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Biomedical subjects

P H David

Publications and source records attributed to P H David.

At least 37 records · Page 2Linked to original sources
PubMed1990

Genetic complexity of Plasmodium vivax parasites in individual human infections analyzed with monoclonal antibodies against variant epitopes on a single parasite protein.

Monoclonal antibodies against variant epitopes of a highly polymorphic protein (PV200) in schizonts of Plasmodium vivax have been used to analyze the variety of genetically distinct populations of parasites present in the peripheral blood of individual P. vivax infections in Sri Lanka. In 9 out of 10 isolates of freshly drawn P. vivax infected blood from different individuals, parasites of only 1 PV200 serotype was found within each individual infection, even though parasites were serotypically distinct between individuals. In 1 isolate parasite population, 3 distinct PV200 serotypes were identified. Thus, most P. vivax infections appeared to consist of a single genetically homogeneous population of parasites within the detection limits of the technique. The prevalence of P. vivax infections in an area of malaria transmission in southern Sri Lanka and the densities of oocysts in mosquitoes fed on P. vivax infected individuals indicated that parasite populations would be transmitted many times before encountering parasites of other origins, and that individual populations would tend to reduce to genetic homogeneity during transmission. These expectations are consistent with the high proportion of genetically homogeneous P. vivax isolates observed.

Adult↗
PubMed1989

Uninfected erythrocytes form "rosettes" around Plasmodium falciparum infected erythrocytes.

The human malaria parasite, P. falciparum, exhibits cytoadherence properties whereby infected erythrocytes containing mature parasite stages bind to endothelial cells both in vivo and in vitro. Another property of cytoadherence, "rosetting," or the binding of uninfected erythrocytes around an infected erythrocyte, has been demonstrated with a simian malaria parasite P. fragile which is sequestered in vivo in its natural host, Macaca sinica. In the present study we demonstrate that rosetting occurs in P. falciparum. Rosetting in P. falciparum is abolished by protease treatment and reappears on further parasite growth indicating that, as in P. fragile, it is mediated by parasite induced molecules which are protein in nature. P. vivax and P. cynomolgi, which are not sequestered in the host, did not exhibit rosetting. Rosetting thus appears to be a specific property of cytoadherence in malaria parasites.

Animals↗
PubMed1989

Continuous monitoring of child mortality from clinic records.

A simple method is described for estimating childhood mortality when mothers are seen at or soon after a birth. The main bias in the method arises from omission of reports from mothers not contacted in the immediate postnatal period. Recent work indicates that the key question on the survival of the preceding born child might be usefully included in household surveys. A comparison between data from Tari, Papua New Guinea on childhood mortality collected as part of a regular community surveillance scheme and the results from asking the supplementary question in clinics indicates that the simpler preceding birth technique appears to give good results quickly and cheaply.

Actuarial Analysis↗
PubMed1988

Plasmodium vivax: karyotype polymorphism of field isolates.

Pulse-field gradient electrophoresis (PFG) has been applied to the karyotype analysis of Plasmodium vivax isolates obtained directly from infected patients in Sri Lanka. Detection of separated chromosomes was performed either by ethidium bromide staining of gels or by hybridization with a telomer specific probe. Each of the 15 different isolates examined exhibited a different chromosome migration pattern, indicating that a high level of polymorphism prevailed in wild populations of P. vivax. Chromosome size variation was further confirmed using a P. vivax chromosome-specific probe which also demonstrated that, in each isolate, the parasite population appeared to be homogeneous. These observations were made directly on parasites from infected blood, without the necessity for culture amplification, indicating that PFG can be used on a large scale for the epidemiological analysis of wild parasite populations.

Animals↗
PubMed1988

Plasmodium vivax: cloning and expression of a major blood-stage surface antigen.

Plasmodium vivax is a highly prevalent malaria pathogen of man; the following report is the first to describe the cloning and expression of a major asexual erythrocytic stage antigen of this species. The screening of a genomic DNA expression library with a monoclonal antibody directed against a 200-kDa surface component (Pv200) of the more mature schizonts of P. vivax led to the selection of a recombinant bacterial clone which produced a fusion protein. Mouse and rabbit immune sera raised against the purified fusion protein recognized the 200-kDa parasite antigen on Western blots and reacted with the surface of segmenters by immunofluorescence. Sequencing of the 1.9-kb P. vivax DNA insert coding for this fusion protein revealed a 45-47% homology at the nucleotide level with the P. falciparum gene of a parasite surface antigen, Pf195, which has been shown to be a promising candidate for a malaria vaccine in primates and in man.

Amino Acid Sequence↗
PubMed1988

Plasmodium vivax malaria: parasite biology defines potential targets for vaccine development.

Although progress in the development of an antimalarial vaccine has been mostly obtained through the study of P. falciparum, significant advances have recently been made in the study of P. vivax, the other major human malarial parasite. Antigens which have been shown to be important in P. falciparum have been characterized and in some cases cloned in P. vivax. Other studies have examined some of the more specific biological characteristics of P. vivax. Among these are studies on components present in caveolae--vesicle complexes of the infected erythrocyte, on the occurrence of delayed hepatic development leading to relapse, or on the Duffy erythrocyte antigen as a key receptor for parasite invasion. Although progress has been made in the short-term in vitro maintenance of P. vivax, the inability to maintain the parasite in continuous culture led to the investigation of wild parasite populations in patients; occurrence of extensive antigenic and karyotype polymorphism was detected in this way, as was a double-blocking and enhancing activity of human antibodies on parasite development in the vector. The association of monoclonal antibodies with DNA recombinant technology allowed the characterization of a number of P. vivax antigens to be made. Among these, an antigen shared between sexual and asexual stages was shown to constitute a target for transmission-blocking immunity. The cloning of an antigen involved in transmission-blocking immunity, along with that of the surface antigen of the sporozoite (CSP) and of a major surface antigen of the invasive merozoite (PV200) constitutes a significant step towards the development of a multivalent recombinant vaccine against P. vivax.

Animals↗
PubMed1988

Rosetting: a new cytoadherence property of malaria-infected erythrocytes.

Plasmodium fragile infection of the toque monkey is a natural host-parasite association in which parasite sequestration occurs as during P. falciparum infection of humans. We have studied parasite sequestration of P. fragile and demonstrated the existence of a new property of cytoadherence of infected erythrocytes, "rosetting," which is defined as the agglutination of uninfected erythrocytes around parasitized erythrocytes. Rosetting in vitro and sequestration in vivo appear simultaneously as the parasite matures. The spleen plays a role in modulating cytoadherence; both sequestration and rosetting, which occur with cloned parasites from spleen-intact animals, are markedly reduced in splenectomized animals infected with parasites derived from the same clone. Sequestration and rosetting can be reversed by immune serum. Protease treatment of infected blood abolishes rosetting; however, if treatment is performed at an early stage of schizogony, rosetting reappears if parasites are allowed to further develop in the absence of protease. These results indicate that with P. fragile in its natural primate host, rosetting and sequestration are related to the presence on the infected erythrocyte surface of a parasite-derived antigenic component, the expression of which is modulated by the spleen.

Animals↗
PubMed1988

Diversity of Plasmodium vivax-induced antigens on the surface of infected human erythrocytes.

Antigens were demonstrated on the surface of Plasmodium vivax schizont-infected erythrocytes by an indirect immunofluorescence test using fresh unfixed infected erythrocytes from acute vivax malaria patients. Surface immunofluorescence was used to show that sera of P. vivax-infected individuals contain antibodies directed against these surface antigens. Thirteen different isolates were screened for reactivity of surface antigens with a panel of 8 heterologous human immune sera and an immune rabbit serum. Surface immunofluorescence was detected in several isolates with some but not all the human sera, and not all sera reacted with the "positive" isolates. These results indicate a high degree of polymorphism of the surface antigens of different P. vivax isolates. Sera from patients who had suffered multiple malaria attacks and the immune rabbit serum (which was raised by immunizing with 7 different isolates) recognized surface antigens on more isolates than sera from patients who had experienced only one attack of malaria, indicating that repeated exposure to the disease confers immunity against a spectrum of variants of a polymorphic malarial antigen(s) prevalent in nature.

Animals↗
PubMed1988

Monoclonal and polyclonal antibodies both block and enhance transmission of human Plasmodium vivax malaria.

Antibodies against gametes of the malarial parasite inhibit the development of the parasite in the mosquito and curtail the transmission of malaria. We now report that a monoclonal antibody against gametes of the human malaria pathogen Plasmodium vivax and antibodies induced during natural infections of P. vivax in humans which suppress infectivity of the parasites to the vector at high concentrations can, at lower concentrations, have the opposite effect and enhance the level of malaria infection in the mosquitoes. Infectivity enhancing effects of up to 12-fold were demonstrated when a transmission blocking monoclonal antibody and immune human sera were diluted, in some undiluted immune human sera, and in the sera of vivax malaria patients during convalescence after drug cure.

Animals↗
PubMed1988

Immunoelectron microscopy of Schüffner's dots in Plasmodium vivax-infected human erythrocytes.

Plasmodium vivax induces morphologic alterations in infected host erythrocytes that are visible by light microscopy in Romanovsky-stained blood smears as multiple brick-red dots. These morphologic changes, referred to as Schüffner's dots, are important in the identification of this species of malarial parasite and have been associated by electron microscopy with caveolavesicle complexes along the erythrocyte plasmalemma. We have produced a monoclonal antibody (MAb A 20) that identifies an antigen in Plasmodium vivax-infected erythrocytes that is associated with the caveola-vesicle complexes of the parasitized host cell. This monoclonal antibody reacts with air-dried P vivax-infected erythrocytes to produce a pattern by the indirect immunofluorescence test (IFT) that is evocative of Schüffner's dots. Immunoelectron microscopy of P vivax-infected human erythrocytes using MAb A 20 confirmed the location of this antigen within vesicles of caveola-vesicle complexes. On Western blots MAb A 20 recognized four polypeptides of 54, 64, 72, and 86 kd. MAb A 20 reacted by IFT with 90% of Sri Lankan isolates and with a Brazilian P vivax isolate, which indicates that the epitope identified by this monoclonal is conserved.

Animals↗
PubMed1987

Antigenic variation of cloned Plasmodium fragile in its natural host Macaca sinica. Sequential appearance of successive variant antigenic types.

The course of infection of Plasmodium fragile in its natural host, the toque monkey Macaca sinica, consists of a primary peak of parasitemia followed by several distinct, successive peaks of lower parasitemia. In the S+ host, the late intraerythrocytic asexual developmental stages of P. fragile induce the expression of antigens on the surface of infected erythrocytes, which could be detected using the technique of surface immunofluorescence. Immunofluorescence using unfixed erythrocytes in suspension has shown that antigens are recognized by immune serum on the surface of the erythrocytes infected with more mature stages of the parasite. These antigens undergo variation, each successive peak of parasitemia being characterized by a different variant antigenic type (VAT). The appearance of the successive VATs occurs in a sequential manner, following the same order in different sets of animals. This constitutes the first example of a sequential expression of antigens in a malaria parasite; it indicates that, in P. fragile, antigenic variation is not the result of random mutations selected by antibody. Parasite-induced antigens on the surface of infected erythrocytes could not be detected in the S- host. However, when nonexpressing parasites from the S- host were transferred by blood passage into a naive S+ animal, they began to express antigens on the surface of infected erythrocytes within two erythrocytic cycles. We have demonstrated that the ability of S- parasites to switch to a particular VAT when passaged into a S+ animal changes during the course of an infection in the S- animal, indicating that, although surface antigens are not expressed, the processes leading to antigenic variation occurs even in the S- host. Antibodies directed against these surface antigens inhibit the growth of intra-erythrocytic parasites. The growth inhibition effects of antibodies are also variant specific, indicating that these variant surface antigens are functionally important for parasite survival.

Animals↗
PubMed1987

Demonstration of antigenic polymorphism in Plasmodium vivax malaria with a panel of 30 monoclonal antibodies.

A panel of 30 monoclonal antibodies was established against asexual erythrocytic stages of Plasmodium vivax and used to investigate the antigenic composition of the parasite. At least 38 different antigenic polypeptides of P. vivax were characterized by the Western blot technique. The possible location of these antigens, as well as their stage and species specificity, was determined on the basis of the staining patterns produced by these antibodies on air-dried parasites in the indirect immunofluorescence test. Immunofluorescence performed with 30 different monoclonal antibodies on 50 different isolates of P. vivax obtained from patients showed that a high level of antigenic polymorphism prevailed in P. vivax. Only six monoclonal antibodies reacted with epitopes that were represented in more than 80% of parasite isolates, and therefore, appeared to be relatively conserved among different isolates. The other 24 monoclonal antibodies reacted with only 20 to 70% of parasite isolates.

Animals↗
PubMed1985

Wright (a + b-) human erythrocytes and Plasmodium falciparum malaria.

We find Wr(a + b-) erythrocytes of donor M. Fr., which appear to carry a rare glycophorin A variant, to be fully susceptible to invasion by nine isolates of Plasmodium falciparum. Thus we fail to confirm the previous publication on the refractoriness of these erythrocytes. In addition the serum of donor M. Fr., which is known to contain anti-Wrb directed against an epitope located on glycophorin A in close proximity to the erythrocyte membrane, was not found to inhibit P. falciparum invasion of blood group O Rh- red blood cells. Despite this, different lines of evidence still indicate that glycophorin A is one of the receptors for erythrocyte invasion by P. falciparum. The Wrb epitope, however, does not appear to represent a distinct receptor site, which is in contrast to previous suggestions.

ABO Blood-Group System↗
PubMed1985

Immunization of monkeys with a 140 kilodalton merozoite surface protein of Plasmodium knowlesi malaria: appearance of alternate forms of this protein.

The merozoite is the invasive stage of the malaria parasite which is released by rupture of the schizont-infected erythrocyte. A monoclonal antibody against a 140 kilodalton (kDa) merozoite surface antigen of Plasmodium knowlesi was used to characterize and to purify this antigen. It was shown by pulse-chase metabolic labeling of mature schizonts that the 140 kDa merozoite antigen was the processed product of a 143 kDa schizont component, and that processing occurred at the time of erythrocyte rupture. Antiserum, prepared by immunizing a rabbit with the 143/140 kDa antigen purified by immunoaffinity chromatography with the monoclonal antibody, strongly inhibited invasion of erythrocytes in vitro; Fab fragments prepared from purified rabbit IgG were inactive at blocking invasion, suggesting that agglutination of merozoites was the mechanism of invasion inhibition. The purified 143/140 kDa antigen was used in Freund's adjuvant to immunize four rhesus monkeys. Two of the immunized animals developed fulminating infections on challenge with 10(4) schizonts, as did the three control animals. The remaining two immunized animals controlled their infections and developed chronic low-grade parasitemias. The animals which were partially protected were those that had developed anti-143/140 kDa antibodies capable of blocking invasion in vitro. Parasites were isolated from the chronic stage of infection (V5 population) and were compared with the original parasite population used for challenge (P population). Inhibition of invasion, immunofluorescence, and immunoprecipitation with anti-143/140 kDa monoclonal antibody, with immune rabbit, and with monkey sera showed that the 143/140 kDa surface antigen had been replaced by multiple cross-reacting alternate antigenic forms of the molecule in the V population. Thus, specific immune response directed against a purified merozoite surface antigen resulted in the replacement of this antigen by variant or mutant forms.

Animals↗
PubMed1984

Perspectives for malaria vaccination.

The need for vaccines to relieve the current global resurgence of malaria is apparent. Immunity is specific for each species of human malaria and for each stage in the life cycle. Once protective immunogens have been identified for one species, the homologous molecules in other species may lead to protection. The usefulness of a particular immunogen will be determined, in part, by its antigenic diversity in the population and the potential for boosting during natural infection. Successful immunization with malarial antigens may require adjuvants to induce effective, long-lived immunity. If different vaccines become available against each stage in the life cycle, then the composition of a particular vaccine may be tailored for different objectives: protection for short periods (for example, during epidemics and for tourists), decrease in disease and death, and malaria eradication.

Animals↗
PubMed1984

Identification of an erythrocyte component carrying the Duffy blood group Fya antigen.

The erythrocyte component carrying the Duffy blood group antigen Fya has been identified as a 35- to 43-kilodalton protein. The protein is degraded by proteases, chymotrypsin, and Pronase, which destroy its antigenicity on intact erythrocytes. Its unusual property of aggregating on being boiled in 5 percent sodium dodecyl sulfate with 5 percent 2-mercaptoethanol distinguishes it from other erythrocyte membrane proteins described to date.

Blood Group Antigens↗
PubMed1984

Processing of a major parasite surface glycoprotein during the ultimate stages of differentiation in Plasmodium knowlesi.

A monoclonal antibody (13C11) was used to investigate the processing of a Plasmodium knowlesi plasma membrane protein during the late stages of schizogony. 13C11 bound to the surface of merozoites, blocked invasion of erythrocytes and immunoprecipitated a 230 kDa glycoprotein from metabolically labelled schizonts. This protein was a major parasite surface component inserted into the membrane of immature schizonts as shown through the study of saponin-freed schizonts which bound 13C11 to their surface (indirect immunofluorescence and immunoelectron microscopy); in addition, the 230 kDa protein on saponin-freed schizonts was susceptible to trypsin treatment. Cleavage of the protein in pulse-chase experiments was followed by immunoprecipitation with 13C11. As schizogony proceeded, the 230 kDa protein was cleaved to 200, 145 and 110 kDa polypeptides. However, this cleavage did not reflect processing but occurred in vitro during detergent extraction and was due to a proteolytic activity which appeared in the parasite during the later stages of schizogony. As schizonts reached maturity and infected erythrocytes lysed, the 230 kDa protein was processed to 75, 57, 50 kDa and 43 kDa polypeptides which were the surface labelled components on purified merozoites immunoprecipitated by 13C11.

Animals↗