RESUMO
BACKGROUND: In the nation-wide double-blind cluster-randomised Finnish Invasive Pneumococcal disease trial (FinIP, ClinicalTrials.gov NCT00861380, NCT00839254), we assessed the indirect impact of the 10-valent pneumococcal Haemophilus influenzae protein D conjugate vaccine (PHiD-CV10) against five pneumococcal disease syndromes. METHODS: Children 6 weeks to 18 months received PHiD-CV10 in 48 clusters or hepatitis B/A-vaccine as control in 24 clusters according to infant 3+1/2+1 or catch-up schedules in years 2009-2011. Outcome data were collected from national health registers and included laboratory-confirmed and clinically suspected invasive pneumococcal disease (IPD), hospital-diagnosed pneumonia, tympanostomy tube placements (TTP) and outpatient antimicrobial prescriptions. Incidence rates in the unvaccinated population in years 2010-2015 were compared between PHiD-CV10 and control clusters in age groups <5 and ≥5 years (5-7 years for TTP and outpatient antimicrobial prescriptions), and in infants <3 months. PHiD-CV10 was introduced into the Finnish National Vaccination Programme (PCV-NVP) for 3-month-old infants without catch-up in 9/2010. RESULTS: From 2/2009 to 10/2010, 45398 children were enrolled. Vaccination coverage varied from 29 to 61% in PHiD-CV10 clusters. We detected no clear differences in the incidence rates between the unvaccinated cohorts of the treatment arms, except in single years. For example, the rates of vaccine-type IPD, non-laboratory-confirmed IPD and empyema were lower in PHiD-CV10 clusters compared to control clusters in 2012, 2015 and 2011, respectively, in the age-group ≥5 years. CONCLUSIONS: This is the first report from a clinical trial evaluating the indirect impact of a PCV against clinical outcomes in an unvaccinated population. We did not observe consistent indirect effects in the PHiD-CV10 clusters compared to the control clusters. We consider that the sub-optimal trial vaccination coverage did not allow the development of detectable indirect effects and that the supervening PCV-NVP significantly diminished the differences in PHiD-CV10 vaccination coverage between the treatment arms.
Assuntos
Proteínas de Bactérias/administração & dosagem , Proteínas de Transporte/administração & dosagem , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/administração & dosagem , Haemophilus influenzae/imunologia , Imunoglobulina D/administração & dosagem , Lipoproteínas/administração & dosagem , Vacinas Pneumocócicas/administração & dosagem , Pneumonia Bacteriana/prevenção & controle , Proteínas de Bactérias/efeitos adversos , Proteínas de Bactérias/imunologia , Proteínas de Transporte/efeitos adversos , Proteínas de Transporte/imunologia , Criança , Pré-Escolar , Método Duplo-Cego , Feminino , Infecções por Haemophilus/imunologia , Vacinas Anti-Haemophilus/efeitos adversos , Vacinas Anti-Haemophilus/imunologia , Humanos , Imunoglobulina D/efeitos adversos , Imunoglobulina D/imunologia , Lactente , Lipoproteínas/efeitos adversos , Lipoproteínas/imunologia , Masculino , Vacinas Pneumocócicas/efeitos adversos , Vacinas Pneumocócicas/imunologia , Pneumonia Bacteriana/imunologia , Vacinas Conjugadas/administração & dosagem , Vacinas Conjugadas/efeitos adversos , Vacinas Conjugadas/imunologiaRESUMO
Rheumatoid arthritis (RA) is a systemic autoimmune disease characterized by joint inflammation, synovial hyperplasia, cartilage degeneration, bone erosion, and pannus. Immunoglobulin D (IgD) plays an important role in autoimmune diseases although the content of it in vivo is low. Increased concentrations of anti-IgD autoantibodies have been detected in many RA patients. IgD-Fc-Ig fusion protein is constructed by connecting human IgD Fc domain and IgG1 Fc domain, which specifically block the IgD/ IgDR pathway and regulate the function of cells expressing IgDR to treat RA. The expression levels of Wnt5A and Frizzled 5 are higher in RA synovial tissue specimens. The complex of Wnt5A-Fzd5-LRP5/6-CTHRC1 promotes the expression of hypoxia inducible factor-1α by activating nuclear factor kappa-B (NF-κB), leading to high expression of VEGF and participating in angiogenesis. VEGF is the strongest angiogenic factor found so far. Here, we aimed to explore whether IgD participates in synovitis by binding to IgDR and regulating the activation of Wnt5A-Fzd5-CTHRC1-NF-κB signaling pathway in fibroblast synovial cells (FLSs), whether IgD-Fc-Ig fusion protein inhibits VEGF production in FLS of CIA and explore mechanism. We found that IgDR is expressed on MH7A and FLS. IgD promotes VEGF expression by activating Wnt5A-Fzd5-CTHRC1-NF-κB signaling pathway in MH7A and FLS. After activation of Fzd5 with Wnt5A, IgD-Fc-Ig reduced VEGF-A level in the culture supernatant of MH7A stimulation by IgD. The expressions of CTHRC1, Fzd5, p-P65 and VEGF in MH7A and FLSs were down-regulated after IgD-Fc-Ig treatment. IgD-Fc-Ig suppressed the combination of CTHRC1 and Fzd5 as well. By using the animal model, we demonstrated that IgD-Fc-Ig suppress ankle CTHRC1 and Fzd5 production resulted in inhibition of index of joint inflammation of CIA rats, which were consistent with vitro results. Conclusively, IgD-Fc-Ig inhibits IgD and Wnt5A-induced angiogenesis and joint inflammation by suppressing the combination of CTHRC1 and Fzd5. Our results show that IgD-Fc-Ig exerts its suppressive effect on IgD and Wnt5A by Wnt5A-Fzd5-CTHRC1-NF-κB signaling pathway.
Assuntos
Artrite Experimental/imunologia , Imunoglobulina D/metabolismo , Proteínas Recombinantes de Fusão/administração & dosagem , Membrana Sinovial/patologia , Sinovite/imunologia , Proteína Wnt-5a/antagonistas & inibidores , Animais , Artrite Experimental/tratamento farmacológico , Artrite Experimental/patologia , Colágeno/administração & dosagem , Colágeno/imunologia , Fibroblastos , Receptores Frizzled/metabolismo , Glicoproteínas/metabolismo , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/genética , Fragmentos Fc das Imunoglobulinas/administração & dosagem , Fragmentos Fc das Imunoglobulinas/genética , Masculino , NF-kappa B/metabolismo , Ratos , Proteínas Recombinantes de Fusão/genética , Transdução de Sinais/efeitos dos fármacos , Transdução de Sinais/imunologia , Membrana Sinovial/efeitos dos fármacos , Membrana Sinovial/imunologia , Sinoviócitos , Sinovite/tratamento farmacológico , Sinovite/patologia , Proteína Wnt-5a/metabolismoRESUMO
Vaccinations with the 10-valent pneumococcal conjugated vaccine (PHiD-CV) started in Iceland in 2011. Protein D (PD) from H. influenzae, which is coded for by the hpd gene, is used as a conjugate in the vaccine and may provide protection against PD-positive H. influenzae We aimed to evaluate the effect of PHiD-CV vaccination on H. influenzae in children, both in carriage and in acute otitis media (AOM). H. influenzae was isolated from nasopharyngeal swabs collected from healthy children attending 15 day care centers in 2009 and from 2012 to 2017 and from middle ear (ME) samples from children with AOM collected from 2012 to 2017. All isolates were identified using PCR for the hpd and fucK genes. Of the 3,600 samples collected from healthy children, 2,465 were culture positive for H. influenzae (68.5% carriage rate); of these, 151 (6.1%) contained hpd-negative isolates. Of the 2,847 ME samples collected, 889 (31.2%) were culture positive for H. influenzae; of these, 71 (8.0%) were hpd negative. Despite the same practice throughout the study, the annual number of ME samples reduced from 660 in 2012 to 330 in 2017. The proportions of hpd-negative isolates in unvaccinated versus vaccinated children were 5.6% and 7.0%, respectively, in healthy carriers, and 5.4% and 7.8%, respectively, in ME samples. The proportion of hpd-negative isolates increased with time in ME samples but not in healthy carriers. The number of ME samples from children with AOM decreased. The PHiD-CV had no effect on the proportion of the hpd gene in H. influenzae from carriage, but there was an increase in hpd-negative H. influenzae in otitis media. The proportions of hpd-negative isolates remained similar in vaccinated and unvaccinated children.
Assuntos
Proteínas de Bactérias/administração & dosagem , Proteínas de Transporte/administração & dosagem , Portador Sadio/microbiologia , Infecções por Haemophilus/microbiologia , Haemophilus influenzae/isolamento & purificação , Imunoglobulina D/administração & dosagem , Lipoproteínas/administração & dosagem , Otite Média/microbiologia , Vacinas Pneumocócicas/administração & dosagem , Proteínas de Bactérias/genética , Proteínas de Transporte/genética , Portador Sadio/prevenção & controle , Criança , Pré-Escolar , Orelha Média/microbiologia , Infecções por Haemophilus/prevenção & controle , Haemophilus influenzae/genética , Humanos , Islândia/epidemiologia , Imunoglobulina D/genética , Lactente , Lipoproteínas/genética , Nasofaringe/microbiologia , Otite Média/prevenção & controle , Vacinas Conjugadas/administração & dosagemRESUMO
PURPOSE: Nontypeable Haemophilus influenzae (NTHi) is a commensal in the human nasopharynx and the cause of pneumonia, meningitis, sinusitis, acute exacerbations of chronic obstructive pulmonary disease and acute otitis media (AOM). AOM is the most common ailment for which antibiotics are prescribed in the United States. With the emergence of new strains of antibiotic-resistant bacteria, finding an effective and broad coverage vaccine to protect against AOM-causing pathogens has become a priority. Mouse models are a cost-effective and efficient way to help determine vaccine efficacy. Here, we describe an NTHi AOM model in C57BL/6J mice, which also utilizes a mouse-adapted H1N1 influenza virus to mimic human coinfection. METHODOLOGY: We tested our coinfection model using a protein vaccine formulation containing protein D, a well-studied NTHi vaccine candidate that can be found in the 10-valent Streptococcus pneumoniae conjugate vaccine. We verified the usefulness of our mouse model by comparing bacterial loads in the nose and ear between protein D-vaccinated and control mice. RESULTS: While there was no measurable difference in nasal bacterial loads, we did detect significant differences in the bacterial loads of ear washes and ear bullae between vaccinated and control mice. CONCLUSION: The results from this study suggest that our NTHi AOM coinfection model is useful for assessing protein vaccines.
Assuntos
Proteínas de Bactérias/imunologia , Proteínas de Transporte/imunologia , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/imunologia , Haemophilus influenzae/imunologia , Imunoglobulina D/imunologia , Lipoproteínas/imunologia , Otite Média/prevenção & controle , Administração Intranasal , Animais , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/genética , Proteínas de Transporte/administração & dosagem , Proteínas de Transporte/genética , Coinfecção/microbiologia , Coinfecção/prevenção & controle , Coinfecção/virologia , Modelos Animais de Doenças , Feminino , Infecções por Haemophilus/microbiologia , Vacinas Anti-Haemophilus/administração & dosagem , Vacinas Anti-Haemophilus/genética , Haemophilus influenzae/genética , Haemophilus influenzae/crescimento & desenvolvimento , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/genética , Vírus da Influenza A Subtipo H1N1/fisiologia , Lipoproteínas/administração & dosagem , Lipoproteínas/genética , Masculino , Camundongos , Camundongos Endogâmicos C57BL , Nariz/microbiologia , Nariz/virologia , Otite Média/imunologia , Otite Média/microbiologia , Vacinas Conjugadas/administração & dosagem , Vacinas Conjugadas/imunologiaRESUMO
BACKGROUND: The immunogenicity, reactogenicity, and safety of the 10-valent pneumococcal non-typeable Haemophilus influenzae protein D conjugate vaccine (PHiD-CV) co-administered with routine childhood vaccines were evaluated among infants from Singapore and Malaysia, where PHiD-CV has been licensed. METHODS: In the primary vaccination phase, 298 infants from Singapore and 168 infants from Malaysia were randomised to receive the Phase III Clinical (Clin) or the Commercial (Com) lot of PHiD-CV at 2, 3, and 5 months of age. In the booster vaccination phase, 238 toddlers from Singapore received one dose of the PHiD-CV Commercial lot at 18-21 months of age. Immune responses to pneumococcal polysaccharides were measured using 22F-inhibition enzyme-linked immunosorbent assay (ELISA) and functional opsonophagocytic activity (OPA) assay and to protein D, using ELISA. RESULTS: Immune responses induced by primary vaccination with the PHiD-CV Commercial lot were non-inferior to the Phase III Clinical lot in terms of adjusted antibody geometric mean concentration (GMC) ratios for each vaccine pneumococcal serotype and protein D. For each vaccine pneumococcal serotype, ≥93.6% and ≥88.5% of infants from Malaysia and Singapore had post-primary vaccination antibody concentrations ≥0.2 µg/mL and OPA titres ≥8, in the Clin and Com groups, respectively. For each vaccine pneumococcal serotype, ≥60.8% and ≥98.2% of toddlers from Singapore had pre- and post-booster antibody concentrations ≥0.2 µg/mL, in the Clin and Com groups, respectively. All children, except one, had measurable anti-protein D antibodies and the primary and booster doses of the co-administered vaccines were immunogenic. The incidence of each grade 3 solicited symptom was ≤11.1% in both study phases. No serious adverse events considered causally related to vaccination were reported throughout the study. CONCLUSIONS: PHiD-CV given as three-dose primary vaccination to infants in Singapore and Malaysia and booster vaccination to toddlers in Singapore was shown to be immunogenic with a clinically acceptable-safety profile.This study has been registered at http://www.clinicaltrials.govNCT00808444 and NCT01119625.
Assuntos
Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/administração & dosagem , Streptococcus pneumoniae/imunologia , Vacinação , Vacinas Conjugadas/administração & dosagem , Anticorpos Antibacterianos/sangue , Anticorpos Antibacterianos/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Transporte/administração & dosagem , Feminino , Humanos , Imunização Secundária , Imunoglobulina D/administração & dosagem , Imunoglobulina D/sangue , Lactente , Lipoproteínas/administração & dosagem , Malásia , Masculino , Polissacarídeos Bacterianos/imunologia , Singapura , Potência de VacinaRESUMO
We characterized cytokine profiles of CD4(+) T-helper (h) cells in adults and young children to ascertain if responses occur to next-generation candidate vaccine antigens PspA, PcpA, PhtD, PhtE, Ply, LytB of Streptococcus pneumonia (Spn) and protein D and OMP26 of non-typeable Haemophilus influenzae (NTHi). Adults had vaccine antigen-specific Th1 and Th2 cells responsive to all antigens evaluated whereas young children had significant numbers of vaccine antigen-specific CD4(+) T cells producing IL-2, (p=0.004). Vaccine antigen-specific CD4(+) T-cell populations in adults were largely of effector (TEM) and/or central memory (TCM) phenotypes as defined by CD45RA(-)CCR7(+) or CD45RA(-)CCR7(-) respectively; however among young children antigen-specific IL-2 producing CD4(+) T cells demonstrated CD45RA(+) expression (non-memory cells). We conclude that adults have circulating memory CD4(+) T cells (CD45RA(-)) that can be stimulated by all the tested Spn and NTHi protein vaccine candidate antigens, whereas young children have a more limited response.
Assuntos
Antígenos de Bactérias/imunologia , Linfócitos T CD4-Positivos/imunologia , Infecções por Haemophilus/prevenção & controle , Vacinas Anti-Haemophilus/imunologia , Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Adulto , Antígenos de Bactérias/administração & dosagem , Proteínas da Membrana Bacteriana Externa/administração & dosagem , Proteínas da Membrana Bacteriana Externa/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/imunologia , Proteínas de Transporte/administração & dosagem , Proteínas de Transporte/imunologia , Pré-Escolar , Citocinas/imunologia , Feminino , Vacinas Anti-Haemophilus/administração & dosagem , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/imunologia , Lactente , Interleucina-2/imunologia , Lipoproteínas/administração & dosagem , Lipoproteínas/imunologia , Masculino , Vacinas Pneumocócicas/administração & dosagem , Porinas/imunologia , Linfócitos T Auxiliares-Indutores/imunologia , Células Th2/imunologiaRESUMO
The pneumococcal polysaccharide protein D-conjugate vaccine (PHiD-CV; Synflorix) contains ten capsular polysaccharide serotypes from the bacterium Streptococcus pneumoniae, eight of which are conjugated to a nonlipidated cell-surface liporotein (protein D) of non-typeable Haemophilus influenzae (NTHi) and two of which are conjugated to either tetanus or diphtheria toxoid. In a three-dose primary vaccination schedule in infants aged <6 months, PHiD-CV elicited high immune responses against all pneumococcal serotypes contained in the vaccine and was noninferior to an approved 7-valent pneumococcal conjugate vaccine (7vCRM) for eight of the ten serotypes (five of the seven common to both vaccines). Moreover, functional antibodies were elicited against all vaccine serotypes in an opsonophagocytic activity (OPA) assay. A fourth booster dose of PHiD-CV during the second year of life elicited an anamnestic response against all ten pneumococcal serotypes, as determined by both antibody concentrations and OPA titers. There were no clinically relevant changes in the immunogenicity of PHiD-CV when coadministered with meningococcal serogroup C conjugate or pentavalent whole cell pertussis combination vaccines, and polio vaccines using two different primary vaccination schedules. 11Pn-PD, an 11-valent prototype of PHiD-CV, demonstrated protection against episodes of acute otitis media (AOM) caused by S. pneumoniae and NTHi in infants aged <27 months. The first occurrence of an episode of AOM caused by pneumococcal vaccine serotypes was reduced by 52.6% in 11Pn-PD vaccinees compared with recipients of a control vaccine (primary endpoint). The tolerability profile of PHiD-CV was generally similar to that of 7vCRM.
Assuntos
Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/imunologia , Vacinas Conjugadas/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Bactérias/imunologia , Proteínas de Transporte/administração & dosagem , Proteínas de Transporte/imunologia , Pré-Escolar , Toxoide Diftérico/administração & dosagem , Toxoide Diftérico/imunologia , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/imunologia , Lactente , Lipoproteínas/administração & dosagem , Lipoproteínas/imunologia , Infecções Pneumocócicas/imunologia , Vacinas Pneumocócicas/administração & dosagem , Vacinas Pneumocócicas/efeitos adversos , Toxoide Tetânico/administração & dosagem , Toxoide Tetânico/imunologia , Vacinas Conjugadas/administração & dosagem , Vacinas Conjugadas/efeitos adversosRESUMO
BACKGROUND: The immunogenicity of the 10-valent pneumococcal nontypeable Haemophilus influenzae protein D-conjugate vaccine (PHiD-CV) was assessed and compared with the 7-valent pneumococcal conjugate vaccine (7vCRM). METHODS: Healthy subjects (1650) were randomized to be vaccinated with 3 doses of PHiD-CV or 7vCRM (Prevenar/Prevnar) at 2-3-4 months of age and a fourth booster dose at 12-18 months. Serotype-specific pneumococcal responses (GlaxoSmithKline's ELISA with 22F-inhibition) and opsonophagocytic activity (OPA) were measured 1 month after primary and booster vaccinations. RESULTS: The primary objective to demonstrate noninferiority of PHiD-CV versus 7vCRM (in terms of percentage of subjects with antibody concentration >or=0.2 microg/mL) for at least 7 of the 10 vaccine serotypes was reached as noninferiority was demonstrated for 8 serotypes. Although, noninferiority could not be demonstrated for ELISA responses against serotypes 6B and 23F, a post-hoc analysis of the percentage of subjects with OPA titers >or=8 suggested noninferiority for the 7 serotypes common to both vaccines including 6B and 23F.Priming of the immune system against all vaccine serotypes was confirmed by robust increases in ELISA antibody levels ( approximately 6.0-17 fold) and OPA titers ( approximately 8-93 fold) after a fourth consecutive dose of PHiD-CV. CONCLUSIONS: PHiD-CV induces ELISA and functional OPA antibodies for all vaccine serotypes after primary vaccination and is noninferior to 7vCRM in terms of ELISA and/or OPA threshold responses. Effective priming is further indicated by robust booster responses.
Assuntos
Anticorpos Antibacterianos/sangue , Proteínas de Bactérias/imunologia , Proteínas de Transporte/imunologia , Imunoglobulina D/imunologia , Lipoproteínas/imunologia , Proteínas Opsonizantes/metabolismo , Infecções Pneumocócicas/prevenção & controle , Vacinas Pneumocócicas/imunologia , Streptococcus pneumoniae/imunologia , Vacinas Conjugadas/imunologia , Proteínas de Bactérias/administração & dosagem , Proteínas de Transporte/administração & dosagem , Feminino , Vacina Pneumocócica Conjugada Heptavalente , Humanos , Esquemas de Imunização , Imunização Secundária , Imunoglobulina D/administração & dosagem , Lactente , Licenciamento , Lipoproteínas/administração & dosagem , Masculino , Fagocitose , Infecções Pneumocócicas/imunologia , Infecções Pneumocócicas/microbiologia , Vacinas Pneumocócicas/administração & dosagem , Sorotipagem , Streptococcus pneumoniae/classificação , Resultado do Tratamento , Vacinação , Vacinas Conjugadas/administração & dosagemRESUMO
Although in vitro development of a Th2 response from naive CD4+ T cells is Stat6 dependent, mice immunized with a goat Ab to mouse IgD have been reported to produce a normal primary IL-4 response in Stat6-deficient mice. Experiments have now been performed with mice immunized with more conventional Ags or inoculated with nematode parasites to account for this apparent discrepancy. The ability of an immunogen to induce a primary in vivo IL-4 response in Stat6-deficient mice was found to vary directly with its ability to induce a strong type 2 cytokine-biased response in normal mice. Even immunogens, however, that induce strong primary IL-4 responses in Stat6-deficient mice induce poor memory IL-4 responses in these mice. Consistent with this, Stat6-deficient CD4+ T cells make relatively normal IL-4 responses when stimulated in vitro for 3 days with anti-CD3 and anti-CD28, but poor IL-4 responses if they are later restimulated with anti-CD3. Thus, Stat6 signaling enhances primary IL-4 responses that are made as part of a type 0 cytokine response (mixed type 1 and type 2) and is required for normal development or survival of Th2 memory cells.
Assuntos
Interleucina-4/biossíntese , Transativadores/fisiologia , Animais , Anticorpos/fisiologia , Anticorpos Monoclonais/administração & dosagem , Complexo CD3/imunologia , Linfócitos T CD4-Positivos/imunologia , Linfócitos T CD4-Positivos/metabolismo , Galinhas , Feminino , Cabras , Imunoglobulina D/administração & dosagem , Memória Imunológica , Injeções Intravenosas , Interleucina-4/antagonistas & inibidores , Enteropatias Parasitárias/imunologia , Masculino , Mastócitos/imunologia , Mastócitos/metabolismo , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C57BL , Camundongos Knockout , Nematospiroides dubius/imunologia , Nippostrongylus/imunologia , Ovalbumina/administração & dosagem , Ovalbumina/imunologia , Receptores de Interleucina-4/deficiência , Receptores de Interleucina-4/genética , Fator de Transcrição STAT6 , Transdução de Sinais/imunologia , Infecções por Strongylida/imunologia , Células Th2/imunologia , Células Th2/metabolismo , Transativadores/deficiência , Transativadores/genética , Trichinella spiralis/imunologia , Triquinelose/imunologiaRESUMO
Injection of mice with a foreign anti-IgD Ab stimulates B and T cell activation that results in large cytokine and Ab responses. Because most anti-IgD-activated B cells die before they can be stimulated by activated T cells, and because IL-4 prolongs the survival of B cells cultured with anti-Ig, we hypothesized that treatment with IL-4 at the time of anti-IgD Ab injection would decrease B cell death and enhance anti-IgD-induced Ab responses. Instead, IL-4 treatment before or along with anti-IgD Ab suppressed IgE and IgG1 responses, whereas IL-4 injected after anti-IgD enhanced IgE responses. The suppressive effect of early IL-4 treatment on the Ab response to anti-IgD was associated with a rapid, short-lived increase in IFN-gamma gene expression but decreased CD4+ T cell activation and decreased or delayed T cell production of other cytokines. We examined the possibilities that IL-4 stimulation of IFN-gamma production, suppression of IL-1 or IL-2 production, or induction of TNF-alpha or Fas-mediated apoptosis could account for IL-4's suppressive effect. The suppressive effect of IL-4 was not reversed by IL-1, IL-2, or anti-TNF-alpha or anti-IFN-gamma mAb treatment, or mimicked by treatment with anti-IL-2Ralpha (CD25) and anti-IL-2Rbeta (CD122) mAbs. Early IL-4 treatment failed to inhibit anti-IgD-induced Ab production in Fas-defective lpr mice; however, the poor responsiveness of lpr mice to anti-IgD made this result difficult to interpret. These observations indicate that exposure to IL-4, while T cells are first being activated by Ag presentation, can inhibit T cells activation or promote deletion of responding CD4+ T cells.
Assuntos
Imunoglobulina E/biossíntese , Imunoglobulina G/biossíntese , Imunossupressores , Interleucina-4/fisiologia , Ativação Linfocitária/imunologia , Linfócitos T/imunologia , Animais , Anticorpos Monoclonais/administração & dosagem , Feminino , Cabras , Imunoglobulina D/administração & dosagem , Imunoglobulina E/sangue , Imunoglobulina G/sangue , Imunossupressores/administração & dosagem , Injeções Intravenosas , Interferon gama/imunologia , Interleucina-1/antagonistas & inibidores , Interleucina-1/biossíntese , Interleucina-2/antagonistas & inibidores , Interleucina-2/biossíntese , Interleucina-4/administração & dosagem , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos MRL lpr , Receptores de Interleucina-2/antagonistas & inibidores , Fator de Necrose Tumoral alfa/fisiologiaAssuntos
Eritroblastose Fetal/prevenção & controle , Imunoglobulina D/administração & dosagem , Isoimunização Rh/prevenção & controle , Eritroblastose Fetal/imunologia , Feminino , Idade Gestacional , Humanos , Imunoglobulina D/sangue , Recém-Nascido , Gravidez , Isoimunização Rh/imunologia , Fatores de RiscoRESUMO
Injection of goat anti-mouse IgD antibodies (GAM IgD) to mice has been shown to induce polyclonal IgG1 and IgE production by B cells and interleukin-4 (IL-4) production by goat Ig-specific T cells. Surface IgD cross-linking also activates B cells to function as antigen-presenting cells (APC). Although the GAM IgD treatment is a well-established system for analysis of B-cell dependent antigen presentation, the influence of GAM IgD treatment on the immune response to irrelevant antigens is not known. To address this issue, we analysed effects of GAM IgD treatment on (1) the mitogen response of freshly isolated T cells, and (2) the listerial antigen-specific response after immunization with viable Listeria monocytogenes, which induces CD4+ interferon-gamma (IFN-gamma) producing protective T cells in normal mice. Spleen CD4+ T cells from the GAM IgD-treated mice produced higher levels of IL-4 but lower levels of IFN-gamma and IL-2 than those from the control mice when they were stimulated with concanavalin A (Con A) in vitro. When spleen T cells were stimulated with listerial antigen 10 days after a low dose (1/20 LD50) of L. monocytogenes infection, CD4+ T cells from the GAM IgD-treated mice showed increased IL-4 production and decreased IFN-gamma and IL-2 production compared with those from the control L. monocytogenes-infected mice. Furthermore, the GAM IgD treatment resulted in a reduction of the survival rate after a high dose (1/2 LD50) of L. monocytogenes infection. These results suggest that treatment of mice with GAM IgD suppresses the T-helper type-1 (Th1)-type T-cell response and induces a Th2-type response against irrelevant antigens, even when they are injected after GAM IgD treatment.
Assuntos
Tolerância Imunológica , Imunoglobulina D/imunologia , Listeriose/imunologia , Células Th1/imunologia , Animais , Anticorpos/imunologia , Antígenos de Bactérias/imunologia , Concanavalina A/imunologia , Citocinas/biossíntese , Feminino , Cabras , Imunoglobulina D/administração & dosagem , Listeria monocytogenes/imunologia , Camundongos , Camundongos Endogâmicos BALB C , Especificidade da Espécie , Taxa de SobrevidaRESUMO
The onset of clinical disease in autoimmune MRL-lpr/lpr mice is preceded by a switch from predominantly IgM production to IgG production. Previous studies have shown that IgG autoantibodies play a central role in the development of life-threatening glomerulonephritis in this strain. Delaying or preventing the switch from IgM to IgG production might therefore be of therapeutic benefit. We previously documented similarities in the B cell repertoire expressed by young MRL-lpr/lpr mice and normal mice treated with the polyclonal activator LPS. Recent in vivo studies indicate that cross-linking membrane IgM or IgD can suppress LPS-dependent IgG production in normal animals. These observations led us to examine whether membrane cross-linking could also lower serum IgG levels in MRL-lpr/lpr mice. Lupus-prone animals were treated with multivalent anti-IgD conjugated to high m.w. dextran. This anti-IgD dextran conjugate was previously shown to reduce IgG production in LPS-stimulated normal animals. Treatment of young lupus-prone MRL-lpr/lpr mice resulted in a significant reduction in the total number of B cells secreting IgG and lower serum titers of IgG anti-DNA and IgG anti-histone autoantibodies. Anti-IgD dextran treatment also delayed the development of glomerulonephritis and improved survival. Thus, anti-IgD dextran interfered with autoantibody-dependent disease progression, perhaps by inhibiting the switch from IgM to IgG autoantibody production.
Assuntos
Linfócitos B/imunologia , Dextranos/administração & dosagem , Imunoglobulina D/administração & dosagem , Lúpus Eritematoso Sistêmico/prevenção & controle , Camundongos Mutantes/imunologia , Fatores Etários , Animais , Formação de Anticorpos , Feminino , Imunoglobulina G/biossíntese , Imunoglobulina M/biossíntese , Região de Troca de Imunoglobulinas , Lúpus Eritematoso Sistêmico/imunologia , Ativação Linfocitária , CamundongosRESUMO
BACKGROUND: Previous work showed that injection of mice with goat anti-mouse IgD antibodies results in increased numbers of Fc epsilon R-positive, non-B, non-T cells in the spleen and Fc epsilon R-positive cells in the bone marrow, and that some of these cells had ultrastructural features of basophils. Fc epsilon R-positive, non-B, non-T cells express virtually all of the capacity of mouse splenic "non-B, non-T cells" to produce interleukin-4 in response to stimulation by cross-linking of Fc epsilon R or Fc gamma R, or by the calcium ionophore, ionomycin. EXPERIMENTAL DESIGN: The present study is a detailed ultrastructural analysis of Fc epsilon R-positive bone marrow cells or Fc epsilon R-positive splenic non-B, non-T cells sorted from mice injected with goat anti-mouse IgD antibody and of Fc epsilon R-positive bone marrow cells or spleen cells pooled from normal mice not injected with goat anti-IgD. RESULTS: Basophils represented the majority (90%) of the granulated cells present in the Fc epsilon R-positive splenic non-B, non-T cells or Fc epsilon R-positive bone marrow cells of goat anti-IgD-injected mice. In contrast, the cytoplasmic granule-containing Fc epsilon R-negative cells sorted from spleen or bone marrow of goat anti-IgD-injected animals contained predominantly a mixture of neutrophils, eosinophils, monocytes and their precursors. Both the Fc epsilon R-positive and -negative preparations contained rare (< 5%) cells with ultrastructural features of very immature mast cells. Basophils were also identified in Fc epsilon R-positive cells sorted from total bone marrow cells or spleen cells of normal mice not injected with goat anti-IgD. CONCLUSIONS: Taken together with data concerning the numbers of Fc epsilon R-positive, non-B, non-T cells in the spleen, and Fc epsilon R-positive B220-negative cells in the bone marrow, these ultrastructural findings indicate that injection of mice with goat anti-IgD results in increased numbers of basophils, particularly in the spleen, that exhibit an 8-fold increase in basophils as a result of injection of goat anti-IgD.
Assuntos
Anticorpos Anti-Idiotípicos/administração & dosagem , Basófilos/ultraestrutura , Células da Medula Óssea , Imunoglobulina D/administração & dosagem , Receptores de IgE , Baço/citologia , Animais , Basófilos/imunologia , Medula Óssea/imunologia , Separação Celular , Feminino , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica , Baço/imunologiaRESUMO
T cell populations derived from naive mice produce very small amounts of interleukin 4 (IL-4) in response to stimulation on anti-CD3-coated dishes. IL-4 production by such cells is mainly found among large- and intermediate-sized T cells and is dependent upon IL-2. Injection of anti-IgD into mice, a stimulus that leads to striking increases in serum levels of IgG1 and IgE, causes a striking increase in the IL-4-producing capacity of T cells. This increase is first observed 4 d after injection of anti-IgD. IL-4 production by T cells from anti-IgD-injected donors is mainly found among large- and intermediate-sized T cells. Small, dense T cells are poor producers of IL-4. The capacity of T cells from anti-IgD-injected donors to produce IL-4 is enhanced by addition of IL-2 and is largely, but not completely, inhibited by neutralization of in situ produced IL-2. These results indicate that the control of IL-4 production in T cells from naive and anti-IgD-injected donors is similar. However, it is possible that a portion of the IL-4-producing activity of T cells from activated donors is IL-2 independent. Although small T cells from naive donors have a very limited capacity to produce IL-4 in response to stimulation with anti-CD3, even in the presence of added IL-2, they can give rise to IL-4-producing cells upon in vitro culture on plates coated with anti-CD3 if both IL-2 and IL-4 are added. This leads to the appearance of IL-4-producing cells within 2 d. When analyzed after 5 d of culture by harvesting and re-exposure to anti-CD3-coated culture wells and IL-2, these cells have increased their IL-4-producing capacity by approximately 100-fold. The development of IL-4-producing cells in response to anti-CD3, IL-2, and IL-4 is not inhibited by interferon gamma (IFN-gamma), nor does IFN-gamma diminish IL-4 production by these cells upon challenge with anti-CD3 plus IL-2.
Assuntos
Interleucina-2/farmacologia , Interleucina-4/biossíntese , Interleucina-4/farmacologia , Linfócitos T/imunologia , Animais , Anticorpos Monoclonais , Células Cultivadas , Replicação do DNA , Feminino , Imunoglobulina D/administração & dosagem , Imunoglobulina D/imunologia , Interferon gama/farmacologia , Cinética , Ativação Linfocitária , Camundongos , Camundongos Endogâmicos BALB C , Proteínas Recombinantes/farmacologia , Linfócitos T/efeitos dos fármacosRESUMO
Using a modification of the splenic focus assay, we analyzed the Salmonella typhimurium-specific B-cell repertoire in salmonella-susceptible BALB/c mice. Although these mice normally succumbed to salmonella infection before antibody was produced, they appeared to have splenic S. typhimurium-specific B-cell precursors that could be activated to differentiate and secrete antibody in a manner which was quantitatively and qualitatively identical to that of salmonella-resistant mouse strains. We also analyzed the primary S. typhimurium-specific B-cell repertoire in BALB/c mice that had been chronically treated with antibodies to immunoglobulin D (IgD) and therefore had no surface IgD-positive B cells. Although the frequency of S. typhimurium-specific precursors in these mice was similar to that of control mice, there was an apparent alteration in the isotype distribution pattern in anti-IgD-treated mice. Control mice generated a significantly greater proportion of IgG-secreting clones than did anti-IgD-treated mice. In addition, a greater proportion of S. typhimurium-specific clones from control mice secreted IgG2 than secreted IgG1, and those clones that secreted IgG2 but not IgM, IgG3, or IgG1 were greater than 20-fold more common in control than in anti-IgD-treated mice. Finally, we analyzed the immune response of control and anti-IgD-treated mice to a live avirulent vaccine, S. typhimurium SL3235. Although both groups were protected after challenge with a live virulent S. typhimurium strain, only the control mice made serum antibodies to this vaccine. Taken together, these results show that (i) salmonella-susceptible BALB/c mice have S. typhimurium-specific B cells, (ii) the S. typhimurium-specific B cells in anti-IgD-treated mice may have a restricted capacity to switch heavy-chain classes, (iii) the similarity observed in the frequency of the S. typhimurium-specific precursors for these two groups of BALB/c mice is not reflected in the serum, and (iv) the failure of anti-IgD-treated mice to generate a serum antibody response to SL3235 in the face of complete protection suggests that this model may be used to study cell-mediated immune mechanisms in the apparent absence of humoral immunity.
Assuntos
Anticorpos Anti-Idiotípicos/administração & dosagem , Anticorpos Antibacterianos/análise , Linfócitos B/microbiologia , Epitopos/imunologia , Imunoglobulina D/imunologia , Salmonella typhimurium/imunologia , Animais , Linfócitos B/classificação , Linfócitos B/imunologia , Feminino , Soros Imunes/análise , Esquemas de Imunização , Imunoglobulina D/administração & dosagem , Isotipos de Imunoglobulinas/classificação , Contagem de Leucócitos , Camundongos , Camundongos Endogâmicos BALB C , Camundongos Endogâmicos C3H , Receptores de Antígenos de Linfócitos B/imunologia , Baço/microbiologiaRESUMO
The authors have analyzed the prevention of Rh-immunization from 1972 to 1983. Results are presented in two six-year periods, i.e. from 1972 to 1977 and from 1978 to 1983. Prevention was applied to all rh-negative women, who have been delivered from a rh-negative baby in their first childbirth (with negative sensibilization tests). Anti D IgG was also applied to all women after their second, third, fourth or subsequent delivery, if they were willing to have more children. Women with Du variant of the Rh factor and having a Rh-positive child were also protected. Preparations containing 250 to 300 micrograms of IgG anti-D were used. During the first period we found rh-negative mothers in 18.41 per cent, in 63.48 per cent of them the newborn was Rh-positive. During the second period 17.89 per cent of our women were rh-negative with 58.45 per cent Rh-positive babies. During the first period, protection was afforded to 60.26 per cent of the rh-negative women with incompatible babies, and in the second period to 79.11 per cent, respectively (P less than 0.05). During the second period, 99.70 per cent of women were protected after their first delivery (except of one case with immunization already during pregnancy), in contrast to the first period, where this percentage amounted only to 84.66 per cent (P less than 0.05). During both periods, a total of 69.51 per cent of the rh-negative women having Rh-positive babies received anti-D-immunization.(ABSTRACT TRUNCATED AT 250 WORDS)
Assuntos
Anticorpos Anti-Idiotípicos/administração & dosagem , Eritroblastose Fetal/prevenção & controle , Imunoglobulina D/imunologia , Imunoglobulina G/administração & dosagem , Feminino , Humanos , Imunização , Imunoglobulina D/administração & dosagem , Recém-Nascido , Gravidez , Fatores de RiscoRESUMO
The frequency of hemolytic disease of fetus and newborn has decreased since introduction of Rh-prophylaxis. In spite of that till now hemolytic disease caused by anti-D antibodies occur. It is probably caused by immunization already during pregnancy. Therefore it seems to be useful to give anti-D-Immunoglobulin (200 micrograms = 1000 IE) in 28-30th week of pregnancy in all d-women with D-husband. This seems especially necessary in pregnancy complications as hemorrhages in 3rd trimester, multiple pregnancies, trauma, and external version of breech. Rh-prophylaxis post partum is to perform in usual manner. As done before 2 time screening for irregular antibodies in pregnancy is necessary. Amniocentesis is often required in presence of antibodies against D, C, c, E, e, K, Fya, Fyb, S, s, U, Jka, Jkb and Dia. When hemolytic disease is confirmed, intrauterine transfusion or preterm delivery has to be performed. Immunization have increasing importance against erythrocyte factors other than D. It should give attention to it in transfusion practice during pregnancy.
Assuntos
Eritroblastose Fetal/prevenção & controle , Anticorpos Anti-Idiotípicos/administração & dosagem , Terapia Combinada , Eritroblastose Fetal/sangue , Feminino , Humanos , Imunoglobulina D/administração & dosagem , Imunoglobulina D/imunologia , Recém-Nascido , GravidezRESUMO
We show that the IgD-induced augmentation of the immune response to trinitrophenylated keyhole limpet hemocyanin can be transferred to syngeneic mice with spleen cells from IgD-injected donors. The augmenting activity is present in the Lyt-1+2-, L3T4+ T cell population and is absent from B cells. The ability of transferred T cells to augment the immune response correlates with the presence of a high frequency of Lyt-1+2- T cells that form rosettes with IgD-coated sheep erythrocytes (T delta cells). Such rosette-forming cells can also be induced by incubation of spleen cells from normal donors in IgD-coated petri dishes. Injection of normal spleen cells exposed to IgD-coated petri dishes together with antigen also augments the immune response of recipients. The existence of a regulatory circuit based upon interactions between T delta cells, antigen, B cell surface IgD, and serum IgD, is proposed.