Peter D Cooper

Australian National University | ANU · Immunology Department

In studying the molecular basis for the potent immune activity of previously described gamma and delta inulin particles and to assist in production of inulin adjuvants under Good Manufacturing Practice, we identified five new inulin isoforms, bringing the total to seven plus the amorphous form. These isoforms comprise the step-wise inulin developme...

We report a novel isoform of β-D-[2 → 1] poly(fructo-furanosyl) α-D-glucose termed delta inulin (DI), comparing it with previously described alpha (AI), beta (BI) and gamma (GI) isoforms. In vitro, DI is the most immunologically active weight/weight in human complement activation and in binding to monocytes and regulating their chemokine production...

Current ideas of the Origin of the Universe favour a Big Bang (Standard Cosmology SC) rather than some Steady State (Infinite Time) variant. Despite that, the SC as it stands is unsafe because each claimed support for the SC has a feasible alternative. In addition, profligate Universe-wide destruction of hydrogen still leaves it in excess. The conc...

Ever since humans realized that they were distinct conscious beings, it seems they wondered about their place in nature. Each pre-literate culture developed its own rationalization of how what it saw around came to be, handed on as oral history, often with a sacred spiritual basis involving discarnate ancestry and becoming an integral part of daily...

Background: Modern living is awash with low-frequency electromagnetic radiation raising concern over health effects, birth defects, and infant cancers especially leukemias. Medical/scientific opinion is ambivalent, especially regarding possible mechanisms of action despite our bodies׳ many electric currents. Aims: Are some cancers induced by mor...

This study combined physical data from synchrotron SAXS, FTIR and microscopy with in-silico molecular structure predictions and mathematical modeling to examine inulin adjuvant particle formation and structure. The results show that inulin polymer chains adopt swollen random coil in solution. As precipitation occurs from solution, interactions betw...

There is an ongoing need for new adjuvants to facilitate development of vaccines against HIV, tuberculosis, malaria and cancer, amongst many others. Unfortunately, the most potent adjuvants are often associated with toxicity and safety issues. Inulin, a plant-derived polysaccharide, has no immunological activity in its native soluble form but when...

There is a need for additional safe and effective human vaccine adjuvants. Advax™ is a novel adjuvant produced from semi-crystalline particles of delta inulin. In animal studies Advax enhanced humoral and cellular immunity to hepatitis B surface antigen (HBsAg) without inducing local or systemic reactogenicity. This first-in-man Phase 1 clinical tr...

We earlier identified a developmental series of seven isoforms/polymorphs of microparticulate inulin by comparing non-covalent bonding strengths. Their pharmaceutical utility lies in the modulation of cellular immunity, exploited as vaccine adjuvants (Advax™) especially for delta inulin (DI). As such particles cannot be sterilized by filtration we...

Active immunotherapy or immunopotentiation against human cancer is an attractive possibility because it suggests a natural treatment in which the body’s own defenses can be mobilized. A treatment modality that could be less mutilating for the patient than many currently employed is offered. If regularly used as an adjunct to these existing treatmen...

Fructose is a significant component in unprocessed food and has become one of the most commonly sweeteners used in food manufacturing. Fructose is also a useful pharmaceutical excipient and derivatives of fructose are exploited as renewable chemical building blocks. Fructose based polysaccharides have extensive pharmaceutical and dietary functions....

Fructose is a significant component in unprocessed food and has become one of the most commonly sweeteners used in food manufacturing. Fructose is also a useful pharmaceutical excipient and derivatives of fructose are exploited as renewable chemical building blocks. Fructose based polysaccharides have extensive pharmaceutical and dietary functions....

The hydrolysis of various carbohydrates was investigated under acidic conditions in real time by (1)H NMR spectroscopy, with a focus on the polysaccharide inulin. Sucrose was used as a model compound to illustrate the applicability of this technique. The hydrolysis of sucrose was shown to follow pseudo first order kinetics and have an activation en...

D-Fructose was analysed by NMR spectroscopy and previously unidentified (1)H NMR resonances were assigned to the keto and α-pyranose tautomers. The full assignment of shifts for the various fructose tautomers enabled the use of (1)H NMR spectroscopy in studies of the mutarotation (5-25°C) and tautomeric composition at equilibrium (5-50°C). The muta...

The role for adjuvants in human vaccines has been a matter of vigorous scientific debate, with the field hindered by the fact that for over 80 years, aluminum salts were the only adjuvants approved for human use. To this day, alum-based adjuvants, alone or combined with additional immune activators, remain the only adjuvants approved for use in the...

alpha-D-glucopyranosyl-[beta-D-fructofuranosyl](n-1)-D-fructofuranoside, commonly referred to as inulin, is a natural plant-derived polysaccharide with a diverse range of food and pharmaceuticalapplications. It is used by the food industry as a soluble dietary fibre and fat or sugar replacement, and in the pharmaceutical industry as a stabiliser an...

Advax is a polysaccharide-based adjuvant that potently stimulates vaccine immunogenicity without the increased reactogenicity seen with other adjuvants. This study investigated the immunogenicity of a novel Advax-adjuvanted Vero cell culture candidate vaccine against Japanese encephalitis virus (JEV) in mice and horses. The results showed that, in...

Host response elicited by photodynamic therapy (PDT) of cancerous lesions is a critical contributor to the clinical outcome, and complement system has emerged as its important element. Amplification of complement action was shown to improve tumour PDT response. In search of a clinically relevant complement activator for use as a PDT adjuvant, this...

There has been a recent resurgence of interest into new and improved vaccine adjuvants. This interest has been stimulated by the need for new vaccines to combat problematic pathogens such as SARS and HIV, and to counter potential bioterrorist attacks. A major bottleneck in vaccine development is the low immunogenicity of purified subunit or recombi...

The adjuvanticity of two gamma inulin/liposomes/Vitamin E combinations was evaluated in the mouse, in contraceptive vaccines with sperm protein extracts or a synthetic HE2 peptide ("Human Epididymis gene product"; residues 15-28) as antigen. The HE2 peptide was not conjugated to a protein carrier, to ensure that the antibodies elicited were specifi...

The adjuvant effect of gamma-inulin, a strong activator of the alternative complement pathway, is well-known, but its exact mechanism is not revealed yet. Here, we show that macrophages, isolated from the peritoneal cavity of gamma-inulin-injected mice and used as antigen-presenting cells, enhance the proliferation of antigen-specific T-cells up to...

Mammals appear to have evolved two types of automatic defence for recognition of microbial invaders. The first type comprises a large variety of cellular receptors on leukocyte surfaces that specifically recognise microbial components such as β-glucans (Czop and Kay, 1991) and lipopolysaccharides (Lynn and Golenbock, 1992). Ligation of such a recep...

Direct activation of the alternative pathway of complement (APC), independently of specific antibody, is well established as a major prime defence against invasion of the body by ‘foreign’ substances [1]. In nature these substances are almost always presented as particles, ranging in size from viruses and bacteria to fungal spores, protozoa or tumo...

Algammulin is a suspension of 1–2 μm ovoids of the immune stimulant gamma inulin (g-1N) in which alum is embedded as a carrier for protein or other anionic antigens. Tests for specific IgG and seroconversion responses in mice immunized with keyhole limpet haemocyanin (KLH) show that the presence of g-IN on the alum has increased its adjuvanticity 6...

Crystallization of inulin with alum forms a fine (1-2 micron) suspension of electron-dense ovoids; the alum is embedded in inulin particles, which are then converted to the immune stimulant polymorphic form, gamma inulin. This very stable hybrid preparation is termed Algammulin. Preferred conditions for its preparation are described. The alum still...

Algammulin is a new vaccine adjuvant comprising a stable suspension of 1-2 microns ovoids of the immune stimulant gamma inulin in which alum (Alhydrogel) is embedded as a protein carrier. Adjuvanticity tests in mice with Algammulin show that the presence of gamma inulin on the alum particles has synergistically enhanced their adjuvanticity for low...

Gamma-inulin (g-IN) is a polymorph identified as the active component of inulin preparations that specifically activates the alternative pathway of complement (APC). The APC is central to many leucocyte functions, including B cell activation. We show here that g-IN, when formulated as a pure, endotoxin-free, fine suspension insoluble at 37 degrees...

Inulin crystallizes in a series of polymorphic forms with different solubility rates, ranging from one instantly soluble at 23 degrees C (beta 0(23) inulin) through a form soluble at 37 degrees C with a half-time of 8 min (alpha 8(37) inulin) to a form virtually insoluble at 37 degrees C (gamma inulin). All forms are interconvertible, the more solu...

Finely divided, insoluble inulin (gamma polymorph), given intraperitoneally (i.p.) to C57BL mice 1-3 days after i.p. B16 melanoma cells, very significantly increased their mean survival time (MST) in low doses (less than or equal to 40 and less than or equal to 100 micrograms/mouse in 50 and 80% of tests, respectively). The gamma inulin was pure an...

Activation of the APC is pointed out as the common factor in all sufficiently studied cancer treatments employing nonspecific, active immunotherapy. This chapter outlines the molecular biology of both APC and classical pathway of complement, summarizes the alternative pathway's biologic activities especially in relation to the C3/C5 convertase C3b,...

Intraperitoneal (i.p.) injection of substances that can ignite the alternative pathway of complement, namely isolated human C3b or C3(H2O), guinea-pig C3(H2O) or cobra venom factor, or conventionally prepared zymosan, will reproducibly and very significantly increase the mean survival time of C57BL mice previously inoculated i.p. with melanoma cell...

Intraperitoneal (i.p.) injection of substances that can ignite the alternative pathway of complement, namely isolated human C3b or C3(H2O), guinea-pig C3(H2O) or cobra venom factor, or conventionally prepared zymosan, will reproducibly and very significantly increase the mean survival time of C57BL mice previously inoculated i.p. with melanoma cell...

Mice (C57BL) succumbed to cultured B16 melanoma cells i.p. with reproducible kinetics and an MST2 of about 26 days. Serum from tumour-bearing or normal mice was treated at 0 degree C with fixed SAC cells and injected i.p. into fresh tumour-bearing mice. If serum was given 7 days or less after B16 inoculation, the MST of the mice was highly signific...

Mice (C57BL) succumbed to cultured B16 melanoma cells i.p. with reproducible kinetics and an MST2 of about 26 days. Serum from tumour-bearing or normal mice was treated at 0°C with fixed SAC cells and injected i.p. into fresh tumour-bearing mice. If serum was given 7 days or less after B16 inoculation, the MST of the mice was highly significantly i...

Mesenchymal cells from primary BMS (baby mouse skin) cultures formed secondary monolayers subject to density-dependent inhibition. The monolayers remained quiescent but in good condition for 6-8 weeks if given weekly medium changes. Exposure of the primary cultures to fluorescent light and/or oxygen produced "altered" cells that gave rise in the se...

When primary baby mouse skin (BMS) cultures were subcultured for 48 hours into media containing 10(-6) to 10(-7) M colchicine or demecolcine, the number of altered cell foci appearing after 3-4 weeks' maintenance at 36 degrees C was substantially enhanced over drug-free controls. This applied whether or not the primary cultures had been irradiated...

Long-term cell lines were readily developed from a proportion of either baby mouse skin (BMS) cultures passing through colchicine-induced crisis or altered-cell foci selected from BMS cultures exposed to light and/or oxygen followed by colchicine. The developing cell lines behaved as though they passed through a continuing, profound genetic reshuff...

The DNAs of wild-type rabbitpox virus (u+.p+), and selected u (white pock, u.p+) and p (white pock, PK-15 cell non-permissive, u.p) mutants were compared by restriction enzyme analysis. The cleavage fragments produced by digestion with HindIII or EcoRI endonucleases were separated by agarose gel electrophoresis. Deletions of approx. 10 x 10(6) were...

Eighteen temperature-sensitive (ts) mutants of rabbitpox virus were examined for defects in synthesis of DNA and protein. Two mutants (ts-3 and ts-16) were defective in DNA synthesis (DNA-), since both incorporated significantly less than wild-type amounts of labelled thymidine into acid-precipitable material when infected cells were incubated at t...

Picornaviruses have long seemed to have many genes. The size of the genome compared with the “average” gene product suggested about ten for poliovirus (Fenner, 1968), which causes at least a dozen synthetic and degenerative changes in the host cell. Many viral polypeptides are found, originally about 14 (Summers et al.,1965) and now more than 34 (A...

Urea is shown to be a useful additive to sodium dodecyl sulfate gels prior to electrophoresis and offers an alternative means for the resolution of some SDS-protein complexes. Two types of effect are described, one of which causes increases in the relative mobilities of certain polypeptides found in poliovirus-infected cells. It is postulated that...

Viral polypeptides associated with the membrane-free replication complex of poliovirus RNA were multiple in nature. The structural protein precursors [VP0, VP1, VP3] predominated, and because they were found in a cytoplasmic component with the same S value and density as the replication complex are likely to be attached to it in vivo. They were not...

Mit Hife von p-[(125)J]Jodphenylisothiocyanat werden nach einer modifizierten Edman-Reaktion die N-terminalen Reste geringer Proteinmengen (2,5-5 n Mol) bestimmt.

The pattern of viral polypeptide synthesis and cleavage in poliovirus-infected cells was shown by autoradiography to be considerably more complex than previously thought. In normal growth, at least 26 distinct polypeptides were found, and various modifications of the cleavage process revealed a total of at least 34. Most of the new polypeptides wer...

Poliovirus proteins were labelled in vivo with [35S]-methionine, and the major products of translation and cleavage were separated by electrophoresis and compared in terms of two-dimensional tryptic peptide maps visualized by autoradiography. The main intermediates p110 and p90 had few or no methionine-labelled sequences in common, but were both co...

Eighteen new ts mutants of poliovirus have been isolated after a variety of mutagenic treatments, and their loci identified in relation to the previous genetic map. The map was only extended by 25%, and the physiological characters of the new isolates corresponded in all aspects tested with those of the previous isolates. Apparently single mutants...

Identification of the N-terminal residue of 2.5-5 nmol of protein was possible by means of a modified Edman reaction and p-iodophenyl[125I] isothiocyanate of high specific activity. The second and third residues could also be identified. Yields of the terminal residue were 25-50%, allowing approximate quantitation of the molar content of protein. U...

The proportion of cells absorbing trypan blue (tb-+ character) can be used to measure the late c.p.e. of wild-type poliovirus (ts-+. tb-+), which was the same at restrictive (39-2 to 39-6 degrees C) or permissive (37 degrees C) temperatures. Of twenty ts mutants, seven showed normal c.p.e. at 37 degrees C but were defective in C.P.E. (TB) AT 39-5 d...

This paper points out certain theoretical problems in DNA synthesis associated with antiprimer transcription and with circularization that could oblige RNA tumor viruses to rely on a polyploid genome. It is suggested that each completed act of reverse transcription may be coupled with an act of genetic recombination aimed at recovering the antiprim...

Some poliovirus-specific protein in infected cell cytoplasm was found to have the same sedimentation coefficient and buoyant density in CsCl as the native 45 S subunits (1.48 g/cc), as viral ribonucleoprotein (1.40 g/cc) and as 60–80 S mono- or oligoribosome/vRNA complexes (1.50 to 1.54 g/cc). Cross-fixation artifacts resulting from glutaraldehyde...

SUMMARY Recombinant and parental poliovirus particles were indistinguishable by rate- zonal and isopycnic sedimentation, and by u.v. inactivation. Sensitive selective procedures, applied to ts + recombinants to detect genetic segregation of one parent, failed to reveal any. Poliovirus genetic recombinant particles thus appear to be conventional vir...

Summary The overall scheme of gene replication and function of a virus, even seen only in outline, represents a compelling criterion for the classification of viruses.Copyright © 1974 S. Karger AG, Basel

Summary Three poliovirus mutants able to form plaques at 37° but not at 33.5° (i.e. cold-sensitive, or cs) were isolated from a mutagenized cs+ stock. Their biochemical lesion has not been defined.Copyright © 1974 S. Karger AG, Basel

By use of [125I] p iodophenylisothiocyanate in a modified Edman procedure, N terminal aspartate, glycine and serine were identified and approximately assayed in the capsid protein of purified poliovirus strain Mahoney. Two separate preparations of virus gave similar results. These three N terminal amino acids were recovered in amounts consistent wi...

SUMMARY Wild-type poliovirus (ts +) rapidly represses synthesis of host cell protein (psr+ character), at permissive (37 °C) or restrictive (39"5 °C)temperatures. A search for ts mutants that were psr+ at 37 °C and psr at 39"5 °C was successful when gene expression was minimized by adding guanidine, depleting cystine and avoiding very high input mu...

SUMMARY A count of specifically labelled tryptic peptides from purified poliovirus (strain Mahoney) showed that the amino acid sequence comprising its structural protein contains at least 2I and 25 uniquely placed residues ofhistidine and methionine, re- spectively. Amino acid analysis after performic acid oxidation showed a content of, respectivel...

Summary A labile subviral particle (‘the equestron’) is proposed as a multifunctional regulator of poliovirus growth, withaffinities for 45s ribosomal subunits and 5’ ends of viral RNA. These suggested properties suffice to explain suppression of hostprotein synthesis, synchronous synthesis of viral RNA and protein, and timing of maturation. The ca...

Attempts to Extend the Genetic MapOrientation of the Genetic MapThe Scale of the Genetic MapImplications for Virion Structure of the Size of Coat Protein GenesViral Inhibitors that Affect Coat ProteinSize of Genes Associated with RNA ReplicationThe Far Left of the MapThe Host-Synthesis Repressor Coded for By PoliovirusA Possible Maturation FactorOt...