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Biomedical subjects

M L Eberhard

Publications and source records attributed to M L Eberhard.

At least 91 records · Page 5Linked to original sources

Guatemalan human onchocerciasis. II. Evidence for IgG3 involvement in acquired immunity to Onchocerca volvulus and identification of possible immune-associated antigens.

Ag-specific isotypic differences in immune response to Onchocerca volvulus Ag were assessed for 778 long term residents of endemic Guatemalan areas by quantitative ELISA with 5-min incubation steps and immunoblot. The study population was separated into five groups based on clinical status: N+F+, N+F-, N-F+, N-F-H+, and N-F-H-, where N = O. volvulus adults (nodule), F = microfiladermia, and H = history of O. volvulus infection. A subset of 44 individuals with high exposure to onchocerciasis from the N-F-H- group were critically evaluated and designated as "putatively immune." IgG1 reactivity to O. volvulus Ag was elevated in the majority of infected persons, but not in putatively immune individuals. Specific IgG3 levels, however, were equally elevated in all groups. The majority of N+F- persons also had elevated IgG1 levels, but they were lower than those found in F+ persons. IgG3 reactivities to a group of antigens at 20 kDa (GP20) were seen in many uninfected persons and some N+F- persons. In contrast, most F+ persons, react to this Ag with IgG1 and not IgG3. A mangabey inoculated with the infectious larval stage of O. volvulus (L3), but showed no signs of infection, began to recognize GP20 at 2 wk postinoculation. Early recognition of GP20 was possibly elicited by the larval stage. Purified nodule Ag from N+F+ individuals contained GP20, however, identical nodule Ag prepared from N+F- individuals did not. These data suggest that GP20 Ag may be common to both uterine microfilaria and the infectious larval stages. The fact that GP20 is predominantly recognized by IgG3 in putatively immune persons and some N+F- persons suggests that this increased IgG3 activity may be important in acquired immunity to onchocerciasis.

Adolescent↗

Maternal filarial infection as risk factor for infection in children.

Familial clustering of filarial infection was investigated through random house-to-house surveys of 643 individuals in Leogane, Haiti, an area with endemic Bancroftian filariasis. Children of infected mothers were 2.4 to 2.9 times more likely to be infected than were those of amicrofilaraemic mothers. Filarial-specific cellular responsiveness in amicrofilaraemic children born to infected mothers was lower than that in amicrofilaraemic children born to amicrofilaraemic mothers. No effect of paternal infection status was seen. The findings show that maternal infection is a risk factor for filarial infection in children and is associated with altered parasite-specific immune reactivity.

Animals↗

Sodium azide: ineffective as a faecal preservative for parasitological diagnosis.

Sodium azide was compared with 10% formalin to evaluate sodium azide's effectiveness as a faecal preservative for intestinal helminths and protozoa. Faecal specimens collected from Haiti were preserved in sodium azide and in 10% formalin and analysed after 1.5, 6.5 and 11.5 weeks by examining direct wet-mount preparations. Sodium azide did not preserve the morphology of either helminths or protozoa as well as 10% formalin did. However, sodium azide prevented embryogenesis of helminth eggs, while some helminth eggs in 10% formalin contained living larvae. Biosafety guidelines regarding the toxicity, reactivity, and disposal of sodium azide were strictly followed. Use of 10% formalin is a significantly better choice than sodium azide for preserving parasites when accurate identification of parasites and biosafety are the main concerns.

Adolescent↗

Evidence of nonsusceptibility to diethylcarbamazine in Wuchereria bancrofti.

This study assessed Wuchereria bancrofti-infected patients who received diethylcarbamazine (DEC) to determined if drug levels were comparable between individuals who continued to harbor circulating microfilariae and those whose blood became clear of parasites, between those with high and low microfilaria counts, and between infected and noninfected persons. Haitian volunteers undergoing treatment for W. bancrofti and two nonendemic controls were enrolled. DEC levels in serum and urine samples were determined using a gas chromatographic method. No correlation was found between pre- and posttreatment microfilarial levels and drug levels. Drug levels were comparable in persons with or without residual microfilaria and in infected and uninfected persons. These results indicate that incomplete drug regimens, differences in serum drug levels, and inadequate drug dosage are not the primary causes of treatment failure and suggest there is a degree of parasite tolerance for DEC.

Animals↗

Antifilarial cellular responses detected in a Haitian pediatric population by use of a microblastogenesis assay.

Previous reports have demonstrated age-related shifts in antifilarial humoral immune responses in 6- to 10-year-old Haitian children; the responses consisted of elevated parasite-specific IgG2 and IgG3 in amicrofilaremic children and elevated IgG4 in microfilaremic children. In this study, the cell-mediated immune responses to soluble adult and microfilarial extracts of Brugia pahangi, determined by use of a microblastogenesis assay, were examined. Capillary blood samples were collected by finger prick from 176 Haitian children in an area with endemic Wuchereria bancrofti. Antigen-specific cellular responsiveness varied as a function of infection status but not age or sex; amicrofilaremic children had significantly greater responses to adult antigens than did microfilaremic children. Significant responses were detected in children less than 2 years of age; thus, correlations observed between filarial antigen-specific responses and infection status are established early in life.

Adolescent↗

Changes in circulating parasite antigen levels after treatment of bancroftian filariasis with diethylcarbamazine and ivermectin.

This study assessed changes in circulating parasite antigen levels after diethylcarbamazine (DEC) and ivermectin treatment of bancroftian filariasis to determine effects of these drugs on adult Wuchereria bancrofti in vivo. Thirty adult Haitians with microfilaremia were treated with 1 mg of ivermectin to reduce counts of microfilariae. Later, subjects were treated with either one or two 200 micrograms/kg doses of ivermectin or with 12 daily 6 mg/kg doses of DEC. Macrofilaricidal activity of these drugs was indirectly monitored by measuring circulating W. bancrofti antigen by EIA. Antigen levels fell by 75% after DEC and by 34% after ivermectin. These results suggest that low-dose ivermectin treatment followed by a standard course of DEC is a more effective macrofilaricidal regimen for W. bancrofti than either of the multidose ivermectin regimens used in this study.

Animals↗

Experimental Onchocerca volvulus infections in mangabey monkeys (Cercocebus atys) compared to infections in humans and chimpanzees (Pan troglodytes).

Three chimpanzees, three mangabey monkeys (Cercocebus atys), and 14 patas monkeys (Erythrocebus patas) were inoculated with L3 Onchocerca volvulus of Guatemalan origin. One chimpanzee and two mangabey monkeys developed antibody activity to at least three different antigens. Both mangabey monkeys recognized a 20 kDa antigen 3.5-5 months post-inoculation, and the monkeys and the chimpanzee developed antibody activity to 14 and 22 kDa antigens 7.5-13 months post-inoculation. One mangabey monkey and the chimpanzee became microfilaria-positive in skin snips at 16 and 21 months post-inoculation, respectively. Antibody activity to the 20 kDa antigen in the mangabey monkeys is noteworthy because of the prominence of this antigen among putatively immune persons living in onchocerciasis-endemic areas. The two mangabey monkeys responded parasitologically in a manner comparable to immune humans. No microfilariae were detected in one monkey and only scant numbers of microfilariae were observed in the second. The mangabey monkey may be a good animal model for the study of onchocerciasis.

Animals↗

Analysis of isotype-specific antifilarial antibody levels in a Haitian pediatric population.

Previous studies of antifilarial antibodies in a pediatric population residing in an area with endemic Wuchereria bancrofti filariasis have demonstrated age related shifts in antifilarial immunity. To further characterize humoral responses in Haitian children, serum samples from 129 patients (3 months-15 years of age) were analyzed by ELISA for isotype-specific antifilarial antibody responses. Age-stratified analysis of geometric mean antibody titers showed significant increases in antibody titers of all isotypes with age in the amicrofilaremic population. Antifilarial IgG1, 2, and 3 levels were higher in amicrofilaremic children than in microfilaremic children, significantly so for IgG2 and IgG3. In contrast, IgG4 antibody levels were higher in microfilaremic subjects than in amicrofilaremic subjects. A multivariate, unconditional, logistic regression model was developed from these data to predict infection status. The model correctly classified 91.6% of the amicrofilaremic subjects, but only 55.6% of the microfilaremic subjects.

Adolescent↗

Comparison of high dose ivermectin and diethylcarbamazine for activity against bancroftian filariasis in Haiti.

This three-phase study was designed to compare high dose ivermectin with a standard diethylcarbamazine (DEC) regimen for patient tolerability, potential to kill adult filaria, and duration of microfilarial suppression in 30 Haitian subjects with Wuchereria bancrofti microfilaremia. All were first given a 1-mg oral dose of ivermectin (phase 1) to reduce microfilaria densities. Participants were randomized into three groups: Group 1 received DEC (6mg/kg per day for 12 days), Group 2 received 200 mcg/kg of ivermectin, and Group 3 received 400 mcg/kg of ivermectin (200 mcg/kg per day for 2 days). All drug regimens were well tolerated with few adverse reactions. Most reactions occurred during phase I and consisted primarily of headache, fever, and myalgia. At the end of phase 1, 27 of 30 (90%) patients were microfilaria negative. During phase 2, four of the six men receiving DEC developed scrotal reactions suggesting killing adult worms; no such reactions were noted in 10 men receiving ivermectin (p less than 0.05). At one-year follow up (phase 3), all treatment groups had less than 10% return to pretreatment microfilaria levels. The mean percent of baseline microfilaria counts were for Group 1, 0.9% (range 0-5%); Group 2, 8.2% (range 0-31%); and Group 3, 3.8% (range 0-25%). Seven individuals in Group 1 were microfilaria-negative, while only one and three individuals were microfilaria-negative in Groups 2 and 3, respectively. These results suggest that DEC causes more damage to the adult worms and greater reduction in microfilaria densities than ivermectin, but that high doses of ivermectin may suppress microfilaremia in lymphatic filariasis for periods much longer than previously reported.

Adolescent↗

Bancroftian filariasis: long-term effects of treatment with diethylcarbamazine in a Haitian population.

Two groups of Wuchereria bancrofti-infected Haitians who had undergone treatment with diethylcarbamazine (DEC) were followed for up to five years after treatment to document the long-term effects of treatment on adult worms and microfilariae and on the recurrence of infection. One group of 69 persons who had received 12 daily treatments had a significant decrease in microfilaria levels until year 4, when a small number of individuals experienced a resurgence of this parasite stage in the peripheral blood. In a second group of 57 persons who had been treated weekly for 12 consecutive weeks, there was a greater reduction in the microfilaria levels following treatment, and for the full four years of followup, these levels remained more depressed than those of the group that received daily treatment. Our results indicate that DEC kills or permanently sterilizes adult W. bancrofti. Furthermore, these results demonstrate conclusively that in Haiti, the use of DEC provides long-term benefits to treated persons, even though they continue to reside in an area with endemic filariasis.

Animals↗

Laboratory diagnosis of filariasis.

In this article, the authors describe how to detect blood-borne and skin-dwelling filarial infections parasitologically. Considerable space is devoted to identifying and separating microfilarial from nonparasite objects, and the evaluation of morphologic features, including size, shape, and internal anatomy is stressed.

Animals↗

Inoculation of ferrets with ten third-stage larvae of Dracunculus insignis.

Infection with Dracunculus insignis was established in 7 of 10 ferrets experimentally inoculated intraperitoneally with 10 third-stage larvae (L3's). Worm recovery and infection rate were comparable to animals inoculated with 50 L3's. This study demonstrates that once infective larvae traverse the gut and pass into the peritoneal cavity, very few larvae are required to establish infection.

Animals↗

A Brugia species infecting rabbits in the northeastern United States.

Brugia sp. microfilariae were observed in more than 60% of wild rabbits collected on Nantucket Island, Massachusetts. The microfilariae measured 294-344 microns in length and had the characteristic subterminal and terminal nuclei observed in other Brugia microfilariae. The microfilaria is similar to those described for Brugia leporis in rabbits in Louisiana. This may be the Brugia species responsible for 21 documented cases of human infection in the northeastern United States.

Animals↗

Chemoprophylactic drug trials for treatment of dracunculiasis using the Dracunculus insignis-ferret model.

Groups of ferrets inoculated with Dracunculus insignis were treated with various anthelminthic compounds to evaluate the potential use of drugs in controlling the human parasite D. medinensis. The three primary compounds tested were diethylcarbamazine (DEC), albendazole (ALBZ), and invermectin (IVER); they were administered in dosages of 60 mg/kg, 10 mg/kg, and 1 mg/kg, respectively. Groups of animals received treatment either once at 60 days after inoculation, once at 60 and 90 days, or for 3 days at either 60 or 90 days postinoculation. The most marked decrease occurred in the animals that received treatment once at 60 and 90 days after inoculation. This was observed for all three drugs tested. Increased dosages, i.e., 3 days of treatment at 60 or 90 days did not result in decreased worm burdens. In no group was there a statistically significant reduction in worm burden when compared with controls. Two other compounds, metrifonate and CGP 6140, were tested in a more limited manner, but again the worm recovery rates were comparable with those in control groups. It would appear that existing drugs commonly used to treat helminthic infections are poor candidates for use in the campaign to eradicate guinea worm disease.

Albendazole↗

Effects of albendazole on Echinococcus multilocularis infection in the Mongolian jird.

Albendazole chemotherapy of larval Echinococcus multilocularis was studied in the Mongolian jird by administration of medicated feed at various concentrations and durations. The effects were evaluated by comparison of treated and control groups in terms of host mortality, larval metastases to the lungs, and final weight and histologic appearance of larval tissue. Viability of larval tissue at necropsy of each animal was tested by inoculation into two noninfected jirds. Albendazole-medicated feed (0.05%-0.10%) significantly inhibited larval growth. Other effects of the drug included larval degeneration and necrosis, inhibition of protoscolex formation, decreased pulmonary metastases, and reduced mortality of hosts. Adverse effects on the parasite correlated significantly with serum albendazole metabolite levels and duration of therapy. However, serum albendazole levels in jirds equal to or exceeding concentrations achieved in humans receiving daily doses of 10 mg/kg of body weight did not kill the parasite.

Albendazole↗

A technique for microfilarial detection in preserved blood using nuclepore filters.

Nuclepore filtration is now the most widely used method of detecting microfilaremia, particularly if microfilariae are few in number. However, this system requires the blood sample to be processed promptly after collection. Using blood from Wuchereria bancrofti-infected patients, 3 solutions were tested to determine whether blood could be held for delayed processing. Of these, we identified 1, 2% formalin/10% Teepol, in which microfilaremic blood can be held for at least 9 mo without deterioration of microfilarial structure or decrease in microfilarial numbers. In addition, this mixture passed easily through a 5-microns filter at all times tested. Examination of more than 300 blood samples held in 2% formalin/10% Teepol showed that this solution can utilize the convenience and sensitivity of membrane filtration while eliminating the need to perform testing immediately after the blood is collected.

Animals↗