RÉSUMÉ
BACKGROUND: Senecavirus A (SVA) causes an emerging vesicular disease (VD) with clinical symptoms indistinguishable from other vesicular diseases, including vesicular stomatitis (VS), foot-and-mouth disease (FMD), and swine vesicular disease (SVD). Currently, SVA outbreaks have been reported in Canada, the U.S.A, Brazil, Thailand, Vietnam, Colombia, and China. Based on the experience of prevention and control of FMDV, vaccines are the best means to prevent SVA transmission. RESULTS: After preparing an SVA inactivated vaccine (CH-GX-01-2019), we evaluated the immunogenicity of the SVA inactivated vaccine mixed with Imject® Alum (SVA + AL) or Montanide ISA 201 (SVA + 201) adjuvant in mice, as well as the immunogenicity of the SVA inactivated vaccine combined with Montanide ISA 201 adjuvant in post-weaned pigs. The results of the mouse experiment showed that the immune effects in the SVA + 201 group were superior to that in the SVA + AL group. Results from pigs immunized with SVA inactivated vaccine combined with Montanide ISA 201 showed that the immune effects were largely consistent between the SVA-H group (200 µg) and SVA-L group (50 µg); the viral load in tissues and blood was significantly reduced and no clinical symptoms occurred in the vaccinated pigs. CONCLUSIONS: Montanide ISA 201 is a better adjuvant choice than the Imject® Alum adjuvant in the SVA inactivated vaccine preparation, and the CH-GX-01-2019 SVA inactivated vaccine can provide effective protection for pigs.
Sujet(s)
Adjuvants immunologiques , Alun , Mannitol/analogues et dérivés , Huile minérale , Acides oléiques , Picornaviridae , Animaux , Souris , Suidae , Vaccins inactivésRÉSUMÉ
Foot-and-mouth disease (FMD) has a high prevalence in cloven-hoofed animals. It is also highly contagious and remains a serious threat to livestock worldwide. Despite the widespread vaccination program in Iran, outbreaks of FMD continue to occur. Vaccination is one of the most effective methods of preventing FMD. The vaccines used in Iran are of the inactivated type and contain several serotypes. Since inactivated vaccines without adjuvants do not induce a high and durable antibody response, it is necessary to use adjuvants. Montanide ISA 206 VG is a mineral oil-based adjuvant that produces a water-in-oil-in-water (w:o:w) emulsion in vaccine preparations. However, a large number of manufacturers in Iran and around the world still use alum adjuvant (with or without saponin) to produce the FMD vaccine. This study used Montanide ISA 206 and alum adjuvants to administer the O2010 serotype of the FMD virus to goats. A total of six goats were divided randomly into three groups. Vaccines were administered subcutaneously twice, at a one-month interval. Blood sampling was done at different times, and the micro-neutralization method was used to measure the neutralizing antibody titer in each serum. Seven days after the second vaccination, the alum group's antibody titer was higher but not statistically significant. However, from the 28th day after the second injection until the end of the study, the Montanide ISA 206 group's antibody titer was significantly higher than that of the alum group. Six months after the second injection, the antibody titer in the ISA 206 group remained at the peak level, while in the alum group, it decreased and reached the minimum protective level. Nine months after the second injection, the antibody titer remained at its peak level in the ISA 206 group, whereas it dropped significantly in the alum group. Based on the findings, ISA 206 VG is capable of generating long-term humoral immunity in goats against the FMD serotype O2010 and could replace aluminum hydroxide adjuvants in FMD vaccine preparations.
Sujet(s)
Adjuvants immunologiques , Hydroxyde d'aluminium , Anticorps neutralisants , Virus de la fièvre aphteuse , Fièvre aphteuse , Maladies des chèvres , Capra , Vaccins antiviraux , Animaux , Hydroxyde d'aluminium/administration et posologie , Hydroxyde d'aluminium/pharmacologie , Virus de la fièvre aphteuse/immunologie , Maladies des chèvres/prévention et contrôle , Fièvre aphteuse/prévention et contrôle , Fièvre aphteuse/immunologie , Adjuvants immunologiques/administration et posologie , Adjuvants immunologiques/pharmacologie , Vaccins antiviraux/immunologie , Vaccins antiviraux/administration et posologie , Anticorps neutralisants/sang , Anticorps antiviraux/sang , Iran , Acides oléiques/administration et posologie , Mannitol/analogues et dérivés , Mannitol/administration et posologieRÉSUMÉ
Streptococcus agalactiae is one of the most important pathogens infecting tilapia worldwide and causes meningoencephalitis, septicemia and high mortalities with considerable losses. Various types of vaccines have been developed against S. agalactiae infection, such as inactivated vaccines, live attenuated vaccines and subunit vaccines. Bacterial ghosts (BGs) are nonliving, empty cell envelopes and have been reported as novel vaccine candidates. Therefore, the main aims of this study were to develop an S. agalactiae ghost vaccine (SAGV) and to evaluate the immune response and protective effect of SAGV against S. agalactiae with two novel adjuvants, Montanide™ ISA 763B VG and Montanide™ GEL02. Nile tilapia, mean weight 50 g, were divided into four groups as follows; 1) fish injected with PBS as control, 2) fish injected with the SAGV alone; 3) fish injected with the SAGV+Montanide™ ISA 763B VG; and 4) fish injected with SAGV+Montanide™ GEL02. Following vaccination, innate immunity parameters including serum lysozyme, myeloperoxidase, catalase, and bactericidal activity were all significantly enhanced. Moreover, specific serum IgM antibodies were induced and reached their highest level 2-8 weeks post vaccination. Importantly, the relative percent survival of tilapia vaccinated against the SAGV formulated with both adjuvants was 80-93%. Furthermore, the transcription of immune-related genes (IgM, TCRß, IL-1ß, IL-8 and TNFα) were up-regulated in tilapia after vaccination, indicating that both cellular and humoral immune responses were induced by these adjuvanted vaccines. In summary, Montanide™ ISA 763B VG and Montanide™ GEL02 can enhance immunoprotection induced by the SAGV vaccine against streptococcosis, demonstrating that both have value as potential adjuvants of fish vaccines.
Sujet(s)
Adjuvants immunologiques/administration et posologie , Cichlides/immunologie , Maladies des poissons/prévention et contrôle , Mannitol/analogues et dérivés , Mannitol/administration et posologie , Infections à streptocoques/prévention et contrôle , Vaccins antistreptococciques/administration et posologie , Streptococcus agalactiae/immunologie , Animaux , Anticorps antibactériens/sang , Catalase/sang , Cichlides/sang , Maladies des poissons/sang , Maladies des poissons/immunologie , Protéines de poisson/sang , Foie/immunologie , Lysozyme/sang , Myeloperoxidase/sang , Rate/immunologie , Infections à streptocoques/sang , Infections à streptocoques/immunologieRÉSUMÉ
Specific link between high fructose uptake and cancer development and progression highlighted fructose transporters as potential means to achieve GLUT-mediated discrimination between normal and cancer cells. The gained expression of fructose-specific transporter GLUT5 in various cancers offers a possibility for developing cancer-specific imaging and bioactive agents. Herein, we explore the feasibility of delivering a bioactive agent through cancer-relevant fructose-specific transporter GLUT5. We employed specific targeting of GLUT5 by 2,5-anhydro-D-mannitol and investigated several drug conjugates for their ability to induce cancer-specific cytotoxicity. The proof-of-concept analysis was carried out for conjugates of chlorambucil (CLB) in GLUT5-positive breast cancer cells and normal breast cells. The cytotoxicity of conjugates was assessed over 24 h and 48 h, and significant dependence between cancer-selectivity and conjugate size was observed. The differences were found to relate to the loss of GLUT5-mediated uptake upon increased conjugate size and hydrophobicity. The findings provide information on the substrate tolerance of GLUT5 and highlight the importance of maintaining appropriate hydrophilicity for GLUT-mediated delivery.
Sujet(s)
Tumeurs du sein/anatomopathologie , Région mammaire/cytologie , Chlorambucil/pharmacologie , Transporteur de glucose de type 5/métabolisme , Mannitol/analogues et dérivés , Antinéoplasiques alcoylants/pharmacologie , Région mammaire/effets des médicaments et des substances chimiques , Région mammaire/métabolisme , Tumeurs du sein/traitement médicamenteux , Tumeurs du sein/métabolisme , Femelle , Humains , Interactions hydrophobes et hydrophiles , Mannitol/métabolisme , Spécificité du substratRÉSUMÉ
The coproduction of polymalic acid (PMA) and liamocins, two important metabolites secreted by Aureobasidium pullulans, from two waste by-products from the xylitol and gluconate industries was investigated in shake flasks and fermentors, confirming that waste xylose mother liquor (WXML) could be utilized as an economical feedstock without any pretreatment. Gluconate could strengthen carbon flux and NADPH supply for the synergetic biosynthesis of PMA and liamocins. High PMA and liamocin titers of 82.9 ± 2.1 and 28.3 ± 2.7 g/L, respectively, were obtained from the coupled WXML and waste gluconate mother liquor (WGML) in batch fermentation, with yields of 0.84 and 0.25 g/g, respectively. These results are comparable to those obtained from renewable feedstocks. Economic assessment of the process revealed that PMA and liamocins could be coproduced from two by-products at costs of $1.48/kg or $0.67/kg (with liamocins credit), offering an economic and sustainable process for the application of waste by-products.
Sujet(s)
Aureobasidium (genre)/croissance et développement , Techniques de culture cellulaire en batch , Gluconates/métabolisme , Malates/métabolisme , Mannitol , Polymères/métabolisme , Xylitol/métabolisme , Mannitol/analogues et dérivés , Mannitol/métabolismeRÉSUMÉ
Adjuvants are the helper substances that increase vaccine efficacy by enhancing the potency and longevity of specific immune responses to antigens. Most existing fish vaccines are presented in the form of oil-based emulsions delivered by intraperitoneal injection. The characterization of their mode of action is a valuable aid to future vaccine development, particularly for the potential identification and stimulation of specific immunological pathways related to the desired protective response. This study characterized the expression of selected immune-related genes in the peritoneal cavity, head kidney and spleen following the administration of two adjuvanted-bacterial vaccines thought to induce humoral (Montanide™ ISA 763A VG) or humoral and cell mediated (Montanide™ ISA 761 VG) immune responses, to determine if differences in responsiveness are readily apparent. The most informative site was the spleen, where Montanide™ ISA 763A VG + bacterin gave rise to upregulation of genes driving T-cell/lymphoid responses, namely IL-2, IL-15 and IL-21. This combined with upregulation of IFNγ1 and IFNγ2, IL-4/13B2, p35A1 and p40 (B1 and C) indicated that the induction of Th1 and possibly Th2 immunity was occurring in fish vaccinated with this adjuvant. Perhaps the most intriguing finding was the lack of a detectable Th1 response in fish given Montanide™ ISA 761 VG + bacterin, suggesting some other arm of the immune system is activated to give protection. Whatever the reason for the different responses detected, it is clear from the present study that the adjuvant used has a major impact on the responses elicited. Since these differences are readily detectable it allows, in principle, their use to help select the most appropriate adjuvants for inclusion into fish vaccines, where the type of response elicited may need to be tailored to a particular pathogen to confer protection.
Sujet(s)
Adjuvants immunologiques/pharmacologie , Aeromonas salmonicida , Vaccins antibactériens/immunologie , Maladies des poissons/prévention et contrôle , Infections bactériennes à Gram négatif/médecine vétérinaire , Mannitol/analogues et dérivés , Oncorhynchus mykiss/immunologie , Animaux , Cytokines/génétique , Cytokines/métabolisme , Régulation de l'expression des gènes/immunologie , Infections bactériennes à Gram négatif/prévention et contrôle , Rein céphalique/métabolisme , Macrophages péritonéaux , Mannitol/pharmacologie , Oncorhynchus mykiss/microbiologieRÉSUMÉ
Montanide ISA 51VG adjuvant has been approved for human clinical application and stimulates cellular and humoral immune responses. Here, HBsAg was formulated in Montanide ISA51VG adjuvant to compare its potency with the Fendrix and HBsAg-alum vaccines. In particular, the long-term humoral response was assessed up to 220 days after the final immunization. BALB/c mice were allocated into six groups. Treatment groups were injected with HBsAg-Montanide ISA51VG, the Fendrix and commercial HBsAg-alum, respectively. Montanide ISA51 VG, Alum and PBS injected mice were considered as control groups. Mice were immunized three times with 2-week intervals on days 0, 14 and 28 by subcutaneous injection. Lymphocyte proliferation was assessed with the BrdU method. IFN-γ, IL-2 and IL-4 cytokines, specific total IgG and IgG1/IgG2a isotypes were assessed using ELISA. The HBsAg-Montanide ISA51VG vaccine resulted in a significant increase in lymphocyte proliferation versus HBsAg-alum and higher IL-2 cytokine production versus the Fendrix. Comparable IL-4 and IFN-γ cytokines responses were observed for these vaccines. Following the first immunization, IgG increased more in HBs-Montanide 51VG group versus the HBs-alum group, while after the second and third shots comparable responses were observed in comparison to the HBs-alum group. Monitoring for 220 days after the final vaccination showed the superiority of HBsAg-Montanide ISA 51VG vaccine versus HBsAg-alum and even the Fendrix vaccine in the induction of long-term antibody responses. This study suggests that HBsAg-Montanide ISA51VG as a novel vaccine formulation can trigger both cellular and long-lasting humoral immune responses more efficiently than conventional HBsAg vaccines.
Sujet(s)
Préparation de médicament/méthodes , Antigènes de surface du virus de l'hépatite B/immunologie , Immunité humorale/immunologie , Mannitol/analogues et dérivés , Acides oléiques/immunologie , Adjuvants immunologiques/administration et posologie , Animaux , Production d'anticorps/effets des médicaments et des substances chimiques , Cytokines/métabolisme , Femelle , Antigènes de surface du virus de l'hépatite B/administration et posologie , Vaccins anti-hépatite B/administration et posologie , Vaccins anti-hépatite B/immunologie , Immunoglobuline G/sang , Injections sous-cutanées , Lymphocytes/effets des médicaments et des substances chimiques , Lymphocytes/métabolisme , Mannitol/administration et posologie , Mannitol/immunologie , Souris de lignée BALB C , Acides oléiques/administration et posologie , TempsRÉSUMÉ
Visceral leishmaniasis (VL) is a neglected tropical disease caused by Leishmania donovani or Leishmania infantum. Currently, the patients are treated with chemotherapeutic drugs; however, their toxicity limits their use. It would be desirable to develop a vaccine against this infection. In this study, we assessed the efficacy of different vaccine formulations at variable time points. Heat-killed (HK) antigen of Leishmania donovani was adjuvanted with two adjuvants (AddaVax and Montanide ISA 201) and three immunizations at a gap of 2 weeks (wk) were given to BALB/c mice. After 2 weeks of the last booster, mice were given challenge infection and sacrificed before challenge and after 4wk, 8wk, and 12 wk post-challenge. Significant protective immunity was observed in all the immunized animals and it was indicated by the notable rise in delayed-type hypersensitivity (DTH) response, remarkably declined parasite burden, a significant increase in the levels of interferon-gamma (IFN-γ), interleukin-12, interleukin-17 (Th1 cytokines), and IgG2a in contrast to infected control mice. Montanide ISA 201 with HK antigen provided maximum protection followed by AddaVax with HK and then HK alone. These findings elaborate on the importance of the tested adjuvants in the vaccine formulations against murine visceral leishmaniasis.
Sujet(s)
Adjuvants vaccinaux/administration et posologie , Antigènes de protozoaire/administration et posologie , Leishmania donovani , Vaccins antileishmaniose/administration et posologie , Leishmaniose viscérale/immunologie , Leishmaniose viscérale/prévention et contrôle , Mannitol/analogues et dérivés , Acides oléiques/administration et posologie , Polysorbates/administration et posologie , Squalène/administration et posologie , Animaux , Anticorps antiprotozoaires/sang , Cytokines/sang , Femelle , Hypersensibilité retardée/immunologie , Immunoglobuline G/sang , Leishmaniose viscérale/parasitologie , Mâle , Mannitol/administration et posologie , Souris de lignée BALB C , Monoxyde d'azote/immunologie , Espèces réactives de l'oxygène/immunologie , Rate/cytologie , Rate/immunologieRÉSUMÉ
DNA vaccines are capable of inducing humoral and cellular immunity, and are important to control bovine herpesvirus 1 (BoHV-1), an agent of the bovine respiratory disease complex. In previous work, a DNA plasmid that encodes a secreted form of BoHV-1 glycoprotein D (pCIgD) together with commercial adjuvants provided partial protection against viral challenge of bovines. In this work, we evaluate new molecules that could potentiate the DNA vaccine. We show that a plasmid encoding a soluble CD40 ligand (CD40L) and the adjuvant Montanide™ GEL01 (GEL01) activate in vitro bovine afferent lymph dendritic cells (ALDCs). CD40L is a co-stimulating molecule, expressed transiently on activated CD4+ T cells and, to a lesser extent, on activated B cells and platelets. The interaction with its receptor, CD40, exerts effects on the presenting cells, triggering responses in the immune system. GEL01 was designed to improve transfection of DNA vaccines. We vaccinated cattle with: pCIgD; pCIgD-GEL01; pCIgD with GEL01 and CD40L plasmid (named pCIgD-CD40L-GEL01) or with pCIneo vaccines. The results show that CD40L plasmid with GEL01 improved the pCIgD DNA vaccine, increasing anti-BoHV-1 total IgGs, IgG1, IgG2 subclasses, and neutralizing antibodies in serum. After viral challenge, bovines vaccinated with pCIgD-GEL01-CD40L showed a significant decrease in viral excretion and clinical score. On the other hand, 80% of animals in group pCIgD-GEL01-CD40L presented specific anti-BoHV-1 IgG1 antibodies in nasal swabs. In addition, PBMCs from pCIgD-CD40L-GEL01 had the highest percentage of animals with a positive lymphoproliferative response against the virus and significant differences in the secretion of IFNγ and IL-4 by mononuclear cells, indicating the stimulation of the cellular immune response. Overall, the results demonstrate that a plasmid expressing CD40L associated with the adjuvant GEL01 improves the efficacy of a DNA vaccine against BoHV-1.
Sujet(s)
Adjuvants immunologiques , Infections à Herpesviridae/médecine vétérinaire , Herpèsvirus bovin de type 1 , Immunogénicité des vaccins , Vaccins à ADN , Vaccins antiviraux/immunologie , Animaux , Anticorps antiviraux , Ligand de CD40/génétique , Bovins , Infections à Herpesviridae/prévention et contrôle , Herpèsvirus bovin de type 1/génétique , Mannitol/analogues et dérivés , Plasmides/génétique , Vaccins à ADN/génétiqueRÉSUMÉ
Opines are low-molecular-weight metabolites specifically biosynthesized by agrobacteria-transformed plant cells when plants are struck by crown gall and hairy root diseases, which cause uncontrolled tissue overgrowth. Transferred DNA is sustainably incorporated into the genomes of the transformed plant cells, so that opines constitute a persistent biomarker of plant infection by pathogenic agrobacteria and can be targeted for crown gall/hairy root disease diagnosis. We developed a general, rapid, specific and sensitive analytical method for overall opine detection using ultra-high-performance liquid chromatography-electrospray ionization quadrupole time-of-flight mass spectrometry (UHPLC-ESI-MS-QTOF), with easy preparation of samples. Based on MS, MS/MS and chromatography data, the detection selectivity of a wide range of standard opines was validated in pure solution and in different plant extracts. The method was successfully used to detect different structural types of opines, including opines for which standard compounds are unavailable, in tumors or hairy roots induced by pathogenic strains. As the method can detect a wide range of opines in a single run, it represents a powerful tool for plant gall analysis and crown gall/hairy root disease diagnosis. Using an appropriate dilution of plant extract and a matrix-based calibration curve, the quantification ability of the method was validated for three opines belonging to different families (nopaline, octopine, mannopine), which were accurately quantified in plant tissue extracts.
Sujet(s)
Arginine/analogues et dérivés , Chromatographie en phase liquide à haute performance/méthodes , Mannitol/analogues et dérivés , Tumeurs végétales , Spectrométrie de masse ESI/méthodes , Agrobacterium , Arginine/analyse , Marqueurs biologiques/analyse , Mannitol/analyse , Maladies des plantes , Racines de plante/composition chimique , Reproductibilité des résultatsRÉSUMÉ
Background: Immune checkpoint blockade with monoclonal antibodies targeting programmed death 1 (PD-1) and its ligand PD-L1 has played a major role in the rise of cancer immune therapy. We have identified naturally occurring self-reactive T cells specific to PD-L1 in both healthy donors and cancer patients. Stimulation with a PD-L1 peptide (IO103), activates these cells to exhibit inflammatory and anti-regulatory functions that include cytotoxicity against PD-L1-expressing target cells. This prompted the initiation of the present first-in-human study of vaccination with IO103, registered at clinicaltrials.org (NCT03042793). Methods: Ten patients with multiple myeloma who were up to 6 months after high dose chemotherapy with autologous stem cell support, were enrolled. Subcutaneous vaccinations with IO103 with the adjuvant Montanide ISA 51 was given up to fifteen times during 1 year. Safety was assessed by the common toxicity criteria for adverse events (CTCAE). Immunogenicity of the vaccine was evaluated using IFNγ enzyme linked immunospot and intracellular cytokine staining on blood and skin infiltrating lymphocytes from sites of delayed-type hypersensitivity. The clinical course was described. Results: All adverse reactions to the PD-L1 vaccine were below CTCAE grade 3, and most were grade 1-2 injection site reactions. The total rate of adverse events was as expected for the population. All patients exhibited peptide specific immune responses in peripheral blood mononuclear cells and in skin-infiltrating lymphocytes after a delayed-type hypersensitivity test. The clinical course was as expected for the population. Three of 10 patients had improvements of responses which coincided with the vaccinations. Conclusion: Vaccination against PD-L1 was associated with low toxicity and high immunogenicity. This study has prompted the initiation of later phase trials to assess the vaccines efficacy. Clinical Trial Registration: clinicaltrials.org, identifier NCT03042793.
Sujet(s)
Antigène CD274/immunologie , Vaccins anticancéreux/administration et posologie , Mannitol/analogues et dérivés , Myélome multiple/traitement médicamenteux , Protéines tumorales/immunologie , Acides oléiques/administration et posologie , Peptides/administration et posologie , Adulte , Sujet âgé , Vaccins anticancéreux/effets indésirables , Femelle , Humains , Mâle , Mannitol/administration et posologie , Mannitol/effets indésirables , Adulte d'âge moyen , Myélome multiple/immunologie , Myélome multiple/anatomopathologie , Acides oléiques/effets indésirables , Peptides/effets indésirables , Vaccins sous-unitaires/administration et posologie , Vaccins sous-unitaires/effets indésirablesRÉSUMÉ
BACKGROUND: Toxoplasma gondii is an obligate intracellular parasite that can infect almost all warm-blooded animals, avian species and humans. Toxoplasmosis is asymptomatic in healthy individuals, whereas it may lead to death in immune suppressed or deficient patients. A vaccine against T. gondii is required to prevent consequences of the infection. The aim of this study is to generate a multivalent recombinant protein vaccine against T. gondii. METHODS: 49 previously discovered antigenic proteins of T gondii were evaluated by their expression level in E. coli and by comprehensive bioinformatics analyses to determine antigenic epitopes. Based on these analyses, six vaccine candidate proteins were selected to generate a hexavalent recombinant protein vaccine adjuvanted with Montanide ISA 50 V. Humoral and cellular immune responses were determined by flow cytometry and ELISA. Vaccinated mice were challenged with T. gondii Ankara strain tachyzoites. RESULTS: In mice vaccinated with hexavalent vaccine, strong total IgG (P < 0.0001) and IgG2a (P < 0.001) responses were induced compared to controls, the ratio of CD4+ and CD8+ T lymphocytes secreting IFN-γ increased, and significantly higher extracellular IFN-γ secretion was achieved compared to the controls (P < 0.001). The survival time of the vaccinated mice increased to 8.38 ± 2.13 days which was significantly higher than controls (P < 0.01). CONCLUSIONS: Altogether, these results show that the hexavalent vaccine which is developed for the first time against T. gondii induced strong and balanced Th1 and Th2 immune responses as well as conferred significant protection against challenge with lethal toxoplasmosis in murine model.
Sujet(s)
Adjuvants immunologiques/pharmacologie , Mannitol/analogues et dérivés , Vaccins antiprotozoaires/pharmacologie , Toxoplasmose/prévention et contrôle , Vaccins à ADN/pharmacologie , Animaux , Test ELISA , Épitopes/génétique , Épitopes/immunologie , Escherichia coli/génétique , Femelle , Immunité cellulaire/effets des médicaments et des substances chimiques , Immunité humorale/effets des médicaments et des substances chimiques , Immunoglobuline G/sang , Mannitol/pharmacologie , Souris , Protéines de protozoaire/génétique , Protéines de protozoaire/immunologie , Vaccins antiprotozoaires/génétique , Vaccins antiprotozoaires/immunologie , Protéines recombinantes/génétique , Protéines recombinantes/immunologie , Toxoplasma/pathogénicité , Toxoplasmose/immunologie , Vaccins à ADN/immunologieRÉSUMÉ
BACKGROUND: Immunogenicity of cancer vaccines is impacted by adjuvants and schedule, but systematic assessments of their effects have not been performed. Montanide ISA-51, an incomplete Freund's adjuvant (IFA), is used in many vaccine trials, but concerns have been raised about negative effects in murine studies. We found in humans that IFA enhances systemic immune responses and that repeat vaccination at one site (same site vaccination (SSV)) creates tertiary lymphoid structures (TLS) in the vaccine site microenvironment (VSME). We hypothesized that vaccination with peptides+IFA+pICLC or SSV×3 with peptides in IFA would create an immunogenic milieu locally at the VSME, with activated dendritic cells (DC), TLS-associated chemokines and a Th1-dominant VSME. METHODS: Biopsies of the VSME were obtained from participants on two clinical trials who were immunized with multiple melanoma peptides (MELITAC 12.1) in adjuvants comprising IFA and/or the TLR3-agonist pICLC. Biopsies were obtained either a week after one vaccine or a week after SSV×3. Controls included normal skin and skin injected with IFA without peptides. Gene expression analysis was performed by RNAseq. RESULTS: VSME samples were evaluated from 27 patients. One vaccine with peptides in pICLC+IFA enhanced expression of CD80, CD83, CD86 (p<0.01), CD40 and CD40L (p<0.0001) over normal skin; these effects were significantly enhanced for SSV with peptides+IFA. Vaccines containing pICLC increased expression of TBX21 (T-bet) but did not decrease GATA3 over normal skin, whereas SSV with peptides in IFA dramatically enhanced TBX21 and decreased GATA3, with high expression of IFNγ and STAT1. SSV with peptides in IFA also reduced arginase-1 (ARG1) expression and enhanced expression of TLR adapter molecules TICAM-1 (TRIF) and MYD88. Furthermore, SSV with IFA and peptides also enhanced expression of chemokines associated with TLS formation. CONCLUSIONS: These findings suggest that SSV with peptides in IFA enhances CD40L expression by CD4 T cells, supports a Th1 microenvironment, with accumulation of activated and mature DC. Increased expression of TLR adaptor proteins after SSV with peptides in IFA might implicate effects of the skin microbiome. Reduced ARG1 may reflect diminished suppressive myeloid activity in the VSME. TRIAL REGISTRATION NUMBER: (NCT00705640, NCT01585350).
Sujet(s)
Adjuvants immunologiques/administration et posologie , Vaccins anticancéreux/administration et posologie , Adjuvant Freund/administration et posologie , Lipides/administration et posologie , Mélanome/thérapie , Tumeurs cutanées/thérapie , Vaccination/méthodes , Adulte , Sujet âgé , Sujet âgé de 80 ans ou plus , Antigènes néoplasiques/immunologie , Arginase/métabolisme , Biopsie , Lymphocytes T CD4+/effets des médicaments et des substances chimiques , Lymphocytes T CD4+/immunologie , Lymphocytes T CD4+/métabolisme , Ligand de CD40/immunologie , Ligand de CD40/métabolisme , Vaccins anticancéreux/immunologie , Essais cliniques de phase I comme sujet , Femelle , Adjuvant Freund/immunologie , Humains , Rappel de vaccin/méthodes , Immunogénicité des vaccins , Injections intralésionnelles , Lipides/immunologie , Mâle , Mannitol/administration et posologie , Mannitol/analogues et dérivés , Mannitol/immunologie , Mélanome/immunologie , Mélanome/anatomopathologie , Adulte d'âge moyen , Acides oléiques/administration et posologie , Acides oléiques/immunologie , Essais contrôlés randomisés comme sujet , Transduction du signal/effets des médicaments et des substances chimiques , Transduction du signal/immunologie , Peau/immunologie , Peau/anatomopathologie , Tumeurs cutanées/immunologie , Tumeurs cutanées/anatomopathologie , Lymphocytes auxiliaires Th1/effets des médicaments et des substances chimiques , Lymphocytes auxiliaires Th1/immunologie , Récepteurs de type Toll/métabolisme , Microenvironnement tumoral/immunologie , Vaccins sous-unitaires/administration et posologie , Vaccins sous-unitaires/immunologie , Jeune adulteRÉSUMÉ
Brucellosis is the most common zoonotic disease worldwide and still there is no vaccine for human use. The commercial animal vaccines also have major problems that limit their use. Therefore, there is a need for an effective Brucella vaccine which is multivalent and produces a good protective immunity with minimal disadvantages. Due to their heterogeneous composition and diverse functions, OMVs are promising acellular vaccine candidates against brucellosis. In the present study, the potential of Poly(I:C) or CpG ODN 1826+ Montanide ISA 70 VG adjuvant formulations were evaluated to enhance the immunity and protection levels conferred by OMVs against Brucella challenge in mice. The results indicated that both vaccine regimens were able to induce strong Th1-biased responses and confer protective levels significantly higher than REV.1 live vaccine. With regard to the results, it is concluded that OMVs in either adjuvant can be introduced as a new vaccine candidate against B. melitensis infection.
Sujet(s)
Adjuvants immunologiques/administration et posologie , Membrane bactérienne externe/immunologie , Vaccin antibrucellique/administration et posologie , Brucellose/prévention et contrôle , Structures de la membrane cellulaire/immunologie , Mannitol/analogues et dérivés , Acides oléiques/administration et posologie , Oligodésoxyribonucléotides/administration et posologie , Poly I-C/administration et posologie , Animaux , Brucella melitensis/effets des médicaments et des substances chimiques , Brucella melitensis/croissance et développement , Cytokines/immunologie , Femelle , Immunoglobuline G/immunologie , Mannitol/administration et posologie , Souris de lignée BALB CRÉSUMÉ
So far, it has been still unknown how liamocins are biosynthesized, regulated, transported and secreted. In this study, a highly reducing polyketide synthase (HR-PKS), a mannitol-1-phosphate dehydrogenase (MPDH), a mannitol dehydrogenase (MtDH), an arabitol dehydrogenase (ArDH) and an esterase (Est1) were found to be closely related to core biosynthesis of extracellular liamocins in Aureobasidium melanogenum 6-1-2. The HR-PKS was responsible for biosynthesis of 3,5-dihydroxydecanoic acid. The MPDH and MtDH were implicated in mannitol biosynthesis and the ArDH was involved in arabitol biosynthesis. The Est1 catalyzed ester bond formation of them. A phosphopantetheine transferase (PPTase) activated the HR-PKS and a transcriptional activator Ga11 activated expression of the PKS1 gene. Therefore, deletion of the PKS1 gene, all the three genes encoding MPDH, MtDH and ArDH, the EST1, the gene responsible for PPTase and the gene for Ga11 made all the disruptants (Δpks13, Δpta13, Δest1, Δp12 and Δg11) totally lose the ability to produce any liamocins. A GLTP gene encoding a glycolipid transporter and a MDR1 gene encoding an ABC transporter took part in transport and secretion of the produced liamocins into medium. Removal of the GLTP gene and the MDR1 gene resulted in a Δgltp1 mutant and a Δmdr16 mutant, respectively, that lost the partial ability to secrete liamocins, but which cells were swollen and intracellular lipid accumulation was greatly enhanced. Hydrolysis of liamocins released 3,5-dihydroxydecanoic acid, mannitol, arabitol and acetic acid. We proposed a core biosynthesis pathway, regulation, transport and secretion of liamocins in A. melanogenum.
Sujet(s)
Ascomycota/génétique , Ascomycota/métabolisme , Voies de biosynthèse/physiologie , Mannitol/analogues et dérivés , Huiles/métabolisme , Transport des protéines/physiologie , Techniques de knock-in de gènes/méthodes , Mannitol/analyse , Mannitol/métabolisme , Huiles/analyseRÉSUMÉ
Agrobacterium tumefaciens pathogens use specific compounds denoted opines as nutrients in their plant tumor niche. These opines are produced by the host plant cells genetically modified by agrobacteria. They are imported into bacteria via solute-binding proteins (SBPs) in association with ATP-binding cassette transporters. The mannityl-opine family encompasses mannopine, mannopinic acid, agropine and agropinic acid. Structural and affinity data on mannopinic acid bound to SBPs are currently lacking while those of the three others mannityl opines are available. We investigated the molecular basis of two pathways for mannopinic acid uptake. MoaA was proposed as the specific SBP for mannopinic acid import in mannityl opines-assimilating agrobacteria, which was validated here using genetic studies and affinity measurements. We structurally characterized the mannopinic acid-binding mode of MoaA in two crystal forms at 2.05 and 1.57â Å resolution. We demonstrated that the non-specific SBP MotA, so far characterized as mannopine and Amadori compound importer, was also able to transport mannopinic acid. The structure of MotA bound to mannopinic acid at 2.2â Å resolution defines a different mannopinic acid-binding signature, similar to that of mannopine. Combining in vitro and in vivo approaches, this work allowed us to complete the characterization of the mannityl-opines assimilation pathways, highlighting the important role of two dual imports of agropinic and mannopinic acids. Our data shed new light on how the mannityl-opines contribute to the establishment of the ecological niche of agrobacteria from the early to the late stages of tumor development.
Sujet(s)
Transport biologique , Protéines de transport , Mannitol/analogues et dérivés , Tumeurs végétales/microbiologie , Transporteurs ABC/métabolisme , Agrobacterium tumefaciens/métabolisme , Protéines bactériennes/composition chimique , Protéines bactériennes/génétique , Protéines bactériennes/métabolisme , Protéines de transport/composition chimique , Protéines de transport/génétique , Protéines de transport/métabolisme , Cristallographie , Gènes bactériens , Interactions hôte-microbes , Mannitol/composition chimique , Mannitol/métabolisme , Oxazines/métabolismeRÉSUMÉ
N-Substituted derivatives of 1,4-dideoxy-1,4-imino-d-mannitol (DIM), the pyrrolidine core of swainsonine, have been synthesized efficiently and stereoselectively from d-mannose with 2,3:5,6-di-O-isopropylidene DIM (10) as a key intermediate. These N-substituted derivatives include N-alkylated, N-alkenylated, N-hydroxyalkylated and N-aralkylated DIMs with the carbon number of the alkyl chain ranging from one to nine. The obtained 33 N-substituted DIM derivatives were assayed against various glycosidases, which allowed a systematic evaluation of their glycosidase inhibition profiles. Though N-substitution of DIM decreased their α-mannosidase inhibitory activities, some of the derivatives showed significant inhibition of other glycosidases.
Sujet(s)
Antienzymes/synthèse chimique , Antienzymes/pharmacologie , Glycosidases/antagonistes et inhibiteurs , Mannitol/analogues et dérivés , Animaux , Antienzymes/composition chimique , Glycosidases/métabolisme , Humains , Iminofuranoses/synthèse chimique , Iminofuranoses/composition chimique , Iminofuranoses/pharmacologie , Concentration inhibitrice 50 , Mannitol/synthèse chimique , Mannitol/composition chimique , Mannitol/pharmacologie , Rats , Tridolgosir/composition chimiqueRÉSUMÉ
This study evaluated plant-based immune-adjuvants from crude extracts of Phoenix dactylifera and Mentha piperita as promising adjuvants for vaccines because of the limited side effects associated with plant extracts. In addition, Montanide™ ISA 201 previously used in vaccines in cattle. Eight different infectious coryza (IC) vaccines were prepared from three serovars [A (W strain and local strain), C (Modesto strain) and B (0222 strain)] with eight Avibacterium paragallinarum vaccines adjuvants formulae using liquid paraffin, Montanide™ ISA 71, Montanide™ ISA 201, and Montanide™ Gel adjuvants, P. dactylifera and M. piperita as immune-stimulants at a concentration of 1 mg and 2 mg incorporated with or without liquid paraffin oil as an adjuvant. These vaccines were applied in a chicken model. After a single immunization, the eight vaccine formulations were evaluated using the ELISA and Microplate agglutination test. Evidence of protection in the immunized birds was based on the results after challenge and bacterial isolation. The incorporation of the crude aqueous extract of P. dactylifera or M. piperita at a concentration of 2 mg in a liquid paraffin oil adjuvanted IC vaccine could be employed as an efficient adjuvant for chicken to IC vaccine to enhance immune responses. Also,Montanide™ ISA 201 may be the best adjuvant to be used to enhance the protective response against Av. paragallinarum. Our results confirm that aqueous extracts of M. piperita leaves and P. dactylifera fruit have immunomodulatory potentials in vivo and elevated serum antibodies against Av. Paragallinarum.
Sujet(s)
Adjuvants immunologiques , Vaccins antibactériens/immunologie , Poulets , Mannitol/analogues et dérivés , Mentha piperita , Acides oléiques , Phoeniceae , Maladies de la volaille/prévention et contrôle , Animaux , Anticorps antibactériens/sang , Immunisation , Mannitol/pharmacologie , Acides oléiques/pharmacologie , Pasteurellaceae/immunologie , Vaccination/médecine vétérinaireRÉSUMÉ
Xylose, the main component of xylan, is the second most abundant sugar in nature after glucose. Consequently, xylose represents an attractive feedstock for the production of value-added compounds such as biosurfactants (BSs), which are produced by various bacteria and yeasts. In this study, we screened and isolated yeast strains that synthesize BSs using xylose as the sole carbon source. We applied matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) to screen for BS-producing yeasts and isolated eight strains as the liamocin producers. Two of the eight strains, AS37 and SK25, were identified as Aureobasidium melanogenum, which is known as black yeasts, by based on 26S ribosomal RNA gene sequences. Both strains produced a wide variety of liamocin structures from not only xylose but also glucose and sucrose. According to the MALDI-TOF MS analysis, signals corresponding to sodium ion adducts of di-, tri-, tetra-, penta- and hexa-acylated C6-liamocins and di-, tri- and tetra-acylated C5-liamocins were detected. In addition, their mono-acetylated form was also detected. The dominant sugar component of liamocins produced by strains AS37 and SK25 is mannitol as estimated by HPLC analysis. This is the first report to describe the screening of liamocins-producing yeasts using xylose as the sole carbon source.
Sujet(s)
Ascomycota/isolement et purification , Ascomycota/métabolisme , Carbone/métabolisme , Mannitol/analogues et dérivés , Mannitol/métabolisme , Xylose/métabolisme , Bactéries , Carbone/ressources et distribution , Mannitol/composition chimique , Techniques microbiologiques/méthodes , Huiles/métabolisme , ARN ribosomique , Spectrométrie de masse MALDIRÉSUMÉ
Given its ability to induce both humoral and cellular immune responses, NY-ESO-1 has been considered a suitable antigen for a cancer vaccine. Despite promising results from early-phase clinical studies in patients with melanoma, NY-ESO-1 vaccine immunotherapy has not been widely investigated in larger trials; consequently, many questions remain as to the optimal vaccine formulation, predictive biomarkers, and sequencing and timing of vaccines in melanoma treatment. We conducted an adjuvant phase I/II clinical trial in high-risk resected melanoma to optimize the delivery of poly-ICLC, a TLR-3/MDA-5 agonist, as a component of vaccine formulation. A phase I dose-escalation part was undertaken to identify the MTD of poly-ICLC administered in combination with NY-ESO-1 and montanide. This was followed by a randomized phase II part investigating the MTD of poly-ICLC with NY-ESO-1 with or without montanide. The vaccine regimens were generally well tolerated, with no treatment-related grade 3/4 adverse events. Both regimens induced integrated NY-ESO-1-specific CD4+ T-cell and humoral responses. CD8+ T-cell responses were mainly detected in patients receiving montanide. T-cell avidity toward NY-ESO-1 peptides was higher in patients vaccinated with montanide. In conclusion, NY-ESO-1 protein in combination with poly-ICLC is safe, well tolerated, and capable of inducing integrated antibody and CD4+ T-cell responses in most patients. Combination with montanide enhances antigen-specific T-cell avidity and CD8+ T-cell cross-priming in a fraction of patients, indicating that montanide contributes to the induction of specific CD8+ T-cell responses to NY-ESO-1.