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Biomedical subjects

D Subrahmanyam

Publications and source records attributed to D Subrahmanyam.

At least 37 records · Page 2Linked to original sources

Protective immune responses with trickle infections of third-stage filarial larvae of Wuchereria bancrofti in mice.

Groups of inbred BALB/c mice were immunized with trickle doses of 20 live third-stage larvae (L3) of Wuchereria bancrofti each subcutaneously or with 150 microg of sonicated microfilarial antigens emulsified in Freund's adjuvant intramuscularly. An antibody response was distinctly seen after seven trickle doses of L3 and following with the sonicated microfilarial immunization. Due to the non-permissive nature of inbred mice to W. bancrofti infections, a novel immunization approach was adopted using appropriate age- and sex-matched controls. The anti-L3 response in terms of antibody-dependent cell-mediated adhesion and killing was assessed in the immunized animals by implanting live L3 in micropore chambers subcutaneously. About 75% L3 W. bancrofti were affected in animals sensitized with seven trickle doses of L3. When sensitizations were continued, as high as 92% of L3 were seen affected with ten trickle doses compared with 27% in age-matched controls. Immunization with sonicated microfilarial antigen affected about 70% of L3 as opposed to only 12% in controls. A positive correlation was observed in the antibody response with protectivity. This method of induction and assessment of the anti-L3 response involving a small set of animals has not only allowed quantification of affected L3 but has also enabled us to visualize larval conditions in immunologically activated animals. The micropore chamber system, would be useful in monitoring the induction of protective immune response against W. bancrofti in inbred mice. Experimentation on large numbers of animals is required to elucidate further the response of mice towards L3 and also to pinpoint the putative protective antigens.

Animals↗

Characterization of a monoclonal antibody against infective larvae of Brugia malayi.

Monoclonal antibodies were produced following immunization of mice with live infective larvae of Brugia malayi. One of these, 46.08.76, is an antibody that promotes adherence of mouse peritoneal macrophages and human peripheral blood leucocytes to the infective larvae of B. malayi and Wuchereria bancrofti, respectively, and kills them. Fresh normal serum, as a source of complement, augments this effect. The same monoclonal antibody conferred 89% protection to jirds (Meriones unguiculatus) against challenge infection of B. malayi stage-three larvae. This monoclonal antibody recognizes antigens of 80,000, 67,000, 52,000 and 36,000 MW proteins present among the antigens of larvae, as detected by an immunoblotting technique. The antibody also reacts with antigens of infective larvae of Litomosoides carinii, Dipetalonema viteae and B. pahangi, but to a smaller extent.

Animals↗

Antifilarials and their mode of action.

Diethylcarbamazine and suramin are the drugs of choice for the control of lymphatic filariasis and onchocerciasis respectively. Benzimidazoles, ivermectin, furapyrimidone, and isothiocyanates and their derivatives emerge as compounds of potential clinical interest as antifilarials. Diethylcarbamazine is predominantly a microfilaricide affecting the neuromuscular system of the parasites and promotes cellular cytotoxicity mediated by immune factors. Suramin is macrofilaricidal, has a narrow therapeutic index and damages the intestinal epithelium of the worms. Benzimidazoles bind to tubulins and inhibit their assembly to microtubules. Ivermectin is an extremely potent microfilaricide in onchocerciasis; it augments immune responses and impairs the neuromuscular function of the parasites, leading to paralysis. Isothiocyanates and their derivatives are both microfilaricidal and macrofilaricidal and affect the energy metabolism of the parasites. Although the precise mode of action of antifilarial drugs is not established, information is available on the responses listed above and also on the effects of the drugs on the carbohydrate and folate metabolism of the parasites.

Animals↗

Complement activation by eggs and microfilariae of filarial parasites.

The complement of fresh normal rat serum was activated by filarial eggs and microfilariae (mf). C3 was deposited on the surface of Litomosoides carinii, Brugia pahangi, Brugia malayi and Dipetalonema viteae as seen by immunofluorescence. Intra-uterine and in vitro-derived mf did not bind C3. In contrast, C3 bound to the blood-derived mf of B. pahangi and B. malayi as well as exsheathed mf of L. carinii and B. malayi. Significant consumption of complement was observed with eggs of all filarial species, as well as sheathed mf of B. pahangi, B. malayi and exsheathed mf of L. carinii and B. malayi. These experiments indicated that complement was activated by filarial parasites via the alternative pathway. The bound complement promoted neutrophil-mediated adherence and cytotoxicity.

Animals↗

Lectin-binding characteristics of Wuchereria bancrofti microfilariae.

The binding of 10 different lectins to the surface of microfilariae of Wuchereria bancrofti has been investigated. Wheat germ agglutinin (WGA) and Helix pomatia lectin (HPA) bound specifically to the sheathed microfilariae indicating the presence of N-acetyl-D-glucosamine and N-acetyl-D-galactosamine respectively on the surface. Exsheathed microfilariae did not react with any of the lectins. Treatment of sheathed microfilariae with proteases resulted in increased binding of WGA and HPA. Such treated microfilariae showed a weak binding of Concanavalin A (Con A), and lectins of lentil (LCH) and of Limulus polyphemus (LPA). Sheathed microfilariae incubated with sera of people living in endemic zones of filariasis but with no apparent evidence of infection (endemic normals), or with sera of chronic elephantiasis patients, or with their respective gamma globulin fractions, bound Con A and LCH. These lectins bound weakly to exsheathed microfilariae under the same conditions. Binding was due to the mannose components of the specific immunoglobulins of the sera which coated the microfilariae. However, microfilariae when incubated with sera or their globulin fractions from non-endemic normals (NEN), or from microfilarial carriers, did not bind Con A and LCH, suggesting that specific immunoglobulins were neither present in NEN sera nor in significant amounts in sera of microfilarial carriers.

Acetylgalactosamine↗

Litomosoides carinii: characterization of surface carbohydrates of microfilariae and infective larvae.

The carbohydrate moieties of microfilariae (Mf) and infective larvae (L3) have been investigated by lectin-binding technique. Mf derived from three sources, namely, uteri (in utero), released in vitro from adults and from blood of rodents infected with Litomosoides carinii were examined by using fluoresceinated lectins. Wheat germ agglutinin (WGA) bound to these Mf and the binding was inhibited by N-acetyl glucosamine. In addition to WGA, Concanavalin A (Con A) and lentil lectin (LCH) bound to in vitro-released and in utero-derived Mf showing the presence of mannose moieties on their surface. In utero-derived Mf also showed binding with the agglutinins of Limulus polyphemus (LPA), peanut (PNA), Ricinus communis (RCA), Helix pomatia (HPA), Soyabean (SBA) and Dolichos biflorus (DBA) but not to that of Ulex europaeus (UEA) indicating the presence of additional carbohydrate molecules like sialic acid, galactose and N-acetyl galactosamine on their sheath. None of the lectins bound to the cuticle of exsheathed Mf. Treatment of blood-derived and in vitro-released Mf with certain proteases exposed additional binding sites for SBA, HPA, Con A and LCH. In case of L3, only PNA bound to the larvae isolated from infective mites Bdellonyssus bacoti, and the binding was inhibited by D-galactose. No such binding of the lectins was seen to the larvae that migrated to the pleural cavity of jirds indicating that there is considerable change on the parasite surface during their migration in the vertebrate host. Sheathed Mf and mite-derived L3 when incubated with immune rat sera, bind Con A and LCH lectins possibly due to the mannose components of the specific immunoglobulins that coat onto the Mf and L3.

Animals↗

Effect of ivermectin on serum dependent cellular interactions to Dipetalonema viteae microfilariae.

The effects of ivermectin (Iv) on the filarial infection has been investigated using Dipetalonema viteae--Mastomys natalensis model system. In vitro, Iv (3 micrograms/ml) had no effect on the microfilariae (mf) in the presence or absence of serum from normal or infected Mastomys (NMS, IMS). However, the sera promoted cell-mediated cytotoxicity with macrophages or eosinophils to the mf when Iv was added at 3 ng/ml. Sera isolated from normal or infected Mastomys after subcutaneous treatment with Iv (100 micrograms/kg) (Iv-NMS, Iv-IMS) also induced cellular cytotoxicity to the mf in vitro. Available evidence suggests that Iv mediates the cellular cytotoxicity by complement activation by alternative pathway. Iv treatment of D. viteae infected Mastomys resulted in an initial mobilization of the mf into circulation followed by the death of both the mf and adults in vivo. There was enhanced antibody level in Iv treated infected animals and antibody dependent cellular cytotoxicity mediated by IgM seems to be predominantly involved in elimination of the mf. The serum containing these antibodies was effective in clearing the circulating mf from Mastomys on passive immunization. Thus, Iv seems to synergize host immune factors against D. viteae infection.

Animals↗

Pharmacokinetics of primaquine in patients with P. vivax malaria.

The pharmacokinetics of primaquine (PQ) and its major carboxylic acid metabolite (PQC) have been studied in seven Indian patients with P. vivax malaria following PQ 15 mg/day p.o. for 14 days. After a single oral dose on Day 1, a mean peak blood concentration of 50.7 ng/ml PQ was attained after 2.3 h, which declined monoexponentially with a half-life of 5.6 h. The mean total body clearance was 37.6 l/h and the volume of distribution was 292 l. The mean renal excretion (0-24 h) of the drug was only 0.54% of the dose and renal clearance was 0.189 l/h. Following chronic administration, none of the pharmacokinetic parameters was affected, and a steady state blood concentration of 2.5-4.2 ng/ml PQ was attained. After the first dose of PQ, PQC had a mean area under the blood concentration - time curve 11-fold higher than that of the parent drug. In contrast to the rapid distribution and elimination of PQ, the metabolite showed a longer mean residence time and accumulation in the body. The mean Cmax and AUC of the metabolite on Day 14 were 48 and 40% higher than the corresponding Day 1 values. The metabolite could not be detected in urine at any time in any patient. PQ and its metabolite did not show any accumulation in blood cells.

Adolescent↗

Brugia malayi: rat cell interactions with infective larvae mediated by complement.

Albino rat macrophages and neutrophils, in the presence of fresh normal rat serum as a source of complement, adhered to and promoted killing of Brugia malayi infective larvae in vitro. Eosinophils, by themselves, were marginally cytotoxic at a high cell-target ratio but promoted cytotoxicity when mixed with macrophages. Eosinophil culture supernatants enhanced the macrophage mediated killing of infective larvae. The complement of fresh normal rat serum was found to act by the alternate pathway. Fresh normal rat serum depleted of alternate pathway complement activity by treatment with zymosan A, or of Factor B by heating at 50 C for 20 min, or of Factor D by passing through Sephadex G75 column, failed to promote cell adherence to the parasite. C3 molecules were detected on the surface of infective larvae by immunofluorescence. There was a significant consumption of complement when Brugia malayi infective larvae were incubated in fresh normal rat serum. Albino rat cells were more potent in inducing cytotoxicity to infective larvae in vitro than those from jird or Mastomys natalensis, which may reflect the greater resistance offered by the rat to B. malayi infection. There was much less cellular infiltration on introduction of Brugia malayi infective larvae into the peritoneal cavity of jirds compared to rats and Mastomys natalensis indicating the greater susceptibility of jirds to intraperitoneally induced infections.

Animals↗

Effect of acute sublethal and chronic administration of DDT (chlorophenotane) on brain lipid metabolism of rhesus monkeys.

Either a single oral dose of 1,1,1-trichloro-2,2-bis-(p-chlorophenyl) ethane (DDT), 150 mg/kg body weight or a long-term chronic dose of the pesticide, 10 mg/kg body weight daily for 100 days was administered to rhesus monkeys and the level of various lipid classes was studied in the brain. A significant decrease was observed in total lipid, unesterified cholesterol and phospholipid (PL) level. Individual PL fractions showed a generalized pattern of reduction, so also did the cholesterol (chol)/PL ratio. Sphingomyelin (SM) registered a significant increase, while no significant alteration was observed in the brain galactolipid and ganglioside level. Lipid lowering effect of DDT was more pronounced in the chronic group. Lipids associated with the myelin sheath were found to be more resistant to pesticide injury, while cholesterol and PL metabolism were more affected.

Animals↗