Abstract: After a birth dose of acellular pertussis (aP) and diphtheria (DT)aP-hepatitis B virus (HBV)-inactivated polio vaccine (IPV)/Haemophilus influenza type b (Hib) at 2, 4, and 6 months, a booster dose of DTaP-HBV-IPV/Hib at 12 to 23 months induced strong anti-pertussis booster responses. Thus, neonatal aP priming did not lead to immune tolerance to pertussis antigens. However, it elicited bystander interference on HBV, Hib, and diphtheria responses.
Abstract: This study compared intramuscular and subcutaneous administration of two doses of measles-mumps-rubella-varicella (MMRV) combination vaccine (Priorix-Tetra, GlaxoSmithKline Biologicals) in children. Healthy children (N = 328) were randomised to receive MMRV either intramuscularly or subcutaneously. Reactogenicity was similar between treatment groups for immediate vaccination pain, vaccination site pain, redness and incidence of fever and rashes. Slightly less vaccination site swelling occurred during days 0-3 of the post-vaccination period after intramuscular administration. Seroconversion rates for all components, 42-56 days post-dose 2, ranged from 99.3% to 100% in the intramuscular group and from 98.6% to 100% in the subcutaneous. Cell-mediated immunity data supported the humoral immunogenicity findings. In summary, the MMRV vaccine is well tolerated and highly immunogenic when administered either subcutaneously or intramuscularly to children in the second year of life.
Abstract: We have reported that cordycepin, an adenosine derivative from the fungus Cordyceps, increased interleukin (IL)-10 expression, decreased IL-2 expression and suppressed T lymphocyte activity. In the present study, we further characterized the regulatory effects of cordycepin on human immune cells. Moreover, a traditional Chinese drug, Cordyceps sinensis (CS) that contains cordycepin, was also investigated. Cytometric Bead Array (CBA) was used to determine the concentrations of IL-1beta, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, TNF-alpha and IFN-gamma in culture of peripheral blood mononuclear cells (PBMCs). The results showed that both cordycepin and CS up-regulated IL-10, IL-1beta, IL-6, IL-8 and TNF-alpha; at the same time, they suppressed phytohemagglutinin (PHA)-induced production of IL-2, IL-4, IL-5, IFN-gamma and IL-12. As compared to cordycepin, CS displayed its regulatory effects on IL-2 and IL-10 in a similar dose-dependent manner even with higher efficiency. The binding activity of transcription factors in a human monocytic cell line THP-1 was tested by the trans-AM method, and a higher binding activity of SP1 and SP3 was observed in cordycepin or CS treated cells compared to the control. These results led to the opinion that cordycepin and CS pleiotropically affected the actions of immune cells and cytokine network in a similar fashion. Cordycepin could be an important immunoregulatory active ingredient in Cordyceps sinensis. In addition, CS may contain substances which possess synergism with cordycepin, as CS showed a higher efficiency in the production of IL-10 and IL-2 than cordycepin. However, merits of these effects in pharmacology and clinical medicine have yet to be proven and the precise mechanism of these immune regulatory actions should be researched.
Abstract: BACKGROUND: PID-ARI.net was one of three infectious disease epidemiological research networks funded by the German Ministry of Education and Research (BMBF). Its objectives were to strengthen the national initiative on infectious diseases epidemiology and to focus on a health care problem of high relevance. PATIENTS AND METHODS: A research network on the epidemiology of ARI in children was formed to generate data on several levels. Key structure was a centrally organized active surveillance system in three areas of Germany from north to south. RESULTS: In the 6 years of funding by the BMBF, an integrated research network with a known population denominator was formed. In the laboratory-based surveillance of up to 19 respiratory pathogens, 18,899 samples were analyzed. The added value is utilization of data on time, place, person and pathogen of a disease episode at several levels - from surveillance and online publication via a website to descriptive, analytical and molecular epidemiology and further specialized projects. Its wide age range including children up to 16 years of age, an extensive panel of pathogens, a known population denominator and the diversity of 3 distant geographical areas should considerably reduce vulnerability due to bias. CONCLUSIONS: Active surveillance systems for ARI are superior to passive systems. If a surveillance system such as the one used in PID-ARI.net is part of a research network which can utilize the data on several levels, the expenditure for such a system should be worthwhile and such a system would be an asset to any health care system.
Abstract: Interleukin (IL)-27, a heterodimer composed of Epstein-Barr virus-induced gene 3 (EBI3) and p28, is an early product of activated dendritic cells (DC). Binding of IL-27 to the WSX-1 receptor initiates Th1 (Thelper 1) responses in naïve T cells. In order to assess the Th1 responses in human neonates with high susceptibility to infectious diseases, expression of EBI3-, p28- and WSX-1-mRNA in response to Toll-like receptor ligands was compared in neonate and adult monocyte-derived (m)DC. Only the combined addition of ligands induced expression of IL-27p28 mRNA. Surprisingly, neonatal mDC produced significantly more IL-27p28 mRNA than that obtained from adults. Furthermore, there was enhanced expression of EBI3 mRNA in cord blood as compared with adult blood. In addition, the secretion of WSX-1 mRNA in neonatal mDC and T cells was also significantly increased. Taken together, these findings indicate that the restricted Th1 responses in human newborns owing to deficient IL-12 production may be compensated for, in part, by enhanced IL-27 secretion.
Abstract: Cell-mediated immune (CMI) responses to an acellular pertussis vaccine administered to 49 subjects, a subset of participants in the National Institutes of Health-funded adult acellular pertussis vaccine efficacy trial, were evaluated and compared with antibody responses to vaccine antigens. Levels of proliferation of and cytokine secretion from lymphocytes cultured in the presence of pertussis toxin, filamentous hemagglutinin, or pertactin were measured before vaccination and 1 month and 1 year after vaccination. Statistically significant increases in lymphocyte stimulation indices and cytokine secretion were noted at both 1 month and 1 year after vaccination. Brisk pertussis antigen-specific immunoglobulin G responses were also noted at 1 month after vaccination, but these responses had declined by nearly 50% at 1 year after vaccination. These studies clearly demonstrate that both cellular and humoral immune responses occur after the administration of acellular pertussis vaccines to adolescents and adults but that the CMI responses are of greater magnitude and longer duration. CMI responses may be a better correlate of long-term protection.
Abstract: Dendritic cells (DC) are thought to be responsible for the reduced ability of human newborns to induce protective T-helper 1 (Th1) immune responses. The key player in Th1 differentiation, interleukin-12 (IL-12), is primarily produced in response to Toll-like receptor (TLR) binding by adult DC but not by neonatal DC. The potential use of various TLR agonist combinations for initiating neonatal monocyte-derived DC to prime Th1 responses was investigated. Single TLR ligands induced maturation only in adult DC; neonatal DC matured with combined targeting of TLR3/TLR8 or TLR4/TLR8, based on the expression of maturation markers. Similarly, the synergistic effects of combined TLR ligands could also be shown with adult and neonatal cytokine production, but different expression patterns were noted. In particular, IL-12p70 was produced by neonatal DC exclusively after combined TLR stimulation. Surprisingly, it was found that supernatants of combined stimulated neonatal DC could induce interferon-gamma production in autologous naïve T cells. Moreover, this interferon-gamma secretion was blocked by anti-IL-12p70 antibodies and increased after addition of recombined IL-12. In conclusion, these findings underline the differences between adult and neonatal DC and might suggest new strategies for promoting newborn Th1 immunity in response to pathogens and vaccine antigens.
Abstract: Booster vaccination with a reduced-antigen-content dTpa, pediatric DTPa or adult Td vaccine in DTPa-primed children aged 4-6 years was evaluated. Immunogenicity and CMI was assessed one month and 3.5 years after vaccination. Symptoms were solicited for 15 days post-vaccination. There were no differences between groups in diphtheria or tetanus seroprotection or pertussis vaccine-response rates. Anti-diphtheria and anti-PRN concentrations were higher after DTPa, but groups differences reduced over time. Non-significant trends toward reduced reactogenicity of dTpa were observed. Many factors influence vaccine choice at pre-school age. The dTpa vaccine was as immunogenic and possibly better tolerated than DTPa at this age.
Abstract: Humoral and cell-mediated immune responses (CMI) were evaluated in subjects 3 and 6 years after primary and booster vaccination with either three-component acellular (Pa) or whole-cell (Pw) vaccines. Low anti-pertussis toxin (PT) antibody levels confirmed the absence of pertussis disease, consistent with ongoing protection. Anti-pertactin (PRN) antibodies, remained at higher levels in Pa-vaccinated subjects. At year 6, CMI responses continued to be present and were higher in Pa-vaccinated than Pw-vaccinated subjects. Long-term protection with Pa vaccines can be expected to be at least as good as that provided by efficacious Pw vaccines.
Abstract: INTRODUCTION: Surveillance systems for acute respiratory infections (ARI) in children currently are often limited in terms of the panel of pathogens and the age range investigated or are only syndromic and at times only active in the winter season. METHODS: Within PID-ARI.net, a research network for ARI in children in Germany, an active, year-round surveillance system was formed in three regions from north to south for population-based analysis. Children from birth to 16 years of age were included and up to 19 noncolonizing airway pathogens were tested for with multiplex RT-PCR. RESULTS: In the 10-year period from July 1996 to June 2006, a total of 18,899 samples were tested. The positive rate increased with the size of the test panel to up to 72.9%. Picornaviruses (35-39%), paramyxoviruses (23-28%) and orthomyxoviruses (5.8-12.5%) comprised the highest fraction. Reoviruses and Legionella pneumophila were not found at all and Chlamydia pneumoniae and Bordetella parapertussis only rarely. Respiratory syncytial virus and parainfluenza virus (PIV) type 3 were anticyclical in rhythmicity with metapneumovirus and PIV1 and PIV2. The age medians per pathogen depended predominantly upon the attack rate and interepidemic intervals. CONCLUSION: Active surveillance systems for ARI are superior to passive systems. They should be pathogen-specific and comprehensive for viruses and bacteria and age ranges. They should be population-based and multilevel to avoid bias. The impact of atypical bacteria in children was highly overestimated in earlier studies.
Abstract: BACKGROUND: Mannose-binding lectin (MBL) is an important factor of the innate immune system, and MBL-initiated complement activation is an important early defense mechanism against various bacterial infections, including invasive meningococcal disease. METHODS: In a pediatric cohort (ages 2-215 months) with invasive meningococcal disease, we investigated the overall and age-stratified frequency of 3 MBL exon 1 variations (C154T, G161A, G170A), previously shown to result in markedly decreased MBL plasma concentrations, by allele specific fluorescent hybridization probe real-time PCR assays and direct sequencing. Healthy age-matched volunteers with the same ethnic background and no history of meningococcal disease served as a control group. RESULTS: The overall frequency of a MBL exon 1 variant genotype was significantly higher in patients than in controls (31.8% vs. 8.2%, P < 0.001). In the patient group with disease onset less than 24 months of age, the prevalence of MBL structural variant genotype was further increased (39.3%; P < 0.001) and most pronounced in children with disease onset less than 12 months of age (57.1%; P < 0.001) when compared with healthy controls. Analysis of clinical severity and outcome revealed no significant difference between patients with wild-type and mutant alleles. CONCLUSIONS: Our data suggest that MBL exon 1 structural variants are significantly associated with susceptibility to childhood meningococcal disease in an age-dependent manner.
Abstract: Peripheral blood cells express the complete non-neuronal cholinergic system. For example synthesis of acetylcholine and nicotinic as well muscarinic receptors have been demonstrated in leucocytes isolated from human peripheral blood. In the present experiments mononuclear cells and granulocytes were isolated from the peripheral blood to investigate content and synthesis of acetylcholine as well as phenotypic functions like respiratory burst, phagocytosis and migration. Mononuclear cells (T-cells and monocytes) contained 0.36 pmol/10(6) cells acetylcholine, whereas acetylcholine content in granulocytes was 100-fold lower. Acetylcholine synthesis amounted to 23.2+/-4.7 nmol/mg protein/h and 2.90+/-0.84 in CD15+ (granulocytes) and CD3+ cells (T-lymphocytes), respectively. Neither atropine (blockade of muscarinic receptors) nor tubocurarine (blockade of nicotinic receptors) exerted an effect on the respiratory burst. Tubocurarine (30 muM), alone or in combination with atropine (1 microM), reduced phagocytosis in granulocytes by 13% and 19%, respectively (p<0.05). Spontaneous transwell migration of granulocytes was doubled by tubocurarine combined with atropine (p>0.05). Also alpha-bungarotoxin (10 microg/ml) enhanced spontaneous granulocyte migration, but hexamethonium (300 microM) was without effect. The present experiments demonstrate a cholinergic modulation of immune functions in peripheral leucocytes under in vitro conditions, i.e. in the absence of a neuronal innervation. Blockade of nicotine receptors (alpha1 muscular subtype) facilitates spontaneous migration of granulocytes.
Abstract: Das Immunsystem reagiert bei Neugeborenen und sehr kleinen Kindern noch anders als bei Erwachsenen. Aber auch Neugeborene können unter ganz bestimmten Bedingungen eine wirksame Immunantwort entwickeln.
Abstract: German adolescents (n=123) without previous pertussis vaccination, no history of pertussis and low IgG-anti-pertussis-toxin (PT) levels received one dose of the Tdap vaccine Boostrix. Blood samples were taken before, and 5-12 days and 29-49 days after vaccination. IgG- and IgA-anti-PT, IgG- and IgA-anti filamentous hemagglutinin, IgG-anti-pertactin, IgG-anti-tetanus-toxin, and IgG-anti-diphtheria-toxin were measured by ELISA. 88.6% of subjects had an immune response to PT, and all vaccinees had an immune response to at least one pertussis antigen 29-49 days after vaccination. IgA-anti-PT and IgA-anti-FHA responses were found in 43 and 81% of subjects, respectively. This study shows that in unvaccinated German adolescents pertussis immunity can be achieved by a single dose of Tdap.
Abstract: Toll-like receptor 4 (TLR4) is required for efficient recognition of bacterial infections. We investigated an association between 2 TLR4 mutations (Asp(299)Gly and Thr(399)Ile) and meningococcal disease in 197 patients and 214 healthy controls by allele-specific real time polymerase chain reaction and direct sequencing. Although the allele frequency was not higher in the overall patient population, a significantly higher frequency in the 40 patients younger than 12 months of age (P = 0.007) was observed. We conclude that TLR4 mutations represent a risk factor for meningococcal disease in this age group.
Abstract: BACKGROUND: Passive immunization with palivizumab is expensive and requires considerable logistic effort. So far 5 monthly injections from November to March are recommended. The RSV season onset and its duration, however, shows considerable variation. In many countries on the northern hemisphere a dual rhythm is described. METHOD: A web-based early warning system within the research network PID-ARI.net is in place since 2002. The surveillance data are published online weekly via www.pid-ari.net. This enables physicians to carry out interventions, like passive immunization for RSV, synchronously with the epidemiology of a given pathogen instead of a rigid schedule. The surveillance of PID-ARI.net is based on a 19 valent multiplex RT-PCR on naso-pharyngeal aspirates. The samples are provided by hospitals and offices in Freiburg, Mainz and Schleswig-Holstein (north, middle, south of Germany). Children with lower airway infections are prospectively enrolled. RESULTS: In the time period from July 1999 to June 2003 with 20 months of recommended palivizumab application, 5 months (25 %) would have been not on target. In two seasons the start of the vaccine campaign would have been too early (waste of two months). In one season the application would have started one month too late and in two seasons the vaccine campaign would have been stopped two months too early leaving the vaccinees on risk for acquiring RSV. CONCLUSIONS: The web-based early warning system of PID-ARI.net is the first, pathogen-specific, comprehensive and fast surveillance-system for airway pathogens in Europe. It facilitates the epidemic-synchronous use of the passive immunization with palivizumab and by this increases its efficiency and should safe costs.
Abstract: Specific functional properties of dendritic cells (DCs) have been suspected as being responsible for the impaired specific immune responses observed in human neonates. To analyze stimulatory requirements for the critical transition from immature, antigen-processing DCs to mature, antigen-presenting DCs, we investigated the effect of different proinflammatory mediators and antigens on phenotype and cytokine secretion of human neonatal DCs derived from hematopoietic progenitor cells (HPCs). Whereas single proinflammatory mediators were unable to induce the maturation of neonatal DCs, various combinations of IFNgamma, CD40L, TNFalpha, LPS and antigens, induced the maturation of neonatal DCs documented by up-regulation of HLA-DR, CD83 and CD86. Combinations of proinflammatory mediators also increased cytokine secretion by neonatal DCs. Especially combined stimulation with LPS and IFNgamma proved to be very efficient in inducing maturation and cytokine synthesis of neonatal DCs. In conclusion, neonatal DCs can be stimulated to express maturation as well as costimulatory surface molecules. However, induction of maturation requires combined stimulation with multiple proinflammatory signals.
Abstract: More than forty cytokines have been extensively researched on the molecular structure, cell signaling and transduction pathway. With respect to cytokine-regulating therapy in immunological imbalance however, the reported results are conflicting because of the pleiotropic functions and the intricate interactions of the cytokine network. In this review, we outline the observations on interleukin-10 (IL-10) upregulatory therapy. Despite varying opinions on its therapeutic effects for different disorders, IL-10 has been considered a potential anti-inflammatory cytokine. Numerous studies support the view that IL-10 shows a strong suppressive effect on Th1 lymphocytes, antigen presenting cells and the production of inflammatory mediators. It is also noticeable that recent research has revealed the relationship between IL-10 induced antigen specific regulatory CD4+ T cells and antigen specific immune tolerance. This specific regulation was mediated in part through IL-10 secretion, because anti-IL-10 treatment reverted the inhibitory effect of regulatory T cell clones. In different models, these cells were shown to inhibit both Th1 and Th2-type inflammatory responses through the secretion of IL-10. With the presence of IL-10, regulatory T cells may induce peripheral immune tolerance. Exogenous administration, transgenic expression and endogenous stimulative agents of IL-10 have been used for a variety of inflammatory diseases, autoimmune diseases and allograft rejection in patients and experimental models. A therapeutic intervention with drug inducing endogenous IL-10 may be more practical than an exogenous administration of IL-10 with transient effect. Although further investigation on gene regulation of IL-10 is necessary, increasing studies have been reported concerning the attempt to develop the agents, which could promote endogenous IL-10 production for the treatment of immunological disorders and inflammatory diseases. With some unclear mechanisms, these agents have strongly upregulated IL-10 production in vitro or in vivo. Reported IL-10 upregulatory agents have shown promising prospects for remission of autoimmune diseases and inflammatory diseases and have even induced antigen specific immune tolerance. It is interesting that the IL-10 upregulatory effect of several traditional immunosuppressive drugs has been detected, e.g. glucocorticoid, which is considered "not more as an immunosuppressive drug but an immune modulating agent". Approximately twenty IL-10 upregulatory agents as instances are described in the present review. In addition, their therapeutic effects in various diseases are discussed.
Abstract: Objectives: Human Toll-like receptor 4 (TLR4) is the major endotoxin-signalling receptor of the innate immune system and is required for efficient recognition of gram-negative bacterial infections. We investigated a possible association between the (2ommon TLR4 mutation Asp299Gly and mortality due to invasive meningococcal disease in children. ~~',
Methods: 197 children (ages 2-215 months, mean 59 months) '.' ~vith clinically and/or microbiologically proven invasive men'ingococcal disease from five different European countries ,vere :~~~~~~(@",(.~tt.:"1!J':415~i'; -:~,: analysed for the Asp299Gly mutation by Lightcycler allele : specific fluorescent hybridisation probe assays and direct ⢠sequencing. Genotyping results were correlated with fatal or none fatal outcome. Statistical analysis was performed applying , Pearsons Chi-Square test (two-sided). '.â¢â¢
Results: The overall allele frequency of the Asp299Gly .........OW<; ,,, ,' . polymorphism was 9.4% (165 Asp/Asp, 27 Asp/Gly, ~ ,;)';:-'~,'",,%" : ."" Gly/Gly) among the analysed patients with meningococcal '. disease. A total of 19 patients (9.6%) succumbed to the .⢠disease. Interestingly the heterozygous TLR4 Asp299Gly mutation was significantly associated with fatal disease outcome: In the none-survivor group 6 out of 19 patients .(31 %) had a heterozygous Asp299Gly polymorphism, ereas , :; this was only the case in 21 out of 178 patients (1 1. ) ' the ~~~.~.~k%ly,)e. ⢠⢠survivor group (p = 0.029). There was no SI nificant ~'~n~.~~f⢠. (p ::: 0.459) difference between the mutation in 0 oz~ â¢gous : state in the survivor (5/178, 2.8%) and none-Ur\ ⢠â¢or group ~" ~."""' , ,,,,'.... (0/19, 0%). ,
Conclusions: Our data suggest that the anal, ~ Tl 4 muta'. tion Asp299Gly is associated with increa_ ortality inion ~ 1 :. I o '-l à CD ine .' heterozygous state and might have an im impact on the clinical course and outcome of meninnr-,.",.~ childhood.
Abstract: The humoral and cellular immune response to inactivated hepatitis A vaccine was investigated dynamically in a time elapse study over 1 year. Fourty-five healthy volunteers, seronegative for anti-HAV, were vaccinated with 1440 enzyme-linked immunosorbent assay units (EU) of formalin-inactivated hepatitis A virus following a 0--6-month schedule. Serum anti-HAV levels and HAV-specific proliferation of peripheral blood mononuclear cells were measured at several time points over a 26- and 28-week period after the first and second injection, respectively. Distinct B and T cell responses were determined within 14 days after primary vaccination. The booster vaccination-induced immediate peak levels for the humoral (anti-HAV GMC=5376mIU/ml) as well as the cellular (median Deltacpm=14173cpm) response.
Abstract: Erstmals wurde im Rahmen einer prospektiven Studie in Deutschland das humane Metapneumovirus (HMPV) nachgewiesen. Der erste Fall wurde in Mainz am 6. Februar 2003 mithilfe der PCR entdeckt. Danach fanden sich prospektiv 41 weitere Fälle in Freiburg, Kiel und Mainz.
Die jeweiligen Amplifikate wurden durch Sequenzierung als HMPV bestätigt. Den Mitarbeitern des vom BMBF geförderten Forschungsnetzwerkes "Atemwegsinfektionen bei Kindern" (www.pid-ari.net) gelang der Nachweis durch die systematische Sur-. veillance von Nasensekret bei Kindern, die wegen einer akuten respiratorischen Infektion (ARI) hospitalisiert wurden. Die HMPV-infizierten Kinder hatten ein breites, unspezifisches Spektrum an klinischen Symptomen, die die oberen oder unteren Atemwege betrafen. Grundlage des Nachweises war eine neu entwickelte Multiplex-RT-PCR, (m-RT-PCR) mit der 16 nichtkolonisierende ARI-Erreger, darunter auch das HMPV, gleichzeitig detektiert werden können. Die neue nl-RT-PCR-Methode erlaubt es, im Rahmen des PID-ARI.Net die erregerspezifische ARI-Diagnose zu stellen, die jeweilige Epidemiologie der Mikroorganismen zu beschreiben und dabei auch neue Erreger wie HMPV zu e.rfassen (1). Die HMPV-Primersequnzen wurden von der Arbeitsgruppe um A. Osterhaus, Rotterdam (Niederlande) zur Verfügung gestellt.
Abstract: Therapeutic options for controlling autoimmune diseases are still very limited. Interleukin-10 has been reported to be a promising approach to therapeutic intervention. In the search for a drug which results in the selective upregulation of interleukin-10, we investigated the immunoregulative effects of cordycepin. We have measured interleukin-10 and interleukin-2 secretion of human peripheral blood mononuclear cells that were incubated with cordycepin and assessed the influence of cordycepin on the expression of interleukin-10 mRNA, the proliferative response and the expression of surface markers on T lymphocytes. In addition, the subsets of interleukin-10-secreting cells, the influence of anti-interleukin-10 neutralizing antibody and cytotoxicity of cordycepin were evaluated. Our results suggest that cordycepin has a significantly upregulative effect on interleukin-10 production and interleukin-10 mRNA expression. Interleukin-10-producing cells included in CD4+, CD8+, CD19+, CD56+ and CD14+ cells. At the same time, cordycepin inhibited phytohaemagglutinin-induced interleukin-2 production and proliferation of peripheral blood mononuclear cells. A restricted T lymphocyte activation was also reflected by a reduced expression of the surface markers CD25, CD45RO, CD54, CD71 and HLA DR. Anti-interleukin-10 neutralizing antibody could not completely block the suppressive effect of cordycepin on production of interleukin-2. Cordycepin in the effective concentration presented slight cytotoxicity but did not increase apoptosis. These results indicate that cordycepin exerts immunoregulative effects. Further research on it may provide an approach for the development of novel immunomodulatory drugs which directly alter the secretion of cytokines.
Abstract: OBJECTIVE: Combination vaccines simplify vaccine administration and have the potential to promote compliance and cost-effectiveness by decreasing the number of injections needed to immunize a child. The objective of this study was to assess the safety and reactogenicity of the diphtheria-tetanus toxoid-acellular pertussis-hepatitis B virus-inactivated polio virus (DTPa-HBV-IPV) vaccine when coadministered with different Haemophilus influenzae type B (Hib) vaccines in comparison with separate, commercially available, control vaccines in a 3-dose primary vaccination series. METHODS: An open-label, randomized, parallel-group study in 5318 infants who were 8 to 16 weeks of age at enrollment was conducted in 90 centers in Germany. The incidence of adverse events that occurred in infants who received the DTPa-HBV-IPV candidate vaccine coadministered with 1 of 4 different Hib vaccines (given in separate sites; groups 1-4) was compared with the incidence that occurred in infants who received commercially available control vaccines (DTPa, Hib, and oral polio virus [OPV] vaccine; group 5) administered separately. The vaccines were given as a 3-dose primary series at 3, 4, and 5 months of age. Infants were assessed for solicited local and general adverse events for 4 days and for unsolicited adverse events for 30 days after each vaccine dose. The primary endpoint was to rule out a 7.5% increase in infants who experienced grade 3 (defined as preventing normal everyday activities unless otherwise specified) solicited local and general adverse events over the 3-dose primary course after the combined DTPa-HBV-IPV vaccine coadministered with Hib as compared with commercially available vaccines. RESULTS: During the 3-dose primary course, 490 of 3029 infants (16.2%) in the pooled DTPa-HBV-IPV vaccine groups and 151 of 744 (20.3%) in the control vaccine group experienced a grade 3 adverse event (rate difference [control minus combination] 4.1%; 90% confidence interval, 1.41-7.13). The lower limit of the 90% confidence interval of the observed difference remained above the prespecified -7.5% limit for noninferiority, thereby meeting the primary endpoint. The incidences of local injection-site reactions were similar for the DTPa-HBV-IPV and DTPa injection sites. Significant differences in the incidence of both local and general adverse events were observed depending on which of the Hib vaccines was coadministered. Infants who received Hib N meningitidis outer-membrane complex protein conjugate vaccine had greater incidences of fever and, to a lesser extent, greater reactions at the Hib injection site than did infants who received other Hib vaccines. CONCLUSIONS: The combination DTPa-HBV-IPV vaccine administered concomitantly with Hib vaccine at separate sites was at least as safe as coadministration of individual DTPa, Hib, and OPV vaccines in terms of the defined endpoints for safety.
Abstract: Background: Decreasing pertussis antibody concentrations after primary
immunization supports the concept of a booster vaccination at 4-6 years. In
previous stucües we have demonstrated that CMI as a specific marker may
help to characterize efficiency of vaccines. Here we present serological and
CMl data prior to and inunecüately after administration of two DTPa
booster vaccinations at the age of 4-6 years.
Materials and methods: 357 infants were primed with four doses of OTPa
during the first two years of life. At the preschool age (4-6 years) these
children received either a OTPa or a dTpa (reduced antigen content)
booster vaccination (SB). Peripheral blood was drawn pre-and 30 days
posrvaccination. The specific T -cell response to pertussis antigens PT, FHA
or PRN was analysed for a subset of subjects (11 = 172). Pertussis specific
IgGl were measured.
Results:· Before the booster vaccination up to 80% of subjects showed a
CMI-response to the pertussis antigens (pT 67%, FHA 80%, PRN 60%).
The antigen specific CM1-respome after the booster exceeded the
prebooster response (Table 1). Pre-booster, pertussis-specific antibody titres
were low. However, booster vaccination induced a vigorous rise of
antibody titres (Table 2).
TJble 1 tllcreast' or n1L'i!nS prebooste-r vs. pO'\[booscer
PT FHA l'RN
dTpa 29 46 24
DTl'. 41 11)) 52
Table 2 GMT, 95% Cl; pre/PG.lt
PT FHA PRN
dTpa .1 / 63.8 70.11763.7 5K.71018.7
DTPa 4.8179.2 59.6/862.2 54.6/1359.5
Conclusion: After booster vaccination of children aged 4-6 years (2-4 years
POSt 4th dose ofDTPa-primary immunization), high frequencies ofpertussis
specific periphereal blood T -cells can be demonstrated, in contrast to low
levels of circulating antibodies. A major increase ofthe proliferative response
postbooster was demonstrated for both (low and high dose) pertussis
vaccines. The fincüngs suggest that T -cell memory prebooster is of sufficient
qualiry to mount immecüate Pertussis specific immune responses. The
presence of prebooster memory T -cells may have to be seen as still high
enough for a sufficient cellul:u response.
Abstract: Nonresponsiveness to HBsAg vaccination is observed in 5-10% of vaccine recipients and is possibly caused by a defect in the T helper cell compartment. The immune response to HBsAg is influenced by genes of the major histocompatibility complex. We have investigated MHC class I and class II antigens in 53 adult responders and 73 nonresponders. Results obtained in this first study were tested in a second study with 56 responders and 62 nonresponders from an infant vaccination trial. In addition, the peripheral Vbeta-chain T-cell receptor repertoire was investigated using monoclonal antibodies and flow-cytometry in 26 adult responders and 38 nonresponders. As previously reported, nonresponsiveness to HBsAg vaccination was associated with DRB1*3 and DRB1*7. In addition, DRB1*13 was significantly increased among vaccine responders (35.2% vs 5.4%;p < 0.0001) suggesting an immune response promoting effect for this allele whereas the closely related allele DRB1*14 was associated with nonresponse in the infant study. There was no evidence for a hole in the T cell receptor Vbeta repertoire. In conclusion, in agreement with results obtained in mice there appears to be a hierarchy of DRB1* genes in the HBsAg immune response. The possible differential association of DRB1*13 and DRB1*14 may allow the identification of differences between responsiveness and nonresponsiveness to a few amino acid differences in the beta1-domain of the class II heterodimer.
Abstract: Impfungen haben wie keine andere medizinische Entdeckung die Epidemiologie von Infektionskrankheiten zugunsten des Menschen verändert. Neben verbesserten Therapieformen und sozioökonomischen Veränderungen ist diese Entwicklung zweifelsohne auf die Anwendung konsequenter ImpfmaÃnahmen zurückzuführen. Trotz dieser Erfolge sind Impfungen gelegentlich umstrittenl Widerstand gegen Impfungen regt sich dort, wo man die "natürliche ErkrankungU der Impfung vorzieht, wegen unerwünschter Nebenwirkungen kein Vertrauen in ImpfmaÃnahmen hat oder deren Notwendigkeit nicht einsieht. Demgegenüber stehen epidemiologische Veränderungen bei Infektionskrankheiten, neue Impfstofftechnologien sowie die Einführung von Kombinationsimpfstoffen den ablehnenden Aspekten gegenüber.
Abstract: To evaluate the reactogenicity and immunogenicity of a Haemophilus influenzae type b conjugate vaccine (HbOC) and of a tricomponent acellular pertussis vaccine (DTaP) when injected simultaneously into either contralateral arms or into contralateral thighs, 110 infants were enrolled to receive three doses of DTaP at 3, 4, and 5 months and two HbOC doses at 3 and 5 months of age. Administration of either of the two vaccines into arms was associated with significantly more local side effects than administration into thighs. There was no difference in geometric mean concentration (GMC) values for any of the four vaccine antigens between subjects who had been vaccinated into arms or thighs. After immunization, all children had protective antibody titers to diphtheria toxin. While post vaccination the mean anti-tetanus toxoid GMC was > or = 1.25 IU/ml, there was no significant rise as compared to the GMC before vaccination. GMCs of antibodies against the various pertussis antigens were similar to those observed before with the same DTaP vaccine. The simultaneous administration of DTaP and HbOC was safe and immunogenic irrespective of the site of vaccine administration, but significantly more local reactions occurred when vaccines were injected into arms.
Abstract: The lack of an adequate immune response to the major polysaccharide of the Haemophilus influenzae type b (Hib) capsule (polyribosyl ribitol phosphate) (PRP) in very young infants (< 18 months) can be overcome by conjugating PRP to a T-cell dependent carrier protein. We studied whether administration of a tetanus-PRP conjugate vaccine reconstituted with a diphtheria-tetanus-acellular pertussis-hepatitis B (DTPa-HBV) vaccine as a three dose primary course at 3, 4 and 5 months of age induced PRP-specific immunological memory, by examining the anti-PRP response to a dose of unconjugated PRP given with the DTPa-HBV booster approximately 1 year later. The unconjugated PRP elicited protective anti-PRP antibody levels (> or = 0.15 microgram/ml) in all but 3 of the 369 vaccinees, including 13 infants who failed to demonstrate a measurable immune response after the primary course. In a sub-cohort of 54 subjects all had anti-PRP levels > or = 0.5 microgram/ml within 7-14 days of the booster showing a rapid anamnestic type response. Both primary and booster responses were predominantly IgGl indicating a T-cell dependent response. The DTPa-HBV components elicited protective anti-diphtheria, anti-tetanus and anti-HBs antibody levels in > or = 98.5% of vaccinees, and immune responses to each of the acellular pertussis vaccine components in 92.3%-97.3% of subjects. CONCLUSION: The tetanus-PRP conjugate vaccine not only elicited a good primary humoral response, but also induced immunological memory so that the infants were able to mount a large and rapid immune response to subsequent exposure to plain PRP, indicating that protection against circulating wild-type Hib had been generated. Successful induction of immunological memory occurred even when there was no measurable humoral anti-PRP response to the primary course. Tetanus-PRP conjugate vaccine can be used in combination with DTPa-HBV vaccine, when administered separately or as a single injection in the same syringe, in primary immunisation schedules at 3, 4 and 5 months of age.
Abstract: The aim of this study was the investigation of the specific cell-mediated (CMI) responses induced by DTaP and to compare these data to immunity after natural infection. The ability of peripheral blood T-lymphocytes to respond to the pertussis related antigens pertussis-toxin (PT), filamentous haemagglutinin (FHA), and Pertactin (PRN) was investigated in 40 children before vaccination and at different times after vaccination with DTaP (Infanrix) by measurement of antigen-specific proliferation, lymphocyte phenotype, cytokine production and expression of activation markers (CD25, HLADR). Similar investigations were performed in children 4-6 weeks after recovery from natural pertussis. DTaP created a specific T-cell-response to PT, FHA and PRN that increased continually, depending on the progress of the vaccination schedule. In contrast to waning antibody titres, CMI was stable even between the post-basic vaccination and the pre-booster period. The magnitude of CMI after DTaP equalled those after natural infection. Measurement of cytokine-pattern showed induction of IFN-tau-producing T-helper-1-cells with lower stimulation of IL 10-producing T-helper-2-cells for DTaP and natural infection. Our data indicate that DTaP-vaccination induces a potent immune response to PT, FHA, and PRN at least equivalent to CMI after natural infection. The finding of a preferential T-helper-1-induction after DTaP and natural infection suggests a role of IFN gamma-activated macrophages in the protective response against B. pertussis-induced disease.
Abstract: Changes in the tubulin-protein and -poly(A) +RNA contents were monitored by means of Western and Northern blot analyses, respectively, during growth and maturation of leaves of a dicotyledonous (tobacco) and monocotyledonous (barley) plant. It was recently argued from immunofluorescence and preliminary biochemical data that the density of microtubular networks and concomitantly the tubulin content are distinctly reduced after cessation of cell growth in leaves [Jung et al., 1993]. The results presented now confirm and extend this view. There appeared to be clear differences between the monocot and the dicot: (1) the loss of tubulin during leaf development was much slower in the dicot than in the monocot leaves (within months instead of days); (2) the degree of loss was more dramatic in the monocot leaf and only very low threshold levels of tubulin were retained in fully differentiated tissues; and (3) the loss of tubulin in the monocot leaf tissue appeared to be correlated with the decrease in the mRNA content, whereas the high level of tubulin-RNA in fully differentiated or even almost senescent dicot leaves indicated a gene expression control at the posttranscriptional level. The comparatively rapid and very distinct tubulin-protein and -RNA disappearance during development of the monocot leaf tissues confirm at the molecular level that differentiation proceeds much faster and is much more determinative in these leaves, as was postulated from histological and physiological data.(ABSTRACT TRUNCATED AT 250 WORDS)
Abstract: Background/Aims: Limited interleukin-12 (IL-12) expression was identified to hamper neonatal immune responses to microbial pathogens. In neonatal dendritic cells we identified increased levels of IL-27 as a potential substitute. We detailed an autocrine mechanism by which IL-27 might compensate for the susceptibility gaps due to the deficiency of IL-12 in neonates.
Methods: Adult volunteers (n=12) donated peripheral blood, cord blood (n=10) was recruited at the obstetrics department. Monocytes from peripheral blood were differentiated to dendritic cells (moDC) within 3 days with GM-CSF and IL-4. Immature moDCs were subjected to IL-27. Gene expression was measured, secreted cytokines were detected using Cytometric Bead ArrayTM, and cell surface marker were analyzed by flowcytometry.
Results: We confirmed propensity of neonatal moDCs for elevated secretion of IL-27, which inhibited IL-27-receptor expression on moDCs. Compared to adults in neonatal moDCs IL-27 stimulated significantly higher levels of inflammatory cytokines (IL-8, IL-6, TNFα) and a concentration dependent maturation (CD80, CD86, HLA-DR, CD38) in neonatal (less in adult) moDCs. Homing factor CCR7 was only detectable in adult moDC. Only neonatal, IL-27-stimulated moDC gene expression screening revealed numerous expressed genes, one third with a capacity to trigger interferon expression.
Conclusions: IL-27 induced an autocrine process in moDCs, self-limiting, as IL-27 down-regulated its receptor. In the presence of IL-27 neonate moDCs showed higher sensitivity to maturate, to stimulate inflammation and to trigger genes connected to interferon gene expression, the latter clearly supporting Th1 responses, otherwise triggered by IL-12. IL-27 may contribute to those Th1 functions needed to combat infectious diseases.
Abstract: Objective:
The initial description of IL-27 indicated that it plays a role in the early regulation of T helper type 1 (Th1) initiation, induces proliferation of naïve CD4 T cells and synergizes with IL-12 in IFN-ï§ production. Furthermore it has broad inhibitory effects on Th2 and Th17 subsets of T cells as well as the expansion of inducible regulatory T cells. As an IL-12-related cytokine IL-27 is frequently present at sites of inflammation that can promote both anti- and pro-inflammatory immune responses. To highlight the mechanism how IL-27 may drive such divergent immune responses in human newborns, we investigated its role in CD4+ T cells in both human cord and adult blood.
Methodology/Principal Findings:
Umbilical cord blood samples were collected from healthy full-term infants born by cesarean sections. Leukocytes from adults were enriched from leukapheresis products. After density centrifugation, the mononuclear cell fraction (PBMC) was washed and cells were magnetically labeled for CD4 Beads and enriched on Separation Columns in a magnetic field, following stimulation with IL-27 for ascertainment of T cell response in vitro.
Our data indicate that addition of IL-27 induced the production of IFNï§ (n= 9-11; p=0,047) in neonatal CD4+ T cells, whereby IL-17 is decreased. Furthermore we noticed enhanced phosphorylation of Stat-1 (n=6-9; p=0,008) in cord blood compared to adult blood. These data suggest that binding of IL-27 to its receptor initiates Th1 responses in naïve T cells of human newborns.
Conclusions/Significance:
Taken together, our results suggest that IL-27 plays a global role in regulating the compensation of the restricted Th1 immune response in human newborns due to the deficient IL-12 production.
Abstract:
Objective:
IL-21, member of the IL-2 cytokine family, is a pleiotropic cytokine that mediates its functions through a heterodimeric receptor, comprising the subunits IL-21R and the common ï§-chain. IL-21 is mostly produced by CD4+ T cells, but molecular mechanisms which regulate IL-21 synthesis remain to be investigated.
In order to assess the effect of IL-21 in the specific context of the neonatal immune response, we analyzed its role in CD4+ T cells in both human cord and adult blood.
Methods:
Umbilical cord blood samples were collected from healthy full-term infants born by cesarean sections. Leukocytes from adults were enriched from leucapheresis products. After density centrifugation, the mononuclear cell fraction (PBMC) was washed and cells were magnetically labelled for CD4+ T cells and enriched in a magnetic field, following stimulation with IL-21 for ascertainment of T cell response in vitro.
Results:
Our data indicate that addition of IL-21 significantly induced the production of IFNï§ (p=0,011; n=15-18;) and IL-10 (p=0,013; n=11) in ï¡CD3/CD28-stimulated, neonatal CD4+ T cells. In contrast adult CD4+ T cells showed only minor differences with or without the addition of IL-21 [during stimulation]. Furthermore we noticed significantly enhanced phosphorylation of Stat-1 (n=6-9; p=0,008) in cord blood compared to adult blood derived CD4+ T cells.
Conclusions:
Taken together, our results indicate a new function for IL-21 that was unanticipated given that IL-21 is required for TH17 production in humans. Here we could demonstrate that IL-21 is a potent inducer of IL-10 and IFNï§, indicating that IL-21 is possibly involved in the generation of IL-10-producing Tr1-like cells. Otherwise IL-21 seems to be implicated in the Th1 immune response and could potentially compensate the restricted Th1 immune response in human newborn
Abstract: Interleukin (IL)-27, a heterodimer comprised of Epstein-Barr virus-induced gene 3 (EBI3) and p28, is expressed early in dendritic cell (DC) activation. Binding of IL-27 to the IL27 receptor moiety WSX-1 has been reported to initiate Th1 responses in naïve T cells. We characterized expression of EBI3-, p28- and WSX-1-mRNA in response to TLR-ligands in human monocyte-derived (m)DC (n=11). We demonstrate that a combined addition of LPS with ssRNA, or, LPS with IFNï§, induced a significantly increased secretion of IL-27p28 mRNA and of EBI3, as compared to single TLR ligand stimulation, or immature DCs and monocytes. Furthermore, exogenous IL-27 added to combined application of TLR ligands did not affected protein secretion IL-6 and IL-8, although slightly increased TNFα, IL-10 and IL-1à secretion, but clearly enhanced IL-12p70 secretion. Exogenous IL-27 did not influence the maturation process of DCs as demonstrated by the expression levels of CD80, CD86 and CD83. In conclusion, these findings indicate, that human dendritic cells support the expression of the Th1 signature cytokine IL-12p70 when receiving combined TLR signals in the presence of IL-27. These data dissect the context of a stimulating effect of IL-27, as opposed to inhibitory activities of IL-27 reported for downstream steps of the adaptive immune response.
Abstract: Purpose: To investigate the extent of cellular and humoral immune alterations in children with
malignancies after cessation of chemotherapy (CTX). Methods: Applying standard immune assays,
cellular and humoral immunity against varicella, mumps, diphtheria, measles, pertussis and tetanus
antigens were measured in 52 children (1-17 years of age) with various malignancies 1 - 96 months after CTX. Using a standardised questionnaire vaccinations before CTX and natural infections were solicited. Results: All children had at least received DT-OPV before entering CTX. In these children the
proliferative response rates against vaccine antigens were significantly reduced post CTX. 1,5 years after CTX serum antibody concentrations were low in 16/19 patients. Revaccination at this point resulted in significant antibody rises. Nevertheless one patient developed rubella 1 year after revaccination. From 29 patients with documented natural infections (22 VZV, 3 pertussis, 5 measles, 1 rubella) before CTX but "non vaccinated status" all had sufficient cellular and humoral immune response 6 months after therapy. Patients with neither previous natural infections nor sufficient vaccination before CTX developed severe clinical disease up to 4 years after CTX (8 chickenpox, 9 zoster, 5 pertussis, 1 mumps, 1 rubella). Conclusion: Children with cancer have persistent humoral and cellular immune deficiencies up to at least 4 years after completion of CTX rendering thern susceptible to vaccine preventable diseases. A prospective multicentre based study was set up to · further evaluate the clinical significance of these findings.
Abstract: Established Pertussis-Specific T-Cellular Immune Responses
Remain Stable after Vaccination with
Multivalent DTaP-Hib Combination Vaccines.
F.ZEPP*, C.U.MEYER, P.SCHMIDTKE, P.HABERMEHL,
M.KNUF, lESKOLA#, M.SLAOUt Children's Hospital University
of Mainz, Gennany; ~ational Public Health Institute, Helsinki,
Finland; +SmithKline Beecham Biologicals, Rixensart, Belgium
Background: In the absence of serological correlates for protection against pertussis,
attention has focused on the role of cell mediated immunity (CMI). Proof 0
etained vaccine efficacy in a mixed fonnulation of combination vaccines require
an extended set of toolsto address comprehensive immunological parameters. W
investigated the CMI to the acellular pertussis components in children receivin
TaP extemporaneously mixed with Hib in the ~ame syringe when given as
ooster. Material & Methods: Study A: 48 infants who had been primed with
TaP and tetanus-PRP (Hib) given separately (DTaPlHib) or combined (DTaP-Hib)
t the age of 3, 4 and 5 month were enrolled. Subjects, 15 - 22 month old, were
oosted with one dose of DTaP and Hib (separate or combined). Study B: 16 indiiduals
primed with DTaP alone at 2 month and DTaP, Hib and IPV (mixed 0
separate) at 4 and 6 month were boosted at the age of 24 month with DTaP and Bi
(mixed or se~arate). Proliferative response of peripheral blood Iymphocytes wa
easured by H-TdR incorporation after 5.5 day cultivation in the presence of PT,
HA, PRN or medium contro!. Results: In both study population (A+B) a signifiant
proliferative T-cell response was documented for all pertussis-antigens. CMI
esponses were seen in subjects who had received either separate or combined adinistration,
irrespective of primary vaccination schedule. For both, separate an
combined administration, post vaccination T-cell proliferation was more than two
fold higher compared with medium controls for 55% (PT) and 80% (FHA) of th
accinees in study A, 94% (PT) and 100% (FHA) of all vaccine es in study B. Proliferativ.
e respo. nses to PRN were less mark.e d. Conclusion: Our data. docum.ent tha
Abstract: Background: Development of new multivalent combination vaccines requires careful
examination of possible interference between the combined vaccine compoents.
Especially for vaccines lacking definite serological correlates of protectio
(e.g. pertussis vaccines), a thorough analysis of emerging interference with immuological
priming is mandatory. We investigated pertussis-specific humoral an
cellular immune responses after vaccination with differently combined multivalen
accines.
Material & Methods: 30 infants received a primary vaccination at 3, 4, and 5
onilis of either DTaP-HBV-HIB or DTaP-HBV-IPV-HIB. Serum and lymphoytes
were isolated before and after vaccination. Pertussis specific proliferation of
ells was measured by 3H_ TdR-uptake after stimulation with pertussis antigens (PT,
HA, PRN). IL-5 and IFNy secretion profile and serum antibodies against pertussis
ntigens were detected with ELISA.
Results: In both groups Immunization induced a significant T cell proliferation fo
all pertussis antigens; rates were c1early higher in subjects who received DTaPBV-
IPV-HIB. In both groups, IFNy secretion was higher than IL-5 indicating
hl-biased immuneresponse. Anti-FHA and anti-PRN titers were also higher in this
roup.
Conclusion: Combined vaccination including IPV may reinforce T cell memory fo
ertussis, a view which IS supported by the correlate levels of cytokine secretion.
n adjuvant effect of IPV on the immunization process may be assumed, based 0
ata from T cell proliferation which parallel serological data.
