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Protection against malaria in Aotus monkeys immunized with a recombinant blood-stage antigen fused to a universal T-cell epitope: correlation of serum gamma interferon levels with protection.

The major surface antigen p190 of the human malaria parasite Plasmodium falciparum contains nonpolymorphic, immunogenic stretches of amino acids which are attractive components for a subunit vaccine against malaria. One such polypeptide, termed 190L, is contained in the 80-kDa processing product of p190, which constitutes the major coat component of mature merozoites. We report here that immunization of Aotus monkeys with 190L gives only poor protection against P. falciparum challenge. However, addition by genetic engineering of a universal T-cell epitope (CS.T3) to 190L improved immunity, and as a result three of four monkeys were protected following challenge infection with blood-stage parasites. Neither antibody against the immunizing antigens or against blood-stage parasites nor the capacity of the monkeys' sera to inhibit in vitro parasite invasion correlated with protection. However, in contrast to sera from nonprotected monkeys, sera from protected animals contained elevated levels of gamma interferon. These results suggest that gamma interferon is directly or indirectly involved in the process of asexual parasite control in vivo.

Animals↗

Sex-determined resistance to Toxoplasma gondii is associated with temporal differences in cytokine production.

Examination of a wide range of inbred mice of diverse genetic backgrounds and major histocompatibility complex haplotypes revealed a dramatic difference in the susceptibilities of males and females to Toxoplasma gondii infection. Female mice were found to be more susceptible to acute infection, as determined by higher mortality levels, than male mice, while those female mice surviving to have chronic infections harbored more cysts in their brains than did surviving males. This phenomenon was therefore investigated in greater depth immunologically in the BALB/K mouse, a strain showing moderate susceptibility to infection with T. gondii. Plasma tumor necrosis factor alpha (TNF-alpha) levels were elevated in both male and female BALB/K mice on days 8 and 10 postinfection, but not thereafter, with males producing significantly higher levels than females. However, it was not until day 12 postinfection that the first deaths occurred, and these were among female mice, indicating that TNF-alpha production was not responsible for mortality. In vitro examination of T. gondii-specific T-cell proliferative responses from day 15 postinfection onwards revealed significantly higher stimulation indices in male mice than in their female counterparts. This difference was most apparent in splenocyte cultures initiated at day 15 postinfection, where complete suppression of proliferation was noted in the splenocytes from female mice but not from male mice. Analysis of tissue culture supernatants from these cultures revealed distinct differences in the kinetics of production as well as the quantities of gamma interferon (IFN-gamma) and interleukin 10 (IL-10) produced. Spleen cells from male mice produced higher levels of IFN-gamma in the early stages of infection than those from female mice. IFN-gamma levels were highest in the supernatants from male splenocyte cultures initiated at day 15 postinfection. Similar levels of IFN-gamma were not obtained from the supernatants of female splenocyte cultures until day 22 postinfection. IL-10 production, on the other hand, peaked at maximal levels in the cell cultures from both sexes initiated at day 22 postinfection. These results suggest that, in male mice, a rapid response to infection with high levels of TNF-alpha and IFN-gamma helps to control parasite multiplication, after which IL-10 production may be important in down regulating these potentially harmful inflammatory mediators. The failure of female mice to respond quickly in terms of T-cell proliferation and IFN-gamma production compared with their male counterparts may account for their poor survival rates and higher cyst burdens.

Animals↗

Soluble factors released by Toxoplasma gondii-infected astrocytes down-modulate nitric oxide production by gamma interferon-activated microglia and prevent neuronal degeneration.

The maintenance of a benign chronic Toxoplasma gondii infection is mainly dependent on the persistent presence of gamma interferon (IFN-gamma) in the central nervous system (CNS). However, IFN-gamma-activated microglia are paradoxically involved in parasitism control and in tissue damage during a broad range of CNS pathologies. In this way, nitric oxide (NO), the main toxic metabolite produced by IFN-gamma-activated microglia, may cause neuronal injury during T. gondii infection. Despite the potential NO toxicity, neurodegeneration is not a common finding during chronic T. gondii infection. In this work, we describe a significant down-modulation of NO production by IFN-gamma-activated microglia in the presence of conditioned medium of T. gondii-infected astrocytes (CMi). The inhibition of NO production was paralleled with recovery of neurite outgrowth when neurons were cocultured with IFN-gamma-activated microglia in the presence of CMi. Moreover, the modulation of NO secretion and the neuroprotective effect were shown to be dependent on prostaglandin E(2) (PGE(2)) production by T. gondii-infected astrocytes and autocrine secretion of interleukin-10 (IL-10) by microglia. These events were partially eliminated when infected astrocytes were treated with aspirin and cocultures were treated with anti-IL-10 neutralizing antibodies and RP-8-Br cyclic AMP (cAMP), a protein kinase A inhibitor. Further, the modulatory effects of CMi were mimicked by the presence of exogenous PGE(2) and by forskolin, an adenylate cyclase activator. Altogether, these data point to a T. gondii-triggered regulatory mechanism involving PGE(2) secretion by astrocytes and cAMP-dependent IL-10 secretion by microglia. This may reduce host tissue inflammation, thus avoiding neuron damage during an established Th1 protective immune response.

Animals↗

Studies of serum total immunoglobulin E concentrations in atopic and non-atopic dogs.

The serum total immunoglobulin E (IgE) concentrations of two groups of atopic dogs and three groups of non-atopic dogs were compared. There was a wide range of concentrations with a high degree of overlap between the groups. The serum total IgE concentrations of a group of 15 non-atopic racing greyhounds were significantly higher than those of all the other groups. Atopic and non-atopic dogs receiving stringent parasite control treatments could not be differentiated on the basis of their serum total IgE concentrations. In the non-atopic dogs there was no correlation between their serum total IgE concentrations and the number of allergen-specific positive results obtained in an ELISA, or between their serum total IgE concentrations and their age.

Aging↗

Trypanosoma cruzi-infected cardiomyocytes produce chemokines and cytokines that trigger potent nitric oxide-dependent trypanocidal activity.

BACKGROUND: The pathogenesis of myocarditis that occurs in Trypanosoma cruzi-infected mice is still poorly understood. Therefore, it is important to know the mediators that trigger leukocyte migration to the heart as well as the cellular source of these possible mediators. In this study, we investigated (1) NO synthase (NOS) induction, (2) NO synthesis, (3) trypanocidal activity, and (4) chemokine and cytokine mRNA expression by isolated cardiomyocytes infected with T cruzi. METHODS AND RESULTS: Mouse cardiomyocytes were isolated, infected with T cruzi, and evaluated for induction of inducible NOS (iNOS), nitrite production, trypanocidal activity, and cytokine and chemokine mRNA expression. We found that T cruzi-infected murine embryonic cardiomyocytes produced nitrite and expressed mRNAs for the chemokines chemokine growth-related oncogene, monokine induced by interferon-gamma, macrophage inflammatory protein-2, interferon-gamma-inducible protein, RANTES, and monocyte chemotactic protein, for iNOS, and for the cytokines tumor necrosis factor (TNF)-alpha and interleukin (IL)-1beta. Separate addition of IL-1beta, interferon-gamma, TNF-alpha or monocyte chemotactic protein, macrophage inflammatory protein-2, and interferon-gamma-inducible protein, to cultured cardiomyocytes resulted in NO production but low trypanocidal activity. However, simultaneous addition of IL-1beta, interferon-gamma, and TNF-alpha or the chemokines to cultures resulted in the induction of iNOS, high levels of nitrite, and a marked trypanocidal activity. The iNOS/L-arginine pathway mediated the latter activity, inasmuch as it was inhibited by treatment with N:(G)-monomethyl-L-arginine. CONCLUSIONS: These results indicate that iNOS activation and the proinflammatory cytokines and chemokines produced by cardiomyocytes are likely to control parasite growth and cell influx, thus contributing to the pathogenesis of chagasic cardiomyopathy seen in T cruzi-infected mice.

Animals↗

CC-chemokine receptors: a potential therapeutic target for Trypanosoma cruzi-elicited myocarditis.

The comprehension of the pathogenesis of Trypanosoma cruzi-elicited myocarditis is crucial to delineate new therapeutic strategies aiming to ameliorate the inflammation that leads to heart dysfunction, without hampering parasite control. The augmented expression of CCL5/RANTES and CCL3/MIP-1alpha, and their receptor CCR5, in the heart of T. cruzi-infected mice suggests a role for CC-chemokines and their receptors in the pathogenesis of T. cruzi-elicited myocarditis. Herein, we discuss our recent results using a CC-chemokine receptor inhibitor (Met-RANTES), showing the participation of CC-chemokines in T. cruzi infection and unraveling CC-chemokine receptors as an attractive therapeutic target for further evaluation in Chagas disease.

Animals↗

Mediation of pyrethroid insecticide toxicity to honey bees (Hymenoptera: Apidae) by cytochrome P450 monooxygenases.

Honey bees, Apis mellifera L., often thought to be extremely susceptible to insecticides in general, exhibit considerable variation in tolerance to pyrethroid insecticides. Although some pyrethroids, such as cyfluthrin and lambda-cyhalothrin, are highly toxic to honey bees, the toxicity of tau-fluvalinate is low enough to warrant its use to control parasitic mites inside honey bee colonies. Metabolic insecticide resistance in other insects is mediated by three major groups of detoxifying enzymes: the cytochrome P450 monooxygenases (P450s), the carboxylesterases (COEs), and the glutathione S-transferases (GSTs). To test the role of metabolic detoxification in mediating the relatively low toxicity of tau-fluvalinate compared with more toxic pyrethroid insecticides, we examined the effects of piperonyl butoxide (PBO), S,S,S-tributylphosphorotrithioate (DEF), and diethyl maleate (DEM) on the toxicity of these pyrethroids. The toxicity of the three pyrethroids to bees was greatly synergized by the P450 inhibitor PBO and synergized at low levels by the carboxylesterase inhibitor DEF. Little synergism was observed with DEM. These results suggest that metabolic detoxification, especially that mediated by P450s, contributes significantly to honey bee tolerance of pyrethroid insecticides. The potent synergism between tau-fluvalinate and PBO suggests that P450s are especially important in the detoxification of this pyrethroid and explains the ability of honey bees to tolerate its presence.

Animals↗

Effects of topoisomerases inhibitors protoberberine on Leishmania donovani growth, macrophage function, and infection.

DNA topoisomerases play a pivotal role in the regulation of cell division. Inhibition of Leishmania spp. topoisomerases represents an alternative to control parasite growth. Cancer research led to the development of several potent topoisomerase inhibitors such as topoisomerase I, topoisomerase II, or both (monobenzimidazole, terbenzimidazole, and protoberberine alkaloid-related compounds) that are effective antitumor agents. In the present study, we evaluated the efficacy of these compounds against Leishmania spp. growth in vitro. Some protoberberine compounds showed pronounced antileishmanial activity and were selected for further analysis in macrophages. These compounds did not affect macrophage viability and only slightly reduced macrophage nitric oxide generation in response to interferon-gamma. Moreover, exposure of infected macrophages to these compounds significantly reduced parasite loads. Collectively, our data suggest that protoberberine-related compounds have powerful antileishmania action and that minor structural variations among them can substantially improve their activity to restrict Leishmania spp. infection in vitro.

Animals↗

Physiological responses of cattle consuming tall fescue to environmental temperature and supplemental phenothiazine.

The influence of supplemental phenothiazine (P) on growth and physiological criteria was studied in parasite-controlled calves consuming endophyte (Acremonium coenophialum)-infected tall fescue (TF). In Exp. 1, nine Angus heifer calves (312 kg) were supplemented with 227 g corn-mineral (CM) mix twice daily and allowed ad libitum access to either high-endophyte (HE) G1-307 (greater than 90% infected) or low-endophyte (LE) Kenhy (less than 1% infected) tall fescue hay, or HE G1-307 plus 2 g/d P in the daily supplement. Calves were kept in temperature-controlled rooms for 12 d at 21 degrees C followed by 7 d at 34 degrees C. In Exp. 2, 48 Angus steer calves (312 kg) were assigned to treatment groups consisting of calves grazing HE Kentucky-31 (57% infected) or LE Johnstone (less than 1% infected) TF, and supplemented daily with either .91 kg of a control CM mix or .91 kg of the CM mix containing 2 g P. The 112-d experiment was initiated on May 4 with BW and rectal temperature (RT) measurements and blood collected at 28-d intervals. In both experiments, calves receiving HE TF had lower (P less than .01) serum prolactin concentrations (PRL) at elevated ambient temperature and lower (P less than .01) serum alkaline phosphatase activities (AP) but higher (P less than .01) RT than calves consuming LE TF regardless of ambient temperature.(ABSTRACT TRUNCATED AT 250 WORDS)

Acremonium↗

IL-12 is required to maintain a Th1 response during Leishmania major infection.

IL-12 initiates Th1 cell development and cell-mediated immunity, but whether IL-12 contributes to the maintenance of a Th1 response is unclear. To address this question, we infected IL-12 p40-/- C57BL/6 mice with Leishmania major, an intracellular protozoan parasite controlled by a cell-mediated immune response, and simultaneously administered IL-12. Whereas untreated p40-/- mice developed an uncontrolled infection, p40-/- mice treated with IL-12 for the first 2 or 4 wk of infection developed a Th1 response and resolved their lesions. However, the induction of this protective Th1 cell response by IL-12 treatment was not associated with long term immunity. We observed that on rechallenge in the absence of IL-12, the mice exhibited a susceptible phenotype. In addition, without rechallenge, lesions in the IL-12-treated p40-/- mice developed several weeks after cessation of IL-12 treatment. In both cases, disease was associated with the loss of a Th1 response and the development of a Th2 response. Our observations are not limited to the C57BL/6 strain, because IL-12 treatment was also unable to provide lasting protection to p40-/- BALB/c mice. Finally, we found that although Th1 cells from healed wild-type C57BL/6 mice adoptively transferred protection to L. major-infected RAG-/- mice, they were unable to protect p40-/- mice. In conclusion, these studies provide the first demonstration that IL-12 is required not only to initiate Th1 cell development but also throughout infection to maintain a Th1 cell response and resistance to L. major.

Adoptive Transfer↗

STAT1 plays a critical role in the regulation of antimicrobial effector mechanisms, but not in the development of Th1-type responses during toxoplasmosis.

The production of IFN-gamma by T cells and the ability of this cytokine to activate the transcription factor STAT1 are implicated in the activation of antimicrobial mechanisms required for resistance to intracellular pathogens. In addition, recent studies have suggested that the ability of STAT1 to inhibit the activation of STAT4 prevents the development of Th1 responses. However, other studies suggest that STAT1 is required to enhance the expression of T-bet, a transcription factor that promotes Th1 responses. To address the role of STAT1 in resistance to T. gondii, Stat1-/- mice were infected with this pathogen, and their response to infection was assessed. Although Stat1-/- mice produced normal serum levels of IL-12 and IFN-gamma, these mice were unable to control parasite replication and rapidly succumbed to this infection. Susceptibility to toxoplasmosis was associated with an inability to up-regulate MHC expression on macrophages, defects in NO production, and the inability to up-regulate some of the IFN-inducible GTPase family of proteins, molecules associated with antitoxoplasma activity. Analysis of T cell responses revealed that STAT1 was not required for the development of a Th1 response, but was required for the infection-induced up-regulation of T-bet. Together these studies suggest that during toxoplasmosis the major role of STAT1 is not in the development of protective T cell responses, but, rather, STAT1 is important in the development of antimicrobial effector mechanisms.

Animals↗

An economic analysis of communal goat production.

The economic impact of different extension messages used was calculated using enterprise budgeting (gross margin analysis). Input data were gleaned from the literature, from participatory appraisals, as well as a field study, spanning 12 months, of small-scale communal goat farming systems in Jericho in the Odi District of North West Province. The number of offspring weaned per annum, as a proportion of does owned, was selected as the desired output for analysis. This study has shown that small-scale communal goat farmers are not adopting or implementing extension messages to improve production capacity. In South Africa the majority of goats are slaughtered in the informal sector. If the informal sector is to be persuaded to market goats commercially through formal channels, then knowledge of the economics of goat farming on communal lands should be provided. The economic aspects of extension messages are probably an important factor in determining acceptance and sustainability yet appear to be seldom investigated. The probable reason for lack of adoption of standard extension messages, which promote improved nutrition, parasite control, vaccination and treatment of goats, was economic. In other words, the so-called 'poor management practices' used by communal farmers appeared to be economically more profitable than the 'good management practices' suggested to increase production. The price of communal goats was not related to their mass. A higher level of inputs would probably have resulted in a heavier kid, however it was established that this would not have influenced the price received as a majority of the goats were slaughtered for ritual purposes where age, colour and sex were more important to the purchaser than body mass. It is standard practice in commercial farming systems to evaluate the economic benefits of all management practices before they are implemented. Production animal veterinarians use veterinary economics to compare different scenarios to control diseases or select management practices in commercial herds. It is suggested that the inputs and outputs of small-scale farming systems should be carefully analysed and that veterinary economics should also be used to evaluate the probable impact of extension messages formulated by veterinarians and animal health technicians.

Animal Husbandry↗

Diphyllobothriasis associated with salmon consumption in Pacific Coast states.

A review of requests for niclosamide ( Niclocide or Yomesan ) made to the Centers for Disease Control Parasitic Diseases Drug Service indicated that an outbreak of diphyllobothriasis occurred in west coast states in 1980, and in the Los Angeles area in 1981. Our investigation of 52 infections that occurred in 1980 implicates fresh salmon (Oncorhynchus spp.) as the host for the Diphyllobothrium parasite in 82% of the persons interviewed. We describe here common symptoms of the infection, methods of fish preparation employed by infected individuals, and epidemiologic data. We suggest that changes in fish marketing practices and the popularity of uncooked or slightly cooked fresh salmon were responsible for this epidemic. We recommend that salmon be either cooked or frozen before it is eaten.

Alaska↗

Use of non-human plasma for in vitro cultivation and antimalarial drug susceptibility testing of Plasmodium falciparum.

Viability, growth rate, and chemotherapeutic susceptibility of the CDC/Indochina III, CDC/Sierra Leone I, and FCR-3 (Subline F-86) isolates of Plasmodium falciparum grown continuously in RPMI 1640 medium supplemented with goat, horse, porcine, bovine, or ovine plasma were evaluated. Results were compared to those obtained from parallel cultures maintained in medium supplemented with non-immune human plasma. Only media supplemented with goat or horse plasma supported significant continuous multiplication of the isolates. Medium supplemented with either ovine or porcine plasma supported continuous multiplication of the CDC/Indochina III isolate, but not the FCR-3 isolate. Medium supplemented with bovine plasma did not support continuous growth of any of the isolates tested. The light microscopic appearance of the isolates during and after continuous culture in medium supplemented with either goat or horse plasma was identical to that of the control parasites maintained in medium supplemented with human plasma. There were no statistically significant differences in the susceptibility to antimalarial drugs of the culture lines maintained in medium supplemented with either human or goat plasma.

Animals↗

Mebendazole.

The broad-spectrum of activity and safety of mebendazole remain, after 5 years of clinical experience, unique features of this anthelmintic. Through microtubular destruction, mebendazole kills helminths by inhibiting glucose uptake into susceptible parasites. The drug's poor absorption does not appear to affect clinical efficacy except, perhaps, in the treatment of systemic helminth infections. Mebendazole is generally considered the drug of choice for trichuriasis and has therapeutic advantages over other anthelmintics in the treatment of enterobiasis and hookworm infections. Although mebendazole is an effective agent against ascariasis, there are preferable alternatives. Among its nonapproved uses, mebendazole shows great promise in the treatment of capillariasis and hydatid disease. Further investigation is needed to establish its role in the treatment of taeniasis, Hymenolepsis nana, strongyloidiasis, trichinosis, and Dipetalonema perstans. Undoubtedly, mebendazole will find its greatest value in the treatment of patients with multiple helminth infections.

Ascariasis↗

Halofantrine in the treatment of uncomplicated falciparum malaria with a three-dose regimen in Papua New Guinea: a preliminary report.

We evaluated the efficacy and safety of halofantrine in 19 patients with acute uncomplicated falciparum malaria. Each patient received oral halofantrine hydrochloride 500 mg every 6 hours for 3 doses (total 1.5 g). In almost all the patients clinical symptoms of malaria and parasitaemia disappeared within 2 and 3 days, respectively, of starting treatment. We observed no recurrence of parasitaemia during 14 days of follow-up. Tolerance to halofantrine was good except for minor and self-limiting gastrointestinal side-effects. Haematological and biochemical indices were not seriously affected. Halofantrine-induced prolongation of Q-T/Q-Tc intervals was the consistent cardiac manifestation in 84% of patients. The Q-T/Q-Tc interval prolongation increased with each dose; it reached a maximum between 18 and 24 hours and thereafter returned to baseline. These preliminary data indicate that, apart from the cardiac side-effects, halofantrine is an effective and safe drug, well tolerated by most of the patients in the study.

Adolescent↗