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K Marsh

Publications and source records attributed to K Marsh.

At least 199 records · Page 11Linked to original sources
PubMed1986

Identification of isolate-specific proteins on sorbitol-enriched Plasmodium falciparum infected erythrocytes from Gambian patients.

We have compared the surface radio-iodinated proteins of uninfected and Plasmodium falciparum-infected erythrocytes from natural infections of human patients. Cryopreserved infected blood from Gambian children with falciparum malaria was thawed, cultured to the middle trophozoite stage, and surface radio-iodinated. Trophozoite-infected cells were enriched about 10-fold on a Percoll gradient newly designed to separate cells based on their differential permeability to sorbitol. Infected blood was radio-iodinated and erythrocytes from the fraction enriched in parasitized cells and uninfected erythrocytes from the same sample obtained from the gradient and compared by SDS-PAGE and autoradiography. In each sample, parasitized erythrocytes contained one or more polypeptides of very high molecular weight (Mr 250 000-300 000) that were not found on uninfected erythrocytes from the same patient. These proteins were isolate-specific in size and number, suggesting that natural isolates contain a variable number of different P. falciparum phenotypes for this surface protein. In addition, these radio-iodinated surface proteins could not be extracted from the host cell membrane by the non-ionic detergent Triton X-100, but were extracted by SDS. The properties of these proteins suggest they are the equivalent for natural infections of the strain-dependent antigen previously described (Leech, Barnwell, Miller & Howard, 1984) on the surface of P. falciparum-infected Aotus erythrocytes. In addition, we observed a second parasite-dependent modification of labelled proteins on infected erythrocytes with the appearance of a new band of Mr 30 000. There were also variations in the pattern of radio-isotope labelled proteins on uninfected erythrocytes from different patients.

Animals↗
PubMed1986

Antigenic diversity and size diversity of Plasmodium falciparum antigens in isolates from Gambian patients. I. S-antigens.

Ring-stage asexual parasites of P. falciparum were collected from six Gambian children and the S-antigens radiolabelled by 3H-glycine uptake during in vitro culture up to rupture of infected cells and merozoite release. Ouchterlony double diffusion of boiled culture supernatants against a panel of adult Gambian sera identified one S-antigen precipitin arc for five isolates and two precipitin arcs for one isolate. Five of the six isolates were serologically distinct. Analysis of S-antigens by comparison of SDS-polyacrylamide gel electrophoresis patterns of heat-treated soluble proteins revealed a more complex pattern of 3H-labelled S-antigens that was different for each isolate. There were between two and six different 3H-labelled bands for each isolate in the size range of molecular weight 137 000 to 285 000. This result confirms the large size range of S-antigens identified with culture adapted P. falciparum. Several bands were relatively weakly labelled with 3H-glycine, suggesting that natural isolates contain one or two predominant S-antigen phenotypes and several other S-antigen phenotypes expressed by minor parasite subpopulations. Immunoprecipitation was performed using a panel of sera from Gambian adults, or, acute and 3 week convalescent sera from the same patients used for S-antigen radiolabelling. Adult sera generally immunoprecipitated some of the S-antigens in each isolate, including antigens that must represent extremely minor parasite subpopulations since they could not be seen in the patterns of non-immunoprecipitated heat-stable proteins. Sera from convalescent children were generally negative on immunoprecipitation, even with the homologous isolate. In one case we observed the acquisition of specific immunoprecipitating antibody to one of the homologous S-antigens during the convalescent period. The antigenic and structural complexity of S-antigens in natural isolates that have not been submitted to the selection pressure of adaptation for in vitro culture is clearly greater than for culture adapted P. falciparum.

Adult↗
PubMed1986

Antigenic diversity and size diversity of P. falciparum antigens in isolates from Gambian patients. II. the schizont surface glycoprotein of molecular weight approximately 200 000.

A panel of monoclonal antibodies has been shown previously to identify both serologically diverse and serologically conserved epitopes on a major polymorphic surface protein of P. falciparum schizonts from culture-adapted isolates. The molecular nature of the antigen recognized by eight of these monoclonal antibodies was studied with three isolates analyzed directly from patients in The Gambia. Malarial (glyco) proteins were labelled by biosynthetic uptake of 3H-glucosamine or 3H-leucine during culture of ring-stage parasites from infected blood to the late-trophozoite/early-schizont stage (26-30 h). Those monoclonal antibodies which reacted positively with an isolate by indirect immunofluorescence also immunoprecipitated a single 3H-leucine or 3H-glucosamine labelled antigen of mol. wt approximately 200 000 from Triton X-100 extracts of the same isolate. Monoclonal antibodies which did not react by indirect immunofluorescence failed to immunoprecipitate this antigen. Although each of the three isolates studied in detail was very similar serologically with the panel of monoclonal antibodies specific for this mol. wt approximately 200 000 antigen, this protein could be distinguished with each isolate on the basis of its apparent size on SDS-polyacrylamide gel electrophoresis. The specifically immunoprecipitated antigen had a mol. wt of 204 000, 197 000 or 202 000, depending on the isolate. Size diversity of this malarial glycoprotein was also detected with seven other Gambian P. falciparum isolates. We conclude that natural isolates of P. falciparum express a major 3H-glucosamine labelled glycoprotein of mol. wt Mr approximately 200 000 which exhibits size diversity and expresses antigenically conserved as well as diverse epitopes as defined by the panel of monoclonal antibodies.

Antibodies, Monoclonal↗
PubMed1986

Inhibition of Plasmodium falciparum growth by IgG antibody produced by human lymphocytes transformed with Epstein-Barr virus.

Supernatants from Epstein-Barr virus (EBV)--stimulated B lymphocytes obtained from two adult Gambians who were partially immune to malaria markedly inhibited the growth of Plasmodium falciparum in vitro (55-95% inhibition). When 22 separate colonies were derived by micromanipulation from one of these primary cultures and their supernatants assayed, the degree of inhibition correlated with levels of IgG fluorescent antibody and total IgG. The inhibitory anti-P. falciparum IgG immunoprecipitated an antigen of mol. wt 195,000, identified as the major schizont surface glycoprotein by dual biosynthetic labelling with 3H-glucosamine or 35S-methionine. Other studies on the analogous schizont surface protein of rodent malarias have shown that this antigen stimulates protective immunity. Production of this inhibitory antibody by adult Gambians may therefore contribute to their immunity to malaria. Human antibodies produced by EBV-stimulated B lymphocytes may be used to identify other important P. falciparum antigens and have potential applications for immunotherapy.

Adult↗
PubMed1986

Parasitized erythrocyte antigens and thrombospondin adhesion in the immunology and pathogenesis of falciparum malaria.

Falciparum-parasitized erythrocytes develop new antigens on the erythrocyte surface. Both antimalarial antibody and thrombospondin bind to surface antigens or ligands on the surface of the infected red cell. Diverse antigens may be involved in parasite evasion of host immunity, but conserved molecules may be required for pathogenesis. There is a high degree of antigenic diversity at the infected erythrocyte surface among isolates. This diversity may underlie the potential of falciparum malaria to repeatedly reinfect the same person. There is also evidence for an antigenically conserved, possibly functionally conserved, determinant on the infected red cell surface. Antibody to a conserved determinant on the infected red cell surface may be associated with immunity to falciparum malaria. A conserved membrane surface determinant may be involved in parasitized erythrocyte sequestration. Thrombospondin binding in vitro is specific to mature trophozoites and schizonts, the same stages which sequester in vivo. Specific adhesion to thrombospondin is common among laboratory strains of falciparum-parasitized erythrocytes bearing knobs and among all isolates of falciparum tested. The numbers of parasitized erythrocytes bound to thrombospondin in vitro increases with parasitemia. Specific binding of parasitized erythrocytes to an endothelial receptor and to thrombospondin, which itself interacts with clotting factors, may play a role in sequestration and vascular obstruction.

Antigens, Protozoan↗
PubMed1985

A vector that replicates as a plasmid and can be efficiently selected in B-lymphoblasts transformed by Epstein-Barr virus.

Epstein-Barr virus (EBV) transforms human B-lymphocytes into proliferating blasts which are efficiently established into cell lines. The viral DNA in these cell lines is usually present as complete, unintegrated plasmid molecules. A cis-acting element of EBV, oriP, permits plasmid maintenance in adherent cells that carry EBV DNA. We constructed a vector, pHEBo, that carries oriP and showed that it is also efficiently maintained as a plasmid when introduced into EBV-transformed B-lymphoblasts. The pHEBo vector carries the coding sequences for the hph gene from Escherichia coli such that it can be expressed in mammalian cells and confers resistance to the antibiotic hygromycin B. Hygromycin B kills EBV-transformed lymphoblasts at concentrations of 50 to 300 micrograms/ml. The combination of oriP plus the expressed hph gene makes pHEBo useful for the stable introduction of genes on plasmids into EBV-transformed lymphoblasts. Because pHEBo is derived from the plasmid pBR322 it can be easily isolated from lymphoblasts by reintroduction into E. coli.

B-Lymphocytes↗
PubMed1982

The inspiratory "squawk" in extrinsic allergic alveolitis and other pulmonary fibroses.

An inspiratory musical sound ("squawk") was recorded in 14 patients with diffuse pulmonary fibrosis. These were divided into two groups: nine patients suffering from extrinsic allergic alveolitis (seven with bird fancier's lung and two with farmer's lung) and five patients with pulmonary fibrosis due to other causes, including rheumatoid disease, Wegener's granulomatosis, systemic sclerosis, and sarcoidosis. Clinical studies and phonopneumographic analysis of 10 consecutive squawks in each patient showed that the sound in the group with extrinsic allergic alveolitis was of shorter duration, occurred later in inspiration, and tended to be of higher frequency than the sound heard in the other group. Inspiratory crackles were present in all patients and in eight a single loud crackle preceded the squawk. We suggest that squawks, like crackles, result from the opening of airways and that the differences between the squawks in the two groups may reflect the size of the affected airways.

Adult↗
PubMedDate not supplied

Antibodies to blood stage antigens of Plasmodium falciparum in rural Gambians and their relation to protection against infection.

Cross-sectional and longitudinal studies were performed in a rural population living in The Gambia to examine the relationship between several in vitro assays of the host immune response to asexual stages of Plasmodium falciparum and protection from malaria in vivo. Assays included an enzyme-linked immunosorbent assay for antibodies to schizont antigens; an indirect immunofluorescence assay for total antiblood-stage antibodies; an immunofluorescence assay on glutaraldehyde-fixed parasites to detect antibodies to antigen Pf 155; an assay for serum inhibition of red blood cell invasion; a micro-agglutination assay to detect antibodies to neo-antigens on the surface of infected red blood cells; and an assay using polymorphonuclear leucocytes to detect antibodies capable of opsonizing schizont infected red blood cells. There were marked differences in the age-related pattern of response for different assays performed on sera obtained at a cross-sectional survey of 280 individuals. Examination of the correlation between the various immune responses and malariometric indices at the population level and at the individual level provided no evidence that any of the in vitro assays were related to protective immunity. The relationship between in vitro measurements of the anti-malarial immune response and protection from clinical episodes of malaria was examined in a group of 134 children aged 11 years and under who were monitored weekly throughout an entire malaria transmission season. The only immune factor to show a consistent protective effect against clinical malaria was the titre of antibodies to neo-antigens on the infected erythrocyte surface (P = 0.01). The same longitudinal techniques were used to examine the effect of two non-immunological factors, sickle cell trait and mosquito net usage, both of which showed significant protection against clinical episodes and malaria.

Age Factors↗
PubMedDate not supplied

Infection with Pneumocystis carinii is prevalent in healthy Gambian children.

Pneumocystis pneumonia is rarely identified in the many immunosuppressed individuals with acquired immune deficiency syndrome (AIDS) and malnutrition in Africa. To test whether infection with Pneumocystis carinii occurs in the continent we conducted a comparative serological study, measuring by enzyme-linked immunosorbent assay antibodies to the parasite in 150 healthy young individuals from both Britain and the Gambian savanna. The prevalence of significant titres of antibody to P. carinii steadily increased with age and included more than 70% of both populations by 8 years of age. Infection with P. carinii is, therefore, common in the Gambia. Thus opportunistic pneumocystis pneumonia may be an important but largely unrecognized disease in the continent, though its impact is probably diminished by the prevalence of fatal tuberculous infection, particularly in the AIDS population.

Adolescent↗
PubMedDate not supplied

The role of shops in the treatment and prevention of childhood malaria on the coast of Kenya.

A community survey of 388 mothers in a rural and peri-urban population surrounding a district hospital on the coast of Kenya revealed that the preferred choice of treatment for childhood febrile illnesses was with proprietary drugs bought over the counter at shops and kiosks (72% of interviews). 67% of the mothers who reported using shops claimed they would buy chloroquine-based drugs. Preventative measures such as mosquito nets were uncommon (6.2%), but the use of commercial pyrethrum mosquito coils was reported more frequently (46.4%). Separate investigations of treatment given to 394 children before presentation at hospital with severe and mild malaria was consistent with the reports in the community of high usage of shop-bought anti-malarials and anti-pyretics. The involvement of the private sector in peripheral health care delivery for malaria is discussed.

Adult↗
PubMedDate not supplied

A single dose of intramuscular sulfadoxine-pyrimethamine as an adjunct to quinine in the treatment of severe malaria: pharmacokinetics and efficacy.

It has been suggested that sulfadoxine-pyrimethamine (SD/PM) may be useful in the treatment of severe malaria since it could enhance the killing of parasites by quinine (QN) and it can be given as a single intramuscular injection. Eighty Kenyan children with severe malaria were allocated at random to receive either intramuscular QN alone (quinine dihydrochloride 20 mg salt/kg as a loading dose, followed by 10 mg salt/kg 12 hourly for a total of 6 doses) or the same QN regimen plus one intramuscular injection of SD/PM (sulfadoxine 25 mg/kg, pyrimethamine 1.25 mg/kg). There was no difference in time to defervescence, aparasitaemia, or 50% reduction in parasitaemia, parasite elimination half-life, or mortality between the 2 groups. In addition, the concentrations of SD and PM were measured in 14 children and of QN in 8 of these children. Concentrations needed to achieve synergy against PM-resistant strains of Plasmodium falciparum were achieved in all of the children with severe malaria within the first hour and maintained for more than 72 h. SD/PM did not perturb the pharmacokinetics of QN.

Acute Disease↗