Influence of antigenic forms and adjuvants on protection against a lethal infection of Aujeszky's disease virus

The influence of antigenic forms and adjuvant types on protection against a lethal infection of Aujeszky's disease virus (ADV) in mice was investigated. Antiviral IgG2a antibody response against particulate (inactivated ADV) and soluble antigen (ADV solubilized with deoxychorate-Na) in approximate order of extent was ISA70>QS-21>positively charged liposome>negatively charged liposome>weak negatively charged liposome>ISA25>lablabside F saponin>aluminum phosphate gel>non adjuvant. Particulate antigen induced higher IgG2a antibody production than soluble antigen. Particulate antigen combined with ISA70, ISA25 or positively charged liposome gave 100, 50 and 40% protection to mice, respectively. In contrast, soluble antigen plus ISA70 conferred 30% protection on mice. Immunogens using the other adjuvants gave 相似文献   

Specific IgG subclass antibody in rubella virus infections

A solid-phase antigen enzyme-linked immunosorbent assay (ELISA) was developed for the detection of rubella-specific IgG subclasses. For rubella-specific IgG1 and IgG3 sera were quantitated in arbitrary units (au) by comparison with standard curves. A concentration of 3 au was taken as that indicating positivity for specific IgG1 and specific IgG3. No sera reactive for specific IgG2 and IgG4 have been found, and thus the assay reagents were controlled by testing dilutions of a standard calibrant serum containing known concentrations of the specific IgG subclasses. Of 105 unselected sera negative for rubella antibody by radial haemolysis (RH), two gave concentrations of specific IgG1 greater than 3 au and both were positive by rubella latex agglutination (LA). The sensitivity of the assay for specific IgG1 was confirmed by examining 25 selected sera negative by RH but reactive by LA. Twenty-one gave concentrations greater than 3 au. None of these 130 was positive for specific IgG3. All 63 sera containing greater than 15 international units rubella antibody by RH from cases of rubella in the remote past contained specific IgG1 and eight contained specific IgG3. In 79 cases of primary rubella, specific IgG1 developed in all cases by day 8. Specific IgG3 became detectable in all cases except one by day 16. Serum taken on day 21 from one case was negative for specific IgG3 but the absence of later sera precluded further investigation. One case had become negative for specific IgG3 by day 56. Sera from 24 cases of rubella reinfection were examined and all contained specific IgG1. In three cases of symptomatic reinfection, specific IgG3 was detectable in two but not in the remaining case. In 2 of the 21 cases of asymptomatic reinfection only a very early or a very late serum was available. Of the remaining 19 cases, 7 had detectable specific IgG3. However, only one of 9 sera collected 30-50 days after contact contained specific IgG3. Thus for the asymptomatic patient for whom other serological tests suggest a recent rubella infection, the failure to detect specific IgG3 in sequential sera collected after contact suggests reinfection rather than primary rubella. The detection of specific IgG3 did not correlate with the presence of specific IgM. Sera collected 6-8 weeks after rubella vaccination had detectable specific IgG1 in 32 of 33 cases and specific IgG3 in 9 of 33. The remaining vaccinee was seronegative.     

Influence of maternal immunity on antibody and T-cell response in mice

The prevalence of maternal antibodies (Abs) and an immature neonate immune system, which is inclined to a T-helper 2 (Th2) response, are factors that counteract active immunization in early life. In a mouse model, the maternal influence on an active immunization of 2-day-old offspring with Sendai virus (SV) envelope proteins was explored. Maternal immunizations were conducted with the same SV antigen preparation as used for offspring immunization, presented in three different formulations, namely micelles of SV (SV-MIC), Al(OH)(3)-adjuvanted SV-MIC (SV-aluMIC) for Th2 and immune stimulation complex (iscom)-adjuvanted SV (SV-ISC) for Th1. An inversely correlation was found between the immunoglobulin G2a (IgG2a) Abs of the mothers and the interleukin 5 (IL-5) levels of the offspring. Although a maternally derived immunity induced by SV-aluMIC suppressed both B- and T-cell responses of the newborn to SV-ISC immunization, significant priming effects of the neonatal immunization on IgG2a Abs and IFN-gamma levels were recorded after reimmunization at adult age.     

Identification of adjuvants that enhance the therapeutic antibody response to host IgE

In the development of a novel vaccine against atopic allergies, we have screened for adjuvants that enhance the therapeutic antibody response against self immunoglobulin E (IgE). The response against self IgE is induced by administration of a vaccine antigen, which contains both self and non-self IgE regions, together with an adjuvant. We evaluated five commonly used adjuvants; Freund's, aluminium hydroxide, ISCOMs, Montanide ISA 51 and Montanide ISA 720, and found that the mineral oil-based adjuvants; Montanide ISA 51 and Freund's induced at least 5-10-fold higher anti-self IgE titers than any of the other candidates. However, with one exception, Alum, the immune responses against the carrier, i.e. the non-self regions, were similar for all adjuvants, indicating that the ability to induce responses against self and non-self antigens differ among adjuvants. The responses against non-self IgE were more than 50-fold higher than antibody responses against self IgE in both the Freund's and Montanide 51-administered animals, indicating that the response against self molecules is markedly inhibited by tolerance-inducing mechanisms. Co-administration of Montanide ISA 51 with immuno-stimulatory substances from bacteria; muramyldipeptide (MDP), monophosphoryl-lipid A (MPL) or a formyl-methionine-containing tripeptide (fMLP), did not elevate the anti-self IgE response. Hence, adjuvants based on pure mineral oil without additional immuno-stimulatory substances appear to be the best adjuvant candidates in therapeutic vaccines aimed at regulating the in vivo levels of self-proteins.     

Frequency of naturally occurring antibody to influenza virus antigenic variants selected in vitro with monoclonal antibody.

Antigenic variants of A/Texas/77 (H3N2) virus were selected in vitro using monoclonal antibody to virus haemagglutinin (HA). The antigenic variants and parental A/Texas/77 viruses were used to to evaluate the frequency of anti-HA antibodies in the sera of children and adults using single-radial-haemolysis (SRH) tests. Twenty to 41% of selected sera from adults, which contained antibody to the parental A/Texas/77 virus, failed to react with the different antigenic mutant viruses. A higher proportion of sera from children (37-58%) failed to react with the antigenic variants. Certain human sera and particularly those of children would appear to possess a more limited antibody repertoire to influenza HA, potentially allowing new antigenic variants to escape neutralization and spread in the community.     

The immune response to infection with vaccinia virus in mice. I. Infection and the production of antibody neutralizing cell-associated and cell-free virus.

The onset, duration and magnitude of antibody responses to a poxvirus infection were examined. Mice were inoculated intravenously with the WR strain of vaccinia virus and developed pocks on their tails. The number of pocks was related to the size of the inoculum. Virus was detectable in the spleen and infected mice were subsequently immune to intravenous and intra-nasal challenge. Sera of infected animals neutralized both cell-free and cell-associated virus. Antibody against cell-free virus appeared first; maximum titres were reached sooner but were lower than those of antibody neutralizing cell-associated virus. Titres remained high for at least 100 days after infection.     

Immunization of mice with P6 of nontypeable Haemophilus influenzae: kinetics of the antibody response and IgG subclasses.

The kinetics of the anti-P6 antibody response was characterized in three strains of mice of different haplotypes (Balb/c; H-2d, C3H/H; H-2k, SJL/J; H-2s). Anti-P6 antibodies were measured on a weekly basis by enzyme-linked immunosorbent assay (ELISA). The primary response peaked 2 or 3 weeks after the initial injection with 40 microg of purified P6. The response remained at a plateau for 8-10 weeks. A maximum titer of 1:1,638,400 was attained and then steadily declined. To study the ability of P6 to generate a recall response, we opted to boost the vaccinated mice with a known subimmunogenic dose of live nontypeable Haemophilus influenzae (NTHI) bacteria. After the anti-P6 antibody titers in the primed animals had stayed at baseline levels for 2 weeks, the mice were injected intraperitonealy with 10(8) cfu of NTHI in sterile saline. This challenge with live NTHI bacteria induced a very rapid and strong secondary antibody response in all mice. Finally, we demonstrated that these murine anti-P6 sera were 100% bactericidal against three strains of NTHI when tested in a complement dependant bactericidal assay.     

Sexual diergism in antibody response to whole virus trivalent inactivated influenza vaccine in outbred mice

An outbred mouse model was used to determine if antibody response to immunization with whole-virus trivalent inactivated influenza vaccine (TIV) differs between the sexes. The antibody response was examined one (serum titer of IgM antibodies), and three and six weeks post-immunization (serum titer of neutralizing and total IgG antibodies and IgG subclass profile). Compared with male in female mice was found (i) the more robust IgM response against all influenza strains included in TIV and (ii) more vigorous neutralizing antibody and total IgG responses against H1N1 influenza virus at both the examined time points post-immunization. The total IgG antibody response against H3N2 and B influenza viruses was comparable between female and male mice three weeks post-immunization, but significantly greater in female mice six weeks post-immunization. The neutralizing antibody response against H3N2 and B influenza viruses did not significantly differ between sexes at both the examined points post-immunization. Finally, three weeks post-immunization subclass profile of IgG specific to the influenza strains included in TIV differed between female and male mice, reflecting the lower titer of IgG1 antibodies in female ones, so that IgG2a (contributing mainly to the total IgG) to IgG1 ratio in mice of this sex was shifted toward the former. In agreement with this shift, compared with male mice, Th1/Th2 balance in female mice was shifted toward Th1, as shown by ELISPOT. Collectively, the results showed influenza virus strain-dependent sexual dimorphism in the magnitude, dynamics and characteristics of antibody response in outbred mice immunized with TIV.     

The antibody response in Legionnaires' disease.

From 22 patients with Legionnaires'' disease, 86 sera were examined for specific serotype 1 IgM and IgG antibodies by the indirect immunofluorescence technique. No antibody was detectable until 8 days or more from the onset of symptoms. When produced the amount was widely variable and remained detectable for periods from less than 34 days to more than 1 year. Intially IgM antibody predominated, ten patients produced only IgM in the first 21 days, six produced only IgM in the first 28 days and three did not produce IgG at any time. One patient, and possibly a second, produced only IgG antibody. Since IgM antibody was still present in one patient after a year it is important not to accept the presence of this as evidence of very recent infection. It is advisable that any type of serological test for L. pneumophila infection should detect the production of both IgM and IgG antibodies.     

The antibody response in Legionnaires' disease.

From 22 patients with Legionnaires' disease, 86 sera were examined for specific serotype 1 IgM and IgG antibodies by the indirect immunofluorescence technique. No antibody was detectable until 8 days or more from the onset of symptoms. When produced the amount was widely variable and remained detectable for periods from less than 34 days to more than 1 year. Intially IgM antibody predominated, ten patients produced only IgM in the first 21 days, six produced only IgM in the first 28 days and three did not produce IgG at any time. One patient, and possibly a second, produced only IgG antibody. Since IgM antibody was still present in one patient after a year it is important not to accept the presence of this as evidence of very recent infection. It is advisable that any type of serological test for L. pneumophila infection should detect the production of both IgM and IgG antibodies.     

Combination adjuvants for the induction of potent, long-lasting antibody and T-cell responses to influenza vaccine in mice

Influenza is controlled by protective titres of neutralizing antibodies, induced with the help of CD4 T-cells, and by antiviral T-cell effector function. Adjuvants are essential for the efficient vaccination of a na?ve population against avian influenza. We evaluated a range of adjuvants for their ability to enhance, in na?ve mice, protective hemagglutination inhibition (HI) titres, which represent the generally accepted correlate of protection, virus-neutralizing titres and T-cell responses to a new generation influenza vaccine produced in cell culture. The selected adjuvants include alum, calcium phosphate (CAP), MF59, the delivery system poly-(lactide co-glycolide) (PLG) and the immune potentiator CpG. MF59 was clearly the most potent single adjuvant and induced significantly enhanced, long-lasting HI and neutralizing titres and T-cell responses in comparison to all alternatives. The combination of alum, MF59, CAP or PLG with CpG generally induced slightly more potent titres. The addition of CpG to MF59 also induced a more potent Th1 cellular immune response, represented by higher IgG2a titres and the induction of a strongly enhanced IFN-gamma response in splenocytes from immunized mice. These observations have significant implications for the development of new and improved flu vaccines against pandemic and inter-pandemic influenza virus strains.     

Response to foot-and-mouth disease vaccines in newborn calves. Influence of age, colostral antibodies and adjuvants

Oil-emulsified (OE) and aqueous (Aq) vaccines were prepared with the same batch of inactivated A24 8345 foot and mouth disease virus (FMDV). Calves born to vaccinated dams did not respond to the Aq vaccine 30 or 90 days post partum. When the OE vaccine was used on a similar group of calves, no responses were elicited up to 21 days post partum. However, calves 30 or more days old responded like adult cattle to the OE vaccine. When the OE vaccine was used in colostral antibody-free calves 3-30 days old, all animals showed good antibody responses but, in calves vaccinated 3 or 7 days post partum, antibodies were detectable only after a considerable period of time. Our results show that both passively acquired colostral antibodies and age are important in the response of very young calves to FMDV oil vaccines. From a practical point of view, in endemic areas where adult cattle are periodically vaccinated, vaccination of calves between 30 and 60 days post partum with OE vaccines would lead to high levels of herd protection.     

Nasal antibody response to mumps virus after vaccination and natural infection.

Nasopharyngeal secretions and sera were collected during an epidemic of mumps in a semi-closed institution. None of the children who received live attenuated mumps vaccine 5 months previously developed any symptoms of mumps. Neutralizing and IgA antibodies against mumps virus were measured by the peroxidase-antiperoxidase method and the fluorescent antibody to membrane antigen method, respectively. The immune responses in nasopharyngeal secretions and serum were studied in four groups (apparently infected, non-apparently infected, non-infected and vaccinated). We found that IgA, which has neutralizing activity, was produced after vaccination as well as after natural infection. It is suggested that local antibody induced by vaccination may contribute to protection against the mumps virus.     

Analogous IgG subclass response to pertussis toxin in vaccinated children,healthy or affected by whooping cough

The study of antigen specific IgG subclass distribution during disease, or during any other natural or artificial immunisation, can provide useful information on the kind of the immune response and the expected levels of protection. This is particularly true for diseases, such as pertussis in which the mechanisms underlying specific defence are still not completely understood. An investigation was therefore performed to evaluate the IgG subclass response to pertussis toxin (PT) in sera from 89 healthy vaccinated children and 131 vaccinated or unvaccinated children convalescent after a confirmed B. pertussis symptomatic infection. Antibody titres were expressed in arbitrary ELISA units/ml, and statistical analyses were performed. In unvaccinated convalescent children IgG1 and IgG3 were prevalent whereas in children immunised with two different acellular pertussis (aP) vaccines, both healthy and convalescent, IgG1, IgG2 and IgG4 antibodies were mainly produced. Maintenance of the same anti-PT antibody response pattern in healthy acellular pertussis vaccine recipients and in vaccinated children who later acquire the disease is an interesting result indicative of the priming effect induced by these vaccines in the direction of a relatively higher Th2 cell-polarisation of the immune response.     

Influence of the incubation temperature and the batch components on the sensitivity of an enzyme-linked immunosorbent assay to detect Aujeszky's disease virus glycoprotein E (gE)

Although licensed batches of an enzyme-linked immunosorbent assay (ELISA) for Aujeszky's disease virus (ADV) were used, and the assays were performed within an ISO/IEC 17025 accredited quality control system, certain routine runs of the ADV ELISA were not validated using the quality system criteria, even when all technical parameters were controlled. Incubation at different temperatures and batch composition were identified as parameters that could result in non-validated assays/runs. Therefore, the effect of incubation temperature and batch composition on the analytical sensitivity of the ELISA was investigated. The World Organisation for Animal Health (OIE) standard reference serum ADV1 was diluted 1:8 and tested in 94 different glycoprotein E ELISA runs performed with different batches and different incubation temperatures. The incubation temperature and batch components had a significant influence on the qualitative result for the OIE standard reference serum. An incubation temperature of at least 22 degrees C was recommended, based on the results of this analysis. Which of the batch components caused these differences in sensitivity was not investigated further.     

The IgG subclass profile of anti-HBs response in vaccinated children and children seroconverted after natural infection

The IgG subclass profiles of anti-HBs antibodies were investigated in 30 children who had recovered from acute hepatitis B and 40 children vaccinated against hepatitis B virus (HBV) with Engerix B. After natural seroconversion the mean geometric value of anti-HBs titres was ca 41-fold lower than at the peak of response in vaccinees, and specific antibodies were highly restricted to IgG1 subclass followed by IgG3 with only a minor contribution of IgG2 and IgG4. Conversely, in children immunized with recombinant HBsAg, IgG1 and IgG3 dominated after two doses of vaccine and 1 month after the third injection but the response was less selective and more variable. One year after vaccination IgG4 anti-HBs antibodies became the second dominating isotype. Significant statistical differences in the profiles of IgG anti-HBs were observed when the age and maturity of humoral response were considered. While children vaccinated below 5 years of age responded mainly with IgG1 and IgG3 subclasses, older children (> 5 years) showed a high individual variability in the specific profiles with a high contribution of IgG4. We concluded that vaccination at a younger age leads to the production of antibody subclasses which are more effective for virus neutralization.     

IgG抗体亲和力试验判定麻疹病例的价值

目的评价麻疹IgG抗体亲和力试验对判定麻疹病例的价值。方法以中国疾病预防控制信息系统2013-2015年天津市麻疹实验室确诊病例和麻疹排除病例为研究对象。回顾性追溯保存的病例血清,开展麻疹IgG抗体亲和力试验,重新对麻疹排除病例进行归类。结果共收集到326例麻疹病例血标本,其中实验室确诊病例267例,排除病例59例,≥20岁病例占92.33%(301/326)。麻疹IgG抗体亲和力试验显示,确诊病例和排除病例中麻疹IgG高亲和力抗体的比例分别为66.95%(158/236)和91.23%(52/57),差异有统计学意义(χ²⁼13.33,P<0.001)。根据判定标准,15.25%(9/59)排除病例被重新判定为麻疹病例,其中8例是高亲和力抗体,有含麻疹成分疫苗(MCV)免疫史,判定为继发性免疫失败病例;1例为低亲和力抗体,有典型的麻疹临床症状,无MCV免疫史。结论麻疹IgG抗体亲和力试验能够提供有参考意义的血清学证据,可以减少麻疹急性期血清学诊断中由于IgM抗体假阴性而造成的错误排除。