Co-doped carbon materials synthesized with polymeric precursors as bifunctional electrocatalysts.

The design of stable and high performance metal free bifunctional electrocatalysts is a necessity in alkaline zinc-air batteries for oxygen reduction and evolution reaction. In the present work co-doped carbon materials have been developed from polymeric precursors with abundant active sites to achieve bifunctional activity. A 3-dimensional microporous nitrogen-carbon (NC) and co-doped nitrogen-sulfur-carbon (NSC) and nitrogen-phosphorus-carbon (NPC) were synthesized using poly(2,5-benzimidazole) as an N containing precursor. The obtained sheet like structure shows outstanding ORR and OER performance in alkaline systems with excellent stability compared to Pt/C catalyst. The doped heteroatom in the carbon is expected to have redistributed the charge around heteroatom dopants lowering the ORR potential and modifying the oxygen chemisorption mode thereby weakening the O-O bonding and improving the ORR activity and overall catalytic performance. The bifunctional activity (ΔE = E j=10 - E 1/2) of an air electrode for NPC, NSC, NC and Pt/C is 0.82 V, 0.87 V, 1.06 V and 1.03 V respectively, and the NPC value is smaller than most of the reported metal and non-metal based electrocatalysts. The ORR (from onset potential) and OER (10 mA cm-2) overpotential for NPC, NSC, and NC is (290 mV, 410 mV), (310 mV, 450 mV) and (340 mV, 600 mV) respectively. In the prepared catalyst the NPC exhibited higher ORR and OER activity (NPC > NSC > NC). The doping of P in NPC is found to have a great influence on the microstructure and therefore on the ORR and OER activity.

Ionophores as Potent Anti-malarials: A Miracle in the Making.

Plasmodium has a complex life cycle that spans between mosquito and human. For survival and pathogenesis it banks upon dynamic alterations in ionic transport across organelle and plasma membrane. Being a fundamental contributor of crucial biological processes in parasite, ionic balance facilitates parasite invasion, augmentation and transmission. Past few decades have witnessed tremendous advancement in understanding the relevance of ionic transit in parasites. Perhaps, not surprisingly, disruption of ionic homeostasis was thought to be detrimental for parasite. Compounds like ionophores are known to facilitate ionic transport across membrane down their electrochemical gradient. Despite continuous effort, malaria treatment is still a challenge particularly due to the development of resistance among parasites against existing therapeutic options. However, repurposing the existing drugs can be advantageous over de novo drug development programs in terms of cost and associated risk factors. Ionophores, being used in coccidiosis have proven to be of significance in the treatment of experimental models of malaria. Several recent reports have highlighted the attractive potential of ionophores such as Monensin, Maduramicin, Valinomycin, etc., that can act against multiple stages of malarial parasite's life cycle. Improved variety of these molecules may help in mitigating the drug resistance problems as well. This review is an attempt to examine the relevant literature and provide insight into the mechanism and prospects of different classes of ionophores as promising anti-malarial potpourri.

Efficacy of Liposomal Monensin on the Enhancement of the Antitumour Activity of Liposomal Ricin in Human Epidermoid Carcinoma (KB) Cells.

The monensin, known to enhance the cytotoxicity of ricin and ricin-based immunotoxins is a very hydrophobic molecule and this limits its administration in optimum doses under in vivo conditions. In order to realise its full potential, monensin was intercalated into various liposomal formulations and its ability to potentiate the cytotoxicity of ricin liposomes in human epidermoid carcinoma (KB) cells was studied. It was observed that ricin cytotoxicity enhancing ability of monensin liposome depends on the surface charge as well as density and chain length of distearoyl phosphatidylethanolamine-methoxy polyethylene glycol present on the surface of liposomal monensin. Maximum potentiation on the cytotoxicity of liposomal ricin was observed by monensin entrapped in neutral liposome (106.5 fold) followed by negatively charged (94.2 fold) and positively charged liposome (90 fold). Studies on the effect of variation of density and chain length of distearoyl phosphatidylethanolamine-methoxy polyethylene glycol showed that neutral monensin liposomes having 2.5 mol% distearoyl phosphatidylethanolamine-methoxy polyethylene glycol with chain length of 2000 exhibits maximum potentiation (117.6 fold) on the cytotoxicity of ricin liposomes when the cellular uptake of monensin liposome was maximum (42.0%) and the zeta potential value on the surface of liposomes was -0.645. The present study has clearly shown that liposomal monensin is very effective in enhancing the cytotoxicity of liposomal ricin in human cancer cells and liposome can be used as in vivo deliver vehicle for monensin to potentiate the cytotoxicity of liposomal ricin to eliminate cancer cells.

A new glycoprotein allergen/antigen with the protease activity from Aspergillus fumigatus.

BACKGROUND: Aspergillus fumigatus is an opportunistic fungus causing allergic and invasive aspergillosis in humans and animals. It secretes an array of complex biologically active glycoprotein antigens and allergens. It is important to identify and characterize probable potential virulent factors playing a major role in the pathogenesis of aspergillosis. METHODS: Using protein purification techniques (lectin affinity chromatography, gel filtration, electroelution and high-pressure liquid chromatography), a major antigen/allergen with a molecular weight of 56 kD (gp56) from A. fumigatus was purified to homogeneity. The protein was characterized by immunoblot, ELISA and protease assays. The N-terminal amino acid sequencing was performed. RESULTS: The gp56 protein showed a single band on silver staining and isoelectric focussing. The protein to carbohydrate ratio was 1.5:1 and gp56 gave a protein band at a molecular weight of 34 kD on enzymatic deglycosylation. It also exhibited IgG and IgE immunobinding with antibodies present in sera of allergic bronchopulmonary aspergillosis patients. The gp56 exhibited protease activity and N-terminal seven-amino acid sequence showed homology with fungal serine proteases. CONCLUSIONS: The gp56 protein by virtue of its proteolytic activity could be one of the virulent factors of A. fumigatus involved in establishing infection in the host along with other factors.

Altered immune response to liposomal allergens of Aspergillus fumigatus in mice.

Aspergillus fumigatus has been implicated as the major pathogenic fungus causing Aspergillus-mediated disorders. It secretes complex glycoprotein antigens and allergens, which induce type I and type III mediated hypersensitivity reactions. The immune response to these allergens/antigens in allergic disorders is characterized by elevated levels of specific IgE, Th2 cytokines and eosinophilia. In the current study, the ability of negatively charged liposomes entrapped with glycoprotein antigens and allergens of A. fumigatus to modulate the immune response was studied. Immune response in mice was evaluated with both free and liposomal formulations. Liposome entrapped glycoprotein antigens/allergens of A. fumigatus elicited a Th1 type response with increased levels of TNF-alpha (5.5-folds), IFN-gamma (four-folds), specific IgG (three-folds) and IgG2a (2.4-folds), low titers of specific IgG1 (2.2-folds decrease) and IgE (three-folds decrease), and decreased peripheral eosinophilia by four-folds in comparison to mice receiving free glycoprotein allergens/antigens of A. fumigatus. Histopathological examination of lung tissue sections clearly indicated reduced eosinophil infiltration in mice immunized with liposomal formulations. These results suggest potential of liposomal formulations for A. fumigatus allergens/antigens for exploration in immunotherapy.

Characterization of Aspergillus fumigatus protein disulfide isomerase family gene.

Aspergillus fumigatus is an opportunistic fungus which causes pulmonary complications in humans and animals. The clinical spectrum observed with A. fumigatus is attributed to the multifunctional nature of its antigens. Lack of understanding on the molecular processes and complexity of the fungus have spurred interest in the identification and characterization of its antigens/allergens with biological activities and virulence functions. For identification of some of these antigens/allergens, a cDNA library of A. fumigatus was screened with antibodies of allergic bronchopulmonary aspergillosis (ABPA) patients. One of the reactive clones was sequenced and observed to have an open reading frame of 1095 nucleotides corresponding to a polypeptide of 364 amino acids. The nucleotide and deduced amino acid sequence showed significant homology with the protein disulfide isomerase (PDI) superfamily. The expressed recombinant fusion protein exhibited specific IgG and IgE binding with antibodies present in ABPA patients' sera. The recombinant protein in vitro catalyzed folding of scrambled RNase. The probable epitopic regions of the deduced amino acid sequence were mapped by algorithmic analysis. This is the first report of isolation of a gene encoding a member of the PDI family from A. fumigatus. The PDI superfamily of proteins may play an important role in the protein folding mechanisms of A. fumigatus antigens/allergens for their interaction with the host.

Tumour targeted delivery of encapsulated dextran-doxorubicin conjugate using chitosan nanoparticles as carrier.

Doxorubicin (DXR) commonly used in cancer therapy produces undesirable side effects such as cardiotoxicity. To minimize these, attempts have been made to couple the drug with dextran (DEX) and then to encapsulate this drug conjugate in hydrogel nanoparticles. By encapsulation of the drug conjugate in biodegradable, biocompatible long circulating hydrogel nanoparticles, we further improved the therapeutic efficacy of the conjugate. The size of these nanoparticles as determined by quasi-elastic light scattering, was found to be 100+/-10 nm diameter, which favors the enhanced permeability and retention effect (EPR) as observed in most solid tumors. The antitumor effect of these DEX-DXR nanoparticles, was evaluated in J774A.1 macrophage tumor cells implanted in Balb/c mice. The in vivo efficacy of these nanoparticles as antitumor drug carriers, was determined by tumor regression and increased survival time as compared to drug conjugate and free drug. These results suggest that encapsulation of the conjugate in nanoparticles not only reduces the side effects, but also improves its therapeutic efficacy in the treatment of solid tumors.

Biodistribution of fluoresceinated dextran using novel nanoparticles evading reticuloendothelial system.

The rapid clearance of circulating nanoparticles from the blood stream coupled with their high uptake by liver and spleen has thus far been overcome by reducing the particle size, and by making the particle surface hydrophilic with poloxamers and poloxamines. We have prepared hydrogel nanoparticles of polyvinylpyrrolidone of a size less than 100 nm diameter with precise size distribution. Since the inner cores of these particles are also hydrophilic, these particles are capable of encapsulating water-soluble compounds. Biodistribution of these particles shows practically negligible (<1%) uptake by the macrophages in liver and spleen, and approximately 5-10% of these particles remain in circulation even 8 h after i.v. injection. Increasing the surface hydrophobicity as well as particle size can increase the RES uptake of these particles. Because of longer residence in blood, the hydrogel nanoparticles have potential therapeutic applications particularly in cancer: the water-soluble cytotoxic agents encapsulated in these particles can be targeted to tumors while minimizing the likelihood of toxicity to reticuloendothelial system (RES).

Biodistribution of amphotericin B when delivered through cholesterol hemisuccinate vesicles in normal and A. fumigatus infected mice.

PURPOSE: This study compared the biodistribution of two amphotericin B formulations in normal and Aspergillus infected mice. Amphotericin B cholesterol hemisuccinate vesicles (ABCV) which reduces the toxicity of amphotericin B and thereby enhances its therapeutic efficacy in a murine model of aspergillosis was compared with conventional amphotericin B deoxycholate suspension (AmB(DOC)). METHODS: ABCV (12 mg/kg wt) and AmB(DOC) (2 mg/kg wt) were intravenously administered to normal and A. fumigatus infected mice. The concentration of amphotericin B in plasma and other organs was determined at different time points. RESULTS: It was observed that ABCV had a significantly different pharmacokinetic profile compared to conventional amphotericin B. In comparison to AmB(DOC) significantly lower levels of amphotericin B were observed in kidneys and plasma, the major target organs of toxicity. Animals receiving ABCV demonstrated high levels of amphotericin B in liver (38% retention till 48 h) and spleen (2.6% retention till 48 h) in comparison to AmB(DOC) (7.3% and 0.21% retention in liver and spleen respectively till 48 h). Biodistribution studies of ABCV in infected mice demonstrated that there was a moderate enhancement in levels of amphotericin B in liver, spleen, lungs and kidneys as compared to normal mice and the plasma levels were reduced. However, such observations were not made after AmB(DOC) administration to infected mice except for kidneys in which there was a marked increase in uptake as compared to normal mice. CONCLUSIONS: Our results suggest that prolonged retention of high concentrations of ABCV in reticuloendothelial system organs is the reason for its reduced toxicity. Enhanced localization of the drug at the infected site may lead to improvement in therapeutic efficacy.

Effect of amphotericin B lipid formulation on immune response in aspergillosis.

The immune response against Aspergillus fumigatus has been studied during infection and therapy in order to understand the mechanism of pathogenesis and the effect of treatment with amphotericin B. With this in view an animal model of aspergillosis was developed in Balb/c mice by intravenous injection of an optimized dose of 3. 6x10(6) A. fumigatus spores. Infection due to Aspergillus was well established by histopathological examination and fungal load in the animal. Lesions and eosinophil infiltration was observed in the infected tissues which indicated the involvement of a Type I hypersensitivity response. Evaluation of serological parameters indicated high levels of interleukin-4 (IL-4) and A. fumigatus specific IgG antibodies. The reduction in fungal load and modulation of immune response in the infected mice was studied following treatment with amphotericin B/cholesterol hemisuccinate vesicles (ABCV). The results clearly indicated significant reduction in the fungal load, disappearance of eosinophils and lesions with the appearance of macrophages and neutrophils in the infected lung tissue, a decrease in IL-4 (fourfold) and a concomitant increase of interferon-gamma (IFN-gamma; twofold) with an improvement in general condition of mice. In the non-treated mice, the rise of IL-4 level indicated the association of T(H)2 cell response with susceptibility to infection while the increase of IFN-gamma in the treated group suggested that T(H)1 cell response may be involved in resistance to Aspergillus infection.

Assessment of targeting potential of galactosylated and mannosylated sterically stabilized liposomes to different cell types of mouse liver.

Galactose and Mannose residues were tagged on the surface of n-glutaryl-phosphatidylethanolamine (NGPE) containing liposomes with and without polyethylene glycol of molecular weight 2000 Da conjugated to distearoyl phosphatidylethanolamine (PEG-2000-DSPE). Biodistribution studies showed that sugar bearing liposomes were cleared more rapidly from circulation than those not bearing the sugar moieties. However, the rate of clearance of glycosylated conventional liposomes was much faster than the sugar bearing sterically stabilized liposomes. Intrahepatic distribution studies showed that a substantial amount of conventional liposomes without sugar residues were taken up by both parenchymal (P) (40%) and non-parenchymal (NP) cells (60%). However, incorporation of PEG-2000-DSPE shifted this uptake slightly in favour of parenchymal cells (47%). While ratio of distribution of galactosylated conventional liposomes to P and NP cells was found to be 74:26, galactosylation of sterically stabilized liposomes further enhanced the affinity of these vesicles towards P cells (P:NP ratio being 93:7). Thus, reduced uptake by Kupffer cells was observed with galactosylated sterically stabilized liposomes as compared to conventional liposomes. Whereas, mannosylation of both the liposomes shifted the distribution towards Kupffer cells in an analogous manner. These findings indicate that sterically stabilized liposomes tagged with galactose residues on their surface are more effective in targeting the entrapped material to hepatocytes as compared to conventional liposomes. This approach can therefore be employed for delivering therapeutic agents like drugs, enzymes, genetic materials, anti-sense oligonucleotides selectively to liver P cells for treatment of hepatic disorders.

Toxicity and therapeutic efficacy of amphotericin B delivered through cholesterol hemisuccinate vesicles in the treatment of experimental murine aspergillosis.

We have studied the toxicity and therapeutic efficacy of amphotericin B in cholesterol hemisuccinate vesicles (ABCV) in the treatment of invasive aspergillosis in Balb/c mice. The toxicity of amphotericin B was significantly reduced when delivered through cholesterol hemisuccinate vesicles as compared to deoxycholate suspension (AmB(DOC)) as evidenced by reduced nephrotoxicity in Balb/c mice, lower in-vitro toxicity to erythrocytes and higher maximum tolerated dose. The latter increased from 2 mg/kg wt for AmB(DOC) to 17 mg/kg wt for ABCV. The vesicles had a mean diameter of 252 nm. In order to evaluate the therapeutic efficacy of ABCV, Aspergillus fumigatus-infected mice were treated with ABCV 2, 4, 8 or 12 mg/kg or AmB(DOC) 1 mg/kg wt. The antifungal activity was highly dependent on the dosage of ABCV on the basis of survival of treated mice and cfu in lung, liver, spleen and kidney. This study found that ABCV had dose-dependent antifungal activity and therapeutic efficacy in the treatment of invasive aspergillosis in Balb/c mice.

A colorimetric estimation of polyethyleneglycol-conjugated phospholipid in stealth liposomes.

The paper describes a colorimetric method for estimation of polyethyleneglycol (PEG)-conjugated phospholipid in either form, free or bound to liposomes. It provides a rapid, highly reproducible, and sensitive tool to detect PEG-coupled phospholipid in amounts as low as 1 microg giving a linear response over a range of 1-100 microg. The method makes use of the biphasic system comprising aqueous ammonium ferrithiocyanate and chloroform, developed by Stewart for estimation of phospholipids. The same system was also applied for quantitation of PEG in PEG-protein conjugates in a recent report. The samples were digested with phospholipase-C, prior to analysis, in order to eliminate the contributions from liposomal phospholipids other than the PEG-conjugated phospholipid. The technique can give valuable information regarding the retention of PEG coating on the surface of the vesicles when employed in combination with markers for the aqueous compartment. In addition, it does not suffer from interference by proteins. This makes it particularly suitable for monitoring the pharmacokinetics of stealth liposomes, using PEG-phospholipid as a lipid probe. Since it does not involve handling hazardous radioisotopes, the suggested technique could even be utilized in clinical trials.

A colorimetric assay for estimation of polyethylene glycol and polyethylene glycolated protein using ammonium ferrothiocyanate.

A colorimetric method for quantitative assay of polyethylene glycol (PEG) described here is based on partitioning of a chromophore present in ammonium ferrothiocyanate reagent from an aqueous to a chloroform phase in the presence of PEG. The method is simple, reproducible, and can detect PEG in amounts as low as 5 microg. It gives a linear response over a range of 5-100 microg. The absence of any interference by proteins makes the assay equally suitable for the estimation of PEG in PEG-protein conjugates. The method was employed to monitor the separation profile of a mixture of free and PEG-5000 coupled to bovine serum albumin during purification through a gel filtration column. In this report we have also demonstrated for the first time an assay method which permits a critical evaluation of pharmacokinetic properties of any PEG-protein conjugate under in vivo conditions.

Potentiation of ricin cytotoxicity by liposomal monensin under in vitro and in vivo conditions.

Effect of monensin, intercalated in liposomes on the cytotoxicities of ricin, Pseudomonas exotoxin A and diphtheria toxin in phagocytic and non-phagocytic cells as well as in mice has been studied. Intercalation does not disturb the integrity of the liposomal bilayer and substantially enhances the cytotoxicities of ricin and Pseudomonas exotoxin A in both phagocytic and non-phagocytic cells while it has no effect on diphtheria toxin. The observed effect is highly dependent on the liposomal lipid composition as well as cell types. The potentiating ability of monensin in neutral vesicle is 2.2-fold higher than in negatively charged vesicles in non-phagocytic cells while no difference was observed in phagocytic cells. Incorporation of stearylamine in liposomes reduces the potentiating effect of monensin. Liposomal monensin has also been found to enhance the cytotoxicity of ricin in mouse in vivo in a dose-dependent manner and is maximal when ricin is injected within 60 min of monensin injection. Liposomal monensin remains in circulation for 2 hr while free monensin remains only for 15 min. Tissue distribution studies reveal that liposomal monensin is present mainly in the liver and spleen which are also the major sites for ricin accumulation. Thus liposome is found to be an effective delivery vehicle for monensin to potentiate the cytotoxicity of immunotoxins or hormonotoxins and could prove useful for selective elimination of cancer cells.

Enhancing potency of liposomal monensin on ricin cytotoxicity in mouse macrophage tumor cells.

Monensin, a carboxylic ionophore, which is known to disrupt intracellular trafficking of proteins was intercalated in liposomes and its effect on the stability of liposomes and cytotoxicities of ricin and Pseudomonas exotoxin A in mouse macrophage tumor cells J774A.1 was studied. Stability of liposomes containing monensin was comparable to liposomes without monensin. The cytotoxicity of ricin and Pseudomonas exotoxin A was significantly enhanced by 1nM liposomal monensin (15.7 and 3.6 fold respectively). The enhancing potency of monensin in neutral and negative vesicles was found to be similar, while it was drastically reduced in positive vesicles. The specific uptake of 125I-gelonin from neutral and negative vesicles was not significantly different, whereas from positive vesicles no uptake was observed. Serum strongly influenced the binding at 4 degrees C of positive vesicles as well as the enhancing potency of monensin in these vesicles. Monensin in neutral and negative vesicles significantly reduced the lag period of ricin action, while in positive vesicles, it had no effect. These studies clearly indicate that liposomes could be used as a delivery vehicle for monensin.

Monensin intercalation in liposomes: effect on cytotoxicities of ricin, Pseudomonas exotoxin A and diphtheria toxin in CHO cells.

Monensin, a carboxylic ionophore was intercalated in liposomes (liposomal monensin) and its effect on cytotoxicities of ricin, Pseudomonas exotoxin A and diphtheria toxin in CHO cells was studied. Intercalation of monensin in liposomal bilayer is found to have no effect on its stability and interaction with cells. Liposomal monensin (1 nM) substantially enhance the cytotoxicities of ricin (62-fold) and Pseudomonas exotoxin A (11.5-fold) while it has no effect on diphtheria toxin. This observed effect is highly dependent on the liposomal lipid composition. The potentiating ability of monensin (1 nM) in neutral vesicles is significantly higher (2.2-fold) as compared to negatively charges vesicles. This ability is drastically reduced by incorporation of stearylamine in liposomes and is found to be dependent on the density of stearylamine as well as on the concentration of serum in the medium. Monensin in liposomes containing 24 mol% stearylamine has a very marginal effect on the cytotoxicity of ricin (7.5-fold) which is further reduced (1.5-fold) in the presence of 20% serum. The uptake of 125I-gelonin from neutral vesicles is significantly higher (approximately 2.0-fold) than that from the negative vesicles. The uptake from positive vesicles is highly dependent on the concentration of stearylamine. The reduction in the lag period (30 min) of ricin action by monensin in neutral and negative vesicle is comparable with free monensin. However, monensin in positive vesicle has no effect on it. These studies have suggested that liposomes could be used as a delivery vehicle for monensin for selective elimination of tumor cells in combination with hybrid toxins.

In vivo potentiation of ricin toxicity by monensin delivered through liposomes.

Monensin, a carboxylic ionophore, which is known to raise intravesicular pH, was intercalated in liposomes and its effect on the toxicity of ricin in mice was studied. The toxicity of ricin in vivo was found to be significantly enhanced by the administration of monensin intercalated in liposomes (liposomal monensin). The observed enhancement of the toxicity of ricin by monensin was highly dose-dependent and was maximal when ricin was injected within 60 min of monensin injection. The survival time was found to be reduced in the range of 8-20 h, depending on the dose of ricin used, by liposomal monensin. Stability of liposomes containing monensin as inferred from the release of entrapped calcein or FITC-dextran under both in vivo and in vitro conditions was comparable to that observed for liposomes without monensin. Liposomal monensin remains in circulation for 2 h and was cleared from the blood stream after 4 h. In contrast, 15 min was required for the clearance of monensin when administered in free form. Studies on the distribution of liposomal monensin and 125I-ricin in various tissues have revealed that monensin is mainly localized in the liver and spleen which are also the major sites for ricin accumulation. Our observation on the substantial enhancement of ricin toxicity in vivo by liposomal monensin strongly supports the potential usefulness of the latter as a potentiating agent in the enhancement of the toxicity of immunotoxin or hormonotoxin for selective elimination of cancer cells.

Liposome immune lysis assay (LILA) for gelonin.

A complement-mediated liposome immune lysis assay using entrapped calcein was developed for a plant toxin gelonin. Gelonin was covalently coupled to DPPE, and then adsorbed on to the surface of liposomes. Such antigen-bearing liposomes when incubated with anti-gelonin antibody in the presence of guinea pig complement undergo lysis. The detection range is from 3 ng to 60 ng. The method was used to monitor isolation of gelonin by affinity chromatography. It was observed that a minor peak in addition to the major one comes with gelonin, shared common epitopes/epitope with gelonin in immunological reaction. This was further confirmed by SDS-gel electrophoresis indicating the former being an isoform of gelonin. A comparative study of the immunocross-reactivity of ricin and ricin A chain with anti-gelonin antibody was carried out. It was found that while ricin A chain cross-reacted extensively with gelonin antibody and intact ricin elicited little or no cross-reactivity. It is suggested that the present LILA may be employed for the detection and quantitation of ricin A chain by this LILA method.

Interaction of gelonin with macrophages: effect of lysosomotropic amines.

The cytotoxic effects of gelonin on various phagocytic and nonphagocytic cells were studied. Peritoneal exudate cells (PEC) are found to be more sensitive to gelonin compared to P388D1 and J774A.1 cells, nonphagocytic cells being the least sensitive. While chloroquine markedly enhances the cytotoxicity of gelonin in macrophages (greater than 100-fold) ammonium chloride confers protection. A higher rate of uptake of 125I-gelonin in PEC (7 times the rate observed in other cells) is probably mediated by an interaction of terminal mannose residues of gelonin with mannose receptors on PEC plasma membrane as inferred from a pronounced inhibitory effect of mannan. In contrast to a pronounced inhibitory effect of mannan on the uptake of gelonin in PEC (7-fold), the cytotoxicity is reduced only by 2-fold. On the other hand, mannan has little or no effect on enhancement of the toxicity of gelonin by chloroquine. The studies have suggested that the internalization of gelonin in PEC may involve two pathways: (a) mannose receptor-mediated endocytosis, which plays a minor role in the intoxication process, and (b) nonspecific fluid phase pinocytosis, which is susceptible to enhancement by chloroquine and has a major role to play in the manifestation of the toxic effect of gelonin. In macrophage-like cells only the latter pathway operates.