Serum IgG Antibody Levels Research Articles

Enhancement of rat tear IgA antibody responses following intranasal immunization with antigen and CpG ODN

Purpose. To determine the effect of unmethylated oligodeoxynucleotides containing bacterial CpG motifs (CpG ODN) on the induction of rat tear IgA antibody responses. Methods. Rats were immunized intranasally with either soluble dinitrophenylated bovine serum albumin (DNP-BSA) or poly(lactide-co-glycolide) (PLG) encapsulated DNP-BSA in combination with CpG ODN. The animals received two immunizations 21 days apart. Following the second immunization, tear, saliva and serum samples were collected for 28 days and analyzed for antigen specific antibodies. Tear IgA, saliva IgA and serum IgG antibody concentrations were determined by ELISA. Results. Co-administration of CpG ODN with either soluble or encapsulated antigen resulted in significantly elevated levels of both tear and salivary IgA antibodies as well as levels of serum IgG antibodies. Microencapsulated DNP-BSA plus CpG ODN elicited higher levels of IgA antibodies in tears than did soluble antigen plus CpG ODN. Conclusions. CpG ODN is an effective mucosal immune modulator for enhancing rat tear IgA antibody responses to both soluble and microencapsulated antigens.

Influence of placental malaria infection and maternal hypergammaglobulinaemia on materno-foetal transfer of measles and tetanus antibodies in a rural west African population.

Placental malaria infection jeopardizes pregnancy outcome, and its influence may also impair the transplacental transfer of some antibodies. Two hundred and thirteen Gambian mother-baby pairs were studied to determine the influence of placental malaria infection and maternal hypergammaglobulinaemia on transplacental transfer of measles and tetanus antibodies in Gambian population. Placental blood and tissue were collected for placental malaria diagnosis. Cord and maternal sera were tested for total IgG concentration by laser nephelometry and for IgG antibody to tetanus toxoid and measles by ELISA. The prevalence of placental malaria infection was 51.1%. Mothers whose placentae were parasitized had a significantly higher mean total serum IgG (22.0 g/L vs 11.3 g/L, p < 0.001) and measles antibody level (4.02 IU/mL vs 1.21 IU/mL, p < 0.01), but not tetanus antibody, than mothers with non-parasitized placentae. Results of multiple regression analysis showed that placental malaria infection and maternal hypergammaglobulinaemia were associated with the reduction of 72% (95% CI 67.84) and 86% (95% CI 76.91) in transplacental transfer of measles antibody respectively but did not influence the transfer of tetanus antibody. It is concluded that the combined influence of placental malaria infection and maternal hypergammaglobulinaemia is significantly associated with the transfer of impaired measles antibody in this population.

Modified immunogenicity of a mucosally administered antigen.

Streptococcus mutans is present in the saliva of most individuals and is modified by salivary components bound to the cells. These saliva-bound S. mutans are swallowed, exposed to high levels of acidity in the stomach, and presented to the common mucosal immune system. Much effort has been directed to identifying the specific S. mutans antigens that the mucosal immune responses are directed against. However, little is known about the host-altered antigenic determinants that the mucosal immune system recognizes. The immunogenicity of gastrically intubated untreated S. mutans cells, cells coated with whole human saliva, cells treated with HCl (pH 2.0), and saliva-coated and acid-treated cells in mice was investigated. Saliva and serum samples were assayed by enzyme linked immunosorbent assay for immunoglobulin A (IgA) and IgG antibodies, respectively, against the untreated or treated S. mutans cells. In general, the levels of salivary IgA and serum IgG antibodies to the antigen against which the mice were immunized were significantly higher (P < or = 0.05). In addition, human saliva and serum samples from 12 subjects were assayed for naturally occurring antibody against the untreated or treated S. mutans cells. In every case, significantly higher reactivity was directed against the saliva-coated and acid-treated cells followed by the saliva-coated S. mutans. These results provide evidence for the altered immunogenicity of swallowed S. mutans in humans by coating native S. mutans antigens with salivary components and/or denaturing surface S. mutans antigens in the acidic environment of the stomach, which would lead to an immune response to modified S. mutans determinants and not to native S. mutans antigens.

Peroral immunization with Helicobacter pylori adhesin protein genetically linked to cholera toxin A2B subunits

Helicobacter pylori is a major cause of gastric-associated diseases. To evaluate the efficacy of a possible vaccine antigen against H. pylori infection, the chimaeric construct adhesin--CTXA2B, derived from H. pylori adhesin genetically coupled to cholera toxin (CTX) subunits A2 and B (CTXA2B), was expressed in Escherichia coli as an insoluble recombinant chimaeric protein. The protein was then purified by denaturation, renaturation and size-exclusion chromatography. The composition of purified adhesin--CTXA2B was verified by SDS/PAGE and Western blotting with antibodies to antigenic components of adhesin and CTXB, and confirmed as a chimaeric protein with G(M1)-ganglioside binding activity and adhesin epitopes by a G(M1)-ELISA developed using antibodies to adhesin. Oral immunization of mice with adhesin--CTXA2B induced higher levels of mucosal IgA and serum IgG antibodies to H. pylori adhesin and to CTXB than in mice immunized with adhesin or CTXA2B alone. Adhesin--CTXA2B was also demonstrated to be a potential protective antigen in a mouse model of H. pylori infection. The immunization of mice with adhesin--CTXA2B protected 62.5% of mice infected with H. pylori SS1 strain, whereas adhesin immunization was not able to confer protection to mice. This protection may be correlated with high levels of mucosal IgA and serum IgG antibodies against H. pylori adhesin. Taken together, the results indicate that the genetically linked CTXA2B acts as a useful mucosal adjuvant, and that the adhesin-CTXA2B chimaeric protein could be a potential component in future H. pylori vaccine development.

Transplacental transfer and age-related levels of serum IgG antibodies to the capsular polysaccharides of Streptococcus pneumoniae types 14 and 19 in Korea.

Little is known about the prevalence of naturally acquired IgG antibodies to the capsular polysaccharides of Streptococcus pneumoniae (pneumococcal IgG) in Korea. In the present study, we investigated transplacental transfer and age-related levels of pneumococcal IgG to provide background seroepidemiologic data for S. pneumoniae in Korea. One hundred thirty eight sera were assayed by ELISA for IgG to pneumococcal polysaccharide capsular serotypes 14 and 19, the predominant serotypes for under 15 yr of age in Korea. The subjects were divided into 7 subgroups according to age. The cord/maternal geometric mean titer of pneumococcal were 4.47+/-5.88/5.21 +/- 5.88 for serotype 14, and 4.68 +/- 5.55/6.55 +/- 6.92 for serotype 1 9 (mean +/- standard deviation, microg/mL). After birth, the geometric mean titers of pneumococcal IgG for serotypes 14 and 19 expressed in microg/mL were 1.18+/-2.12 and 1.41+/-2.17 in the 0-6 months group, 0.27+/-0.19 and 0.69+/-0.93 in 7-12 months, 0.21+/-0.22 and 0.64+/-1.32 in 1-2 yr, 0.69+/-0.78 and 2.65+/-2.46 in 3-6 yr, 2.52+/-2.72 and 8.29+/-4.24 in 7-10 yr, respectively. In conclusion, reduced transplacental transfer and very low serum concentrations of pneumococcal IgG may contribute to the susceptibility of neonates, infants, and young children to S. pneumoniae infection.

Role of antibody response in outcome of antibiotic-associated diarrhoea

There are approximately 18 000 reports of antibioticassociated Clostridium difficile diarrhoea per year in England and Wales. 1 Communicable Disease Surveillance CentreClostridium difficile in England and Wales: 1999. Commun Dis Rep CDR Weekly. 2000; 10: 135 Google Scholar Over 80% of cases occur in inpatients aged over 65 years, 1 Communicable Disease Surveillance CentreClostridium difficile in England and Wales: 1999. Commun Dis Rep CDR Weekly. 2000; 10: 135 Google Scholar and those most at risk are aged greater than 75 years with severe co-existing illness. 2 Kyne L Merry C O'Connell B Kelly A Keane C O'Neill D Factors associated with prolonged symptoms and severe disease due to. Clostridium difficile. Age Ageing. 1999; 28: 107-113 Crossref PubMed Scopus (112) Google Scholar This iatrogenic disease can cause major disruption to hospital activity and prolong admission by an average of 21 days. 3 Wilcox MH Cunniffe JG Trundle C Redpath C Financial burden of hospital-acquired Clostridum difficile infection. J Hosp Infect. 1996; 34: 23-30 Summary Full Text PDF PubMed Scopus (218) Google Scholar Administration of broadspectrum antibiotics, notably second and third generation cephalosporins disrupts the pattern of the normal colonic bacterial flora and enables colonisation with C difficile, which is highly prevalent in hospitals. Colonisation with C difficile may result in gut clearance of the normal flora, symptomless carriage of C difficile, diarrhoea, colitis, pseudomembranous colitis, and, in some cases, death. Those strains that are pathogenic cause disease by producing potent exotoxins, toxin A and/or toxin B. Recurrent episodes of C difficile infection seem to be due to re-infection rather than relapse, and perhaps some strains are more virulent than others. 4 Wilcox MH Fawley WN Settle CD Davidson A Recurrence of symptoms in Clostridium difficile infection – relapse or reinfection?. J Hosp Infect. 1998; 38: 93-100 Summary Full Text PDF PubMed Scopus (160) Google Scholar Some patients, however, remain symptom-free despite toxin production. Why only about half of those patients colonised by C difficile develop diarrhoea has long been a mystery, 5 Kyne L Warny M Qamar A Kelly CP Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A. N Engl J Med. 2000; 342: 390-397 Crossref PubMed Scopus (758) Google Scholar as has why some of these people experience multiple symptomatic episodes. 3 Wilcox MH Cunniffe JG Trundle C Redpath C Financial burden of hospital-acquired Clostridum difficile infection. J Hosp Infect. 1996; 34: 23-30 Summary Full Text PDF PubMed Scopus (218) Google Scholar , 4 Wilcox MH Fawley WN Settle CD Davidson A Recurrence of symptoms in Clostridium difficile infection – relapse or reinfection?. J Hosp Infect. 1998; 38: 93-100 Summary Full Text PDF PubMed Scopus (160) Google Scholar Simplistically, the possible explanations for these observations can be grouped into pathogen and/or host factors. Association between antibody response to toxin A and protection against recurrent Clostridium difficile diarrhoeaA serum antibody response to toxin A, during an initial episode of C difficile diarrhoea, is associated with protection against recurrence. Full-Text PDF

Non-urease producing Helicobacter pylori in chronic gastritis.

Helicobacter pylori infection is the commonest cause of gastritis. Different patterns of immune response to H. pylori infection and characteristics of bacteria are considered to contribute to clinical outcomes. To determine characteristics of the host H. pylori relationship in subjects with non-ulcer dyspepsia and a histological diagnosis of gastritis. Thirty-five subjects with chronic gastritis undergoing endoscopy (mean age 53 years, range 24-82, 14 male and 21 female) were studied, none of whom was on nonsteroidal anti-inflammatory drugs or antibiotics. H. pylori infection was determined by rapid urease test (CLOtest), culture, antibody and RT-PCR for Ure C, Cag A and 26 kDa gene and histology. Cytokine production of mucosal IL-6 and IL-8 were measured by ELISA. Fifteen subjects were positive by CLOtest and/or bacterial culture. In these subjects histology showed numerous helical forms of H. pylori (Group I). Nine subjects were negative by CLOtest, bacterial culture, and mRNA for urease C fragment, but positive by PCR for the 26 kDa protein encoding gene. Histology in these subjects showed the presence of either coccoid forms (four), or scant helical forms (two), or mixed coccoid/helical forms (three) (Group II). Eleven subjects were negative by all methods of detection (Group III). IgG and IgA antibody levels in serum (p<0.05) and gastric tissue culture supernatant (p<0.001) were significantly higher in Group I than those in Group II or III. There were significant differences in the IgG serum and IgA supernatant antibody levels (p<0.01 and p<0.05) when Group II was compared to Group III. Supernatant IL-6 levels were significantly higher in Group I (p<0.01) than those from Groups II and III. IL-8 levels were higher in Group I (p<0.01) and Group II (p<0.05) when compared to Group III. 'H. pylori-negative' gastritis can be associated with a non-urease producing form of H. pylori, with a reduction in both local and systemic antibody levels and mucosal pro-inflammatory cytokines.

Polarization of Porphyromonas gingivalis-specific helper T-cell subsets by prior immunization with Fusobacterium nucleatum.

Antigen-specific T-cell clones were obtained from mice immunized with Fusobacterium nucleatum ATCC 10953 and/or Porphyromonas gingivalis 381. 10 BALB/c mice per group were immunized with F. nucleatum followed by P. gingivalis, or with P. gingivalis alone by intraperitoneal injection of viable microorganisms. Spleen T cells were isolated and stimulated in vitro with viable P. gingivalis cells to establish P. gingivalis-specific T-cell clones. T-cell phenotypes and cytokine profiles were determined along with T-cell responsiveness to F. nucleatum or P. gingivalis. Serum immunoglobulin G antibody titers to F. nucleatum or P. gingivalis were also determined by enzyme-linked immunosorbent assay. All the T-cell clones derived from mice immunized with F. nucleatum followed by P. gingivalis demonstrated Th2 subsets, while those from mice immunized with P. gingivalis alone demonstrated Th1 subsets based on the flow cytometric analysis and cytokine profiles. All T-cell clones from both groups were cross-reactive to both P. gingivalis and F. nucleatum antigens. Phenotypes of T-cell clones were all positive for CD4. Mean post-immune serum IgG antibody levels to F. nucleatum or P. gingivalis were significantly higher than the pre-immune levels (P < 0.05, P < 0.01, respectively). There were no significant differences in the antibody titers between the two groups. It was concluded that P. gingivalis-specific T cells initially primed by cross-reactive F. nucleatum antigens were polarized to Th2 subset, while T cells stimulated with P. gingivalis alone maintained the profile of Th1 subset.

Induction of secretory immunity with bioadhesive poly (D,L-lactide-co-glycolide) microparticles containing Streptococcus sobrinus glucosyltransferase.

The effect of mucosal delivery of Streptococcus sobrinus glucosyltransferase (GTF) in bioadhesive poly (D,L-lactide-co-glycolide) (PLGA) microparticles on induction of salivary IgA and serum IgG antibody responses was measured in Sprague-Dawley rats. Preparations of GTF/PLGA/gelatin microparticles, or PLGA/gelatin microparticles or GTF in alum, were administered four times at weekly intervals by intranasal or intragastric routes. Two subcutaneous injections of GTF in PLGA/gelatin microparticles or in alum were given to separate groups of rats. Significant elevations in salivary IgA antibody levels to S. sobrinus GTF were observed only in the groups immunized intranasally 28 days after immunizations were begun. Five of six rats given the GTF microparticles intranasally had positive salivary IgA antibody responses to GTF, and the mean salivary IgA antibody level of this group was 30-fold higher than any other mucosally or systemically immunized group. Salivary IgA responses in the GTF-microparticle group remained significantly higher than all other mucosally immunized groups for at least 10 weeks after the primary immunization. All rats in this group demonstrated aspects of anamnesis following a more limited secondary course of intranasal administration. Intranasal administration of GTF in microparticles also induced a serum IgG response to GTF in some rats. After secondary intranasal GTF microparticle administration, several rats had sustained serum IgG antibody levels that were within the range of sera from rats subcutaneously injected with GTF in microparticles or in alum. Thus intranasal delivery of GTF-containing bioadhesive microparticles induced the highest and longest lasting salivary immune response of any mucosal or systemic route or vehicle tested and could be expected to be a useful method for induction of mucosal immunity.

Asymptomatic carriage of Clostridium difficile and serum levels of IgG antibody against toxin A.

Clostridium difficile infection can result in asymptomatic carriage, mild diarrhea, or fulminant pseudomembranous colitis. We studied whether antibody responses to C. difficile toxins affect the risks of colonization, diarrhea, and asymptomatic carriage. We prospectively studied C. difficile infections in hospitalized patients who were receiving antibiotics. Serial stool samples were tested for C. difficile colonization by cytotoxin assay and culture. Serum antibody (IgA, IgG, and IgM) levels and fecal antibody (IgA and IgG) levels against C. difficile toxin A, toxin B, and nontoxin antigens were measured by an enzyme-linked immunosorbent assay (ELISA). Of 271 patients, 37 (14 percent) were colonized with C. difficile at the time of admission, 18 of whom were asymptomatic carriers. An additional 47 patients (17 percent) became infected in the hospital, 19 of whom remained asymptomatic. The baseline antibody levels were similar in the patients who later became colonized and those who did not. After colonization, those who became asymptomatic carriers had significantly greater increases in serum levels of IgG antibody against toxin A than did the patients in whom C. difficile diarrhea developed (P<0.001). The adjusted odds ratio for diarrhea was 48.0 (95 percent confidence interval, 3.4 to 678) among patients with colonization who had a serum level of IgG antibody against toxin A of 3.00 ELISA units or less, as compared with patients with colonization who had a level of more than 3.00 ELISA units. We find no evidence of immune protection against colonization by C. difficile. However, after colonization there is an association between a systemic anamnestic response to toxin A, as evidenced by increased serum levels of IgG antibody against toxin A, and asymptomatic carriage of C. difficile.

Localized sinus inflammation in a rabbit sinusitis model induced by Bacteroides fragilis is accompanied by rigorous immune responses

We evaluated inflammatory and immune responses against Bacteroides fragilis in a rabbit sinusitis model. Bacteroides was inoculated into the left maxillary sinus, and inflammatory (histology, cell number/cytology, lactose dehydrogenase, and apoptosis) and immune responses in the sinus, airway, and peripheral blood (PB) were determined for up to 4 weeks. In the inflamed sinus, the lactose dehydrogenase level was markedly elevated, with neutrophilic infiltration, severe tissue inflammation, and increased apoptosis. Low-grade tissue inflammation was present in the contralateral and sham-operated sinuses, but other parameters remained unchanged, and so did those in the airway and PB in the inoculated rabbits. Serum IgG antibody levels increased rapidly, were highest at 3 weeks, and began to decline at 4 weeks. Cellular immune responses (proliferation and interferon-γ mRNA expression) against Bacteroides were detected in the PB of all inoculated rabbits. Vigorous immune responses against Bacteroides may have localized but failed to terminate inflammation in the sinus, indicating importance of microenvironmental factors. (Otolaryngol Head Neck Surg 1999;120:869-75.)

Intranasal immunization confers protection against murine Pneumocystis carinii lung infection.

To evaluate the feasibility of mucosal immunization against Pneumocystis carinii (Pc) experimental infection, female BALB/c mice were intranasally immunized three times with soluble Pc antigens plus cholera toxin fraction B (Pc-CTB); control groups received either Pc antigen, CTB, or phosphate-buffered saline (PBS) alone. Two weeks after the last immunization, five animals from each group were sacrificed, and cellular and humoral immune responses were evaluated. The remaining five mice were CD4 depleted using a monoclonal antibody against mouse CD4 and inoculated with viable Pc. Significantly higher specific lymphoproliferative responses from tracheobronchial lymph node cells, immunoglobulin M (IgM) and IgG antibody levels in serum, and bronchoalveolar lavage (BAL)-derived IgA antibody concentrations were observed in the Pc-CTB group of mice relative to control groups (P < 0.01). Five weeks after challenge, no Pc organisms were observed in the lung smears of the Pc-CTB group, while the animals receiving antigen, adjuvant, or PBS had progressively higher numbers of Pc microorganisms. By Western blot analysis, a strongly reactive 55- to 60-kDa antigen was recognized by BAL IgA and by serum IgG. In summary, mucosal immunization elicited specific cellular and humoral immune responses and protected against Pc lung infection after immunosuppression.

Human IgG subclass responses in relation to serum bactericidal and opsonic activities after immunization with three doses of the Norwegian serogroup B meningococcal outer membrane vesicle vaccine

Ten adult volunteers, with low prevaccination levels of serum IgG antibodies against meningococcal antigens (<1 μg ml −1), received three doses of the Norwegian group B meningococcal outer membrane vesicle (OMV) vaccine intramuscularly at weeks 0, 6 and 46. Anti-OMV IgG subclass responses were measured and compared with serum bactericidal activity (SBA) and opsonic activity against the vaccine strain 44/76. All vaccinees showed an IgG1 antibody response after each vaccine dose. The vaccine-induced median serum IgG1 antibody levels were 16, 17 and 18 μg ml −1 2–6 weeks after the first, second and third dose, respectively. Three vaccinees showed a weak IgG3 response after the first dose, whereas 8 and 9 showed a response after the second (median=10 μg ml −1) and third dose (median=10 μg ml −1), respectively. Low levels of anti-OMV IgG2 antibodies were found, whilst specific IgG4 antibodies were only detected for one vaccinee. The vaccine induced at least a fourfold increase in SBA titre in 8 vaccinees after the first dose, in 9 vaccinees after 2 doses and in all vaccinees after 3 doses. A positive correlation was found between IgG1 subclass antibody levels and SBA ( r=0.62, P<0.0001). Elevated opsonophagocytic activity, measured as respiratory burst (RB), was observed in all vaccinees after one vaccine dose and usually increased after 2 and 3 doses. A strong positive correlation was found between IgG1 antibody levels and RB ( r=0.76, P<0.0001). In conclusion, we have shown that systemic meningococcal OMV vaccination in adult vaccinees mainly induced IgG1 antibodies which correlated with bactericidal and opsonic activity, but also a considerable amount of IgG3 antibodies, which, in contrast to the IgG1 response, was induced only after 2 or 3 vaccine doses and declined more rapidly.

The stability and immunogenicity of a protein antigen encapsulated in biodegradable microparticles based on blends of lactide polymers and polyethylene glycol

Protein-loaded microparticles were produced from blends of poly(ethylene glycol) (PEG) with poly( l-lactide) (PLA) homopolymer or poly( dl-lactide co-glycolide) copolymers (PLG) using a water-in oil-in oil method. The stability of ovalbumin (OVA) associated with microparticles prepared using PEG and 50:50 PLG, 75:25 PLG and PLA, respectively, was analysed by SDS-PAGE and quantified by scanning densitometry following incubation in PBS at 37°C for up to 1 month. Fragmentation and aggregation of OVA was detected with all 3 formulations. The extent of both processes correlated with the degradation rate of the lactide polymer used and decreased in the order PLA<75:25 PLG<50:50 PLG. Extensive degradation of the PLG/PEG microparticles also occurred over 4 weeks whereas the use of PLA/PEG blends resulted in a stable microparticle morphology and much reduced fragmentation and aggregation of the associated protein. Following a single sub-cutaneous immunisation, high levels of specific serum IgG antibody were elicited by OVA associated with the PLA/PEG particles. Injection of OVA associated with the 75:25 PLG/PEG microparticles resulted in very low levels of specific antibody. A higher response was induced by the 50:50 PLG/PEG formulation but there was very large inter-animal variation in this group. Antibody levels elicited by all 3 formulations were significantly higher than those elicited by a single injection of soluble OVA. Analysis of antigen specific IgG1 and IgG2a antibody subtype levels also revealed the greater efficacy of the PLA/PEG microparticles as an adjuvant system. The use of PLA/PEG microparticles shows improved protein loading and delivery capacity while maintaining a high level of stability of the associated protein. These results indicate a strong correlation between the stability of microencapsulated antigen and the magnitude of the immune response following sub-cutaneous immunisation.

Modulation of mucosal and systemic immunity by intranasal interleukin 12 delivery

Interleukin-12 (IL-12) is an important mediator of both cell-mediated and humoral immunity. We have now utilized a non-invasive intranasal (i.n.) delivery system to evaluate the ability of IL-12 to modulate both mucosal and systemic components of the immune system. Mice immunized i.n. with dinitrophenyl conjugated to ovalbumin (DNP–OVA) in combination with cholera toxin B subunit and IL-12 were found to have elevated levels of IFN- γ and IL-10 mRNA transcripts in both lungs and spleens compared with mice not receiving IL-12. In addition, expression of lung IL-5 mRNA was inhibited. Analysis of bronchoalveolar lavage fluid after IL-12 treatment revealed a significant increase in IgG2a and unaltered IgG1 and IgA anti-OVA antibody levels. Serum IgG2a, IgG2b and IgG3 anti-DNP antibody levels were significantly increased by IL-12 given i.n., while serum IgG1 antibody levels were suppressed, results that are similar to those seen after systemic antigen plus IL-12 administration. Delivery of IL-12 i.n. also enhanced faecal IgG2a and suppressed IgA levels, in contrast to parenteral treatment which increased both faecal IgG2a and IgA antibody expression. These results provide evidence that i.n. IL-12 treatment can effectively modulate antigen-specific immune responses and enhance immunization strategies for mucosal vaccines.

Induction of protective immunity for Bordetella pertussis by nasal inoculation of pertussis vaccine in mice

Although nasal vaccination has emerged as an interesting alternative to intramuscular or oral vaccination, knowledge is scarce about the immune responses after such immunization. In the present study, we inoculated purified Pertussis Toxin (PT) and Filamentous Haemagglutinin (FHA) with or without adjuvant (kayexalate), or Diphtheria acellular Pertussis Tetanus (DaPT) combined vaccine to mice intranasally three times every four weeks to investigate the references of the immunoresponses between nasal and intramuscular vaccination. The levels of pertussis specific serum IgG antibodies (Abs) and secretory IgA Abs in the nasal wash were measured by ELISA, and cytotoxic T cell activities were examined by proliferative response, and compared with the result from intramuscular inoculation. We also studied the efficacy of adjuvant in the nasal vaccination. The intramuscular inoculation of pertussis vaccine induced serum IgG antibodies and cellular immunity against PT and FHA, but did not induce local IgA antibodies. On the other hand, the nasal inoculation induced both serum and local antibody responses. Moreover, it also induced significant cellular immunity to pertussis antigen. In nasal vaccination, the inoculation with adjuvant was superior to inoculation without adjuvant for the induction of both humoral and cellular immunity.

Natural IgM and IgG Antibodies to Thomsen-Friedenreich (T) Antigen in Serum of Patients with Gastric Cancer and Blood Donors: Relation to Lewis (a,b) Histo-blood Group Phenotype

A possible association of serum anti-T IgM and IgG antibody levels with Lewis blood-group phenotype was investigated in 168 blood donors and 132 gastric cancer patients using ELISA with synthetic T-disaccharide-polyacrylamide conjugate as antigen. The donors of Le(a-b+) phenotype showed the highest anti-T IgM level irrespective of ABO(H) blood group. A significant decrease in anti-T IgM in serum was observed among cancer patients of Le(a-b+) phenotype: 95% of weak responders versus 17.5% for related groups of donors (p < 10(-6)). In contrast, no significant difference between patients and donors was found for Le(b-) individuals. Thus, a level of natural anti-T antibodies in serum of blood donors and its decrease in patients with gastric cancer are related to Le(a,b) phenotype. This should be taken into account where anti-T antibody level in the serum is used as a tumour marker or for monitoring patients during cancer immunotherapy with mucin-type vaccines.

Analysis of serum antibodies to alpha-streptococci in patients with tonsil-related pustulosis palmaris et plantaris

To determine the systemic immune response to alpha-streptococci (Str. sanguis, Str. salivarius and Str. mitis) and beta streptococcus (Str. pyogenes T12) in patients with tonsil related pustulosis palmaris et plantaris (PPP), we measured serum antibody levels to whole cell body antigens of Streptococcus (Str.) sanguis, Str. salivarius, Str. mitis or Str. pyogenes by enzyme-linked immunosorbent assay (ELISA). The serum IgG antibody levels to alpha-streptococci, Str. sanguis and Str. salivarius were significantly higher in patients with PPP (n = 44) than in patients with recurrent tonsillitis (RT: n = 25) and in healthy adults (n = 17). Serum antibody IgG level to Str. pyogenes was not different among the 3 groups. The IgA antibody levels against any Streptococcus strain were not different among the 3 groups. The IgM antibody levels to Str. pyogenes were significantly higher in patients with RT than in patients with PPP. In western blot analysis, the serum IgG antibodies against 25-27 kDa protein from whole cell body of Str. sanguis, Str. salivarius and Str. mitis were found more frequently in patients with PPP than in healthy adults. However, the western blot profile in Str. pyogenes was not different between PPP and healthy adults. No significant difference was seen in the western blot profile of IgM or IgM antibodies to any streptococcal whole cell bodies. These data suggest that systemic hyper immune response to alpha-streptococci may be present in patients with tonsil-related PPP and the 25-27 kDa protein of the organism may be the target for this immunologic abnormality.

Antibodies specific to outer membrane antigens of Moraxella catarrhalis in sera and middle ear effusions from children with otitis media with effusion

Objective: recent studies have shown that bacterial DNA is present in a significant percentage of middle ear effusions, suggesting that persistent bacterial infection may be more important in pathogenesis and recurrence of otitis media with effusion (OME) than previously considered. Although Moraxella (M.) catarrhalis is one of the most common pathogens of otitis media, relatively little is known about immune response to the organism. The objective of the present study is to investigate how systemic and local immune activities against M. catarrhalis may be associated with severity of OME. Methods: the antibody levels specific to outer membrane antigens of M. catarrhalis in sera and middle ear effusions (MEEs) from 59 children with OME were measured by enzyme-linked immunosorbent assay. Their ages ranged from 1 to 12 years with a median 5.0 years. The children were followed 1 year prospectively and classified into two groups with or without recurrent/persistent OME according to severity of OME during the follow-up 1 year. Results: serum IgG, IgM, and IgA antibodies specific to outer membrane antigens of M. catarrhalis were detected in all samples and the median levels were 35, 0.93, and 1.2 μg/ml respectively. The MEE IgG, IgM, IgA, and secretory IgA antibodies were detected in over 95% samples tested and the median levels were 371, 158, 20, and 50 ng/mg total protein respectively. A comparison between acute and subacute/chronic phases revealed that the median levels of MEE IgG and IgM antibodies were higher at the acute phase (692 vs. 340, P=0.06; 35 vs. 10, P=0.02, respectively); while the MEE secretory IgA antibody level was increased at the subacute/chronic phase (74 vs. 35, P=0.02). Either serum or MEE IgG antibody level was significantly lower in recurrent/persistent OME group than that in nonrecurrent/non-persistent OME group (13 vs. 43 μg/ml, P=0.009; 238 vs. 577 ng/mg protein, P=0.006, respectively). Conclusions: these data provide additional information on the immunologic aspects of children with OME. Decreased serum and MEE IgG antibody levels specific to outer membrane antigens of M. catarrhalis may lead to failure to eliminate this organism, resulting in persistent and/or recurrent appearance of MEE.

HSP-derived Peptides Inducing Uveitis and IgG and IgA Antibodies

Heat shock protein (HSP) 65 kD-derived peptides, which specifically stimulate T cells from patients with Behçet's disease (BD), are capable of inducing uveitis in rats. Mycobacterial HSP 65 kD and BD-specific peptides were injected into Lewis rats and the development of uveitis was monitored clinically and histologically, and IgG and IgA antibodies were measured using an enzyme-linked immunosorbent assay (ELISA). Rats immunized with HSP peptides that developed uveitis showed significantly higher serum IgG antibody levels to peptide 311–326 (P<0.05) and the corresponding homologous human peptide 336–351 (P<0.01) than rats without uveitis. Significant increases in serum IgA antibodies in rats with uveitis were also observed in those immunized with peptides 111–125, 311–326 and 336–351 (P<0.05). Rats injected with HSP 65 kD showed a rise in IgG antibody levels to peptides 111–125, 154–172 and 311–326 and to a lesser extent, a rise in IgA antibody level to peptide 311–326. HSP showed almost complete inhibition of binding of IgG antibodies to HSP 65 kD, but peptides 111–125, 154–172, 311–326 and 336–351 showed inhibition to a lesser extent in a competitive assay. These results suggest that increases in IgG and IgA antibody levels to specific peptides within HSP, develop in rats with uveitis. The T and B cell epitopes responsible for the development of ocular disease in rats immunized with HSP-derived peptides, appear to be similar or identical to those found in patients with the ocular type of BD.