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At least 19 recordsLinked to original sources
PubMed1997

Plasmodium yoelii sporozoite infectivity varies as a function of sporozoite loads in Anopheles stephensi mosquitoes.

Mechanisms by which Plasmodium sporozoites survive and maintain their infectivity within the salivary glands of mosquitoes are unknown. In this study we establish a relationship between the number of sporozoites present in the salivary glands of individual mosquitoes (sporozoite load) and sporozoite infectiousness (or "quality") as measured by infections in BALB/c or ICR mice. When Plasmodium yoelii-infected Anopheles stephensi mosquitoes were each allowed to feed on a single mouse, we noted that sporozoites from mosquitoes with higher sporozoite loads were more infectious in 13 of 30 (43%) mice. In a second experiment, we inoculated mice with known numbers of sporozoites from individual mosquitoes. Eleven of 18 (61%) and 16 of 18 (89%) mice that received 25 and 100 sporozoites, respectively, became infected. For inoculations using 100 sporozoites, again we noted that sporozoites from mosquitoes with higher sporozoite loads were more infectious to mice. In a third and final experiment, the overall infectiousness of sporozoites from individual mosquitoes was evaluated first by allowing individual mosquitoes to feed on individual mice and then by intravenous inoculations of 100 sporozoites in a second mouse. There was a significant difference in host infections as a function of sporozoite loads in 14 of 19 (74%) mice. Analysis of the feeding times for infected versus noninfected mosquitoes did not show a significant difference between the 2 groups. The mean total feeding times for 50 infected and 45 noninfected An. stephensi mosquitoes were 306 (standard deviation [SD] = +/-230) and 441 (SD = +/-273) sec, respectively. Further, among infected An. stephensi mosquitoes there was no difference in probing times between cohorts that transmitted infectious sporozoites to mice and cohorts that failed to transmit infectious sporozoites. Our findings that sporozoite load influences sporozoite infectiousness or quality suggest that this may be an important factor in malaria parasite transmission.

Animals↗
PubMed1987

Experimental vaccination of chicks with Plasmodium gallinaceum sporozoites. I. Circumsporozoite proteins are expressed by sporozoites recovered from both salivary glands and midguts of mosquitoes.

Immunogenicity of Plasmodium gallinaceum sporozoites for chicks and their in vitro reactivity with normal and specific immune sera were studied. Two sporozoite populations recovered from experimentally infected Aedes fluviatilis were used: sporozoites from salivary glands and sporozoites from midgut oocysts. Populations seven to nine days old of sporozoites recovered from salivary glands were infective for all chicks until the chicks were three weeks old; however, sporozoites recovered from midguts containing oocysts infected these chicks only if isolated on days 8-9, but not on day 7 after the mosquitoes' infective blood meal. Infectivity of the sporozoites was lost after exposure to ultraviolet (UV) light (30 min) or X-rays (13 krad). Inactivated sporozoites from both sources proved highly immunogenic to chicks that were immunized by several intravenous or intramuscular injections. These parasites elicited a strong humoral immune response in the chicks, as measured by the circumsporozoite precipitation (CSP) reaction. The levels of the CSP antibodies were similar with sporozoites from both sources, there being no detectable differences in the percentage of reactive sporozoites or the intensity of the CSP reaction with sera containing antibodies to either sporozoites from salivary glands or sporozoites from oocysts. These results provide the first evidence that avian malaria sporozoites express the circumsporozoite protein that has been extensively characterized in mammalian malaria (rodent, simian, human sporozoites). Furthermore, we observed that the yields of sporozoites obtained from mosquito midguts, on days 8 and 9 of the P. gallinaceum infection, were at least twice as great as those obtained by salivary gland dissection, even 20 days after a blood meal.(ABSTRACT TRUNCATED AT 250 WORDS)

Aedes↗
PubMed1999

Irradiated sporozoites prime mice to produce high antibody titres upon viable Plasmodium berghei sporozoite challenge, which act upon liver-stage development.

C57BL6 mice were protected against Plasmodium berghei sporozoite challenge by immunization with live 12 krad dose-irradiated sporozoites, but not by 20 krad dose-irradiated sporozoites. Immunization with 12 krad irradiated sporozoites generated low levels of antibody reactive to liver-stage parasites (titres of 1/100). Inoculation of as few as 100 live P. berghei sporozoites induced complete host protection accompanied by a very quick and high boost of antibody titres up to 1/4000. This sporozoite challenge-drive antibody boost was absent in mice immunized by 20 krad sporozoites and in non-protected, and non-immunized mice. Antibody was mainly liver-stage (LS) specific and due to an increase of IgG2a and IgG2b. The in vitro effect of pre- and post-challenge sera upon either sporozoite invasion or LS development was assessed in Hep-G2 cultures. Both were found to have a strong effect upon LS development even at 1/2500 dilution, and conversely a low effect upon invasion. These results suggest that sporozoites irradiated at doses that induce protection are able to prime T-cells which, upon challenge by non-irradiated sporozoites, provide help to B-lymphocytes to trigger the production of high titres of anti-LS antibodies that can inhibit LS development in vitro.

Animals↗
PubMed1990

Plasmodium sporozoite-host cell interactions during sporozoite invasion.

In vitro and in vivo studies were performed to clarify the nature of some interactions between Plasmodium berghei sporozoites and rodent host cells. Videomicroscopic observations were made on in vitro interactions between sporozoites and cultured host cells (rodent peritoneal macrophages, W138 human lung fibroblasts, and HepG2 human hepatoma cells). The results showed a diversity of dynamic interactions and sporozoite activities, including active sporozoite penetration, often followed by sporozoite escape, killing of invaded macrophages, phagocytosis of sporozoites by macrophages, and elaboration of a weblike structure by the sporozoite, following its escape from cells. Other studies were performed to examine host cell inflammatory responses to challenge sporozoites in the livers of rats that had been previously immunized with radiation-attenuated sporozoites. The results showed a variety of focal inflammatory infiltrates in the livers of the challenged animals. The relevance of these observations to the problems of sporozoite invasion into natïve and immunized animals is discussed.

Animals↗
PubMed1993

Plasmodium falciparum sporozoite immunization protects against Plasmodium berghei sporozoite infection.

Irradiated sporozoites are generally thought to elicit protective immune responses that are parasite stage and species specific. But immunization with Plasmodium falciparum sporozoites delivered by the bite of infected mosquitoes protects an average of 60% mice from Plasmodium berghei sporozoite infection. Protection appears to be specific as P. falciparum sporozoite-immunized mice protected against P. berghei remain susceptible to Plasmodium yoelii sporozoite infection. Passively transferred immunoglobulin from P. falciparum sporozoite-immunized mouse serum protected naive mice against challenge with P. berghei sporozoites, indicating that cross-protection is mediated, at least in part, by anti-sporozoite antibody. Antibody-mediated protection may not be due to cross-reaction between P. falciparum and P. berghei CS proteins because mice immunized with the recombinant P. falciparum CS protein repeat vaccine candidate, R32tet32, remain susceptible to P. berghei sporozoite infection.

Animals↗
PubMed1986

Plasmodium berghei sporozoite invasion is blocked in vitro by sporozoite-immobilizing antibodies.

A monoclonal antibody directed against the circumsporozoite protein on the surface of Plasmodium berghei sporozoites inhibited sporozoite motility in vitro. These immobilized sporozoites could adhere to but not invade target cultured cells. Other sporozoite-immobilizing agents also inhibited sporozoite invasion into cultured cells and did not prevent sporozoite adherence. These results indicate that sporozoite invasiveness is associated with sporozoite motility. Thus, the immobilizing effect of this antibody could explain its functional activity against sporozoite invasion in vivo.

Adhesiveness↗
PubMed1993

Induction of hepatic inflammatory response by Plasmodium berghei sporozoites protects BALB/c mice against challenge with Plasmodium yoelii sporozoites.

BALB/c mice are about 2,000 times less susceptible to sporozoites of Plasmodium berghei than to Plasmodium yoelii. Associated with this is the innate cellular response mounted after injection with P. berghei. Host inflammatory cells do not normally attack P. yoelii during their development as exoerythrocytic forms (EEFs) in the liver. We used P. berghei sporozoites to induce host inflammation that might act against developing P. yoelii EEFs. Mice injected with P. berghei sporozoites followed 1 hr later with P. yoelii had a 58% reduction in P. yoelii EEFs. To establish whether this was due to events that occurred before vs. after invasion of hepatocytes by P. yoelii sporozoites, mice received P. yoelii sporozoites that were allowed to invade for 1 hr before subsequent injection with P. berghei; these mice showed minimal reduction in P. yoelii EEFs. Thus, most of the deleterious effects of P. berghei sporozoites appear to have been directed against P. yoelii sporozoites prior to their invasion of hepatocytes. Plasmodium yoelii that had already invaded were relatively unaffected. Further timing experiments showed that this effect was induced only by viable P. berghei sporozoites, which may thus induce rapid changes in sinusoid physiology leading to host resistance against P. yoelii sporozoites.

Animals↗
PubMed1987

Plasmodium falciparum: sporozoite boosting of immunity due to a T-cell epitope on a sporozoite vaccine.

Plasmodium falciparum: Sporozoite boosting of immunity due to a T-cell epitope on a sporozoite vaccine. Experimental Parasitology 64, 64-70. The impact of a malaria sporozoite vaccine may be enhanced if protective immunity elicited by the vaccine is boosted by natural exposure to sporozoites. For this to occur, a helper T lymphocyte epitope present on the vaccine must be shared by sporozoites. These studies show that T cells from mice immunized with R32tet32, the Plasmodium falciparum sporozoite vaccine candidate, recognize an epitope of less than or equal to 7 amino acids derived from the circumsporozoite protein repeat region of R32tet32, as well as an epitope on the tet32 fusion protein tail of R32tet32. Exposure of R32tet32 immunized animals to P. falciparum sporozoites elicits a significant secondary antibody response which suggests that humans who are immunized and respond to this vaccine may be boosted by field exposure to sporozoite infected mosquitoes.

Animals↗
PubMedDate not supplied

Invasion of different cell types by sporozoites of Eimeria species and effects of monoclonal antibody 1209-C2 on invasion of cells by sporozoites of several apicomplexan parasites.

Sporozoites of avian Eimeria species differed markedly in their ability to invade cells in vitro. Invasion by E. tenella and E. adenoeides was significantly greater in baby hamster kidney (BHK) and chicken cecal cell (CC) cultures than in primary chicken (PCK) or turkey kidney (PTK) cell cultures. Moreover, invasion of BHK cell cultures by E. adenoeides was significantly greater than that of other Eimeria species, and invasion by E. acervulina sporozoites was significantly lower. Monoclonal antibody 1209-C2 (MAb 1209-C2) reacted by immunofluorescent labeling (IFA) with refractile bodies of sporozoites of 5 species of Eimeria and Caryospora bigenetica, but not with sporozoites of Toxoplasma gondii, Hammondia hammondi, or Cryptosporidium parvum, which have no refractile bodies. The MAb also cross-reacted with formalin-fixed BHK, CC, turkey cecal (TC) cells, and PTK. Pretreatment of BHK cells with MAb 1209-C2 significantly reduced invasion of the cells by sporozoites of E. tenella, E. acervulina, E. meleagrimitis, and C. bigenetica, but did not alter invasion by T. gondii, C. parvum, or H. hammondia. Apparently, reactivity of MAB 1209-C2 with the sporozoites was required for inhibition of invasion despite the fact that the inhibition resulted from pre-treatment of the host cell. Conversely, although MAb 1209-C2 also reacted moderately with PTK and TC cells, pre-treatment of these cell cultures with the MAb did not inhibit invasion by either MAB 1209-C2-reactive or -nonreactive parasites. Collectively, the data indicated that refractile body antigens of sporozoites of Eimeria and Caryospora, which are recognized by MAb 1209-C2, may function in cellular invasion, but also suggest that cellular invasion is probably not mediated by interactions between the conserved epitopes in sporozoites and cultured host cells that are recognized by the MAb.

Animals↗
PubMed1988

Human malaria transmission studies in the Anopheles punctulatus complex in Papua New Guinea: sporozoite rates, inoculation rates, and sporozoite densities.

Malaria sporozoite rates and inoculation rates were measured over periods up to 25 months in the different anopheline species biting humans in 13 villages in Madang Province, Papua New Guinea. Analysis of three members of the Anopheles punctulatus complex, 68,458 An. farauti, 36,779 An. koliensis, and 11,667 An. punctulatus caught in landing catches was made using monoclonal antibody based ELISAs to detect sporozoites of Plasmodium falciparum and P. vivax. Sporozoite rates ranged from 0%-5.5% in An. farauti, 0.2%-3.8% in An. koliensis, and 0%-3.3% in An. punctulatus. In addition, over 3,000 An. longirostris were analyzed and sporozoites were not detected in this species. No significant differences were observed between the three vector species in the densities of P. falciparum sporozoites (geometric mean 2,320). However, the geometric mean P. vivax sporozoite density was significantly higher in An. punctulatus (350) than in either An. koliensis (160) or An. farauti (150). An. koliensis was less susceptible to infections of P. falciparum or P. vivax than either An. farauti or An. punctulatus. Variations in average sporozoite and inoculation rates were found among different villages, despite their close geographic proximity. Sporozoite and inoculation rates varied greatly within a village over time, but malaria transmission was perennial with a higher transmission during the wet season by An. koliensis and An. punctulatus.

Animals↗
PubMed1990

Differences in susceptibility among mouse strains to infection with Plasmodium berghei (ANKA clone) sporozoites and its relationship to protection by gamma-irradiated sporozoites.

Three inbred mouse strains, C57BL/6 (H-2b), A/J (H-2a), and BALB/c (H-2d), and 1 outbred strain, CD-1, demonstrated differences in susceptibility to iv challenge with the ANKA clone of Plasmodium berghei. Mice were challenged with 100, 1,000, or 10,000 sporozoites, then evaluated daily beginning on day 4 for patency. CD-1 mice were further evaluated at challenge doses of 12,500, 25,000, and 50,000 sporozoites. C57BL/6 mice were the easiest to infect, with 90% becoming infected with 100 sporozoites. The outbred strain CD-1 was the most difficult to infect, requiring a challenge dose of 25,000 sporozoites/mouse in order to achieve a 100% infection rate. Mouse strains also demonstrated differences in their ability to be protected by intravenous immunization with gamma-irradiated sporozoites. A/J mice needed a minimum of 3 doses of irradiated sporozoites for protection against a challenge with 10,000 sporozoites. In contrast, BALB/c mice immunized with a single dose of 1,000 irradiated sporozoites are protected against a 10,000 sporozoite challenge. These data suggest that both infectivity and protection are genetically restricted and that susceptibility to infection may be inversely related to protection.

Animals↗
PubMed1991

Inhibitory activity against Plasmodium vivax sporozoites induced by plasma from Saimiri monkeys immunized with circumsporozoite recombinant proteins or irradiated sporozoites.

Two Saimiri monkey vaccine trials have been conducted comparing four recombinant Plasmodium vivax circumsporozoite proteins and irradiated sporozoites. Only a small number of animals immunized with certain recombinants or irradiated sporozoites became fully protected against sporozoite challenge. Preimmunization and postimmunization plasma samples obtained from 30 monkeys on the day of challenge were tested in an in vitro assay based on sporozoite development into exoerythrocytic stages in primary cultures of Saimiri monkey hepatocytes. The percentage of inhibition was determined by comparison of the number of exoerythrocytic stages developing from sporozoites preincubated with a preimmunization and a postimmunization plasma sample of each animal. The plasma samples of the day of challenge of nearly all the immunized animals had a variable, but significant inhibitory effect, when compared with the corresponding preimmunization sample. We found no correlation between the degree of in vitro inhibition of liver stage development, and the in vivo protection against sporozoite challenge of individual animals. The variable results of the incubation of sporozoites with "normal" plasma of different animals indicates that the in vitro results were affected by plasma factors unrelated to anti-sporozoite antibodies.

Animals↗
PubMed1982

The role of the Kupffer cell in the infection of rodents by sporozoites of Plasmodium: uptake of sporozoites by perfused liver and the establishment of infection in vivo.

The uptake of Plasmodium yoelii nigeriensis sporozoites by isolated perfused rat liver was very rapid and efficient. 67% of the initial load was removed from the perfusion media in the first passage through the liver, and 95% after 15 min of perfusion. Much of the uptake was explained by mechanical trapping in the liver. Up to 75% of the sporozoite load was retained after 15 min both by heat killed liver and liver cooled to 4 degrees C, therefore at least 20% of the sporozoite uptake in perfused normal livers was due to a biologically active process. In perfused normal livers, non-infective (heat-killed or trypsin-treated) sporozoites were taken up with an efficiency equal to infective sporozoite controls. However, a reduction in Kupffer cell number and activity, induced by silica treatment, resulted in a very significant decline in uptake of infective sporozoites by the perfused liver--and a parallel fall in the successful infection of the host by inoculated sporozoites in vivo. Since silica treatment produced no significant detectable pathological changes in hepatocytes, and infected blood passage results in a normal parasitaemia in silica treated animals it was concluded that the Kupffer cell was a component of the natural route of infection of the mammalian host by the majority of the infecting population of sporozoites of Plasmodium yoelii nigeriensis.

Animals↗
PubMed1995

A conserved peptide sequence of the Plasmodium falciparum circumsporozoite protein and antipeptide antibodies inhibit Plasmodium berghei sporozoite invasion of Hep-G2 cells and protect immunized mice against P. berghei sporozoite challenge.

Minutes after injection into the circulation, malaria sporozoites enter hepatocytes. The speed and specificity of the invasion process suggest that it is receptor mediated. The region II sequence of Plasmodium falciparum circumsporozoite (CS) protein includes a nonapeptide (WSPCSVTCG) which is highly conserved in all of the CS proteins sequenced to data, including the one from Plasmodium berghei. We have found that two peptides based on the P. falciparum region II sequence, P18 (EWSPCSVTCGNGIQVRIK) and P32 (IEQYLKKIKNS ISTEWSPCSVTCGNGIQVRIK), significantly inhibited P. berghei sporozoite invasion into Hep-G2 cells in vitro. This inhibition was enhanced if either peptide was preincubated with Hep-G2 cells prior to sporozoite invasion. We confirm that region II is a sporozoite ligand for the hepatocyte receptor; moreover, despite the few differences between P. falciparum and P. berghei region II sequences around the nonapeptide sequence (66% homology), the functional characteristics of the motif sequences are not affected. Since the conserved motifs represent a crucial sequence involved in Plasmodium sporozoite invasion of hepatocytes, antibodies to region II should inhibit sporozite invasion into hepatocytes. Indeed, we found that polyclonal antibodies generated to the P. falciparum-based peptide P32 inhibited P. berghei sporozoite invasion of Hep-G2 cells. Furthermore, inbred mice (C57BL/6) immunized with P32 were protected against a lethal challenge of P. berghei sporozoites. Our results suggest that the conserved region II of the CS protein contains crucial B- and T-cell epitopes, that such peptide sequences from the human malaria parasite P. falciparum can be screened in the P. berghei rodent model, and, finally, that region II can be considered useful as one of the components of a malaria vaccine.

Amino Acid Sequence↗
PubMed1990

Reduction in cell entry of Eimeria tenella (Coccidia) sporozoites by protease inhibitors, and partial characterization of proteolytic activity associated with intact sporozoites and merozoites.

The role of proteases in the invasion of host cells by Eimeria tenella (Wisconsin strain) was studied in vitro. Protease inhibitors were used to treat sporozoites before inoculation or were applied to cultured chicken kidney cells before infection. The inhibitors antipain, leupeptin, aprotinin, L-1-tosylamide-2-phenyl-ethyl chloromethyl ketone (TPCK), or N-alpha-p-tosyl-L-lysine chloromethyl ketone (TLCK) reduced parasite invasion to 16-66% of control after treatment of cultured cells or sporozoites with 5- or 50-micrograms/ml concentrations of inhibitors in the culture medium. Phenylmethylsulfonyl fluoride (PMSF) reduced invasion to 32-57.7% at concentrations of 1-4 mM. The optimum pH for hydrolysis of azocasein by intact sporozoites or merozoites was determined over a range of pH 5.0 to pH 9.0. Sporozoites were highly active over a broad range from pH 5.5 to pH 9.0, with an apparent optimum at pH 8.0. Merozoites had a much lower specific activity with pH optima at 7.0 and 8.5. The protease activity of sporozoites or merozoites could be inhibited completely by the addition of 50 micrograms/ml of leupeptin, TPCK, or TLCK or of 4 mM PMSF. Antipain inhibited proteases of sporozoites but not of merozoites. Pepstatin had little effect on either sporozoites or merozoites. The results suggest that parasite proteases of Eimeria may be necessary for invasion of host cells.

Animals↗
PubMed1997

Time lapse video microscopy and ultrastructure of penetrating sporozoites, types 1 and 2 parasitophorous vacuoles, and the transformation of sporozoites to tachyzoites of the VEG strain of Toxoplasma gondii.

Videomicroscopy and transmission electron microscopy were used to study the interaction of Toxoplasma gondii sporozoites with cultured cardiopulmonary artery endothelial, embryonic bovine tracheal and Madin-Darby bovine kidney cells. No moving junction or exocytosis of rhoptries, micronemes, and dense granules was detected during the initial penetration of sporozoites into cultured cells, whereas constriction of the sporozoite and partial exocytosis of rhoptries occurred during movement of the sporozoite from the first parasitophorous vacuole (PV1) into the second vacuole (PV2). The PV1 was unusually large, lacked a tubulovesicular membrane network (TMN), and had an indistinct parasitophorous vacuolar membrane (PVM). Comparatively, the PV2 was small, had a distinct PVM, contained a well-developed TMN, and was surrounded by numerous host cell mitochondria. Sporozoites that passed completely through cells carried with them an envelope of host cell membranes and cytoplasm. Cultured cells occasionally endocytosed sporozoites that were enveloped by host cell material. After formation of the PV2, sporozoites replicated by endodyogeny to form tachyzoites.

Animals↗
PubMed2005

Transgenic Plasmodium berghei sporozoites expressing beta-galactosidase for quantification of sporozoite transmission.

Malaria transmission occurs during a blood-meal of an infected Anopheles mosquito. Visualization and quantification of sporozoites along the journey from the mosquito midgut, where they develop, to the vertebrate liver, their final target organ, is important for understanding many aspects of sporozoite biology. Here we describe the generation of Plasmodium berghei parasites that express the reporter gene lacZ as a stable transgene, under the control of the sporozoite-specific CSP promoter. Transgenic sporozoites expressing beta-galactosidase can be simply visualized and quantified in an enzymatic assay. In addition, these sporozoites can be used to quantify sporozoites deposited in subcutaneous tissue during natural infection.

Animals↗
PubMed2005

The novel Plasmodium gallinaceum sporozoite protein, Pg93, is preferentially expressed in the nucleus of oocyst sporozoites.

To study gene expression differences between oocyst and salivary gland sporozoites, cDNA libraries previously constructed from the two sporozoite populations of the avian malaria parasite, Plasmodium gallinaceum, were used in a subtractive hybridization protocol to isolate Pg93, a novel oocyst sporozoite gene. Pg93 encodes a putative approximately 76 kDa translated protein that was predicted to localize to the nucleus. Transcriptional analysis indicates that Pg93 is preferentially expressed in oocyst sporozoites versus salivary gland sporozoites. Immunolocalization assays confirm both the nuclear prediction and transcriptional analysis, suggesting that Pg93 is a nuclear protein. BLAST sequence analysis indicates that Pg93 represents a novel gene that has significant homology with a Plasmodium falciparum hypothetical protein and translated Plasmodium knowlesi and Plasmodium vivax nucleotide sequences. This is the first characterization of a Plasmodium nuclear protein that shows preferential expression in one sporozoite population as compared with the other population.

Amino Acid Sequence↗