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HIV-1 Tat-coated nanoparticles result in enhanced humoral immune responses and neutralizing antibodies compared to alum adjuvant.

HIV-1 Tat has been identified as an attractive target for vaccine development and is currently under investigation in clinical trials as both a therapeutic and preventative vaccine for HIV-1. It is well known that protein based vaccines produce poor immune responses by themselves and therefore require adjuvants to enhance immune responses. We have previously reported on the use of anionic nanoparticles (NPs) for enhancing cellular and humoral immune responses to Tat (1-72). The purpose of this study was to further evaluate the immune response of HIV-1 Tat (1-72) coated on anionic nanoparticles compared to alum using various doses of Tat (1-72). Nanoparticles were effective at generating comparable antibody titers at both 1 and 5 microg doses of Tat (1-72), whereas the antibody titers significantly decreased at the lower dose of Tat (1-72) using alum. Anti-sera from Tat (1-72) immunized mice reacted greatest to the N-terminal and basic regions of Tat, with the NP groups showing stronger reactivity to these regions compared to alum. Moreover, the anti-sera from all Tat (1-72) immunized groups contained Tat-neutralizing antibodies and were able to significantly inhibit Tat-mediated long terminal repeat (LTR) transactivation.

AIDS Vaccines↗

Immunogenicity and lower dose requirement of polymer entrapped tetanus toxoid co-administered with alum.

Low adjuvanticity of microparticles based vaccine formulation necessitates the use of alum along with particles to elicit improved antibody titers from single point immunization. It was observed that antibody response from immunization with admixture of alum and polymer entrapped antigen was dependent on particle size, amount of antigen released during burst phase and dose of microencapsulated antigen. In the animals immunized with polymer entrapped tetanus toxoid (TT) very large particles (50-150 microm) did not elicited high antibody titers where as microparticles in the range 2-8 microm exhibited remarkable improvement in the antibody response. Very small size particles (<2 microm) were also not as effective as 2-8 microm size particles for generation of antibody response. Presence of alum improved the immune response by adsorbing the burst released antigen from the particle surfaces. Role of alum in potentiation of immune response from polymer entrapped TT was highly significant at lower dose regimes. Polymer entrapped TT as little as 0.1 Lf when immunized along with alum generated antibody responses superior to those elicited by 10 Lf soluble antigen. Immunization with admixture of particles and alum generated two to three times higher antibody titer than that observed from immunization with similar single doses of alum adsorbed TT. Single point immunization of admixture of particles entrapped TT and alum generated sustained long lasting antibody responses comparable to two divided doses of alum-adsorbed antigen. Superiority of single dose polymeric formulation in comparison to two divided doses of alum adsorbed TT was more evident at lower doses of TT immunization. This reflected the profound synergistic effect of both the adjuvants at lower doses. Affinity of antibodies generated from single point immunization was comparable to that achieved with two doses of alum adsorbed TT immunization. Particle alone elicited more of IgG2a type antibody where as immunization with admixture of alum and particles improved the overall antibody response and more of Th2 type response.

Alum Compounds↗

Surrogate markers of immunity to Leishmania major in leishmanin skin test negative individuals from an endemic area re-visited.

BACKGROUND: In the screening of vaccine candidates it is important to select candidates that evoke immune responses associated with protection. Valid surrogate markers against human leishmaniasis are still lacking. METHODS: A controlled injection of live Leishmania known as leishmanization, (LZ), was used to evaluate vaccine (alum-precipitated autoclaved Leishmania major with BCG) efficacy and more accurately define surrogate markers of immunity to leishmaniasis in humans. Cellular immune responses to this artificial infection were monitored in the volunteers prior to and 9 months post infection. Comparisons were made between those who developed a lesion after infection and those who did not. RESULTS: In the volunteers monitored there was no significant difference in LST, IFNgamma production, or source of IFNgamma between those who developed a lesion and those who did not after LZ, with the exception that ulcer development was associated with an enhanced number of IFNgamma secreting CD4(+) CD45RA(-) (memory) T cells. DISCUSSION: Ulcer development following LZ was lower than anticipated by a pilot study (47% versus 78%) using the same stabilate several years earlier. While this may be an effect of low viability/virulence of the LZ inocula, alternative explanations are also possible. The IFNgamma responses in the study subjects were significantly lower compared to volunteers with previous history of cutaneous leishmaniasis. The findings raise the possibility that the selection of LST-negative volunteers in an endemic area may bias the study towards potentially non/low L. major-reactive volunteers.

Adjuvants, Immunologic↗

RLJ-NE-299A: a new plant based vaccine adjuvant.

Alum has been in use since long as an adjuvant for vaccines. However, its use as a vaccine adjuvant offers limitation in supporting cell mediated response. Therefore, a new plant based product RLJ-NE-299A from Picrorhiza kurroa reported for its immunostimulatory activity, has been explored for its potential as an alternative adjuvant. In order to compare the adjuvant activity with alum, antigen-specific immune responses were evaluated following immunization with a formulation containing hepatitis B surface antigen (HBsAg) adjuvanted with RLJ-NE-299A and alum in mice. The adjuvant RLJ-NE-299A up-regulated remarkably the expression of Th1 cytokines IL-2, IL-12, IFN-gamma, TNF alpha and Th2 cytokine IL-4 in lymph node cell cultures after 2 weeks of primary immunization with HBsAg. Further, the levels of both immunoglobulins IgG2a (Th1) and IgG1 (Th2) subtypes increased profoundly in blood sera of mice immunized with HBsAg/RLJ-NE-299A. The results indicated that RLJ-NE-299A has strong potential to increase both cell mediated and humoral immune responses and is capable of sustaining the total antigen-specific antibody response. Besides, the RLJ-NE-299A provides a signal to gear up both CD4 helper cells (Th1 and Th2) and CD8 cells populations, which may have important implications for vaccination against hepatitis B virus. Variable doses of RLJ-NE-299A (0.312-40 microg) containing vaccine antigen (HBsAg) were well tolerated with optimum T cell response at 2.5 microg/ml. Not only this, the adjuvant was also able to induce cellular immune responses to HBsAg as evidenced by Th1 and Th2 cytokines upregulation, which enabled mice to overcome the unresponsiveness to antigen HBsAg encountered with alum-adjuvanted vaccine in otherwise non-responding mice population. The study presents evidence that the HPLC standardized fraction RLJ-NE-299A, is an adjuvant of choice over alum in improving and maintaining the improved immune status against HBsAg, and may also prove useful adjuvant candidate with other vaccine antigens, too.

Adjuvants, Immunologic↗

Immunogenicity of a receptor-binding domain of SARS coronavirus spike protein in mice: implications for a subunit vaccine.

We studied the immunogenicity of an anti-SARS subunit vaccine comprised of the fragment of the SARS coronavirus (SARS-CoV) spike protein amino acids 318-510 (S318-510) containing the receptor-binding domain. The S protein fragment was purified from the culture supernatant of stably transformed HEK293T cells secreting a tagged version of the protein. The vaccine was given subcutaneously to 129S6/SvEv mice in saline, with alum adjuvant or with alum plus CpG oligodeoxynucleotides (ODN). Mice immunized with the adjuvanted antigen elicited strong antibody and cellular immune responses; furthermore, adding the CpG ODN to the alum resulted in increased IgG2a antibody titers and a higher number of INF-gamma-secreting murine splenocytes. Mice vaccinated with S318-510 deglycosylated by PNGase F (dgS318-510) showed a lower neutralizing antibody response but had similar numbers of INF-gamma-producing cells in the spleen. This finding suggests that carbohydrate is important for the immunogenicity of the S318-510 protein fragment and provide useful information for designing an effective and safe SARS subunit vaccine.

Alum Compounds↗

Poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) is a potent enhancer of mixed Th1/Th2 immune responses in mice immunized with influenza virus antigens.

We investigated the ability of a novel polyphosphazene polyelectrolyte, poly[di(sodium carboxylatoethylphenoxy)phosphazene] (PCEP) to enhance antigen-specific immune responses. BALB/c mice were immunized once subcutaneously with either bovine serum albumin (BSA) or influenza virus X:31 antigen alone, or in combination with PCEP, or either of the adjuvants poly[di(sodium carboxylatophenoxy)phosphazene] (PCPP) and alum. Both PCEP and PCPP significantly enhanced serum antigen-specific total IgG, IgG1 and IgG2a antibody titers, and these responses were highest in PCEP-immunized mice. Alum induced only a modest enhancement of antibody responses. Reducing the dose of X:31 antigen by 25-fold had no effect on antibody responses in mice immunized with PCPP and PCEP, but resulted in reduced titers in those immunized with alum. Analysis of X:31 antigen-specific cytokines revealed that alum and PCPP were associated with a predominantly IL-4 response. In contrast, PCEP was associated with production of both IFNgamma and IL-4. We conclude that PCEP is a potent enhancer of antigen-specific Th1 and Th2 immune responses and is a promising adjuvant for vaccine applications.

Adjuvants, Immunologic↗

Adjuvant-enhanced antibody responses to recombinant proteins correlates with protection of mice and monkeys to orthopoxvirus challenges.

Recombinant proteins are being evaluated as smallpox and monkeypox vaccines because of their perceived safety compared to live vaccinia virus. Previously, we demonstrated that three or more injections of a Ribi-type adjuvant with a combination of three proteins from the outer membranes of intracellular (L1 protein) and extracellular (A33 and B5 proteins) forms of vaccinia virus protected mice against a lethal intranasal challenge with vaccinia virus. Here, we compared several adjuvants and found that QS-21 and to a lesser extent alum+CpG oligodeoxynucleotides accelerated and enhanced neutralizing antibody responses to a mixture of L1 and A33 proteins, provided the highest ratio of IgG2a to IgG1 isotype response, and protected mice against disease and death after only two immunizations 3 weeks apart. In addition, monkeys immunized with recombinant vaccinia virus proteins and QS-21 developed neutralizing antibody to monkeypox virus and had reduced virus load, skin lesions, and morbidity compared to the non-immunized group following monkeypox virus challenge.

Adjuvants, Immunologic↗

Return sludge employed in enhancement of color removal in the integrally industrial wastewater treatment plant.

In this study, the recirculation of chemical sludge and integrated sludge were employed for enhancement of color removal in an integrally industrial wastewater treatment plant. The jar test was conducted for simulating chemical coagulation process with different coagulants, aluminum sulfate and polyaluminum chloride (PACl), after different activated sludge systems (with air or high-purity oxygen). The results showed that with sludge recirculation the process of coagulation for the color removal has up to 35% enhancement in comparison with no sludge recirculation. Meanwhile, the color removal enhancement of coagulant aluminum sulfate was approximately 2 to 3-times the color removal enhancement of coagulant PACl at the optimum ratio of return sludge. In coagulation process with influent color of 1220 true color unit (TCU), the optimum dosage of return sludge was 3010mg/L, when coagulant PACl or aluminum sulfate was employed. In coagulation process with influent color of 536 TCU, the optimum dosages of return sludge were 2340 and 4680mg/L using PACl and aluminum sulfate, respectively.

Alum Compounds↗

Floc morphology and cyclic shearing recovery: comparison of alum and polyaluminum chloride coagulants.

This study investigated the floc formation and re-aggregation potential for alum, polyaluminum chloride (PACl) and a blend of these coagulants. Bench-scale testing included floc morphology characterization for well-developed floc, post-shear floc, and non-settleable (filter influent) floc. Different applications of coagulants were observed to produce non-settleable floc that was morphologically different. The alum treatment had a decrease in average floc size from coagulated to non-settleable conditions, whereas the PACl and PACl/alum treatments resulted in similar sized floc between these processes. Zeta potential distribution measurements showed that the alum treatment resulted in a negative shift from coagulated to non-settleable conditions whereas the PACl and PACl/alum treatments had no significant shift. A photometric dispersion analyzer (PDA) was employed to compare the differences between coagulant treatments with respect to shear induced aggregate breakup and recovery. The PDA allowed a dynamic monitoring of initial floc aggregation and measured the degree of recovery from cyclic shearing. The degree of recovery from shearing was greatest for the PACl/alum treatment likely as a result of increased collision efficiency due to more effective charge neutralization.

Alum Compounds↗

Re-use of water treatment works sludge to enhance particulate pollutant removal from sewage.

This paper attempted to study the feasibility of reusing water treatment works sludge ("alum sludge") to improve particulate pollutant removal from sewage. The main issues focused upon were: (1) the appropriate dosage of the alum sludge, (2) the appropriate operating conditions, and (3) the possible mechanisms for enhancement by adding alum sludge. Actual alum sludge and sewage were applied to a series of jar tests conducted under various conditions. It has been found that both the SS and COD removal efficiencies could be improved by the addition of the alum sludge, which was mainly attributed to the removal of relatively fine particles with a size of 48-200 microm. The appropriate dosage of the alum sludge was determined to be 18-20 mg of Al/L. Increasing the mixing speed or reducing the floc size of the alum sludge enhanced the SS and COD removal and the dispersed alum sludge could remove particulate contaminants with smaller size than the raw sewage. ToF-SIMS evidence revealed that the aluminum species at the surface of the alum sludge were effectively utilized for improving the SS and COD removal. It was postulated that the sweep flocculation and/or the physical adsorption might play key roles in the enhancement of particulate pollutant removal from sewage.

Adsorption↗

Bench-scale testing of a magnetic ion exchange resin for removal of disinfection by-product precursors.

The objective of this research was to compare enhanced coagulation with anion exchange for removal of disinfection by-product (DBP) precursors (i.e. natural organic matter (NOM) and bromide). Treatment with a magnetic ion exchange resin (MIEX((R))) was the primary focus of this study. Raw waters from four utilities in California were evaluated. The waters had low turbidity, low to moderate organic carbon concentrations, a wide range of alkalinities, and moderate to high bromide ion concentrations. The treated waters were compared based on removal of ultraviolet (UV) absorbance, dissolved organic carbon (DOC), trihalomethane formation potential (THMFP), and haloacetic acid formation potential (HAAFP). The results indicated that treatment with MIEX is more effective than coagulation at removing UV-absorbing substances and DOC. Treatment with MIEX and treatment with MIEX followed by coagulation yielded similar results, suggesting that coagulation of MIEX-treated water does not provide additional removal of organic carbon. MIEX treatment reduced the THMFP and HAAFP in all waters, and did so to a greater extent than coagulation. Treatment with MIEX was most effective in raw waters having a high specific UV absorbance and a low anionic strength. Following MIEX treatment, subsequent chlorination resulted in a shift to the more brominated THM and HAA species as compared to chlorination of the raw water. MIEX also removed bromide to varying degrees, depending on the raw water alkalinity and initial bromide ion concentration.

Acetates↗

Sequential polymer dosing for effective dewatering of ATAD sludges.

Dewatering problems associated with the sludge from autothermal thermophilic aerobic digestion (ATAD) of sludge, result in large chemical conditioning costs for effective dewatering. A variety of chemical conditioners were used to improve dewatering, but none of them were able to dewater the sludge as desired at acceptable conditioning doses. It was found that during the digestion process, chemical precipitation of divalent cations occurred. Some of the ATAD sludge colloids were found to have a positive zeta potential and these were thought to be the precipitated divalent cations. Sequential polymer dosing using either iron or cationic polymer, followed by anionic polymer, was found to improve dewatering. The use of anionic polymer is essential and allows the use of smaller amounts of iron or cationic polymer for effective dewatering. The use of the less expensive anionic polymer along with cationic polymers has the potential to make the use of the ATAD process more economical.

Alum Compounds↗

Inactivation of particle-associated viral surrogates by ultraviolet light.

This study investigated whether colloid-sized particles can enmesh and protect viruses from 254-nm ultraviolet (UV) light and sought to determine the particle characteristics (e.g. size, chemical composition) that are most relevant in causing a protective effect. Two viral surrogates (MS2 coliphage and bacteriophage T4), three types of particles (kaolin clay, humic acid powder, and activated sludge), two coagulants (alum and ferric chloride), two filtration conditions (none and 0.45 microm), and two UV doses (40 and 80 mJ/cm2 for MS2 coliphage; 2 and 7 mJ/cm2 for bacteriophage T4) were considered in a series of bench-scale UV collimated beam experiments. Transmission electron microscopy was used to qualitatively confirm the phage particle-association after coagulation. Humic acid and activated sludge floc particles shielded both viral surrogates to a statistically significant degree (with >99% confidence) relative to particle-free control conditions, while the kaolin clay particles provided no significant protection. The results of the study suggest that particles <2 microm in diameter are large enough to protect viruses from UV light and that particulate chemical composition (e.g. UV-absorbing organic content) may be a critical factor in the survival of particle-associated viruses during UV disinfection.

Alum Compounds↗

Laboratory investigation of aluminum solubility and solid-phase properties following alum treatment of lake waters.

Water samples from two southern California lakes adversely affected by internal nutrient loading were treated with a 20 mg/L dose of Al3+ in laboratory studies to examine Al solubility and solid-phase speciation over time. Alum [Al2(SO4)3 . 18 H2O] applications to water samples from Big Bear Lake and Lake Elsinore resulted in a rapid initial decrease in pH and alkalinity followed by a gradual recovery in pH over several weeks. Dissolved Al concentrations increased following treatment, reaching a maximum of 2.54 mg/L after 17 days in Lake Elsinore water and 0.91 mg/L after 48 days in Big Bear Lake water; concentrations in both waters then decreased to <0.25 mg/L after 150 days. The solid phase was periodically collected and analyzed using X-ray diffraction (XRD), differential scanning calorimetry-thermogravimetric analysis (DSC-TGA), scanning electron microscopy (SEM), and surface area analyses to investigate the nature of the reaction products and crystallinity development over time. Poorly ordered, X-ray amorphous solid phases transformed over time to relatively well-ordered gibbsite, with strong diffraction peaks at 4.8 and 4.3 A. XRD also indicated the formation of a second (possibly aluminosilicate) crystalline phase after 150 days in Lake Elsinore water. Surface areas also decreased over time as crystals reordered to form gibbsite/microcrystalline gibbsite species. DSC-TGA results suggested that the initially formed amorphous Al(OH)3 underwent transformation to >45% gibbsite. These results were supported by geochemical modeling using Visual MINTEQ, with Al solubility putatively controlled by amorphous Al(OH)3 shortly after treatment and approaching that of microcrystalline gibbsite after about 150 days. These findings indicate that Al(OH)3 formed after alum treatment undergoes significant chemical and mineralogical changes that may alter its effectiveness as a reactive barrier to phosphorus release from lake sediments.

Alum Compounds↗

Evaluation of water treatment sludges toxicity using the Daphnia bioassay.

Alum and ferric chloride sludges from two water treatment plants (WTPs) were analyzed regarding their physicochemical characteristics and toxicity to Daphnia similis. Experiments were carried out in the dry and rainy seasons. Acute and chronic toxicity was measured using survival and reproduction as measurement endpoints. No acute toxicity of the sludge was observed in 48 h exposure. Ferric chloride sludge caused chronic toxicity, demonstrated by low fecundity and some mortality, while alum sludge caused chronic toxicity characterized by low fecundity. Some sludge characteristics varied between samplings, including turbidity, solids contents, N, P and metal (Al and Fe) concentrations. These variables and the increase of chemical oxygen demand (COD) were identified as the main cause of degradation of the receiving waters. However, no relationship was observed between these variables and degree of toxicity. It is apparent from these results that water treatment sludges may be toxic and therefore may impair receiving waters. Alum sludge was less toxic than ferric chloride sludge.

Alum Compounds↗

Submerged microfiltration membrane coupled with alum coagulation/powdered activated carbon adsorption for complete decolorization of reactive dyes.

Even the presence of very low concentrations of dyes (1mgL(-1)) in the effluent is highly visible and is considered aesthetically undesirable. It must be removed from wastewater completely. This study systematically evaluates the performance of adsorption (three kinds of powdered activated carbons), coagulation (AlCl3.6H2O) and membrane (submerged hollow fiber microfiltration) processes individually in treating two kinds of reactive dyes (Orange 16 and Black 5) and then using a hybrid process with combined coagulation-adsorption-membrane treatment system. Adsorption capacity and kinetics of Orange 16 were much higher and faster than those of Black 5. The dye removal efficiency by coagulation was highly dependent on dye concentration and solution pH. The hybrid process performance was far more superior that individual process in removing both kinds of dyes. It was evident that the combined coagulation-adsorption-membrane process has a great potential application for complete reactive dye removal, production of high-quality treated water and allows the reduction in the use of coagulant and adsorbent.

Adsorption↗

Influence of aging on phosphorus sorption to alum floc in lake water.

Alum is often added to eutrophic lakes to limit the release of phosphorus from sediments. This study quantified the effect of age and extent of crystallization on the phosphate (PO4-P) sorption capacity of alum floc. Aluminum hydroxide flocs were formed from alum addition at a dose of 25 mg/L of Al3+ to Big Bear Lake waters returned to the laboratory; flocs were then aged for 4, 20, 50, 120, and 180 days in the treated lake waters. The physical and mineralogical properties of the alum floc were characterized using surface area and thermal analyses. Phosphate sorption to the floc was evaluated using filtered lake water and NaCl/NaHCO3 solutions spiked with PO4-P concentrations ranging from 0 to 100 mg/L. The Langmuir model provided reasonable fits to data (r2 = 0.97-1.00), from which sorption constants and sorption maxima were determined. Phosphate sorption decreased with increased floc age and crystallinity and decreased surface area. Phosphate sorption maximum of the alum floc aged for 6 months was about 50% lower than freshly precipitated floc, while the binding constant, Kads, decreased approximately 65% over this same time period.

Adsorption↗

A pilot-scale evaluation of magnetic ion exchange treatment for removal of natural organic material and inorganic anions.

The objective of this research was to evaluate a magnetic ion exchange process (MIEX) for the removal of natural organic material (NOM) and bromide on a continuous-flow pilot-scale basis under different operating conditions and raw water characteristics. The most important operating variable was the effective resin dose (ERD), which is the product of the steady-state resin concentration in the contactor and the regeneration ratio. The raw water employed in this study had a moderate concentration of ultraviolet (UV)-absorbing substances and dissolved organic carbon (DOC), and a low turbidity, alkalinity, and concentration of competing anionic species. Experiments were conducted using the ambient raw water and raw water spiked with bromide, chloride, and sulfate. Substantial removal of UV-absorbing substances and DOC was achieved at ERDs as low as 0.16mL/L. Moderate bromide removal was achieved, depending on the ERD. Increasing the sulfate concentration resulted in decreased removal of UV-absorbing substances, DOC, and bromide. Consistent results were observed between the continuous-flow pilot plant tests and batch equilibrium studies.

Adsorption↗