Liposomes as drug carriers in leishmaniasis and malaria

Experimental studies suggest that liposomes could substantially improve the performance of anti-leishmanial drugs in the chemotherapy of visceral leishmaniasis. In this article, Carl Alving discusses the potential for overcoming resistance to antimonial drugs; for 'buffering' the toxicity of drugs, and for drug delivery under conditions where hospitalization is impossible or inconvenient. Liposomes can also be used experimentally to reduce the toxicity and increase the efficacy of parenterally-administered primaquine in the treatment of sporozoite-induced murine malaria.     

Development and evaluation of tripalmitin emulsomes for the treatment of experimental visceral leishmaniasis

The antifungal and antileishmanial agent amphotericin B (AmB) was formulated in tripalmitin based nanosize lipid partices (emulsomes) for macrophage targeting for the treatment of visceral leishmaniasis (VL). Emulsomes were modified by coating them with macrophage-specific ligand (O-palmitoyl mannan, OPM). The antileishmanial activity of AmB (0.5 and 1?mg/kg) was investigated in-vivo against VL by the inhibition of parasitic load in the spleen of L. donovani infected hamsters after intraperitoneal injections of AmB-Doc (Mycol), plain emulsomes (TPEs) and OPM coated emulsomes (TPEs-OPM). The formulations were found to be less effective at the dose of 0.5?mg/kg. At the dose of 1?mg/kg, formulation TPEs-OPM eliminated intracellular amastigotes of L. donovani within splenic macrophages more efficiently (62.76?±?3.54 % parasite inhibition) than the formulation TPEs (42.68?±?2.36 % parasite inhibition) (P?相似文献   

Chemotherapy of experimental visceral leishmaniasis in the opossum

Visceral leishmaniasis is a severe chronic disease of people and animals. The disease is caused by several subspecies of a protozoal organism, Leishmania donovani. If not treated, visceral leishmaniasis is often fatal. The most commonly used chemotherapeutic agents to treat the disease are pentavalent antimonials, which can be toxic, must be administered by parenteral routes, and are sometimes ineffective. In this study, meglumine antimoniate, a pentavalent antimony, was compared with WR 6026, an 8-aminoquinoline derivative, as to antileishmanial efficacy. The results indicate that either of these 2 drugs are effective in the suppression of amastigotes in the liver and spleen of the opossum. Despite the marked parasite suppression in the liver and spleen of the infected opossums, the experimental disease was fatal in all of the infected opossums, regardless of the therapy.     

Sugar receptor mediated drug delivery to macrophages in the therapy of experimental visceral leishmaniasis

Methotrexate (MTX) conjugate of a neoglycoprotein, mannosyl bovine serum albumin, containing an average of 30 moles of MTX per mole of neoglycoprotein was taken up efficiently by murine peritoneal macrophages through cell surface mannosyl receptors. The conjugate strongly inhibits the growth of Leishmania donovani inside macrophages, with 50% inhibitory dose of 0.11 micrograms/ml MTX, which makes it 100 times more active than free MTX (50% inhibitory dose of 12.1 micrograms/ml). MTX conjugated to BSA or other non-specific neoglycoproteins like galactose-BSA and glucose-BSA have leishmanicidal effects comparable to free MTX. Moreover, in a murine model of experimental visceral leishmaniasis, the drug conjugate reduced the spleen parasite burden by more than 85% in a 30 day model whereas the same concentration of free drug caused little effect. The results demonstrate that neoglycoproteins may be useful as carriers for receptor mediated drug delivery to treat macrophage associated diseases.     

A novel mechanism for an old drug: amphotericin B in the treatment of visceral leishmaniasis

Visceral leishmaniasis (VL) is caused by various species of the genus Leishmania. Internalization of Leishmania into host cells is facilitated by a large number of receptors, and therefore no panacea is available for the treatment of leishmaniasis. We previously demonstrated the requirement of host membrane cholesterol in the entry of Leishmania into macrophages by cholesterol depletion using methyl-β-cyclodextrin (MβCD). We recently showed that leishmanial infection is inhibited upon sequestration of host membrane cholesterol using amphotericin B (AmB), considered as the best existing drug against VL. The reason for the antileishmanial activity of AmB is generally believed to be its ability to bind ergosterol in parasite membranes. Our recent results offer the opportunity to reexamine the mechanism behind the effectiveness of current AmB-based therapeutic strategies to treat leishmaniasis. We propose here a novel mechanism in which the effectiveness of AmB treatment could be partly based on its ability to sequester cholesterol in the host membrane, thereby abrogating macrophage-parasite interaction.     

Anemia in experimental visceral leishmaniasis in hamsters.

Experimental infection of hamsters with Leishmania donovani caused visceral leishmaniasis in which hematological changes occurred. The infected hamsters were anemic and reticulocyte counts were high. No significant change in the serum erythropoietin level was noted. Red cell membrane Na(+)-K(+)-ATPase and acetylcholinesterase activities increased. Osmotic fragility of the erythrocytes from infected animals increased. The level of 2,3-diphosphoglycerate of the red cells increased with the degree of anemia.     

Radio-attenuated leishmanial parasites as immunoprophylactic agent against experimental murine visceral leishmaniasis

The present study intends to evaluate the role of radio-attenuated leishmania parasites as immunoprophylactic agents for experimental murine visceral leishmaniasis. BALB/c mice were immunized with gamma (γ)-irradiated Leishmania donovani. A second immunization was given after 15 days of first immunization. After two immunizations, mice were infected with virulent L. donovani promastigotes. Protection against Kala-azar (KA) was estimated from spleen and liver parasitic burden along with the measurement of nitrite and superoxide anion generation by isolation of splenocytes and also by T-lymphocyte helper 1(Th1) and T-lymphocyte helper 2(Th2) cytokines release from the experimental groups. It was observed that BALB/c mice having prior immunization with radio-attenuated parasites showed protection against L. donovani infection through higher expression of Th1 cytokines and suppression of Th2 cytokines along with the generation of protective free radicals. The group of mice without prior priming with radio-attenuated parasites surrendered to the disease. Thus it can be concluded that radio-attenuated L. donovani may be used for.     

Treatment of experimental visceral leishmaniasis with lymphokine encapsulated in liposomes

Highly susceptible mice were infected with Leishmania donovani chagasi and were treated with supernatants, free or encapsulated in liposomes, from concanavalin A-stimulated or unstimulated mouse spleen cell cultures. Treatment consisted of multiple i.v. injections beginning 2 days before to 2 days after infection. Mice treated with lymphokine-rich supernatants encapsulated in liposomes had significantly fewer liver parasites than the control groups, demonstrating in vivo activity of lymphokine against an infectious organism.     

A long term experimental study of canine visceral leishmaniasis

Previous studies on Leishmania infantum and the canine immune response are derived mainly from short-term studies. To date, there have been no longitudinal studies that perform a serial analysis of the intensity of infection in conjunction with immunological parameters and clinical signs in Leishmania-infected dogs. For this purpose, six dogs were infected experimentally by the i.v. route and were monitored for 1 year. Clinical, immunological (humoral and cellular response) and parasitological (parasitaemia) parameters were evaluated monthly. Four dogs developed clinico-pathological signs compatible with leishmaniasis, whereas two dogs showed few abnormalities during the study. Evaluation of clinical, immunological and parasitological parameters showed that the intensity of Leishmania infection in blood samples, as indicated by the amount of Leishmania DNA, was correlated significantly with IgG, IgG1, IgG2, IgA, and IgM concentrations and with clinical signs. Parasitaemia and Leishmania-specific cell-mediated immunity were inversely correlated. Moreover, higher quantities of Leishmania DNA were detected in the liver, spleen, lymph node, skin and bone marrow of dogs exhibiting clinical signs than those exhibiting few such signs. These findings suggest that progressive disease in experimental canine leishmaniasis is associated with specific T-cell unresponsiveness and unprotective humoral responses which allow the dissemination and multiplication of L. infantum in different tissues.     

Water-soluble polymer–drug conjugates for combination chemotherapy against visceral leishmaniasis

There is a need for new safe, effective and short-course treatments for leishmaniasis; one strategy is to use combination chemotherapy. Polymer–drug conjugates have shown promise for the delivery of anti-leishmanial agents such as amphotericin B. In this paper, we report on the preparation and biological evaluation of polymer–drug conjugates of N-(2-hydroxypropyl)methacrylamide (HPMA), amphotericin B and alendronic acid. The combinatorial polymer–drug conjugates were effective anti-leishmanial agents in vitro and in vivo, but offered no advantage over the single poly(HPMA)–amphotericin B conjugates.     

Activation of invariant NKT cells exacerbates experimental visceral leishmaniasis

We report that natural killer T (NKT) cells play only a minor physiological role in protection from Leishmania donovani infection in C57BL/6 mice. Furthermore, attempts at therapeutic activation of invariant NKT (iNKT) cells with α-galactosylceramide (α-GalCer) during L. donovani infection exacerbated, rather than ameliorated, experimental visceral leishmaniasis. The inability of α-GalCer to promote anti-parasitic immunity did not result from inefficient antigen presentation caused by infection because α-GalCer–loaded bone marrow–derived dendritic cells were also unable to improve disease resolution. The immune-dampening affect of α-GalCer correlated with a bias towards increased IL-4 production by iNKT cells following α-GalCer stimulation in infected mice compared to naïve controls. However, studies in IL-4–deficient mice, and IL-4 neutralisation in cytokine-sufficient mice revealed that α-GalCer–induced IL-4 production during infection had only a minor role in impaired parasite control. Analysis of liver cell composition following α-GalCer stimulation during an established L. donovani infection revealed important differences, predominantly a decrease in IFNγ⁺ CD8⁺ T cells, compared with control-treated mice. Our data clearly illustrate the double-edged sword of NKT cell–based therapy, showing that in some circumstances, such as when sub-clinical or chronic infections exist, iNKT cell activation can have adverse outcomes.     

Efficacy of Leishmania donovani ribosomal P1 gene as DNA vaccine in experimental visceral leishmaniasis

The acidic ribosomal proteins of the protozoan parasites have been described as prominent antigens during human disease. We present here data showing the molecular cloning and protective efficacy of P1 gene of Leishmaniadonovani as DNA vaccine. The PCR amplified complete ORF cloned in either pQE or pVAX vector was used either as peptide or DNA vaccine against experimentally induced visceral leishmaniasis in hamsters. The recombinant protein rLdP1 was given along with Freund’s adjuvant and the plasmid DNA vaccine, pVAX-P1 was used alone either as single dose or double dose (prime and boost) in different groups of hamsters which were subsequently challenged with a virulent dose of 1 × 10⁷L.donovani (MHOM/IN/DD8/1968 strain) promastigotes by intra-cardiac route. While the recombinant protein rLdP1 or DNA vaccine pVAX-P1 in single dose format were not found to be protective, DNA vaccine in a prime-boost mode was able to induce protection with reduced mortality, a significant (75.68%) decrease in splenic parasite burden and increased expression of Th1 type cytokines in immunized hamsters. Histopathology of livers and spleens from these animals showed formation of mature granulomas with compact arrangement of lymphocytes and histiocytes, indicating its protective potential as vaccine candidate.     

Macrophages, pathology and parasite persistence in experimental visceral leishmaniasis

Many infectious diseases are associated with parasite persistence, often restricted to certain tissue sites, yet the determinants of such persistence are poorly understood. Infection with the protozoan parasite Leishmania donovani has proved a useful experimental tool to address how immune responses can be differentially effective in clearing parasites from different tissues and, conversely, it might also provide a good model for understanding the basis of parasite persistence. This article reviews recent studies on the determinants and consequences of persistent parasite infection in the spleen and suggest that some of the messages to emerge could have important implications for the study of a broad range of infectious diseases.     

Macrophage specific drug delivery in experimental leishmaniasis

Macrophage-specific delivery systems are the subject of much interest nowadays, because of the fact that macrophages act as host cells for many parasites and bacteria, which give rise to outbreak of so many deadly diseases(eg. leishmaniasis, tuberculosis etc.) in humans. To combat these deadly diseases initially macrophage specific liposomal delivery system were thought of and tested in vivo against experimental leishmaniasis in hamsters using a series of indigenous or synthetic antileishmanial compounds and the results were critically discussed. In vitro testing was also done against macrophages infected with Leishmania donovani, the causative agent for visceral leishmaniasis. The common problem of liposome therapy being their larger size, stability and storage, non-ionic surfactant vesicles, niosomes were prepared, for their different drug distribution and release characteristics compared to liposomes. When tested in vivo, the retention capacity of niosomes was found to be higher than that of liposomes due to the absence of lipid molecules and their smaller size. Thus the therapeutic efficacy of certain antileishmanial compounds was found to be better than that in the liposomal form. The niosomes, being cheaper, less toxic, biodegradable and non-immunogenic, were considered for sometime as suitable alternatives to liposomes as drug carriers. Besides the advent of other classical drugs carriers(e.g. neoglycoproteins), the biggest challenge came from polymeric delivery vehicles, specially the polymeric nanoparticles which were made of cost effective biodegradable polymers and different natural polymers. Because of very small size and highly stable nature, use of nanoparticles as effective drug carriers has been explored in experimental leishmaniasis using a series of antileishmanial compounds, both of indigenous and synthetic origin. The feasibility of application in vivo, when tested for biological as well as for other physicochemical parameters, the polymeric nanoparticles have turned out to be the best and thus may be projected for effective use in the clinics.     

A comparison of experimental visceral leishmaniasis in the opossum, armadillo and ferret

Visceral leishmaniasis is a severe, chronic protozoal disease of humans and animals. Although chemotherapeutic agents are available for the treatment of this disease, problems such as drug toxicity, drug ineffectiveness and drug resistance of the parasite are responsible for treatment failures. To determine whether a drug is a potential antileishmanial agent, screening tests are performed using in vitro and in vivo models. Subsequently, a study using an appropriate animal model is performed to clearly determine the efficacy of a drug against Leishmania. Due to current public concerns regarding the use of companion animals in addition to the high costs of obtaining and maintaining these animals for research use, conventional animal models used in these chemotherapy studies, notably the dog and monkey, are becoming less acceptable. Therefore, new, less expensive and more accessible animal models are needed for the study of antileishmanial compounds. In this study, the armadillo, ferret and opossum were evaluated as possible new animal models for visceral leishmaniasis. The marked body weight loss, hepatomegaly, splenomegaly, large amastigote densities and the microscopic lesions observed in the infected opossums indicated that the opossum was more susceptible to visceral leishmaniasis than the armadillo or ferret.     

Selectins ligand decorated drug carriers for activated endothelial cell targeting

New active particulate polymeric vectors based on branched polyester copolymers of hydroxy-acid and allyl glycidyl ether were developed to target drugs to the inflammatory endothelial cell surface. The hydroxyl and carboxyl derivatives of these polymers allow grafting of ligand molecules on the polyester backbones at different densities. A known potent nonselective selectin ligand was selected and synthesized using a new scheme. This synthesis allowed the grafting of the ligand to the polyester polymers, preserving its binding activity as assessed by docking simulations. Selectin expression on human umbilical cord vascular endothelial cells (HUVEC) was induced with the pro-inflammatory bacterial lipopolysaccharide (LPS) or with the nonselective inhibitor of nitric oxide synthase L-NAME. Strong adhesion of the ligand decorated nanoparticles was evidenced in vitro on activated HUVEC. Binding of nanoparticles bearing ligand molecules could be efficiently inhibited by prior incubation of cells with free ligand, demonstrating that adhesion of the nanoparticles is mediated by specific interaction between the ligand and the selectin receptors. These nanoparticles could be used for specific drug delivery to the activated vascular endothelium, suggesting their application in the treatment of diseases with an inflammatory component such as rheumatoid arthritis and cancer.     

Blood monocytes: differing effector role in experimental visceral versus cutaneous leishmaniasis

Irrespective of the tissue infected or the strain involved, all Leishmania species selectively parasitize and replicate within the resident tissue macrophage. Henry Murray here discusses the role of a second mononuclear phagocyte, the blood monocyte, which is also attracted to leishmanial lesions, but which appears to play quite different roles in experimental visceral versus cutaneous infection: in visceral disease, the monocyte is a critical host defense effector cell, in cutaneous disease, it may, paradoxically, serve to perpetuate intracellular infection.     

Albumin microspheres as carriers for the antiarthritic drug celecoxib

The present study investigates the preparation of celecoxib-loaded albumin microspheres and the biodistribution of technetium-99m (^99mTc)-labeled celecoxib as well as its microspheres after intravenous administration. Microspheres were prepared using a natural polymer BSA using emulsification chemical cross-linking method. The prepared microspheres were characterized for entrapment efficiency, particle size, and in vitro drug release. Surface morphology was studied by scanning electron microscopy. Biodistribution studies were performed by radiolabeling celecoxib (CS) and its microspheres (CMS) using^99mTc and injecting arthritic rats intravenously. The geometric mean diameter of the microspheres was found to be 5.46 μm. In vitro release studies indicated that the microspheres sustained the release of the drug for }6 days. Radioactivity measured in different organs after intravenous administration of celecoxib solution showed a significant amount of radioactivity in the liver and spleen. In case of celecoxib-loaded microspheres, a significant amount of radioactivity accumulated in the lungs. No significant difference (P>.1) in the radioactivity was observed between the inflamed joint and the noninflamed joint following intravenous injection of^99mTc-CS. However, in case of the microspheres (CMS), the radioactivity present in the inflamed joint was 2.5-fold higher than in the noninflamed joint. The blood kinetic studies revealed that celecoxib-loaded albumin microspheres exhibited prolonged circulation than the celecoxib solution.     

Evidence-based diagnostic algorithm for visceral leishmaniasis in Bangladesh

Evidence-based diagnostic algorithm is highly recommended for the visceral leishmaniasis (VL). This cross-sectional study was performed in Bangladesh to evaluate VL diagnostic tools including serology, buffy coat smear microscopy for LD body and various DNA-based techniques using buffy coat in 100 confirmed VL cases and 100 controls. The performance of tools against spleen smear (gold standard) was evaluated using kappa coefficient. Diagnostic precision and other inherent indicators were considered for index scoring (IS) of performance of tools using factor analysis. A diagnostic algorithm was formulated based on the IS and availability of the tools at different health care facilities of Bangladesh. A high level of agreement (kappa ≥  0.80) was observed for all the diagnostic tools. The highest kappa coefficients were found for rK39 RDT and rK39 ELISA (0.95), followed by ssuRNA-PCR (0.94), Buffy coat smear (0.93), rK28 ELISA (0.92), rK28 RDT (0.89), LAMP (0.89), Mini-exon PCR (0.86), ITS1 (0.85), and ITS2 PCR (0.80). rK39 RDT was found to be the best diagnostic test (IS: 1.7) followed by rK28 RDT (IS: 1.5), buffy coat smear microscopy (IS: 0.5), rK39 & rK28 ELISA (IS: 0.3), ssuRNA-PCR (IS: ‐0.7) and LAMP, Mini-exon, ITS1, & ITS2 PCR (IS: ‐0.9). rK39 RDT has been proposed as the best option for primary health care facilities, while buffy coat smear microscopy was found to be a good adjunct for confirmation of serology-positive cases and proposed for secondary and tertiary facilities. ssuRNA-PCR or LAMP can be an alternate confirmation tool only applicable to the tertiary facilities.