Identification of Estrogen Receptor Modulators as Inhibitors of Flavivirus Infection.

Flaviviruses such as Zika virus (ZIKV), dengue virus (DENV), and West Nile virus (WNV) are major global pathogens for which safe and effective antiviral therapies are not currently available. To identify antiviral small molecules with well-characterized safety and bioavailability profiles, we screened a library of 2,907 approved drugs and pharmacologically active compounds for inhibitors of ZIKV infection using a high-throughput cell-based immunofluorescence assay. Interestingly, estrogen receptor modulators raloxifene hydrochloride and quinestrol were among 15 compounds that significantly inhibited ZIKV infection in repeat screens. Subsequent validation studies revealed that these drugs effectively inhibit ZIKV, DENV, and WNV (Kunjin strain) infection at low micromolar concentrations with minimal cytotoxicity in Huh-7.5 hepatoma cells and HTR-8 placental trophoblast cells. Since these cells lack detectable expression of estrogen receptors-α and -ß (ER-α and ER-ß) and similar antiviral effects were observed in the context of subgenomic DENV and ZIKV replicons, these compounds appear to inhibit viral RNA replication in a manner that is independent of their known effects on estrogen receptor signaling. Taken together, quinestrol, raloxifene hydrochloride, and structurally related analogues warrant further investigation as potential therapeutics for treatment of flavivirus infections.

Advax augments B and T cell responses upon influenza vaccination via the respiratory tract and enables complete protection of mice against lethal influenza virus challenge.

Administration of influenza vaccines via the respiratory tract has potential benefits over conventional parenteral administration, inducing immunity directly at the site of influenza exposure as well as being needle free. In this study, we investigated the suitability of Advax™, a stable particulate polymorph of inulin, also referred to as delta inulin, as a mucosal adjuvant for whole inactivated influenza vaccine (WIV) administered either as a liquid or dry powder formulation. Spray freeze-drying produced Advax-adjuvanted WIV powder particles in a size range (1-5 µm) suitable for inhalation. The physical and biological characteristics of both WIV and Advax remained unaltered both by admixing WIV with Advax and by spray freeze drying. Upon intranasal or pulmonary immunization, both liquid and dry powder formulations containing Advax induced significantly higher systemic, mucosal and cellular immune responses than non-adjuvanted WIV formulations. Furthermore, pulmonary immunization with Advax-adjuvanted WIV led to robust memory B cell responses along with an increase of lung localization factors i.e. CXCR3, CD69, and CD103. A less pronounced but still positive effect of Advax was seen on memory T cell responses. In contrast to animals immunized with WIV alone, all animals pulmonary immunized with a single dose of Advax-adjuvanted WIV were fully protected with no visible clinical symptoms against a lethal dose of influenza virus. These data confirm that Advax is a potent mucosal adjuvant that boosts vaccine-induced humoral and cellular immune responses both in the lung and systemically with major positive effects on B-cell memory and complete protection against live virus. Hence, respiratory tract immunization, particularly via the lungs, with Advax-adjuvanted WIV formulation as a liquid or dry powder is a promising alternative to parenteral influenza vaccination.

Dengue Virus Induces Increased Activity of the Complement Alternative Pathway in Infected Cells.

Severe dengue virus (DENV) infection is associated with overactivity of the complement alternative pathway (AP) in patient studies. Here, the molecular changes in components of the AP during DENV infection in vitro were investigated. mRNA for factor H (FH), a major negative regulator of the AP, was significantly increased in DENV-infected endothelial cells (EC) and macrophages, but, in contrast, production of extracellular FH protein was not. This discord was not seen for the AP activator factor B (FB), with DENV induction of both FB mRNA and protein, nor was it seen with Toll-like receptor 3 or 4 stimulation of EC and macrophages, which induces both FH and FB mRNA and protein. Surface-bound and intracellular FH protein was, however, induced by DENV, but only in DENV antigen-positive cells, while in two other DENV-susceptible immortalized cell lines (ARPE-19 and human retinal endothelial cells), FH protein was induced both intracellularly and extracellularly by DENV infection. Regardless of the cell type, there was an imbalance in AP components and an increase in markers of complement AP activity associated with DENV-infected cells, with lower FH relative to FB protein, an increased ability to promote AP-mediated lytic activity, and increased deposition of complement component C3b on the surface of DENV-infected cells. For EC in particular, these changes are predicted to result in higher complement activity in the local cellular microenvironment, with the potential to induce functional changes that may result in increased vascular permeability, a hallmark of dengue disease.IMPORTANCE Dengue virus (DENV) is a significant human viral pathogen with a global medical and economic impact. DENV may cause serious and life-threatening disease, with increased vascular permeability and plasma leakage. The pathogenic mechanisms underlying these features remain unclear; however, overactivity of the complement alternative pathway has been suggested to play a role. In this study, we investigate the molecular events that may be responsible for this observed alternative pathway overactivity and provide novel findings of changes in the complement system in response to DENV infection in primary cell types that are a major target for DENV infection (macrophages) and pathogenesis (endothelial cells) in vivo Our results suggest a new dimension of cellular events that may influence endothelial cell barrier function during DENV infection that could expand strategies for developing therapeutics to prevent or control DENV-mediated vascular disease.

Erratum to "Molecular Responses of Human Retinal Cells to Infection with Dengue Virus".

[This corrects the article DOI: 10.1155/2017/3164375.].

Molecular Responses of Human Retinal Cells to Infection with Dengue Virus.

Recent clinical reports indicate that infection with dengue virus (DENV) commonly has ocular manifestations. The most serious threat to vision is dengue retinopathy, including retinal vasculopathy and macular edema. Mechanisms of retinopathy are unstudied, but observations in patients implicate retinal pigment epithelial cells and retinal endothelial cells. Human retinal cells were inoculated with DENV-2 and monitored for up to 72 hours. Epithelial and endothelial cells supported DENV replication and release, but epithelial cells alone demonstrated clear cytopathic effect, and infection was more productive in those cells. Infection induced type I interferon responses from both cells, but this was stronger in epithelial cells. Endothelial cells increased expression of adhesion molecules, with sustained overexpression of vascular adhesion molecule-1. Transcellular impedance decreased for epithelial monolayers, but not endothelial monolayers, coinciding with cytopathic effect. This reduction was accompanied by disorganization of intracellular filamentous-actin and decreased expression of junctional molecules, zonula occludens 1, and catenin-ß1. Changes in endothelial expression of adhesion molecules are consistent with the retinal vasculopathy seen in patients infected with DENV; decreases in epithelial junctional protein expression, paralleling loss of integrity of the epithelium, provide a molecular basis for DENV-associated macular edema. These molecular processes present potential therapeutic targets for vision-threatening dengue retinopathy.

A Reevaluation of the Effectiveness of Homoeoprophylaxis Against Leptospirosis in Cuba in 2007 and 2008.

OBJECTIVES: In 2007 and 2008, Cuban health officials conducted large homoeoprophylaxis interventions against rising rates of leptospirosis caused by extensive hurricane damage. Published results showed that the interventions were highly successful, but some questions regarding possible confounders were raised. The objective of this research was to assess the influence of potential confounders on initial results. DESIGN: Weekly leptospirosis reporting data entries for 9 years were checked to ensure data consistency. Some errors in weekly reports for 2000-2008 were discovered, and corrected, and the changes incorporated in this analysis. The corrected data was reanalyzed to investigate the impact of potential confounders. RESULTS: New analyses of the timing and extent of vaccination and chemoprophylaxis in 2007 and 2008 and changes in leptospirosis notifications were presented. CONCLUSIONS: The results support the previous conclusions that homoeoprophylaxis can be used to effectively immunize people against targeted infectious diseases such as leptospirosis.

Large-scale application of highly-diluted bacteria for Leptospirosis epidemic control.

BACKGROUND: Leptospirosis is a zoonotic disease of major importance in the tropics where the incidence peaks in rainy seasons. Natural disasters represent a big challenge to Leptospirosis prevention strategies especially in endemic regions. Vaccination is an effective option but of reduced effectiveness in emergency situations. Homeoprophylactic interventions might help to control epidemics by using highly-diluted pathogens to induce protection in a short time scale. We report the results of a very large-scale homeoprophylaxis (HP) intervention against Leptospirosis in a dangerous epidemic situation in three provinces of Cuba in 2007. METHODS: Forecast models were used to estimate possible trends of disease incidence. A homeoprophylactic formulation was prepared from dilutions of four circulating strains of Leptospirosis. This formulation was administered orally to 2.3 million persons at high risk in an epidemic in a region affected by natural disasters. The data from surveillance were used to measure the impact of the intervention by comparing with historical trends and non-intervention regions. RESULTS: After the homeoprophylactic intervention a significant decrease of the disease incidence was observed in the intervention regions. No such modifications were observed in non-intervention regions. In the intervention region the incidence of Leptospirosis fell below the historic median. This observation was independent of rainfall. CONCLUSIONS: The homeoprophylactic approach was associated with a large reduction of disease incidence and control of the epidemic. The results suggest the use of HP as a feasible tool for epidemic control, further research is warranted.

Mucosal immunization using proteoliposome and cochleate structures from Neisseria meningitidis serogroup B induce mucosal and systemic responses.

Most pathogens either invade the body or establish infection in mucosal tissues and represent an enormous challenge for vaccine development by the absence of good mucosal adjuvants. A proteoliposome-derived adjuvant from Neisseria meningitidis serogroup B (AFPL1, Adjuvant Finlay Proteoliposome 1) and its derived cochleate form (Co, AFCo1) contain multiple pathogen-associated molecular patterns as immunopotentiators, and can also serve as delivery systems to elicit a Th1-type immune response. The present studies demonstrate the ability of AFPL1and AFCo1 to induce mucosal and systemic immune responses by different mucosal immunizations routes and significant adjuvant activity for antibody responses of both structures: a microparticle and a nanoparticle with a heterologous antigen. Therefore, we used female mice immunized by intragastric, intravaginal, intranasal or intramuscular routes with both structures alone or incorporated with ovalbumin (OVA). High levels of specific IgG antibody were detected in all sera and in vaginal washes, but specific IgA antibody in external secretions was only detected in mucosally immunized mice. Furthermore, antigen specific IgG1 and IgG2a isotypes were all induced. AFPL1 and AFCo1 are capable of inducing IFN-gamma responses, and chemokine secretions, like MIP-1alpha and MIP-1beta. However, AFCo1 is a better alternative to induce immune responses at mucosal level. Even when we use a heterologous antigen, the AFCo1 response was better than with AFPL1 in inducing mucosal and systemic immune responses. These results support the use of AFCo1 as a potent Th1 inducing adjuvant particularly suitable for mucosal immunization.

Ensayo de inmunogenicidad y toxicidad local del cocleato de Neisseriameningitidis en ratas Sprague Dawley / Study of immunogenicity and local toxicity of the Neisseria meningitidis cochleate in Sprague Dawley rats

Los estudios de tolerancia local para productos vacunales resultan fundamentales como requisitos regulatorios para los estudios preclínicos. En este ensayo se utilizaron 48 ratas Sprague Dawley, de ambos sexos, con un peso corporal de 180-220 g, suministradas por el Centro Nacional para la Producción de Animales de Laboratorio, y una duración de 17 días. Las ratas fueron distribuidas en 5 grupos: 3 con diferentes concentraciones de cocleatos, uno sin inocular y otro que recibió el diluente del producto en prueba. A estas se les realizaron las observaciones clínicas necesarias, tales como: mediciones de incremento de peso, consumo de alimentos y agua, así como la evaluación anatomopatológica en los dos tiempos de sacrificio, uno a los 12 y otro a los 17 días; se enfatizó en el estudio histopatológico de encéfalo y las fosas nasales en tres niveles de su extensión. Se hicieron investigaciones inmunológicas consistentes en la determinación de IgG en suero, saliva y líquido cefalorraquídeo. Los resultados arrojaron un incremento del peso. En el consumo de agua y alimento no se observaron alteraciones clínicas, la anatomopatología evidenció discretos cambios inflamatorios que guardaron una relación directa con el aumento de las concentraciones del producto, los títulos de IgG en la saliva y suero de las ratas inoculadas con los cocleatos difirieron de forma significativa (P < 0,05) respecto a los grupos controles, el líquido cefalorraquídeo resultó negativo a la presencia de anticuerpos específicos IgG. Se concluye que los cocleatos aplicados por esta vía en ratas resultaron inmunogénicos e inocuos para el referido producto(AU)

Local Tolerance studies for vaccine products are a main link in the chain of Regulatory Requirements for toxicological preclinical studies. Mainly considering that the use of mucosal road seems to be a current tendency due to the advantages it offers. Sprague Dawley rats of both sexes weighing 100-120 g (reception weight) provided by CENPALAB with the corresponding certificates of sanitary and genetic quality were used in this test. Local tolerance study lasted 17 days. Rats were distributed in 5 groups (3 groups with different cochleate concentrations, one without inoculation and another one that received the diluents of the product in test. Measurement of weight increase, food and water consumption, as well as the anatomopathologic studies in the two times of sacrifice, at 12 and 17 days were performed. Main histopathological studies were carried out in brain and nasal nostrils in three levels of their extension (anterior, medium and posterior areas). In addition, immunological assays such as the determination of IgG in serum, saliva and cerebrospinal fluid (CSF) were carried out. Results showed a sustained weight increase. There were no clinical alterations during water and food consumption, while the existence of discreet acute inflammatory changes was found by anatomopathological studies, which were directly related to the increase of the product concentration. The presence of IgG antibodies in saliva and in sera of the rats inoculated with the cochleates showed significant differences (P <0.05) compared to the non inoculated control group and to the animals inoculated with the diluent. CSF studies resulted negative regarding the presence of specific IgG antibodies. The existence of a direct relation among the concentrations of the product in test and the inflammatory processes in the medium and posterior levels of the nasal nostril was proven, therefore it is considered that cochleates applied by this route and in the concentrations ...(AU)

AFCo1, a meningococcal B-derived cochleate adjuvant, strongly enhances antibody and T-cell immunity against Plasmodium falciparum merozoite surface protein 4 and 5.

BACKGROUND: Whilst a large number of malaria antigens are being tested as candidate malaria vaccines, a major barrier to the development of an effective vaccine is the lack of a suitable human adjuvant capable of inducing a strong and long lasting immune response. In this study, the ability of AFCo1, a potent T and B cell adjuvant based on cochleate structures derived from meningococcal B outer membrane proteoliposomes (MBOMP), to boost the immune response against two Plasmodium falciparum antigens, merozoite surface protein 4 (MSP4) and 5 (MSP5), was evaluated. METHODS: Complete Freund's adjuvant (CFA), which is able to confer protection against malaria in animal MSP4/5 vaccine challenge models, was used as positive control adjuvant. MSP4 and 5-specific IgG, delayed-type hypersensitivity (DTH), T-cell proliferation, and cytokine production were evaluated in parallel in mice immunized three times intramuscularly with MSP4 or MSP5 incorporated into AFCo1, synthetic cochleate structures, CFA or phosphate buffered saline. RESULTS: AFCo1 significantly enhanced the IgG and T-cell response against MSP4 and MSP5, with a potency equivalent to CFA, with the response being characterized by both IgG1 and IgG2a isotypes, increased interferon gamma production and a strong DTH response, consistent with the ability of AFCo1 to induce Th1-like immune responses. CONCLUSION: Given the proven safety of MBOMP, which is already in use in a licensed human vaccine, AFCo1 could assist the development of human malaria vaccines that require a potent and safe adjuvant.

AFCo1, a meningococcal B-derived cochleate adjuvant, strongly enhances antibody and T-cell immunity against Plasmodium falciparum merozoite surface protein 4 and 5

BACKGROUND: Whilst a large number of malaria antigens are being tested ascandidate malaria vaccines, a major barrier to the development of an effective vaccine is the lack of a suitable human adjuvant capable of inducing a strong and long lasting immune response. In this study, the ability of AFCo1, a potent T and B cell adjuvant based on cochleate structures derived from meningococcal B outer membrane proteoliposomes (MBOMP), to boost the immune response against two Plasmodium falciparum antigens, merozoite surface protein 4 (MSP4) and 5 (MSP5), was evaluated. METHODS: Complete Freund's adjuvant (CFA), which is able to confer protection against malaria in animal MSP4/5 vaccine challenge models, was used aspositive control adjuvant. MSP4 and 5-specific IgG, delayed-type hypersensitivity (DTH), T-cell proliferation, and cytokine production were evaluated in parallel in mice immunized three times intramuscularly with MSP4 or MSP5 incorporated intoAFCo1, synthetic cochleate structures, CFA or phosphate buffered saline. RESULTS: AFCo1 significantly enhanced the IgG and T-cell response against MSP4 and MSP5, with a potency equivalent to CFA, with the response being characterized by bothIgG1 and IgG2a isotypes, increased interferon gamma production and a strong DTH response, consistent with the ability of AFCo1 to induce Th1-like immune responses. CONCLUSION: Given the proven safety of MBOMP, which is already in use in a licensed human vaccine, AFCo1 could assist the development of human malaria vaccines that require a potent and safe adjuvant(AU)

Teratoma quístico maduro con transformación maligna: a propósito de un caso / Mature cystic teratoma with malignant transformation: a purpose of a case

Paciente femenina de 26 años de edad, quien es referida en posoperatorio de ooforectomía derecha, cuya biopsia reporta “teratoma quístico maligno, patrón epitelial tipo adenoescamoso”. Es reintervenida con laparotomía estadificadora donde se evidencia hallazgos de plastrones tumorales en peritoneo pélvico, mesocolon y epiplón, además múltiples adenopatías pre aórticas, ausencia de ovario derecho, y ovario izquierdo con tumor quístico de 6 cm de diámetro, adherido a la trompa ipsilateral y el piso pélvico. La biopsia definitiva reporta “adenocarcinoma moderadamente diferenciado con diferenciación escamosa infiltrando ovario izquierdo, pared de la trompa uterina izquierda, parametrios derecho, izquierdo, piso pélvico, peritoneo y epiplón inframesocólico”. Recibe quimioterapia adyuvante con esquema de paclitaxel y carboplatino por 4 ciclos. Al mes de terminar el tratamiento adyuvante presenta sangrado genital, evidenciándose lesión en cúpula vaginal cuya biopsia reporta “carcinoma epidermoide moderadamente diferenciado”. Se planifica tratamiento con doxorrubicina, ifosfamida y mesna combinado con radioterapia externa pélvica y braquiterapia complementaria.

We report the case of 26 year old female patient who is referred after undergoing right oophorectomy. The pathology of the surgical specimen was reported as “malignant cystic teratoma with adenosquamous epithelial pattern”. Staging laparotomy is performed. The surgical findings were: tumor implants in pelvic peritoneum, mesocolon, omentum. paraortic lymphadenopaties. Left ovary with 6 cm cystic tumor on its surface firmly adhered to the ipsilateral fallopian tube and pelvic wall. Absence of right ovary. Pathology reports “moderately differentiated adenocarcinoma with squamous differentiation infiltrates left ovary, left fallopian tube, right and left parametria, pelvic wall and inframesocolic omentum”. Adjuvant chemotherapy based on paclitaxel carboplatin for 4 cycles. One month after ending adjuvant treatment refers vaginal bleeding. After detailed gynecologic evaluation, an exophytic lesion on the vaginal vault is found. The biopsy reports “moderately differentiated squamous carcinoma”. Combined therapy based on chemotherapeutic agents (doxorubicin, iphosphamide and mesna) and external radiotherapy was scheduled.

Epidemeología y factores de riesgo: en pacientes con adenocarcinoma de endometrio / Risk factors and epidemeology: in patients with endometrial adenocarcinoma

El adenocarcinoma de endometrio es causa importante de morbilidad y mortalidad, correlacionado con aumento en uso de terapia de reemplazo hormonal en posmenopáusicas. Estudio descriptivo corte transversal; se determinaron características epidemiológicas, factores de riesgo en pacientes evaluadas entre enero 1992 diciembre 2005. En 129 pacientes: mayoría entre 70- 80 años. Menarquía antes 10 años 5, de 10 a 12 años 45, 13 a 15 años 48, > de 15 años 7 y desconocida en 24. Premenopáusicas 18 pacientes, posmenopáusicas 90, desconocido 21. Nuligestas 21, de 1 a 5 gestas 64, de 6 a 10 20, más de 10 gestas 10, desconocidas 14. Hipertensión 35, diabéticos 10, tabáquicos 18, obesidad 5, alcohólicos 5, cáncer 3, otros 8, desconocidos 21, sin antecedentes 34. Antecedentes oncológicos familiares: cáncer de mama 9, cáncer de endometrio 2, otros cánceres ginecológicos 10, cáncer de vías digestivas 14, otros cánceres 12, desconocidos 21 sin antecedentes 78. Estadio clínico 38, quirúrgico 81, no se clasificaron 10 casos. Estadio clínico: Ia 2 Ib 5 II 21 III 5, IVa 1 IVb 4. Estadio quirúrgico: Ia 10, Ib 28, Ic 15, IIa 6, IIb 3, IIIa 8, IIIb 5, IVa 1 y IVb 3. Tipo histológico: endometrioide 115, seroso-papilar 7, adenoescamoso 2, células claras 2, células pequeñas 1, indiferenciado 2, grado histológico: Bien diferenciado 75, moderadamente diferenciado 40 poco diferenciado 17. Realizar más estudios y compararlos con otras series.

Neoplasias infiltrantes de vulva / Infiltrant vulvar neoplasia

El cáncer de vulva es una enfermedad de mujeres en edad avanzada, también ha sido observado en mujeres premenopáusicas. Generalmente es del tipo carcinoma de células escamosas, aunque se presentan otros tipos histológicos. La cirugía es la terapéutica de elección por ser más eficaz, incluye exéresis amplias, vulvectomías parciales, simples, radicales (esta última con vaciamiento inguino femoral) y exenteraciones pélvicas. La radioterapia y poliquimioterapia son terapéuticas generalmente adyuvantes al tratamiento quirúrgico. Trabajo de investigación de tipo retrospectivo, descriptivo con estudios de distribución de frecuencias. El universo correspondió a 36 pacientes ingresados al servicio de ginecología oncológica del Instituto de Oncología “Dr. Miguel Pérez Carreño” de la ciudad de Valencia, Venezuela, entre los años de 1995 y 2006, y con diagnóstico de neoplasia infiltrante de vulva. De la totalidad de las pacientes evaluadas se obtuvo que la mayor incidencia de esta patología se encontraba entre las pacientes mayores de 71 años con 36,11 por ciento. La histología más frecuente fue el carcinoma epidermoide. Más de un 75 por ciento de las pacientes ingresó en estadios avanzados. El tratamiento más realizado fue la vulvectomía radical con un 63,8 por ciento. Conclusión: En base a los datos obtenidos se sugiere lograr mejoras en los métodos de pesquisa para así lograr diagnósticos en etapas más tempranas, de esta manera evitar cirugías más agresivas que repercuten en un aumento de la morbi-mortalidad en este grupo de pacientes, y así lograr una mejor calidad de vida.

Motor impairment and neuronal damage following hypothermia in tropical amphibians.

Although the induction of mild to moderate cerebral hypothermia in mammals can have neuroprotective activity, some deleterious effects have been described when inducing deep hypothermia during cooling of the brain. In the spinal cord, rapid deep cooling can induce seizure activity accompanied by release of the excitatory neurotransmitters, glutamate and aspartate. We used cold-sensitive tropical amphibians as a model to determine (a) the critical temperature inside the central nervous system necessary to induce seizures during rapid cooling; (b) the survival rate during slow deep cooling of the whole animal; and (c) whether deep cooling can cause neuronal cell damage. Seizures induced by deep rapid (or=30 min) deep cooling of the whole animal (12 h at 2-3 degrees C), around 70% of animals died. Spinal reflexes were enhanced when temperatures within the spinal cord reached between 9.0 degrees C and 11.6 degrees C. A fivefold increase in blood glucose level was observed during slow deep cooling. Recovery after slow deep cooling was accompanied by motor impairment and the main histological findings were condensation of the cytoplasm and nuclear pyknosis. Severe neuronal cell damage was characterized by swelling, vacuolated cytoplasm with distended neuronal bodies. These results indicate that deep cooling can easily induce neuronal cell damage in the central nervous system of cold-sensitive animals. They also warn us to the potential sequels associated with the use of deep brain cooling as a neuroprotective strategy.

Bacterial derived proteoliposome as ideal delivery system and cellular adjuvant.

We explored the potential of a proteoliposome (PL) from the outer membrane of N. meningitidis B, as an immunopotentiator and as a vector for antigen delivery to dendritic cells (DC). DC were incubated with PL resulting in up-regulation of MHC-II, CD40, CD80, and CD86 expression and production of TNFalpha and IL12(p70). Ovoalbumin (OVA) was incorporated within PL (PL-OVA). PL-OVA presented OVA-specific peptides to CD4+ and CD8+ OVA-specific T-cell hybridomas. PL exerts an immunomodulatory effect on DC and is a general system to deliver antigens for presentation to CD4+ and CD8+ T-cells possibly implicated in the induction CD8+ cytotoxic T lymphocytes (CTLs) responses.

Proteoliposome derived cochleate as novel adjuvant.

Cochleate structures (CS) consist in a highly stable lipid structures that have been reported to be a good antigen delivery system. The incorporation of pathogen associated molecular pattern (PAMP) from bacterial membranes into CS became in a promising approach to develop adjuvants, particularly mucosal adjuvants. Therefore, we prepare CS from proteoliposome (PL) obtained from Neisseria meningitidis B (PLCS) and evaluated it for its capability to stimulate the immune system as well as the adjuvant activity. The ability of PLCS to induce Thl polarization was also explored. The results and the easy capability for new antigen incorporation on CS support its use as adjuvant for immunization with a large variety of pathogen derived antigens and different routes of immunization.

The adjuvant potential of synthetic alkylglycerols.

Alkylglycerols (AGs) have shown immune stimulant and adjuvant activity in many studies, but natural sources are not so accessible and their extraction from them is very complicated. Therefore, a group of chemists at IFAL have synthesized AG analogs. The aim of this work was to evaluate the adjuvant potential of different synthetic AGs. A mix of ovoalbumin (Ova) and AGs increase anti-Ova IgG antibodies production in sera of immunized mice. The predominant subclass was IgG1 although higher levels of IgG2a were observed as the carbon chain length of AGs increased. AGs also induced the production of IL-12 and nitric oxide (NO) in the U937 human histiocyte and J774 mouse macrophage cell lines, respectively. These results indicate that synthetic AGs are effective adjuvants for the standardized antigen, Ova.

Bacterial derived proteoliposome for allergy vaccines.

One current approach in developing anti allergic vaccines is the use of potent adjuvants, capable of inducing Th1 or T regulatory cells. Proteoliposomes (PL) could be a suitable adjuvant. Purified Dermatophagoides siboney (Ds) allergens were mixed with PL and adsorbed into Al(OH)3 and evaluated in mice. The Th1/Th2 responses were measured at classes, subclasses, cytokines, and DTH levels. Anti Ds response was deviated to a Thl pattern, with the production of IgG2a and gamma1FN. A positive DTH response and a dramatic decrease of specific IgE and IL5 were not detected. The low dose was more effective than high dose. These results clearly support the potential use of PL as possible adjuvants for anti-allergic vaccines.