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Biomedical subjects

A Saul

Publications and source records attributed to A Saul.

At least 127 records · Page 7Linked to original sources
PubMed1991

Role of single amino acids in the recognition of a T cell epitope.

T cell epitopes can be defined by the use of synthetic peptides, which when added to APC efficiently mimic naturally processed Ag. Free peptide is thought to bind to cell-surface MHC glycoproteins and the TCR then recognizes the resulting complex. The specificity of a tetanus toxin-specific human Th cell clone was investigated using a complete replacement set of peptides in which every amino acid within the minimal T cell epitope was replaced by each of the 19 alternative genetically coded amino acids. Within the minimal epitope, found to be YSYFPSVI (tetanus toxin 593-600), a small number of substitutions could be made without significant loss of activity, defined as substitutions giving peptides whose activity fell within +/- 3 SD of the mean parent response. Y593 could be substituted with F, W, M, L, V, and I; S594 with G and T; Y595, F596, and P597 with no other amino acids; S598 with A; V599 with S, and I600 with L. Rank ordering of the substitutions allowed a precise description to be made of MHC and/or TCR interaction with each amino acid side chain within the epitope. Simplified theoretic calculations based on this study indicate that class II T cell recognition has a specificity greater than 1 in 10(8). Competition experiments indicate that Y595, F596, P597, and I600 are critical for binding of this epitope to its restricting element, HLA DR4Dw14.

Amino Acid Sequence↗
PubMed1991

Inhibition of HLA B8-restricted recognition by unrelated peptides: evidence for allosteric inhibition.

A panel of synthetic peptides representing human lymphocyte antigen (HLA) B8, other class I and class II restricted T cell epitopes and two B cell epitopes, were all able to compete with recognition of a HLA B8 restricted epitope by a cytotoxic T cell clone. Competition was obtained when the competitor peptides were added either before or after the target epitope. The target epitope also had a slow off rate, implicating allosteric inhibition. The presence of non-specific, allosteric binding sites may interfere with experiments attempting to define immunologically relevant MHC binding specificities.

Allosteric Site↗
PubMed1991

Comparative immunogenicity of free and carrier-conjugated peptides derived from the constant regions of a polymorphic malarial surface antigen.

One peptide (L7) representing the entire constant N-terminal region, and two peptides (L5 and L6) representing the entire C-terminal constant region of the variable merozoite surface antigen MSA2, were synthesised by solid-state (tBOC) chemistry. Mice were immunised with the peptides alone and conjoined to the carrier protein diphtheria toxoid (DT) using the hetero-bifunctional reagent maleimidocaproyloxysuccinimide (MCS). Immune response was evaluated against the peptide itself by ELISA and against the intact protein MSA2 by immunoblotting and immunofluorescence assay (IFA). Whereas all peptides elicited a strong specific antipeptide response when administered as conjugates only L7 and L6 elicited an anti-peptide response in the absence of carrier. L5 and L7 conjugates elicited sera reactive with the intact MSA2, whereas only L7 elicited such specificity in the absence of carrier.

Amino Acid Sequence↗
PubMed1991

Identification of a common Plasmodium epitope (CPE) recognised by a pan-specific inhibitory monoclonal antibody.

A Plasmodium falciparum genomic expression library was screened with a monoclonal antibody produced from mice infected with Plasmodium yoelii. Eleven unique clones were isolated all of which contained the sequence NKND, IKND or KKND. This sequence was confirmed as the epitope of M26-32 by testing a series of overlapping peptides and the allowable substitutions determined by testing the binding of M26-32 to peptides containing all possible single amino acid replacements of NKND. Potential epitopes of M26-32 occur in many plasmodial proteins and this is consistent with the large number of proteins recognised in these parasites by Western blotting. Since this monoclonal antibody shows marked in vitro inhibition of P. falciparum growth, these data suggest that an anti-malarial vaccine may be produced by targeting such common plasmodial epitopes without necessarily identifying the corresponding antigens.

Amino Acid Sequence↗
PubMed1991

7H8/6, a multicopy DNA probe for distinguishing isolates of Plasmodium falciparum.

A method using DNA fingerprinting to distinguish Plasmodium falciparum isolates has been developed using a clone, 7H8/6, isolated by screening a genomic expression library of the Malayan Camp strain with a monoclonal antibody. Unlike other P. falciparum DNA probes that contain internally repetitive sequences, this probe contains a single full-length open reading frame and lacks any repetitive sequences. Chromosome mapping located the hybridizing sequences to most chromosomes. 7H8/6 sequences are located at sites near the subtelomeric regions, although they are further from the telomeres than the subtelomeric repetitive sequences rep20. The probe gave multiple polymorphic bands on Southern blots of genomic DNA allowing all unrelated isolates tested to be readily distinguished. Hybridization patterns of parent and progeny from the genetic cross of two P. falciparum clones showed that most bands were inherited independently and no new bands were generated during the cross. This probe is useful for distinguishing isolates of P. falciparum and also for analyzing relationships between closely related isolates.

Animals↗
PubMed1991

Fine specificities of monoclonal antibodies against the Plasmodium falciparum circumsporozoite protein: recognition of both repetitive and non-repetitive regions.

The fine specificities of 6 monoclonal antibodies (MoAbs) raised against the circumsporozoite (CS) protein of the human malaria parasite, Plasmodium falciparum, were defined by their binding to a series of overlapping octapeptides corresponding to the 7G8 variant of the CS protein. The precise specificities of the MoAbs to the immunodominant NANP repeat region were elucidated by their binding to all possible 4, 5, 6, 7 and 8 amino acid peptides in this region. All 6 MoAbs recognized the NANP repeats. In addition all MoAb bound to nonrepetitive sites with 4 of the 6 MoAbs recognizing known functional sites outside the repeat region including sites required for T cell recognition and hepatocyte invasion. Antibody pressure may therefore be responsible for generating the epitope variation observed at T cell sites. The multiple specificities for all the MoAbs suggests that the repeat region may act as an internal immunological 'smokescreen' by competing more effectively for antibody binding compared to single epitope copy functional sites located outside the repeat region.

Amino Acid Sequence↗
PubMed1991

The development of a malaria vaccine.

This paper is a progress report and summarises R & D by the consortium to develop a malaria vaccine. The research strategies and research progress are discussed. The business arrangements and the roles of each partner are given.

Antigens, Protozoan↗
PubMed1990

Human erythrocyte Band-3 has an altered N terminus in malaria-resistant Melanesian ovalocytosis.

There is a high prevalence of the erythrocyte polymorphism ovalocytosis associated with reduced susceptibility to malaria in Papua New Guinea. The major erythrocyte integral membrane protein, Band-3, showed markedly increased phosphorylation in whole cells or isolated ghosts from ovalocytic individuals. The cytoplasmic domain of the ovalocyte Band-3 was found to be approx. 3 kDa larger than the normocytic protein. The N-terminal sequence of the ovalocytic Band-3 was different from the reported sequence for human Band-3, suggesting that the increased size results from an N-terminal extension. Since this is the region of Band-3 which is phosphorylated and interacts with the red cell cytoskeleton, it is likely that this alteration in ovalocytic Band-3 is the underlying cause of the diverse alterations in ovalocytic cells including increased phosphorylation, increased membrane rigidity, decreased agglutinability by blood group antibodies and refractoriness to invasion by malarial parasites.

Anion Exchange Protein 1, Erythrocyte↗
PubMed1990

Chemical characterization of the parasitophorous vacuole membrane antigen QF 116 from Plasmodium falciparum.

On the basis of amino acid sequencing and immunological cross-reactivity, the Plasmodium falciparum parasitophorous vacuole antigens QF116 and exp-1/CRA are apparently identical. The epitope recognized by an inhibitory monoclonal antibody directed against QF116 is located proximal to the C-terminus of the protein. The QF116 protein is processed during maturation by the cleavage of a 22-amino-acid signal peptide and acylated as measured by labeling with myristic acid.

Acylation↗
PubMed1990

Refractoriness of erythrocytes infected with Plasmodium falciparum gametocytes to lysis by sorbitol.

Unlike erythrocytes infected with mature asexual parasites of Plasmodium falciparum, those infected with gametocytes are not lysed by 5% sorbitol solutions. This observation was used to devise a method for producing synchronized cultures of gametocytes, free of asexual stage parasites. The refractoriness to sorbitol suggests that the major anion transport pathway, which appears in the membrane of erythrocytes infected with asexual stage parasites, is not present in cells infected with gametocytes.

Animals↗
PubMed1990

Peptide vaccines derived from a malarial surface antigen: effects of dose and adjuvants on immunogenicity.

Peptides P2122 (CKNNNSTNSGI) and P513 (CSQRSTNSAST) containing an epitope of a malarial surface antigen (MSA2) recognised by inhibitory monoclonal antibodies were conjugated to diphtheria toxoid (DT) protein and formulated with various gel-based and water in oil emulsion adjuvants in vaccine trials in mice and rabbits. The P2122-DT construct was effective in raising antibodies reactive with both the immunising peptide and the native antigen. Effective adjuvanticity as measured by the titre of the anti-peptide or anti-protein response in mice varied in the order: Algammulin, Montanide ISA 50 greater than or equal to Freund's adjuvant, Montanide ISA 708, 721, 70 much greater than alum, Squalene Arlacel greater than SAF-1. A similar order of adjuvant efficacy: Freund's greater than alum greater than Squalene Arlacel greater than SAF-1, was observed in rabbits.

Adjuvants, Immunologic↗
PubMed1990

Variable linking region immunogenicity using malarial peptide carrier protein conjugates of defined composition.

Thirty-five overlapping peptides from both conserved and variable parts of the N-terminal region of a malarial merozoite surface antigen (MSA2) were synthesised using solid phase chemistry. All peptides were synthesised with an added N-terminal cysteine and purified by reverse-phase HPLC to facilitate coupling to a carrier protein diphtheria toxoid (DT) using the hetero-bifunctional reagent maleimidocaproloxysuccinimide (MCS). Mice were immunised with these peptide-DT conjugates using Freund's complete adjuvant (FCA). The immune response of these mice was tested against peptide hapten, carrier protein (DT) and the linking region itself, using enzyme-linked immunoassay (ELISA). Although there was great variation in the immune response to each part of the immunogen construct, no significant correlation could be seen between each set of responses.

Amino Acid Sequence↗
PubMed1990

Analysis of the sequences flanking the translational start sites of Plasmodium falciparum.

The 5' and 3' regions adjacent to the initiation codon in 22 Plasmodium falciparum sequences were examined. A 5' consensus sequence (AAAA/ATG) was found. Although P. falciparum non-translated DNA is A-rich, A occurred significantly more frequently in the 4 positions preceding the initiation ATG than in adjacent non-translated DNA, suggesting that this consensus sequence has functional significance in the initiation of translation. This region has similarities with the equivalent sequences in yeast and Drosophila but differs markedly from that in vertebrates. No significant bias in nucleotide frequencies was found 3' to the initiation codon.

Animals↗
PubMed1990

Itraconazole in common dermatophyte infections of the skin: fixed treatment schedules.

Itraconazole is an effective medication against the most common dermatophytoses. It has been shown to be more active than griseofulvin and ketoconazole. Ease of use, affinity for keratinized tissues, lack of toxicity, continued activity after discontinuation, and the possibility of using fixed schedules are advantages of itraconazole. The fixed schedules indicated by pharmacokinetics and clinical studies are one 100 mg capsule daily for 15 days in cases of tinea corporis and tinea cruris and the same dosage for 30 days in cases of tinea pedis and tinea manuum. These fixed treatments have some limitations, and they are not recommended for treating tinea capitis and tinea unguium. The drug is well tolerated.

Antifungal Agents↗
PubMed1990

Variations in malaria transmission rates are not related to anopheline survivorship per feeding cycle.

Anopheline survivorship, vectorial capacity, and mosquito infection probability estimates from mosquito infection rates were determined 4 times in 1 year in a Papua New Guinea village. Estimates of survivorship over the length of the extrinsic incubation period differed significantly during the year. However, survivorship per feeding cycle, individual mosquito vectorial capacity, and mosquito infection probability did not vary significantly. Estimates of these parameters were then compared to estimates of survivorship, individual vectorial capacity, and mosquito infection probability in mosquito populations in other villages in the study area. Since survivorship per feeding cycle did not vary significantly among the mosquito populations in these villages, changes in malaria transmission potential can be better gauged from estimates of survivorship over the length of the extrinsic incubation period. However, as measurements of relative inoculation rates are easier to perform and have been related to parasite prevalences in children in this area, estimates of inoculation rates are a preferred option for estimating malaria transmission in the Madang area of Papua New Guinea.

Animals↗