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Persistence of cellular and humoral response to synthetic peptides from defined Plasmodium falciparum antigens.

The cellular and humoral immune responses to synthetic peptides reproducing the repeat sequences of two major vaccine candidates (circumsporozoite protein and Pfl55/RESA) were investigated in two groups of African subjects according to the length of their stay outside endemic areas. The relation between the lymphoproliferative response and the antibody levels to these antigens was studied. The results confirm the existence of T-cell epitopes within the repeat sequences of the CS protein and the Pfl55/RESA capable of inducing lymphocyte proliferation. Cellular response to all studied peptides was more frequently observed in individuals living in France for less than one year than in individuals living in France for a longer time. T-cell proliferation in the presence of the tetrapeptide and of the octapeptide from the C-terminus repeat of Pfl55/RESA was related, with an immunodominance of the tetrapeptide over the octapeptide. Cellular responses to the CS protein repeat and to the 11-amino-acid peptide from the Pfl55/RESA N-terminus were the longest lasting after termination of exposure. In a given individual, cellular and humoral responses were not related for any peptide studied.

Adolescent↗

Isoenzyme studies on Theileria (Protozoa, Sporozoa). Enzyme activity associated with the erythrocytic stage.

Bovine blood containing piroplasms of Theileria parva, as well as non-infected blood, was lysed and subjected to iso-electric focussing. Staining for 13 different enzymes revealed parasite-associated bands of glucose phosphate isomerase (GPI) activity, not of any of the other enzymes. There were no variations between individual donor animals in the host cell GPI bands and these bands did not interfere with the recognition of the parasite-associated bands, so that purification of the piroplasms was unnecessary. Blood from cattle infected with T. mutans also gave parasite-associated bands of GPI, but no such bands were seen in zymograms of blood from cattle infected with a Theileria sp. from Japan. Dependent on the level of parasitaemia, up to four parasite-associated bands were found in one strain of T. parva and up to three in two other strains. Among the disadvantages of using piroplasm material for the study of isoenzymes of T. parva is the fact that animals often die before their parasitaemia is sufficiently high, and that some strains never give rise to a high parasitaemia.

Animals↗

The influence of various bovine sera on the maintenance of Theileria parva lymphoblastoid cell culture.

Theileria parva infected lymphoblastoid bovine cells were grown in a medium based on HEPES-buffered RPMI 1640 with glutamine and antibiotics, supplemented with bovine serum. There were no significant differences in growth rate, viability, and percentage of infected cells when the substrate contained 10 or 20 per cent of either commercially available newborn calf serum of serum prepared from adult non-infected Friesian cattle or of serum prepared from a Friesian calf immunised against East Coast fever and having a high titre of antibodies to T. parva antigen in the indirect fluorescent antibody test. If studies showing that newborn calf serum gives results in the establishment and maintenance of T. parva cell culture similar to those of foetal calf serum are confirmed, this finding could mean an appreciable saving in the cost of in vitro work on this parasite.

Animals↗

Theileria orientalis in Iran.

A benign species of Theileria of cattle in northern Iran proved to be indistinguishable from T. orientalis in the indirect fluorescent antibody test as well as in the morphology of its piroplasms. It was transmissible transstadially by Haemaphysalis punctata.

Animals↗

Cultivation of Theileria. I. Attempts to complete the cycle of Theileria parva in vitro.

Microschizonts and free merozoites developed in bovine lymphoblastoid cell cultures containing macroschizonts of 6 different strains of Theileria parva. Clean bovine red cells were added to the cultures, which were incubated in various ways. No penetration of red cells by merozoites was observed, not even when cultures in diffusion chambers were introduced into the peritoneal cavity of non-infected cattle.

Animals↗

Cultivation of Theileria. II. Attempts to cultivate erythrocytic stages of Theileria mutans.

Blood from calves with a parasitaemia of Theileria mutans was cultivated in vitro in various ways. No significant increase in the percentage of infected red cells was observed, but the percentage of dividing forms of the parasite rose sharply during the first two days of incubation. The conditions of the cultures were apparently unsuitable for the invasion of erythrocytes.

Animals↗

Stage-specific antigenicity in Theileria annulata: a case report.

Blood from sick cattle in Bahrain transmitted piroplasms of Theileria annulata to a splenectomized calf. Larvae of Hyalomma anatolicum anatolicum were infected on the calf and, after moulting, induced clinical theileriosis, associated with numerous schizonts, in the same calf. The animal was cured by specific treatment. Antigenic differences thus shown between piroplasms on the one hand, and sporozoites and schizonts on the other hand, were confirmed in the indirect fluorescent antibody test, as a significant titre to T. annulata piroplasm antigen developed after the inoculation of blood, but to schizont antigen only after the infective ticks had induced the appearance of schizonts.

Animals↗

Theileria taurotragi in Zambia.

Theileria sp. (Bwengwa) of low virulence was isolated by feeding R. appendiculatus ticks collected from the field on a susceptible calf and subsequently transmitted between cattle by R. appendiculatus ticks- Theileria sp. (Bwengwa) was shown to be T. taurotragi on parasitological, clinical and serological grounds. T. taurotragi is the fourth Theileria sp. shown to be present in Zambia.

Animals↗

Comparative pathogenicity of Theileria annulata strains.

Twenty three susceptible crossbred calves were inoculated with five Indian strains of Theileria annulata, collected from natural cases of tropical theileriosis at Ludhiana, Hissar, Jaipur, Uruli-Kanchan and Bangalore. All the strains produced clinical disease with typical symptoms and lesions of acute theileriosis. The strains were considered highly virulent and equally pathogenic.

Animals↗

Statistical analysis of the exon-intron structure of higher and lower eukaryote genes.

Statistics of the exon-intron structure and splicing sites of several diverse eukaryotes was studied. The yeast exon-intron structures have a number of unique features. A yeast gene usually have at most one intron. The branch site is strongly conserved, whereas the polypirimidine tract is short. Long yeast introns tend to have stronger acceptor sites. In other species the branch site is less conserved and often cannot be determined. In non-yeast samples there is an almost universal correlation between lengths of neighboring exons (all samples excluding protists) and correlation between lengths of neighboring introns (human, drosophila, protists). On the average first introns are longer, and anomalously long introns are usually first introns in a gene. There is a universal preference for exons and exon pairs with the (total) length divisible by 3. Introns positioned between codons are preferred, whereas those positioned between the first and second positions in codon are avoided. The choice of A or G at the third position of intron (the donor splice sites generally prefer purines at this position) is correlated with the overall GC-composition of the gene. In all samples dinucleotide AG is avoided in the region preceding the acceptor site.

Algorithms↗

The microsporidian spore invasion tube. IV. Discharge activation begins with pH-triggered Ca2+ influx.

The microsporidian spore extrusion apparatus activates with a calcium influx from Spraguea lophii spore wall/plasma membrane; this influx requires preconditioning with an extrasporular shift in medium pH to the alkaline in the presence of the polyanions mucin or polyglutamate. Undischarged S. lophii spores display calcium bound to the wall/plasma membrane with a characteristic calcium-chlorotetracycline fluorescence; this fluorescence declines significantly during spore discharge. S. lophii spores do not discharge when spore wall/plasma membrane calcium is removed with EGTA. Extrasporular mucin or polyglutamate and a pH shift to the alkaline appear to be necessary preconditions for the triggering of the influx of spore wall/plasma membrane-bound 45Ca2+. Ionophore A-23187 also effectively activates spore discharge without other extrasporular polyanions. Micromolar concentrations of the calcium antagonists lanthanum or verapamil prevent spore discharge, and micromolar concentrations of calmodulin inhibitors chlorpromazine and trifluroperazine prevent spore discharge. Calmodulin, visualized with a calmodulin antibody and a peroxidase conjugate, is localized particularly on the plasma membrane and the polaroplast membranes of the extrusion apparatus.

Animals↗

The entry of Theileria parva sporozoites into bovine lymphocytes: evidence for MHC class I involvement.

We have examined the process of Theileria parva sporozoite entry into susceptible bovine lymphocytes and have begun to identify one of the possible molecular interactions involved in the process. The entry process involves a defined series of events and we have used a number of experimental procedures in combination with a method of quantitation to examine various aspects of this process. T. parva sporozoites are nonmotile organisms and the initial sporozoite-lymphocyte interaction is a chance event which can occur at 0-2 degrees C. All subsequent stages in the process are temperature dependent, require the participation of live intact sporozoites and host cells, and involve some cytochalasin-inhibitable rearrangement of the host cell surface membrane or cytoskeleton. Sporozoite entry can be inhibited by antibodies (mAbs) reactive with major histocompatibility complex (MHC) class I molecules (IL-A 19, IL-A 88) and with beta 2 microglobulin (B1G6), whereas mAbs reactive with MHC class II molecules (IL-A 21, J 11), and a common panleucocyte surface antigen, (IL-A 87; a bovine equivalent of CD 11a) have no effect. These results indicate that MHC class I molecules play a role in the process of T. parva sporozoite entry into bovine lymphocytes although as yet the precise role has not been determined. Once internalized within the lymphocyte, a process that takes less than 3 min at 37 degrees C, the sporozoite rapidly escapes from the encapsulating host cell membrane; a process which occurs concurrently with the discharge of the contents of the sporozoite rhoptries and microspheres. The intracytoplasmic parasite is covered by a layer of sporozoite-derived fuzzy material to which host cell microtubules rapidly become associated.

Actins↗