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[
Acta Trop,
1998]
Evaluation of antifilarial activity of new potential agents in vivo is extremely time consuming and uneconomic. In the present study effort has been made to develop an in vitro screening method using Acanthocheilonema viteae, a subcutaneously dwelling rodent filariid with anaerobic metabolic characteristics like human filariids, W. Bancrofti/Brugia malayi as test parasite. Motility test and tetrazolium (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide, MTT) based colorimetric assay were used as parameters in in vitro assay. Results showed that 92.3% of compounds (in vivo active) could be picked up in the in vitro assay when both adults and microfilarae (mf) were used simultaneously. Mf and adult stages separately detected, respectively, 84.6 and 69.2% of in vivo active compounds. The adults and mf separately and both the life stages together exhibited, respectively, 80.0, 50.0 and 80.0% false positive results in the in vitro test with in vivo inactive compounds. It is felt that mf stage when used in in vitro test using motility and MTT assays as parameters would be useful in primary screening of new potential filaricides.
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[
Folia Parasitol (Praha),
1992]
The chemotherapeutic efficacy of mebendazole given in combination with Freund's complete adjuvant (FCA) against Brugia malayi in multimammate rat was evaluated. Animals treated with mebendazole, orally at 200 mg/kg x 5 consecutive days along with FCA given subcutaneously (s.c.) on day -10, day 0 and day +15 of the drug treatment killed 48.51% of the adult worms. This drug given alone at the same regimen and by the same route showed only 18.7% mortality rate on adults. Mebendazole given intraperitoneally along with FCA given s.c., however, was four times more efficacious as filaricide than mebendazole alone. Nevertheless, the animals receiving FCA alone also revealed 23.5% mortality rate of adult worms. The animals receiving a combination therapy or FCA alone showed significant increase in antibody titre to the filariae which however decreased in the later stages. No enhancement of antibody level could be detected in animals treated with mebendazole alone. The non-specific immunopotentiation induced by FCA appeared to play a major role in enhancing the activity of mebendazole.
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[
Acta Trop,
1998]
Investigations on various aspects of human filariasis using target filarial parasite, Brugia malayi is jeopardised to a great extent due to its prolonged incubation period and poor harvest from the existing experimental animal models. To obviate these difficulties it was contemplated to establish B. malayi infection in immunosuppressed Mastomys coucha. Cortisone, a well known immunosuppressant, was used at 10 mg/kg dose level subcutaneously in two courses each of 5 days duration. The first course was administered 1 week before and the second, a week after infective exposure. Mastomys were exposed either with 100 or 200 infective larvae (L3) each. Untreated age-matched animals were also exposed simultaneously. The minimum prepatent period was observed to be 90.7 days in immunosuppressed animals exposed to 200 L3. The course of microfilaraemia in immunosuppressed and control animals was identical up to 180 days of observation period. However, the adult worm recovery from the former group of Mastomys was higher. It is surmised that exposure with B. malayi L3 in immunosuppressed Mastomys would be of great advantage in getting larger harvests of adult worms of B. malayi.
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[
Acta Trop,
1998]
Investigations on various aspects of human filariasis using target filarial parasite, Brugia malayi is jeopardised to a great extent due to its prolonged incubation period and poor harvest from the existing experimental animal models. To obviate these difficulties it was decided to establish B. malayi infection in immunosuppressed Mastomys coucha. Cortisone, a well-known immunosuppressant, was used at 10-mg/kg dose level subcutaneously in two courses each of 5 days duration. The first course was administered 1 week before and the second, 1 week after infective exposure. Mastomys were exposed either with 100 or 200 L3 each. Untreated age-matched animals were also exposed simultaneously. The minimum prepatent period was observed to be 90.7 days in immunosuppressed animals exposed to 200 L3. The course of microfilaraemia in immunosuppressed and control animals was identical up to 180 days of observation period. However, the adult worm recovery from the former group of mastomys was higher. It is surmised that exposure with B. malayi L3 in immunosuppressed mastomys would be of great advantage in getting larger harvests of adult worms of B. malayi.
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[
Folia Parasitol (Praha),
1997]
The fate of intraperitoneally inoculated infective third-stage larvae (L3) of the nematode Brugia malayi Lichtenstein and the status of the peritoneal macrophage function were investigated in the susceptible rodent hosts Mastomys natalensis Roberts and Meriones unguiculatus Milne-Edwards (jird). Jirds and M. natalensis were inoculated intraperitoneally with 125 and 250 L3 and the worm burden and peritoneal macrophage function in the two species were compared at different days post-inoculation (DPI). None of the infected M. natalensis had adult worms in the peritoneal cavity; very few degenerating L3 surrounded by peritoneal cells were recovered 7 and 15 DPI. In contrast, all the infected jirds showed the parasite in different stages of development and the worm burden at different days PI was more in 250 L3 dose group than in 125 L3 dose group. The phagocytic function of peritoneal macrophages of normal M. natalensis was twice higher than that of jirds. This function was found significantly suppressed in both host species at 15 DPI; at 35 DPI, the activity was still at this low level in the jird, while that in M. natalensis reverted to uninfected age- and sex-matched control levels. These findings demonstrate that the peritoneal environment of M. natalensis is not conducive to the development of B. malayi and this is probably related to high macrophage activity in the peritoneum of this host compared to that found in the jird.
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[
Parasitol Res,
1989]
Litomosoides carinii-, Acanthocheilonema viteae- or Brugia malayi-infected Mastomys natalensis were sensitised against sheep red blood cells (SRBC) on various occasions after infection to determine the effect of filarial infections on the immune response to a non-filarial antigen. The phagocytic activity of the reticuloendothelial system (RES) was controlled in vivo by the elimination of 51Cr-labelled SRBC. Antibody titres against SRBC (agglutinating and lytic antibodies) were similar to those of uninfected controls in L. carinii- or B. malayi-infected Mastomys sensitised during prepatency or early patency up to 90 days post infection (p.i.) but were reduced in animals sensitised during patency. A significant inverse correlation existed between anti-SRBC antibody titres and microfilaraemia levels. In contrast, A. viteae-infected Mastomys showed reduced humoral anti-SRBC responses at the end of prepatency, whereas the response tended towards normal with increasing parasitaemia. Delayed-type hypersensitivity (DTH) against SRBC was measured as footpad swelling after sensitisation by the s.c. or i.v. route and intraplantar challenge. DTH reactions were reduced during prepatency in all infections after s.c. sensitisation. During patency, 24-h reactions were similar to those of age-matched controls but the swelling persisted 24 or 48 h longer than in the latter. In A. viteae infections, even enhanced 24-h reactions were found during patency. Histological investigations did not reveal differences in the type of cell infiltrations between infected and control animals. After i.v. sensitisation with SRBC, L. carinii- and A. viteae-infected animals showed weaker DTH reactions than the controls, independent of the period after infection. In the case of B. malayi infections, DTH reactions were similar to those of controls during early prepatency, whereas reduced DTH responses were observed later than 50 days p.i. As shown in L. carinii-infected animals, depressed DTH reactions after i.v. sensitisation did not depend on an altered expression phase but rather on an altered regulation during the inductive phase of the response: increases in the sensitising SRBC doses that caused decreasing DTH reactions in uninfected animals led to enhanced reactions in infected animals. Phagocytosis of i.v. injected 51Cr-labelled SRBC was enhanced during prepatency in L. carinii infection and during patency in all infections.
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[
Parasitology,
1999]
Humoral immune responses of the Indian leaf monkey (Presbytis entellus) experimentally infected with Brugia malayi and exhibiting disease manifestations were studied. Microfilaraemia, filaria-specific IgG and circulating immune complexes (CICs) were determined in the monkeys at different time-points after inoculation of B. malayi 3rd-stage larvae. Sera were analysed for recognition pattern of adult parasite antigen molecules by immunoblotting. More than 60% of the infected monkeys developed episodic or persistent limb oedema with or without fever and with low or no microfilaraemia. While both CIC and filaria specific IgG levels were comparable in animals showing no disease symptoms (asymptomatics) and some animals showing symptoms (symptomatics), IgG levels peaked during pre-patent stage in symptomatics and during latent stage in asymptomatic animals. However, some of the symptomatic animals showed a low level of filaria-specific IgG as compared to asymptomatic and other symptomatic animals. The immunoblot analysis showed non-reactivity of 17 and 55 kDa antigens with sera of symptomatic animals. The results thus suggest that humoral immune responses as measured in the present study do not precede the development of the manifestations. However, 2 non-reactive antigen molecules identified by symptomatic sera need further study to establish their possible involvement, if any, in the development of acute disease manifestations in this model.
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[
J Parasitol,
1999]
To investigate the cell-mediated immune (CMI) responses of the host during the development of acute filarial disease manifestations, we studied the sequential changes in CD4+ and CD8+ T-cell subsets, leukocyte migration inhibition (LMI) response to Brugia malayi adult worm antigen, and concanavalin-A (ConA) and filarial antigen-induced lymphocyte transformation (LT) in the Indian leaf monkey (Presbytis entellus)-B. malayi model. Filarial infection was established in monkeys by subcutaneous inoculations of infective larvae (L3) (700-1,250 L3/monkey) in multiple doses, and the infected monkeys were categorized as symptomatic (Sym) and asymptomatic (Asym) depending on whether or not acute clinical manifestations were shown by them. In Sym monkeys, LMI response to homologous adult parasite antigen was significantly suppressed as compared to Asym monkeys. In Asym monkeys, LMI response varied among the animals; 2 showed an increase throughout the study period and 2 showed suppression at different time points. When compared with Asym monkeys, CD8+ T cells in Sym monkeys showed a trend of significant increase after day 180 postinoculation (PI). CD4+ T cells remained within the normal range till day 300 (PI), after which they showed a marginal increase. ConA-stimulated LT was suppressed in Asym monkeys from day 60 PI. Antigen-stimulated LT was unresponsive in both Asym and Sym animals. Thus, the host's LT response to ConA is suppressed in Asym animals, and alteration in CD8+ T-cell number and LMI response in Sym monkeys may be involved in the development of the acute disease manifestations in this model.
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[
J Assoc Physicians India,
1987]
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[
Magn Reson Imaging,
1999]
Metabolite mapping of human filarial parasite, Brugia malayi was carried out in vitro as well as in situ in host Mastomys coucha by 31P nuclear magnetic resonance (NMR) spectroscopy. Detection of parasites by visualizing contrast spots due to pathologic changes was observed by 1H magnetic resonance imaging (MRI). Major metabolites of adult B. malayi observed by 31P-NMR spectroscopy were of sugar phosphates (SP), phosphomonoesters (PME), glycerophosphoryl-ethanolamine (GPE), -choline (GPC), phosphoenolpyruvate (PEP), inorganic phosphate (Pi), nucleoside diphosphosugar and nucleotides-mono, -di and -tri phosphates. PEP and GPC were present in high concentration; PEP being the major energy reservoir and GPC the major phospholipid in this species of filaria. The 31P NMR spectra of testis of mastomys, showed seven major peaks of SP, PME, phosphocreatine (PCr), phosphodiesters (PDE), Pi, and nucleotides di- and tri-phosphates. The 31P-NMR spectra of testis of B. malayi infected animal also consisted of seven major peaks with significant decrease in the SP and PME peak showing changes in the carbohydrate and lipid metabolism of filaria infected testis. Thus, in vivo 31P MRS provided a non-invasive assessment of tissue bioenergetics and phospholipid metabolism.