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Biomedical subjects

W E Collins

Publications and source records attributed to W E Collins.

At least 181 records · Page 10Linked to original sources

Genetic selection of a Plasmodium-refractory strain of the malaria vector Anopheles gambiae.

The anopheline mosquito is the target in most malaria control programs, primarily through the use of residual insecticides. A mosquito was studied that is refractory to most species of malaria through a genetically controlled mechanism. A strain of Anopheles gambiae, which was selected for complete refractoriness to the simian malaria parasite Plasmodium cynomolgi, also has varying degrees of refractoriness to most other malaria species examined, including the human parasites P. falciparum, P. ovale, and P. vivax for which this mosquito is the principal African vector. Furthermore, the refractoriness extends to other subhuman primate malarias, to rodent malaria, and to avian malaria. Refractoriness is manifested by encapsulation of the malaria ookinete after it completes its passage through the mosquito midgut, approximately 16 to 24 hours after ingestion of an infective blood meal. Fully encapsulated ookinetes show no abnormalities in parasite organelles, suggesting that refractoriness is due to an enhanced ability of the host to recognize the living parasite rather than to a passive encapsulation of a dead or dying parasite. Production of fully refractory and fully susceptible mosquito strains was achieved through a short series of selective breeding steps. This result indicates a relatively simple genetic basis for refractoriness. In addition to the value these strains may serve in general studies of insect immune mechanisms, this finding encourages consideration of genetic manipulation of natural vector populations as a malaria control strategy.

Animals↗

Evaluation of serological cross-reactivity between antibodies to Plasmodium and HTLV-III/LAV.

Serum samples from 460 patients with existing or previous Plasmodium infections, high antimalarial antibody titres, and no apparent risk of exposure to human T-lymphotropic virus type III/lymphadenopathy-associated virus (HTLV-III/LAV) were assayed for HTLV-III/LAV antibody; only 1 sample, from a 21-year-old African woman, was strongly reactive by enzyme-linked immunosorbent assay (ELISA) and positive by western blot. Conversely, no sample from 100 HTLV-III/LAV-positive American homosexual men was strongly reactive for antibodies to the four Plasmodium species that infect human beings by an indirect fluorescent antibody technique, or for antibodies to Plasmodium falciparum by an ELISA technique. Thus, exposure to Plasmodium does not result in HTLV-III/LAV seropositivity, and HTLV-III/LAV antibodies are not strongly cross-reactive with malarial antigens.

Acquired Immunodeficiency Syndrome↗

Specific detection of Plasmodium falciparum malaria by a molecularly cloned DNA probe.

A highly repeated DNA sequence from Plasmodium falciparum was cloned and used as a probe in molecular hybridization to detect malaria. Our results indicate that the probe is specific to P. falciparum but not to other species of Plasmodium and is extremely sensitive. As little as a 20 pg parasite DNA, which is equivalent to about 1000 parasites can be detected. The cloned DNA can be used as a diagnostic tool to follow the course of infection of falciparum malaria.

Animals↗

Inhibition of development of exoerythrocytic forms of malaria parasites by gamma-interferon.

A specific DNA probe was used to study the effect of recombinant rat, mouse, and human gamma-interferon (gamma-IFN) on the course of sporozoite-induced malaria infections. In mice and rats infected with sporozoites of Plasmodium berghei, mouse and rat gamma-IFN's strongly inhibited the development of the exoerythrocytic forms in the liver liver cells of the hosts, but not the development of the erythrocytic stages. The degree of inhibition of the exoerythrocytic forms was proportional to the dose of gamma-IFN administered, but was independent of the number of sporozoites used for challenge. A 30 percent reduction in the development of exoerythrocytic forms in rat liver was achieved when 150 units (about 15 nanograms of protein) of rat gamma-IFN were injected a few hours before sporozoite challenge; the reduction was 90 percent or more with higher doses of gamma-IFN. The effect was less pronounced if the gamma-IFN was administered 18 hours before or a few hours after challenge. Human gamma-IFN also diminished the parasitemia in chimpanzees infected with sporozoites of the human malaria parasite Plasmodium vivax. The target of gamma-IFN activity may be the infected hepatocytes themselves, as shown by in vitro experiments in which small doses of the human lymphokine inhibited the development of exoerythrocytic forms of Plasmodium berghei in a human hepatoma cell line. These results suggest that immunologically induced interferon may be involved in controlling malaria infection under natural conditions.

Animals↗

Stage-specific and species-specific antigens of Plasmodium vivax and Plasmodium ovale defined by monoclonal antibodies.

Monoclonal antibodies (MAbs) were produced against the asexual blood stages of Plasmodium vivax and Plasmodium ovale and used to define antigens of plasmodial parasites in an indirect fluorescent antibody assay. The anti-P. vivax MAbs produced two distinct patterns in the indirect fluorescent antibody assay. Four patterns were found with the anti-P. ovale MAbs. Species-specific epitopes were defined for P. vivax and P. ovale; epitopes shared among all four species of human malaria parasites were also defined. Some of the anti-P. vivax MAbs reacted only with mature stages, and others reacted with all asexual stages. No asexual blood-stage specificity could be found with the anti-P. ovale antibodies. Five of the anti-P. vivax MAbs and three of the anti-P. ovale MAbs also reacted with sporozoites.

Animals↗

Concentration of Plasmodium ovale- and Plasmodium vivax-infected erythrocytes from nonhuman primate blood using Percoll gradients.

Plasmodium vivax and Plasmodium ovale schizont-infected erythrocytes were separated from peripheral blood by centrifugation using discontinuous Percoll (colloidal silica) gradients. Infected Aotus monkey or chimpanzee blood was diluted and placed on a discontinuous gradient containing 30%, 40%, 45%, and 50% Percoll (v/v in media) layers before centrifugation at 1,450 X g. Parasitized erythrocytes were concentrated to greater than 95% schizont-infected cells in two bands that contained an average of one leukocyte per 500 infected cells. Mononuclear cells and trophozoites were isolated in another band and noninfected red cells, ring-infected cells, and granulocytes were pelleted to the bottom. The yield of parasitized erythrocytes ranged from 50% to close to 100% of the estimated number of infected cells in the original whole blood. Use of this Percoll procedure results in a high yield of concentrated parasitized erythrocytes and separation of these cells from host white blood cells.

Animals↗

Observations on early and late post-sporozoite tissue stages in primate malaria. IV. Pre-erythrocytic schizonts and/or hypnozoites of Chesson and North Korean strains of Plasmodium vivax in the chimpanzee.

In a continuing reexamination of plasmodial tissue stages within the context of the hypnozoite theory of malarial relapse, 2 strains of Plasmodium vivax with distinct and disparate relapse characteristics in humans were studied in chimpanzees. Following intravenous inoculation of massive numbers of salivary gland sporozoites, both the frequently relapsing Chesson strain and a North Korean strain characterized by predominantly delayed relapses exhibited relapse patterns and antimalarial sensitivity in the splenectomized chimpanzee essentially indistinguishable from those seen in humans. Examination of hepatic biopsies obtained at 7 and 10 days after infection revealed both pre-erythrocytic (pre-e) schizonts and hypnozoites in tissue obtained from the animal infected with the Chesson strain, but only rare hypnozoites (no pre-e schizonts) at 7 days in the animal infected with the North Korean strain. These findings, combined with the comparability of relapse behavior--which indicates the suitability of the chimpanzee as a model for the natural (human) host-parasite relationship--are essentially as predicted by the hypnozoite theory, despite the small numbers of tissue forms seen. Pre-erythrocytic schizogony of the Chesson strain in the liver was essentially indistinguishable from that of other strains studied, also underlining the suitability of this model system for tissue stage studies of P. vivax.

Animals↗

Adaptation of the Indochina I/CDC strain of Plasmodium falciparum to the squirrel monkey (Saimiri sciureus).

The Indochina I/CDC strain of Plasmodium falciparum was linearly passaged in squirrel monkeys (Saimiri sciureus) of 3 phenotypes. Splenectomized monkeys of Guyanan and Peruvian type developed high density parasitemias, but considerably lower than the mean peak parasitemia (greater than 10(6)/mm3) in Bolivian phenotype squirrel monkeys. Spleen-intact Bolivian and Peruvian squirrel monkeys all developed potentially lethal infections after linear passage of parasites from Saimiri and Aotus. For the evaluation of induced immunity to P. falciparum, the Indochina I/CDC strain in Saimiri will be a valuable model system.

Animals↗

Infection of mosquitoes with Plasmodium vivax from chimpanzees using membrane feeding.

Six splenectomized chimpanzees were infected with the Chesson or the North Korean strains of Plasmodium vivax. Heparinized blood taken from the animals was fed to approximately 45,000 mosquitoes using parafilm membranes. High-level mosquito infections were obtained with the blood from 4 animals. One animal infected mosquitoes only at a very low level. The other chimpanzee failed to produce a parasitemia high enough to warrant mosquito feeding.

Animals↗

Studies of comparative infectivity of fifteen strains of Plasmodium vivax to laboratory-reared anopheline mosquitoes, with special reference to Anopheles culicifacies.

The Sattoki strain of species A of the taxon Anopheles culicifacies Giles was infected with 15 different strains of Plasmodium vivax from Asia, New Guinea, and Central and South America. A comparison of the relative infectivity indicated a marked variation for the different strains of P. vivax when compared to Anopheles freeborni mosquitoes.

Animals↗

Infection of Aotus azarae boliviensis monkeys with different strains of Plasmodium falciparum.

Eleven strains of Plasmodium falciparum from Asia, Africa, and Central America were inoculated into a total of 58 splenectomized Aotus azarae boliviensis monkeys. Eight of the strains produced high-level parasitemias, whereas 3 (2 from Honduras and 1 from Zaire) produced only low-level parasitemias. Mosquito infections were only obtained during the first 2 linear passages of the Santa Lucia strain from El Salvador. The results indicate that this species of Aotus monkey is highly susceptible to infection with strains of P. falciparum from different geographic areas, and therefore may be useful for a number of chemotherapeutic or immunologic studies. Its usefulness for mosquito infection studies is very limited.

Africa↗

The sporogonic cycle of Plasmodium reichenowi.

Plasmodium reichenowi, a malarial parasite of the chimpanzee, was infective to Anopheles freeborni, Anopheles quadrimaculatus, Anopheles stephensi, Anopheles maculatus, Anopheles dirus, and Anopheles culicifacies mosquitoes. Anopheles gambiae and Anopheles albimanus were not infected. Mean oocyst diameters of P. reichenowi were smaller than those of the other chimpanzee parasite, Plasmodium schwetzi. Sporozoites were present in the salivary glands of An. freeborni at 15 days when held at 25 to 26 C.

Animals↗

Class II positive human dermal fibroblasts restimulate cloned allospecific T cells but fail to stimulate primary allogeneic lymphoproliferation.

Recently, a number of laboratories have shown that gamma interferon (IFN-gamma) is a potent modulator of HLA class II antigen expression in a variety of cell types ranging from classical antigen presenting cells to those not expected to participate in physiological antigen presentation such as fibroblasts. In order to examine the role of HLA class II expressing fibroblasts in antigen presentation, we established dermal fibroblast (FIB) strains from five HLA typed donors. After optimal preculture with IFN-gamma, class II positive FIB were fully competent to restimulate proliferative responses of two DR specific T cell clones and one DP specific T cell line. However, they failed to elicit strong primary allogeneic proliferation from fully DR mismatched fresh PBMC. This failure was not due to a direct suppressive effect of FIB and could not be corrected by exogenous IL1 or by factors contained in conventional mixed leukocyte culture supernatants.

Antigen-Presenting Cells↗

Monoclonal antibodies produced against sporozoites of the human parasite Plasmodium malariae abolish infectivity of sporozoites of the simian parasite Plasmodium brasilianum.

We have used a sporozoite neutralization assay to define the biological relevance of the cross-reactivity of two monoclonal antibodies, raised against sporozoites of the human parasite Plasmodium malariae (Uganda 1/CDC), with sporozoites of the simian parasite Plasmodium brasilianum (Colombian). In vitro incubation of each of these two monoclonal antibodies with sporozoites of P. brasilianum totally abolished the infectivity of these parasites for Saimiri sciureus. Using Western blot analysis and one of the P. malariae monoclonal antibodies, we identified two sporozoite proteins characteristic of the Colombian isolate of P. brasilianum with apparent molecular weights of 56,000 and 66,000. The same monoclonal antibody identified two proteins in an extract of the Peruvian isolate of P. brasilianum with apparent molecular weights of 59,000 and 69,000.

Animals↗