharide (PS) PCV23 vaccine at age 24 mo, and exit; and MMR antibodies at entry, 1–10 weeks after MMR at age 12 mo, age 24 mo and exit. Results:Of the 193 subjects in BABY HUG, T cell subsets were available for 91 HU and 88 placebo (PL) subjects. At entry, there were no significant differences in absolute lymphocyte count (ALC), or % or absolute T cell subsets between the HU and PL groups. At age 24 mo, both groups had an age-related decline in ALC, with significant differences between the HU and PL groups in ALC (5740 vs. 7323/mm3; p=0.003), absolute CD4 cells (1912 vs. 2247/mm3; p=0.022) and memory CD4 cells (392 vs. 487/mm3; p=0.003). The same pattern was seen at exit for ALC (4836 vs. 5764/mm3; p=0.015), absolute CD4 cells (1510 vs. 1747/mm3; p=0.043) and memory CD4 cells (360 vs. 444/mm3; p=0.003).IgG Ab levels to Pnu antigens were measured at entry [after 3 doses of PS/protein conjugate (PC) PCV7 containing Pnu PS type 26 (6B), but not 51 (7F)], before and after immunization with PS PCV23 vaccine at 24 mo, and at exit, with no significant differences between the HU and PL groups in mean Ab levels at any time point for either serotype (Fig 1&2). [Display omitted] For those immunized with MMR prior to study treatment, there were no differences in % immune for measles, mumps, or rubella between the HU and PL groups at any time point. For 40 HU and 38 PL subjects immunized after starting study treatment, there was a smaller % of HU-treated children with protective response to measles vaccine 1 – 10 wks after immunization [HU 71.4% (10/14) vs. PL 100% (24/24); p=0.014]. Response to mumps [HU 78.6% (11/14) vs. PL 91.7% (22/24)] and rubella [HU 66.7% (10/15) vs. PL 92% (23/25)] vaccines showed similar trends, but no statistical significance. The HU vs. PL difference in failure of response to at least one vaccine virus was of similar significance (p = 0.021) to that of measles alone. By age 24 mo, there were no significant differences in response to MMR (89.7 to 100%). Mean Ab levels to measles and rubella were also significantly lower in the HU group 1–10 wks after immunization (p=0.005 and p=0.011, respectively). Conclusions:HU given to infants and young children with SCD causes a reduction in ALC, CD4 T cell and memory T cell counts. Additional monitoring of HU-treated children with SCD for these and related T cell parameters is warranted until the biological effects on immunity are known with certainty.Response to Pnu vaccine was not affected by HU therapy, suggesting that there is no increased risk for Pnu sepsis in infants and children with SCD treated with HU, consistent with the clinical results in the BABY HUG trial.Although the sample was small, a delay in achieving protective measles Ab levels was seen in the HU group, and possibly for mumps and rubella. This delay cannot be estimated precisely, due to study design, but Ab levels to all 3 viruses were similar at exit, indicating that effective immunization can be achieved despite HU use. For endemic or local epidemics of measles, mumps, rubella or other pathogens requiring immunization, adherence to recommended accelerated immunization schedules will be especially important for children with SCD treated with HU. Disclosures:Off Label Use: Off-label use of hydroxyurea in infants and children. Casella:Adventrx: Consultancy, Research Funding.

Human Antibody Response to Anopheles Saliva for Comparing the Efficacy of Three Malaria Vector Control Methods in Balombo, Angola

Human antibody (Ab) response to Anopheles whole saliva, used as biomarker of Anopheles exposure, was investigated over a period of two years (2008–2009), in children between 2 to 9 years old, before and after the introduction of three different malaria vector control methods; deltamethrin treated long lasting impregnated nets (LLIN) and insecticide treated plastic sheeting (ITPS) - Zero Fly®) (ITPS-ZF), deltamethrin impregnated Durable (Wall) Lining (ITPS-DL – Zerovector®) alone, and indoor residual spraying (IRS) with lambdacyhalothrin alone. These different vector control methods resulted in considerable decreases in all three entomological (82.4%), parasitological (54.8%) and immunological criteria analyzed. The highest reductions in the number of Anopheles collected and number of positive blood smears, respectively 82.1% and 58.3%, were found in Capango and Canjala where LLIN and ITPS-ZF were implemented. The immunological data based on the level of anti-saliva IgG Ab in children of all villages dropped significantly from 2008 to 2009, except in Chissequele. These results indicated that these three vector control methods significantly reduced malaria infections amongst the children studied and IRS significantly reduced the human-Anopheles contact. The number of Anopheles, positive blood smears, and the levels of anti-saliva IgG Ab were most reduced when LLIN and ITPS-ZF were used in combination, compared to the use of one vector control method alone, either ITPS-DL or IRS. Therefore, as a combination of two vector control methods is significantly more effective than one control method only, this control strategy should be further developed at a more global scale.

B Cell Memory to a Serogroup C Meningococcal Conjugate Vaccine in Childhood and Response to Booster: Little Association with Serum IgG Antibody

The maintenance of adequate serum Ab levels following immunization has been identified as the most important mechanism for individual long-term protection against rapidly invading encapsulated bacteria. The mechanisms for maintaining adequate serum Ab levels and the relationship between Ag-specific memory B cells and Ab at steady state are poorly understood. We measured the frequency of circulating serogroup C meningococcal (MenC)-specific memory B cells in 250 healthy 6- to 12-y-old children 6 y following MenC conjugate vaccine priming, before a booster of a combined Haemophilus influenzae type b-MenC conjugate vaccine and then 1 wk, 1 mo, and 1 y after the booster. We investigated the relationship between circulating MenC-specific memory B cell frequencies and Ab at baseline and following the booster vaccine. We found very low frequencies of circulating MenC-specific memory B cells at steady state in primary school-aged children and little association with MenC IgG Ab levels. Following vaccination, there were robust memory B cell booster responses that, unlike Ab levels, were not dependent on age at priming with MenC. Measurement of B cell memory in peripheral blood does not predict steady state Ab levels nor the capacity to respond to a booster dose of MenC Ag.

Retinoic Acid Attenuates Rheumatoid Inflammation in Mice

Retinoic acid is the active vitamin A derivative and is well-known to have diverse immunomodulatory actions. In this study, we investigated the impact of all-trans retinoic acid (ATRA), a biologic key metabolite of vitamin A, on the development of arthritis and the pathophysiologic mechanisms by which ATRA might have antiarthritic effects in animal model of rheumatoid arthritis (RA; collagen-induced arthritis [CIA] in DBA/1J mice). We showed that treatment with ATRA markedly suppressed the clinical and histologic signs of arthritis in the CIA mice. It reduced the expression of IL-17 in the arthritic joints. Interestingly, Foxp3(+) regulatory T cells were markedly increased and IL-17-producing CD4(+) T cells (Th17 cells) were decreased in the spleens of ATRA-treated mice. In vitro treatment with ATRA induced the expression of Foxp3 and repressed the IL-17 expression in the CD4(+) T cells in mice. ATRA suppressed the production of total IgG and IgG2a in splenocytes that were stimulated by LPS. It also reduced serum levels of total IgG and IgG2 anti-collagen Abs and germinal center formation in CIA mice. In addition, the ATRA-treated mice showed decreased osteoclast formation in arthritic joints. Moreover, ATRA downregulated the expression of receptor activator of NF-κB ligand, the leading player of osteoclastogenesis, in the CD4(+) T cells and fibroblast-like synoviocytes from patients with RA. Furthermore, ATRA prevented both human monocytes and mice bone marrow-derived monocytes/macrophage cells from differentiating into osteoclasts. These data suggest ATRA might be an effective treatment modality for RA patients.

A single cidofovir treatment rescues animals at progressive stages of lethal orthopoxvirus disease

BackgroundIn an event of a smallpox outbreak in humans, the window for efficacious treatment by vaccination with vaccinia viruses (VACV) is believed to be limited to the first few days post-exposure (p.e.). We recently demonstrated in a mouse model for human smallpox, that active immunization 2–3 days p.e. with either VACV-Lister or modified VACV Ankara (MVA) vaccines, can rescue animals from lethal challenge of ectromelia virus (ECTV), the causative agent of mousepox. The present study was carried out in order to determine whether a single dose of the anti-viral cidofovir (CDV), administered at different times and doses p.e. either alone or in conjunction with active vaccination, can rescue ECTV infected mice.MethodsAnimals were infected intranasally with ECTV, treated on different days with various single CDV doses and monitored for morbidity, mortality and humoral response. In addition, in order to determine the influence of CDV on the immune response following vaccination, both the "clinical take”, IFN-gamma and IgG Ab levels in the serum were evaluated as well as the ability of the mice to withstand a lethal challenge of ECTV. Finally the efficacy of a combined treatment regime of CDV and vaccination p.e. was determined.ResultsA single p.e. CDV treatment is sufficient for protection depending on the initiation time and dose (2.5 – 100 mg/kg) of treatment. Solid protection was achieved by a low dose (5 mg/kg) CDV treatment even if given at day 6 p.e., approximately 4 days before death of the control infected untreated mice (mean time to death (MTTD) 10.2). At the same time point complete protection was achieved by single treatment with higher doses of CDV (25 or 100 mg/kg). Irrespective of treatment dose, all surviving animals developed a protective immune response even when the CDV treatment was initiated one day p.e.. After seven days post treatment with the highest dose (100 mg/kg), virus was still detected in some organs (e.g. lung and liver) yet all animals survived, suggesting that efficacious single CDV treatment requires a potent immune system. The combination of CDV and vaccination provided no additional protection over CDV alone. Yet, combining CDV and vaccination maintained vaccination efficacy.ConclusionsAltogether, our data substantiate the feasibility of single post-exposure antiviral treatment to face orthopoxvirus infection.

Immunological consequences of intermittent preventive treatment against malaria in Senegalese preschool children

BackgroundIntermittent preventive treatment in children (IPTc) is a promising strategy to control malaria morbidity. A significant concern is whether IPTc increases children's susceptibility to subsequent malaria infection by altering their anti-Plasmodium acquired immunity.MethodsTo investigate this concern, IgG antibody (Ab) responses to Plasmodium falciparum schizont extract were measured in Senegalese children (6 months-5 years old) who had received three rounds of IPTc with artesunate + sulphadoxine-pyrimethamine (or placebo) at monthly intervals eight months earlier. Potential confounding factors, such as asexual malaria parasitaemia and nutritional status were also evaluated.ResultsFirstly, a bivariate analysis showed that children who had received IPTc had lower anti-Plasmodium IgG Ab levels than the non-treated controls. When epidemiological parameters were incorporated into a multivariate regression, gender, nutritional status and haemoglobin concentration did not have any significant influence. In contrast, parasitaemia, past malaria morbidity and increasing age were strongly associated with a higher specific IgG response.ConclusionsThe intensity of the contacts with P. falciparum seems to represent the main factor influencing anti-schizont IgG responses. Previous IPTc does not seem to interfere with this parasite-dependent acquired humoral response eight months after the last drug administration.

Secretory-IgA Antibodies Play an Important Role in the Immunity to Streptococcus pneumoniae

This study was designed to investigate whether secretory-IgA (S-IgA) Abs induced by a pneumococcal surface protein A (PspA)-based nasal vaccine are necessary for prevention of streptococcal colonization. Mice nasally immunized with PspA plus a plasmid expressing Flt3 ligand (pFL) cDNA as a mucosal adjuvant showed significantly higher levels of PspA-specific S-IgA and IgG Ab responses in both plasma and nasal washes when compared with naive mice. Although IgA(-/-) mice given nasal PspA plus pFL had significantly high levels of PspA-specific IgG Abs, high numbers of CFUs were detected in nasal washes and nasal passages. In contrast, vaccinated wild-type mice showed essentially no bacteria in the nasal cavity. Further, a nasal vaccine consisting of PspA plus pFL effectively reduced pre-existing Streptococcus pneumoniae in the nasal cavity. These results show that PspA-based vaccine-induced specific S-IgA Abs play a necessary role in the regulation of S. pneumoniae colonization in the nasal cavity.

Usefulness of anti-cyclic citrullinated peptide antibody and rheumatoid factor to detect rheumatoid arthritis in patients with systemic sclerosis

The purpose of this study was to determine the prevalence of anti-CCP antibodies (anti-CCP Abs) and to assess associations between the presence of anti-CCP Ab and arthritis or arthralgia in SSc patients. Serum samples were obtained from 146 SSc patients. Anti-CCP Ab, anti-agalactosyl (AG) IgG Ab, IgM-RF, IgG-RF and MMP-3 were determined, respectively. The presence of anti-CCP Ab was found in 18/146 (12%) patients with SSc. Elevated levels of anti-AG IgG Abs, IgM- and IgG-RFs were observed in 50/146 (34%), 17/146 (12%) and 4/146 (3%), respectively. Serum anti-CCP Ab levels were significantly elevated in SSc-RA overlap patients compared with SSc patients with or without arthralgia (P < 0.05 or P < 0.001, respectively). Serum MMP-3 levels did not correlate with the presence of arthritis or arthralgia but were significantly associated with modified Rodnan total skin thickness score. In SSc-RA overlap patients, 10/11 (91%) patients were positive for two or more RA-related Abs. The serum titre of anti-CCP Ab is higher in SSc-RA overlap patients than in SSc patients with or without arthralgia. The finding of high titres of anti-CCP Abs and the elevated levels combinatory with other RA-related Abs may help to define the diagnosis of SSc-RA overlap. MMP-3 might be a better marker to assess skin involvement rather than joint involvement in SSc patients.

Disordered Lymphoid Purine Metabolism Contributes to the Pathogenesis of Persistent Borrelia garinii Infection in Mice

Extracellular ATP and adenosine are important regulators of immune responses; however, contribution of purinergic signaling to host defense during persistent microbial infections remains obscure. Lyme borreliosis is a common arthropod-borne infection caused by Borrelia burgdorferi sensu lato. In this study, we investigated whether lymphoid purinergic signaling contributes to the mechanisms by which borreliae species evade the immune system and trigger joint inflammation. Intracutaneous inoculation of Borrelia garinii to C3H/He mice induced symptomatic infection manifested in elevated levels of borrelia-specific IgG Abs, persistent spirochete dissemination into the tissues and joint swelling, as well as approximately 2- to 2.5-fold enlargement of draining lymph nodes with hyperplasia of B cell follicle area and L-selectin shedding from activated T lymphocytes. Purine catabolism was also activated in lymph nodes but not spleen and blood of infected C3H/He mice within the first 4 postinfection weeks, particularly manifested in transient upregulations of adenosine triphosphatase/ectonucleoside triphosphate diphosphohydrolase and ecto-5'-nucleotidase/CD73 on CD4(+)CD8(+) T lymphocytes and adenosine deaminase activity on B220(+) B lymphocytes. Compared with borrelia-susceptible C3H/He strain, lymphocytes from C57BL/6 mice displayed markedly enhanced adenosine-generating capability due to approximately three times higher ratio of ecto-5'-nucleotidase to adenosine deaminase. Borrelia-infected C57BL/6 mice efficiently eradicated the inoculated spirochetes at more chronic stage without any signs of arthritis. Strikingly, deletion of key adenosine-generating enzyme, ecto-5'-nucleotidase/CD73, was accompanied by significantly enhanced joint swelling in borrelia-infected CD73-deficient C57BL/6 mice. Collectively, these data suggest that insufficient basal adenosine level and/or pathogen-induced disordered lymphoid purine homeostasis may serve as important prerequisite for promotion of inflammatory responses and further host's commitment to persistence of bacterial infection and arthritis development.

A rice-based oral cholera vaccine induces macaque-specific systemic neutralizing Abs but does not influence natural intestinal immunity (46.4)

Abstract We previously showed that oral immunization of mice with a rice-based vaccine expressing cholera toxin (CT) B subunit (MucoRice-CT-B) induced CT-specific immune responses with toxin-neutralizing activity in both systemic and mucosal compartments. Here, we examined whether the rice-based vaccine can induce CT-specific Ab responses in nonhuman primates. Orally administered MucoRice-CT-B induced high levels of CT-neutralizing serum IgG Abs in the 3 cynomolgus macaques we immunized. Although the Ab level gradually decreased, detectable levels were maintained for at least 6 months, and high titers were rapidly recovered after an oral booster dose of the rice-based vaccine. In contrast, no serum IgE Abs against rice storage protein were induced even after multiple immunizations. Interestingly, the macaques harbored naturally acquired intestinal secretory IgA (SIgA) Abs that were capable of reacting with CT and homologous heat-labile enterotoxin (LT) produced by enterotoxigenic E. coli (ETEC) and that had toxin-neutralizing activity. The SIgA Abs were present in macaques 1 month to 29 years old, and the level was not enhanced after oral vaccination with MucoRice-CT-B or after subsequent exposure to the native form of CT. These results show that oral MucoRice-CT-B can effectively induce CT-specific, neutralizing, serum IgG Ab responses even in the presence of pre-existing, naturally occurring CT- and LT-reactive SIgA Abs in nonhuman primates.

Mucosal S-IgA Abs is required for complete immunity to Streptococcus pneumoniae (46.6)

Abstract We investigate whether pneumococcal surface protein A (PspA)-specific secretory (S)-IgA antibodies (Abs) induced by nasal vaccine is necessary for the prevention of streptococcal colonization and their clearance. IgA deficient (IgA-/-) and control mice were nasally immunized four times at weekly intervals with PspA plus flt3 ligand plasmid (pFL). One week after the last immunization, PspA-specific S-IgA and IgG Ab as well as Th1- and Th2-type cytokine responses by CD4+ T cells were determined. Further, mice were challenged with Streptococcus pneumoniae (EF3030) via the nasal route. Five days after challenge, nasal washes (NWs) and nasal passages (NPs) were collected and the numbers of bacterial colonies were determined. Mice nasally immunized with PspA plus pFL showed significantly increased levels of Th2 cell-mediated PspA-specific S-IgA and IgG Ab responses in NWs and plasma when compared with naïve mice. Thus, these vaccinated wild type mice showed essentially no bacteria in the nasal cavity. Although IgA-/- mice given nasal PspA plus pFL had significantly high levels of PspA-specific IgG Abs, high numbers of bacteria were detected in NWs and NPs. Further, nasal PspA plus pFL effectively reduced pre-existing S. pneumoniae in the nasal cavity of wild-type, but not IgA-/- mice. These results show that PspA-specific S-IgA Abs play important roles in the prevention of S. pneumoniae colonization and clearance. Supported by NIH grants AG025873 and DE12242.

Immunoadsorption in SLE: Three different high affinity columns are adequately effective in removing autoantibodies and controlling disease activity

Introduction Pathogenic autoantibodies (Abs) are a hallmark of SLE and their rapid removal is beneficial in active SLE. Immunoadsorption (IAS) is effective in removing serum levels of all classes of immunoglobulin (Ig), immune complexes (IC) and anti-dsDNA Abs and appears superior to plasmapheresis with respect to side effects. IAS can be performed with different columns, which use different ligands to bind their target. In particular, high affinity columns are in the focus of interest. Their ligands are either sheep IgG directed against human Ig (Ig-column, Ig-Therasorb(®)), or staphylococcal Protein A (ProtA-column, Immunosorba(®)), or the synthetic peptide Gam146 (GAM-column, Globaffin(®)). In our experience Ig-columns have been effective in treating active renal SLE. However, no analysis has so far been published on which column type should be preferred in treating SLE patients. Patients and methods Among our SLE patients maintained on prolonged IAS therapy, we identified those with stable renal SLE and low to moderate disease activity who were successfully treated by using Ig-columns. Six of these patients were switched to ProtA-columns, keeping the rest of the protocol and the medication constant. In addition, two patients were switched from Ig- to GAM-columns. Results All types of columns significantly lowered the serum levels of IgG, IgM, and anti-dsDNA Abs. Disease activity was constantly low before and after the switch, as were parameters of renal function. In addition, patients with highly active disease were effectively treated when ProtA- (n=6) or GAM-columns (n=1) were used as first-line extracorporeal treatment. Conclusion Our data demonstrate that all columns are adequately effective in controlling key parameters of SLE. Thus, it is not the type of the ligand, but only the outcome, i.e. the successful removal of Ig, IC, and (auto-) Abs that is required for controlling SLE activity.

A Rice-Based Oral Cholera Vaccine Induces Macaque-Specific Systemic Neutralizing Antibodies but Does Not Influence Pre-Existing Intestinal Immunity

We previously showed that oral immunization of mice with a rice-based vaccine expressing cholera toxin (CT) B subunit (MucoRice-CT-B) induced CT-specific immune responses with toxin-neutralizing activity in both systemic and mucosal compartments. In this study, we examined whether the vaccine can induce CT-specific Ab responses in nonhuman primates. Orally administered MucoRice-CT-B induced high levels of CT-neutralizing serum IgG Abs in the three cynomolgus macaques we immunized. Although the Ab level gradually decreased, detectable levels were maintained for at least 6 mo, and high titers were rapidly recovered after an oral booster dose of the rice-based vaccine. In contrast, no serum IgE Abs against rice storage protein were induced even after multiple immunizations. Additionally, before immunization the macaques harbored intestinal secretory IgA (SIgA) Abs that reacted with both CT and homologous heat-labile enterotoxin produced by enterotoxigenic Escherichia coli and had toxin-neutralizing activity. The SIgA Abs were present in macaques 1 mo to 29 years old, and the level was not enhanced after oral vaccination with MucoRice-CT-B or after subsequent oral administration of the native form of CT. These results show that oral MucoRice-CT-B can effectively induce CT-specific, neutralizing, serum IgG Ab responses even in the presence of pre-existing CT- and heat-labile enterotoxin-reactive intestinal SIgA Abs in nonhuman primates.

Impact of child malnutrition on the specific anti-Plasmodium falciparum antibody response

BackgroundIn sub-Saharan Africa, preschool children represent the population most vulnerable to malaria and malnutrition. It is widely recognized that malnutrition compromises the immune function, resulting in higher risk of infection. However, very few studies have investigated the relationship between malaria, malnutrition and specific immunity. In the present study, the anti-Plasmodium falciparum IgG antibody (Ab) response was evaluated in children according to the type of malnutrition.MethodsAnthropometric assessment and blood sample collection were carried out during a cross-sectional survey including rural Senegalese preschool children. This cross-sectional survey was conducted in July 2003 at the onset of the rainy season. Malnutrition was defined as stunting (height-for-age <-2 z-scores) or wasting (weight-for-height <-2 z-scores). The analysis was performed on all malnourished children in July (n = 161, either stunted, n = 142 or wasted, n = 19), pair-matched to well-nourished controls. The IgG Ab response to P. falciparum whole extracts (schizont antigens) was assessed by ELISA in sera of the included children.ResultsBoth the prevalence of anti-malarial immune responders and specific IgG Ab levels were significantly lower in malnourished children than in controls. Depending on the type of malnutrition, wasted children and stunted children presented a lower specific IgG Ab response than their respective controls, but this difference was significant only in stunted children (P = 0.026). This down-regulation of the specific Ab response seemed to be explained by severely stunted children (HAZ ≤ -2.5) compared to their controls (P = 0.03), while no significant difference was observed in mildly stunted children (-2.5 < HAZ <-2.0). The influence of child malnutrition on the specific anti-P. falciparum Ab response appeared to be independent of the intensity of infection.ConclusionChild malnutrition, and particularly stunting, may down-regulate the anti-P. falciparum Ab response, both in terms of prevalence of immune responders and specific IgG Ab levels. This study provides further evidence for the influence of malnutrition on the specific anti-malarial immune response and points to the importance of taking into account child malnutrition in malaria epidemiological studies and vaccine trials.

Innate Immunity Mediated by MyD88 Signal Is Not Essential for Induction of Lipopolysaccharide-Specific B Cell Responses but Is Indispensable for Protection against Salmonella enterica serovar Typhimurium infection

Salmonella organisms are Gram negative and facultative anaerobic bacteria that cause typhoid fever in humans. In this study, we evaluated LPS-specific adaptive immunity in innate immune-deficient mice after oral administration of attenuated Salmonella enterica serovar Typhimurium (S. Typhimurium) strains. Of interest, identical levels of LPS-specific IgG and IgA Abs were elicited in the systemic (i.e., serum and spleen) and mucosal (i.e., fecal extract and small intestine) compartments of wild-type, TLR4(-/-), and MyD88(-/-) mice following oral vaccination with recombinant attenuated S. Typhimurium (RASV). Depletion of CD4(+) T cells during RASV vaccination completely abrogated the generation of LPS-specific Abs in MyD88(-/-) mice. In addition, mRNA expression levels of a B cell-activating factor of the TNF family were significantly increased in the spleens of MyD88(-/-) mice after oral administration, implying that T cell-independent B cell switching might be also enhanced in the MyD88 signal-deficient condition. Of most interest, orally vaccinated MyD88(-/-) mice that possessed high levels of LPS-specific IgG and IgA, which had a neutralizing effect against Salmonella, died earlier than nonvaccinated wild-type mice following lethal oral challenge with virulent Salmonella species. These results suggest that innate immunity mediated by MyD88 signal is dispensable for induction of LPS-specific Ab responses following oral administration of attenuated Salmonella strains but indispensable for efficient protection.

Disruption of Allergenic Activity of the Major Grass Pollen Allergen Phl p 2 by Reassembly as a Mosaic Protein

The recognition of conformational epitopes on respiratory allergens by IgE Abs is a key event in allergic inflammation. We report a molecular strategy for the conversion of allergens into vaccines with reduced allergenic activity, which is based on the reassembly of non-IgE-reactive fragments in the form of mosaic proteins. This evolution process is exemplified for timothy grass pollen-derived Phl p 2, a major allergen for more than 200 million allergic patients. In a first step, the allergen was disrupted into peptide fragments lacking IgE reactivity. cDNAs coding for these peptides were reassembled in altered order and expressed as a recombinant mosaic molecule. The mosaic molecule had lost the three-dimensional structure, the IgE reactivity, and allergenic activity of the wild-type allergen, but it induced high levels of allergen-specific IgG Abs upon immunization. These IgG Abs crossreacted with group 2 allergens from other grass species and inhibited allergic patients' IgE binding to the wild-type allergen. The mosaic strategy is a general strategy for the reduction of allergenic activity of protein allergens and can be used to convert harmful allergens into safe vaccines.

Humoral immune responses to MUC1 in women with a BRCA1 or BRCA2 mutation

Introduction Breast cancer patients with early disease and a natural humoral response to MUC1 have a favourable prognosis, suggesting a possible role of MUC1 antibodies (ab) in controlling haematogenous tumour dissemination and outgrowth. The aim of the study was to evaluate humoral immune responses to MUC1 in women at hereditary high risk of breast cancer to investigate whether this immune response could play a role in the prevention of disease. Materials and methods CA15.3 (U/mL), and IgG and IgM ab to MUC1 (arbitrary units per mL, Arb-U/mL) were measured in serum samples obtained from 422 women at hereditary high risk of breast/ovarian cancer, of whom 127 BRCA1/2 carriers, attending the Familial Cancer Clinic of the VU University Medical Centre, and from 370 age-matched healthy controls. Serum samples obtained from women who developed breast cancer ( N = 12) or breast cancer recurrence ( N = 17), and from women who underwent prophylactic mastectomy ( N = 12) and had no breast lesions were also tested. Results CA15.3 ranked significantly higher in mutation carriers than in controls ( P = 0.03). MUC1 IgG ab levels ranked significantly lower in BRCA1/2 mutation carriers than in controls ( P = 0.003). MUC1 IgG levels were not significantly different ( P = 0.53) between women who developed primary breast cancer (median 0.72 Arb-U/ml, range 0.52–2.44 Arb-U/ml) and women who underwent prophylactic mastectomy and had no breast lesions (median 1.04 Arb-U/ml, range 0.43–2.88 Arb-U/ml). Conclusion Serum levels of natural IgG ab to MUC1 are lower in BRCA1/2 mutation carriers than in healthy controls. Furthermore, in contrast to previous results in women with sporadic breast cancer, no elevated MUC1 IgG ab were seen in women at hereditary high risk who developed breast cancer. Prophylactic immunotherapy with MUC1 substrates may be a strategy to reduce the risk of breast cancer in BRCA1/2 mutation carriers, strengthening tumour immune surveillance.

A single intranasal immunization with inactivated influenza virus and α-galactosylceramide induces long-term protective immunity without redirecting antigen to the central nervous system

alpha-Galactosylceramide (alpha-GalCer), originally isolated from a marine sponge, was known to activate natural killer T (NKT) cells through CD1d-mediated Ag presentation and induce Th1 and/or Th2 immunity. In this study, we evaluated the nasal adjuvanticity of alpha-GalCer when co-administered with formalin-inactivated influenza virus A/PR/8/34 (PR8) in BALB/c mice. A single nasal immunization of inactivated PR8 and alpha-GalCer induced brisk levels of PR8-specific IgG and IgA Abs in serum and lung washes. Antigen-specific Ab responses lasted for 3 months, providing protective immunity against challenge with live PR8. In addition, mice given alpha-GalCer also exhibited cellular immune responses including cytotoxic T lymphocyte (CTL) generation. Because it did not redirect Ags into brain, alpha-GalCer would likely pose no risk if administered as a nasal adjuvant. These results suggest for the first time that a single nasal immunization of inactivated virus and alpha-GalCer is a safe and effective means of preventing influenza infection.

Immunogenicity and protective efficacy offered by a ribosomal-based vaccine from Shigella flexneri 2a

Shigellosis is a major form of bacillary dysentery caused by Shigella infection. Shigella ribosome-based vaccines (SRV), considered among the potent vaccine candidates, are composed of O-antigen and ribosome isolated from S. flexneri 2a. To investigate the immunogenicity and protective efficacy of SRV, mice were vaccinated with SRV via the intranasal (i.n.) route. Interestingly, robust levels of Shigella-derived LPS-specific IgG and IgA Abs and antibody-forming cells were elicited in systemic and mucosal compartments following two i.n. administrations of SRV. Groups of mice receiving i.n. SRV developed milder pulmonary pneumonia upon challenge with virulent S. flexneri 2a than did those receiving parenteral SRV. We further found that the MyD88-dependent TLR2 signal partially mediates SRV-induced mucosal immunity, with the exception of TLR4- and TLR5-governed innate immunity. Most importantly, polymeric immunoglobulin receptor knockout (pIgR-/-) mice, which lack secretory IgA Ab, were afforded less protective efficacy than were wild-type mice. It can be concluded then that SRV is immunogenic and provides protective efficacy in mice. It can also be surmised that a mucosal SRV vaccine would be particularly relevant in targeting shigellosis, which provokes inflammation in the human colon.

Is there any relationship between toxoplasma infection and reactive arthritis?

The diagnosis of reactive arthritis is a challenging clinical problem in daily practice. Although there are many triggering infectious agents for reactive arthritis, Toxoplasmosis, a worldwide parasitic infection has not been reported. We investigated the serologic evidence of Toxoplasma gondii ( T. gondii ) infection in patients with newly diagnosed reactive arthritis after six weeks of the onset of the first symptom but no demonstrable triggering agent for reactive arthritis. Clinical controlled study. We screened serologically the serum toxoplasma IgM and IgG antibody (Ab) titers which revealed toxoplasma infection in 50 patients with reactive arthritis (40 female, 10 men) and no demonstrable triggering agent and control subjects (32 female, 8 male). SPSS 10.0 software package program was used. The mean age of the patients and controls was similar (41.3+/- 12.0 vs. 39.6+/-11.8 years) respectively. The prevalence of IgG Ab titers of T. gondii in patients and controls were found to be 52% and 47.5%, respectively. Mean serum Toxoplasma IgG Ab levels were found to be 16.5+/-14.5 IU/ml, and 16.9+/-13.8 IU/ml in patients and control subjects respectively ( P> 0.05). We did not find any Toxoplasma IgM Ab titer demonstrating the acute or sub-acute infection in the serum of patients or controls. Although past Toxoplasma infection was prevalent in both groups, we did not find any subject with acute Toxoplasma infection in patients with newly diagnosed reactive arthritis and healthy controls. Despite the fact that our study group was small, we suggest that T. gondii does not seem to be a triggering agent for reactive arthritis and past infection may be a coincidental finding.