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Biomedical subjects

G Gregoriadis

Publications and source records attributed to G Gregoriadis.

At least 55 records · Page 3Linked to original sources
PubMed1994

The immunological adjuvant and vaccine carrier properties of liposomes.

Animal immunization studies by numerous laboratories have shown that liposomes promote humoural and cell-mediated immunity to a wide spectrum of bacterial, protozoan and viral antigens as well as tumour cell antigens, venoms and allergens. Adjuvanticity depends on liposomal structural characteristics which determine vesicle fate in vivo and, thus, the mode of antigen interaction with antigen-presenting cells. Adjuvanticity is further promoted by receptor mediated targeting of liposomes to macrophages, or the presence of other adjuvants including cytokines. The immunoadjuvant function of liposomes is supplemented by their ability to act as a carrier for co-entrapped B and T-cell epitopes, thus eliminating the need for a carrier protein. Recently, a technique has been developed for the entrapment of live or attenuated microbial vaccines into giant liposomes under conditions which retain their viability. Liposomes containing microbial vaccines (together with other soluble antigens or cytokines) could be used as carriers of vaccines in cases where there is a need to prevent interaction of vaccines with maternal antibodies or preformed antibodies to vaccine impurities.

Adjuvants, Immunologic↗
PubMed1994

Entrapment of cyclodextrin-drug complexes into liposomes: potential advantages in drug delivery.

A novel concept in drug delivery discussed here, takes advantage of certain properties of the drug "containers" cyclodextrins and liposomes to combine them into a single system thus circumventing problems associated with both systems. The concept, entailing entrapment of water-soluble cyclodextrin-drug inclusion complexes in liposomes, would allow accommodation of insoluble drugs in the aqueous phase of vesicles. This would potentially increase the drug to lipid mass ratio to levels above those attained by conventional drug incorporation into the lipid phase, enlarge the range of insoluble drugs amenable to encapsulation to include, for instance, membrane destabilizing agents, allow targeting of complexes to specific sites and reduce toxicity. In the present work, soluble inclusion complexes of hydroxypropyl-beta-cyclodextrin with dehydroepiandrosterone, retinol and retinoic acid were prepared and entrapped into multilamellar liposomes by the dehydration-rehydration procedure. Complex-containing liposomes were then exposed to blood plasma. Results show that complex entrapment into liposomes depends on the lipid composition used. Nearly all of the cyclodextrin and considerable portions of the drugs were found to remain associated with the carrier in the presence of plasma.

Animals↗
PubMed1993

Liposomes as vaccine carriers. Incorporation of soluble and particulate antigens in giant vesicles.

Giant liposomes (mean diameter 5.5 microns) composed of egg phosphatidylcholine or distearoyl phosphatidylcholine, phosphatidyl glycerol, cholesterol and triolein were prepared by a double emulsion technique. They were then mixed with model particulate (killed Bacillus subtilis, and killed Bacille Calmette-Guérin) and soluble (tetanus toxoid) vaccines and freeze-dried. Rehydration of the powder resulted in the generation of vesicles of similar mean diameter and diameter range, containing up to 27% (mean value) of the materials used for entrapment. Separation of entrapped from non-entrapped material was carried out by sucrose gradient centrifugation (B. subtilis and BCG) or centrifugation at 600 x g (toxoid). Light microscopy of liposomes containing B. subtilis labelled with fluorescein isothiocyanate revealed the presence of bacteria in individual vesicles which, in separate studies, were also found to entrap latex particles (0.5 and 1.0 micron diameter). Bacteria-containing liposomes could be freeze-dried in the presence of trehalose with most (83-87%) of the entrapped material recovered within the vesicles on reconstitution with saline. Liposomes were also shown to retain quantitatively their content of B. subtilis and, to a lesser extent, toxoid in the presence of mouse plasma at 37 degrees C and in situ after intramuscular injection into mice, for up to 24 h. Since liposomes are known (Gregoriadis, G. (1990) Immunol. Today 11, 89) to act as immunological adjuvants and vaccine carriers, giant vesicles containing microbes (live or attenuated if needed since the conditions of entrapment are mild) and, when appropriate, soluble antigens, could be used as multiple vaccines to ensure simultaneous presentation of antigens to immunocompetent cells.

Adjuvants, Immunologic↗
PubMed1993

Coupling of ligands to liposomes independently of solute entrapment: observations on the formed vesicles.

Bovine serum albumin (BSA), employed as a model ligand, was covalently linked (about 16% of the amount used) to small unilamellar vesicles (SUV) composed of phospholipid, cholesterol and N-(p-aminophenyl)stearylamide (APSA) (molar ratios 1:1:0.05). SUV with bound BSA were then used to generate dehydration-rehydration vesicles (DRV) in the presence of tetanus toxoid and/or carboxyfluorescein (CF). Nearly all of the SUV-bound BSA (about 15% of the original amount) was recovered in the multilamellar DRV formed, with a considerable proportion (42-62%) of the ligand becoming available on the outer bilayers. This apparent spatial reorientation of BSA within DRV also caused the entrapped toxoid to shift to some extent to the liposomal surface. There was no significant difference in the z average mean size between DRV with and without coupled BSA (543 and 555 nm diameter, respectively). Percent number diameter distribution data revealed that 71.2 (BSA-free) and 76.4% (BSA-containing DRV) of the vesicles had diameters of about 300-440 and 330-420 nm, respectively. However, in terms of percent mass diameter distribution, 69.5% (BSA-free) and 65.2% (BSA-containing DRV) of the mass was in vesicles with corresponding ranges of diameter of 1381-2975 and 1086-2840 nm. Vesicle size heterogeneity in both preparations was confirmed by freeze-fracture electron microscopy which also indicated that structures with or without bound BSA, were mostly vesicular of the multilamellar type. Judging from CF latency values, ligand-bearing DRV were stable on incubation with blood plasma at 37 degrees C for 24 h. Stability was, however, reduced significantly when the amount of ligand bound was excessive. The present approach allows for the coupling of ligands to and the entrapment of antigens and other labile solutes in liposomes independently, thus avoiding potential damage of such solutes by the coupling reagents.

Cholesterol↗
PubMed1993

Polysialic acids: potential in drug delivery.

A number of bacterial polysialic acids were injected intravenously into mice. Half-lives (up to 40 h) in the blood circulation were dependent on the polysialic acid used, increased by deacylation of their phospholipid moiety, decreased with shorter chain derivatives and appeared to be dose independent. A model drug (fluorescein) covalently coupled to a polysialic acid was found to assume the half-life of its carrier. Results suggest that intact or deacylated polysialic acids and shorter chain derivatives can be used to augment the half-lives of drugs, small peptides, proteins and drug delivery systems in the blood circulation, thus prolonging their pharmacological action.

Animals↗
PubMed1993

The subclass IgG responses of mice to influenza surface proteins formulated into liposomes.

Unprimed mice and mice primed by prior infection with an H1N1 subtype of influenza virus were used to assess the total and subclass IgG serum antibody responses to influenza virus A/Sichuan/2/87 (H3N2) surface haemagglutinin and neuraminidase proteins incorporated into four different formulations of liposomes. Only one of these liposome preparations, DSPC(B), induced greater total IgG, and subclass IgG1 and IgG2a antibody levels, in sera from both primed and unprimed mice than the aqueous A/Sichuan surface preparations alone administered at equivalent levels of haemagglutinin protein. The same DSPC(B) liposome formulation of A/Sichuan antigens was also the only preparation found to elicit levels of IgG2b and IgG3 subclass antibodies above baseline values in these animals.

Animals↗
PubMed1993

Immunopotentiation of local and systemic humoral immune responses by ISCOMs, liposomes and FCA: role in protection against influenza A in mice.

The immunogenicity and protective efficacy of an influenza A subunit vaccine preparation administered to mice in an aqueous form, or presented as immunostimulatory complexes (ISCOMs), liposomes or with Freund's complete adjuvant (FCA), were assessed in comparative studies with live infectious virus. Both intranasal and parenteral routes of administration were assessed. An enzyme-linked immunosorbent assay (ELISA) was used to measure nasal wash and serum antibody responses in groups of unprimed mice, while protection was determined by the recovery of homologous influenza virus from mouse nasal washes and lung homogenates following challenge infection by the intranasal route. The results showed that parenteral administration of the influenza antigen preparations induced variable levels of both local and systemic antibodies at weeks 3, 7 and 22 postimmunization. Although the overall greatest levels of antibody and protection were elicited in mice following live virus infection, formulation of influenza surface haemagglutinin (HA) and neuraminidase (NA) proteins into ISCOMs elicited high and persistent antibody responses and provided relatively good protection of the upper and lower respiratory tracts of these animals. The results also show a relatively poor effect of the subunit antigen preparations in promoting humoral immune responses and protection irrespective of the nature of their presentation, when given by the intranasal route.

Adjuvants, Immunologic↗
PubMed1993

Liposomes in drug delivery. Clinical, diagnostic and ophthalmic potential.

Liposomes (phospholipid-based vesicles) have been investigated since 1970 as a system for the delivery or targeting of drugs to specific sites in the body. Because of their structural versatility in terms of size, composition, surface charge, bilayer fluidity and ability to incorporate almost any drug regardless of solubility, or to carry on their surface cell-specific ligands, liposomes have the potential to be tailored in a variety of ways to ensure the production of formulations that are optimal for clinical use. This includes controlled retention of entrapped drugs in the presence of biological fluids, controlled vesicle residence in the blood circulation or other compartments in the body, and enhanced vesicle uptake by target cells. Accumulated in vivo evidence, particularly in areas such as cancer chemotherapy, antimicrobial therapy, vaccines, diagnostic imaging and the treatment of ophthalmic disorders has indicated clearly that some liposome-entrapped drugs and vaccines exhibit superior pharmacological properties to those observed with conventional formulations. Such work has encouraged the application of liposomes in the treatment of diseases in humans. A large number of trials in patients with cancer or infections suggest that certain liposomal drug formulations are likely to prove clinically useful.

Adjuvants, Immunologic↗
PubMed1993

Fat soluble vitamins in liposomes: studies on incorporation efficiency and bile salt induced vesicle disintegration.

A systematic investigation has been undertaken to study the possible role of liposomes in facilitating the absorption of fat soluble vitamins (FSV) after oral administration in situations where FSV absorption is impaired. Conditions for the optimal incorporation of vitamins A, D3 and E individually or in combination, into multilamellar liposomes composed of egg phosphatidylcholine, dipalmitoyl phosphatidylcholine or distearoyl phosphatidylcholine (total vitamin to phospholipid molar ratios of 0.5:1, 0.75:1 and 0.9:1) have been studied. When two or three FSVs were incorporated simultaneously, FSV molar ratios used were based on the daily allowances (RDA) recommended for human males by the US Food and Nutrition Board. [Monitoring of FSV incorporation was carried out by the assay of 3H radioactivity (vitamins D3 or E, incorporated either individually or together with vitamin A), fluorometrically (vitamin A) or by an HPLC method developed in the present study for the assay of all three FSV simultaneously.] Incorporation yield of FSV used alone or in combination was found to decrease with increasing total vitamin to phospholipid molar ratio and to broadly depend on the phospholipid used. Although a vitamin to phospholipid molar ratio of 0.5:1 was found optimal in terms of percent vitamin incorporation (up to about 94% of the amount used), greater absolute amounts of FSV were recovered in most of the formulations with higher molar ratios. Moreover, when two or three FSV were incorporated simultaneously, amounts recovered with liposomes were reasonably similar to the RDA used initially.(ABSTRACT TRUNCATED AT 250 WORDS)

Bile Acids and Salts↗
PubMed1993

Liposome-entrapped T-cell peptide provides help for a co-entrapped B-cell peptide to overcome genetic restriction in mice and induce immunological memory.

We have investigated the possibility of a T-cell epitope peptide providing help for a B-cell epitope peptide when both peptides are co-entrapped in the same liposomes. Epitope models used were a 28 amino acid peptide from the S region of the hepatitis B surface antigen (HBsAg) (subtype adw) containing an H-2s Th-cell epitope, and a 33 amino acid peptide from the pre-S1 region of the HBsAg (subtype adw) designed to exclude an adjacent H-2s T-cell epitope, the latter (pre-S1) peptide being recognized by SJL (H-2s) mice as a B-cell epitope. SJL(H-2s) mice were immunized twice intramuscularly with S or pre-S1 peptide alone, co-entrapped in the same liposomes or entrapped in separate liposomes which were mixed before injection. Analysis of sera for anti-peptide IgG1 antibodies revealed that the Th-cell peptide provided help for the pre-S1 peptide only when the two peptides were co-entrapped in the same vesicles. This helper effect was found to correlate with the ability of S peptide (co-entrapped with the pre-S1) to stimulate T-cell proliferation in vitro. There was no IgG1 response against pre-S1 peptide in mice immunized with a mixture of the free peptides or a mixture of separately entrapped peptides. A helper effect, albeit much weaker, was also observed in mice immunized with the two peptides emulsified in incomplete Freund's adjuvant. Antisera from mice immunized with both peptides co-entrapped in liposomes were found to bind to full length (pre-S1 containing) recombinant HBsAg. Moreover, binding values were much higher than those seen with antisera from animals immunized with the liposomal S peptide above, presumably because of full access of anti-pre-S1 antibodies to the pre-S1 region of the rHBsAg. It is concluded that liposomes could serve not only as an immunological adjuvant for peptides but also as a carrier for Th- and B-cell epitopes thus eliminating the need for covalent linkage to a carrier protein.

Adjuvants, Immunologic↗
PubMed1992

Liposomes enhance the immunogenicity of reconstituted influenza virus A/PR/8 envelopes and the formation of protective antibody by influenza virus A/Sichuan/87 (H3N2) surface antigen.

Reconstituted influenza virus (A/PR/8 strain) envelopes (RIVE) and influenza virus (A/Sichuan/87 (H3N2) strain) surface antigens were entrapped in dehydration-rehydration vesicles (DRV liposomes) composed of egg phosphatidylcholine (PC) or distearoyl phosphatidylcholine (DSPC DRV) and equimolar (32 mumol) cholesterol. Entrapment values for RIVE were 31.2 (PC) and 29.4% (DSPC DRV) of the material used. Corresponding entrapment values for the A/Sichuan/87 strain antigens were 40.7 and 39.3%. Balb/c mice injected intramuscularly with PC or DSPC DRV liposomes containing 0.1 and 1.0 microgram RIVE exhibited primary (higher dose only) and secondary responses (IgG1) which were significantly higher than those obtained in mice injected with identical amounts of non-entrapped RIVE. Significantly higher secondary responses were also observed for the IgG2a and IgG2b subclasses. In experiments designed to assess the effectiveness of DRV liposomes as a carrier of influenza virus antigens in a potential vaccine, hamsters were immunized intramuscularly with 0.1, 0.5 and 5.0 micrograms of free or liposome-entrapped influenza A/Sichuan/87 surface antigens. Results showed increased haemagglutination inhibition (HI) antibody levels in terms of both primary (0.5 and 5.0 micrograms doses) and secondary (all doses) responses in the sera of animals treated with the liposomal formulations. DSPC compared with PC DRV exhibited greater adjuvanticity when the lower doses of antigens were used.

Adjuvants, Immunologic↗
PubMed1992

Antibodies against phosphatidylinositol and inositol monophosphate specifically inhibit tumour necrosis factor induction by malaria exoantigens.

The active component of the exoantigens of malarial parasites which stimulates macrophages to secrete tumour necrosis factor (TNF) has been shown to depend upon a phospholipid, the activity of which was blocked by phosphatidylinositol (PI) and inositol monophosphate (IMP) in competitive inhibition studies. Antisera made against the exoantigens of Plasmodium yoelii, which inhibited their induction of TNF, were found by an ELISA assay to contain antibody against several other phospholipids. However, the inhibitory antibody was removed specifically by adsorption with liposomes containing PI, but not other phospholipids. Furthermore, PI was the only phospholipid in non-liposomal form which induced the production of inhibitory antisera. Mice immunized with IMP, but not inositol, also produced inhibitory antisera. When incorporated into liposomes several other phospholipids did give rise to inhibitory antibodies but, in contrast to the antisera against parasite exoantigens, PI and IMP, the inhibitory activity was removed by adsorption with heterologous phospholipid liposomes, suggesting that it was directed against a common determinant, presumably the phosphate ester head group. Inhibitory antibodies in the antisera tested were predominantly IgM and titres were not increased after repeated injections. Antisera raised against PI, IMP or the cross-reacting phospholipid liposomes also inhibited TNF secretion by macrophages stimulated by exoantigens of the human parasites P. falciparum and P. vivax, but not by bacterial lipopolysaccharide. These findings confirm our conclusion that exoantigens from these different species contain phosphate bound to inositol in their TNF-inducing moiety.

Animals↗
PubMed1991

Influence of surface hydrophilicity of liposomes on their interaction with plasma protein and clearance from the circulation: studies with poly(ethylene glycol)-coated vesicles.

Well-defined liposome systems have previously established the influence of size, surface charge lipid composition and surface ligands, on in vivo fate and behaviour of model compounds entrapped in liposomes. In the present study, preformed liposomes which quantitatively retain aqueous markers were covalenty coupled via dipalmitoylphosphatidyl-ethanolamine, to the hydrophilic polymer, monomethoxypoly(ethylene glycol) (MPEG 5000). Such liposomes retain the coating in the presence of plasma, and appear to adsorb plasma components more slowly than liposomes without the polymer, shown using an aqueous two-phase partitioning technique. MPEG-coupled liposomes were cleared from the blood circulation up to 30% more slowly than liposomes without MPEG after intravenous administration to mice, despite the unmodified liposomes being of a composition and size shown previously to favour achievement of maximum half-life. It is suggested that the polymer acts as a surface barrier to plasma factors which otherwise bind to liposomes in the blood and accelerate vesicle removal.

1,2-Dipalmitoylphosphatidylcholine↗
PubMed1991

Overview of liposomes.

Many of the problems associated with conventional drug therapy may be circumvented by the use of delivery systems which, in a variety of ways, will optimize drug action. Among the systems being investigated, liposomes hold considerable promise. Versatility in both the structural characteristics of liposomes and their ability to accommodate a wide range of drugs has contributed to the design of appropriate formulations with optimal pharmacological actions. Areas in which such formulations have been already applied successfully experimentally or clinically, include cancer chemotherapy, antimicrobial therapy and vaccines.

Animals↗