Long-term follow-up of treatment with diethylcarbamazine on anti-filarial IgG4: dosage, compliance, and differential patterns in adults and children.

We have followed a population in an area endemic for Brugia malayi for three years after intensive treatment with diethylcarbamazine (DEC). Microfilariae were cleared from the circulation within four months in all eligible study participants (n = 60). There appeared to be a strong correlation between the maximum reduction in specific IgG4 and the number of days drug was taken under supervision (p = 0.41, P < 0.001), indicating that high total dosage of DEC is necessary for optimal reduction of active infection. In individuals with good compliance (at least 180 mg/kg of body weight, n = 34), we observed variable IgG4 patterns. All pre-treatment IgG4+ children (9-14 years old) and 40% of the IgG4+ adult population (> or = 15 years old) showed a gradual decrease in anti-filarial IgG4; 53% of these showed complete clearance of worm burden by the end of the study. In contrast, another group of male IgG4+ adults showed IgG4 patterns that started to increase between nine months and two years after treatment, indicating either a partial efficacy of DEC that allowed recovery of resident adult worms or reinfection.

Anti-filarial and total IgG4 and IgE antibody levels are correlated in mothers and their offspring.

In mothers who suffer from helminth infections or allergic diseases, prenatal sensitization with antigens/allergens is suspected to bias the immune system of the offspring towards a Th2-type response. To investigate this at the antibody level, we collected 113 blood samples on filter paper from a paediatric population aged 3 months to 10 years and their mothers, who resided in an area endemic for brugian filariasis in Indonesia. The results showed that antibody levels in children were strongly correlated with maternal antibody levels. However, for anti-filarial IgG4 and IgE this relationship was manifested directly after birth, whereas for total antibody levels a positive correlation could be detected only with children aged > or = 2 years. To investigate the influence of paternal antibody on progeny, specific IgG4 was determined in a different set of samples from 229 children and both of their parents. Interestingly, the influence of paternal IgG4 became apparent only after the age of 4 years. In contrast, maternal antibody levels were already correlated to levels produced by their offspring at a young age (3 months onwards). Taken together, it appears that children can become sensitized to parasite antigens in utero, allowing them to produce Th2-dependent specific IgG4 and IgE antibodies at a young age, whereas with increasing age, the influence of environmental factors, shared in households, such as filarial transmission and other helminth infections, becomes dominant.

Regulation of anti-filarial IgE by infection pressure.

In lymphatic filariasis, specific IgG4 responses to the parasite and their relationship with infection have been studied extensively, but only a few studies have concentrated on anti-filarial and total IgE. Here we have investigated the role of filarial infection pressure on production of IgE by considering length of exposure (age), filarial endemicity and parasitological status. Antibody levels were determined in 366 individuals, who were resident in 3 villages in South-Sulawesi, Indonesia, with varying degrees of filarial transmission intensity, as indicated by the prevalence of Brugia malayi microfilaraemia (0.7%, 9% and 32%, respectively). Anti-filarial IgE levels were significantly lower in the low transmission village than in the areas with intermediate and high filarial transmission; however, in the latter village a remarkable suppression of specific IgE was found. Microfilaria-positive individuals showed elevated levels of total IgE, but suppression of specific IgE, which has been reported before. Taken together, these observations suggest that 2 opposing mechanisms regulate anti-parasite IgE expression: increasing experience of filarial infection stimulates specific IgE, but antibody levels become specifically suppressed when microfilariae or adult worms develop. Using a simple mathematical model, we illustrate how anti-filarial IgE increases with parasite antigen up to a threshold level, but levels off and becomes down-regulated after the threshold is exceeded.

Adults acquire filarial infection more rapidly than children: a study in Indonesian transmigrants.

To dissociate the influence of host age from length of exposure on acquisition of filarial infection, we examined the development of microfilaraemia and anti-filarial IgG4 in all ages of a naive population that became suddenly exposed to Brugia malayi as a result of transmigration. Responses in 247 transmigrants, who had settled for periods of several months up to 6 years in their new homesteads, were compared with those of 133 life-long residents. As shown in earlier studies, anti-filarial IgG4 increased with age in the indigenous populations, whose age is equivalent to length of exposure. However, by examining transmigrants, it became clear that development of specific IgG4 was influenced by age, since levels of this antibody were consistently higher in transmigrant adults than in transmigrant children, despite an equal length of exposure to filarial infection. Examining microfilaraemia, it was confirmed that infection establishes more rapidly in adults than in children.

The development of specific IgG4 and IgE in a paediatric population is influenced by filarial endemicity and gender.

We set out to study how anti-filarial IgG4 and IgE, which have been studied extensively in adult populations, are influenced by gender and by the degree of filarial endemicity during childhood. Development of specific IgG4 and IgE was examined in 502 children aged 3 months to 12 years, who were resident in 3 villages in South-Sulawesi with microfilaria prevalences of 6, 23 and 42 %. Specific IgG4 and IgE could be detected as early as 18 months after birth, in low amounts, and increased to levels comparable to those produced by adults at the age of 3 years. A higher prevalence of anti-filarial IgG4 in boys, indicating higher filarial infection compared to girls, became apparent after the age of 7. The specific IgG4 response was strongly influenced by the degree of filarial endemicity and production of this antibody was considerably delayed in the low transmission village. With respect to IgE, it was noted that specific IgE was consistently higher in boys from infancy onwards indicating a predisposition for high IgE production in males. The influence of filarial endemicity was less profound on IgE thaon on IgG4. In conclusion, reactivity to filarial antigens begins early in life and is differentially influenced by gender and transmission intensity.

Strong association of interleukin-6 and lipopolysaccharide-binding protein with severity of adverse reactions after diethylcarbamazine treatment of microfilaremic patients.

To assess the involvement of inflammatory mediators in the development of adverse reactions in filarial patients undergoing treatment, 29 microfilaremic subjects were treated with diethylcarbamazine (DEC). Before and at serial time points after initiation of treatment, plasma levels of inflammatory mediators and DEC were measured, and adverse reactions were recorded. Patients experienced no or mild, moderate, or severe adverse reactions. Increasing pretreatment microfilarial counts were associated with escalating severity of adverse reactions. Plasma concentrations of DEC were not different among patients suffering from varying degrees of illness. Interleukin (IL)-6, IL-10, lipopolysaccharide-binding protein (LBP), and soluble tumor necrosis factor receptors (sTNF-Rs) increased after treatment. IL-6 and LBP, however, showed the strongest association with adverse reactions. Increasing levels of these molecules were closely correlated with the mounting severity of adverse reactions, which raises the possibility that they play an important role in systemic inflammation that arises after DEC treatment of filarial patients.

Clustering of Brugia malayi infection in a community in South-Sulawesi, Indonesia.

The present study was undertaken in village Karondang in South-Sulawesi, Indonesia, to investigate the influences of genetic, household and environmental factors on Brugia malayi infection. Infection status was determined by measuring both microfilariae in night blood and anti-filarial IgG4, as a marker for detection of active filarial infection. A total of 171 residents participated in the study; familial relationships between subjects were registered to construct pedigrees and distances between households were measured. The data were analysed using a test statistic for familial aggregation. For distribution of microfilariae over the study population a genetic influence on infection susceptibility was favoured over the household and environmental effects. For anti-filarial IgG4, all 3 clustering models gave significant results, suggesting that genetic, household and/or environmental factors influence specific IgG4 antibodies.

Adverse reactions following diethylcarbamazine (DEC) intake in 'endemic normals', microfilaraemics and elephantiasis patients.

This paper reports on adverse reactions following a 12-day course of 6 mg/kg diethylcarbamazine (DEC) therapy in brugian filariasis patients in Indonesia. Microfilaria-positive individuals (n = 26), 'endemic normals' (n = 12) and elephantiasis patients (n = 17) were included in the study. Fever, headache and body aches started between 2 and 24 h after DEC intake. Adverse reactions were categorized into 'no or mild', 'moderate' or 'severe' depending on the total reaction score. Four microfilaraemic individuals (15.4%) suffered from severe adverse reactions and their pre-treatment microfilarial levels (geometric mean, GM = 3060 mf/10 mL) were significantly higher than in the 5 microfilaraemic individuals (19.2%) suffering from moderate reactions (GM = 1268 mf/10 mL) and in the 17 microfilaraemic patients (65.4%) who experienced no or mild reactions (GM = 6 mf/10 mL)(P < 0.001 and P < 0.001, respectively). Endemic normals showed no or mild adverse reactions. No or mild adverse reactions were also recorded in all but 2 elephantiasis patients after DEC intake. Two elephantiasis patients with moderate reactions had high levels of circulating microfilariae at pre-treatment (2097 and 7375 mf/10 mL). Concentrations of DEC were measured in plasma, but could not explain the differences in the severity of adverse reactions.

Anti-filarial IgG4 in men and women living in Brugia malayi-endemic areas.

To assess whether antifilarial IgG4 can be used to study various epidemiological facets of filarial infections, we studied this isotype in 238 individuals resident in areas endemic for brugian filariasis, focusing on the differences between men and women. In the study area, the prevalence of microfilariae was 6.7% and the prevalence of antifilarial IgG4 was 49.2%. All microfilariae carriers were positive for antifilarial IgG4, whereas a proportion of the endemic normals (94/208) and clephantiasis patients (7/14) had IgG4 antibodies to filarial antigens. Data were analysed as a function of gender in distinct clinical groups and stratified for age. The prevalence of microfilariae was higher in males in all age groups, as reflected in significantly higher antifilarial IgG4 antibody levels compared to females. The prevalence of IgG4 increased to reach a plateau at the age of 30 years in both males and females. These results indicate that antifilarial IgG4 antibodies can reflect the differences in the extent of infection in males and females as measured by microfilarial counts, and that this parameter can be used for epidemiological assessments of filarial infection.

Inflammatory cytokines following diethylcarbamazine (DEC) treatment of different clinical groups in lymphatic filariasis.

In an earlier study in Indonesia we reported on adverse reactions to diethylcarbamazine (DEC) in brugian filariasis patients identified as microfilaraemics (n = 26), endemic normals (n = 11) and elephantiasis patients (n = 17). To assess the link between adverse reactions and cytokines we have now analysed an array of inflammatory mediators in plasma samples collected during the same study. Pre-treatment levels of interleukin (IL)-6 and soluble tumour necrosis factor receptor 75 (sTNF-R75) were higher in elephantiasis patients compared to microfilaraemics and endemic normals, indicating the presence of an ongoing inflammation in patients with chronic disease. After initiation of treatment, the levels of IL-6 and LPS-binding protein (LBP) were consistently and significantly higher in microfilaraemics who suffered most from adverse reactions compared with endemic normals and elephantiasis patients. In microfilaraemics the levels of sTNF-R75 increased after treatment to reach levels recorded in elephantiasis patients. IL-6 increased early, concurrent with the development of adverse reactions and peaked by 24 h post treatment. The levels of LBP and sTNF-R75 in microfilaraemics also increased to peak, later than IL-6, at 32 h post DEC therapy. Although changes were recorded in IL-8 and IL-10 levels in some individuals, no significant differences were found between the 3 clinical groups. These results demonstrate that intake of DEC leads to an increase in a selected number of inflammatory mediators in the group of filarial patients who suffer most from adverse systemic reactions.

A filter paper technique for the detection of anti-filarial IgG4 in lymphatic filariasis.

In a previous study performed in south Sulawesi (Sulawesi Selatan), Indonesia, we established that the immunoglobulin G4 (IgG4) enzyme-linked immunosorbent assay (ELISA) is a suitable community diagnostic method and that it can distinguish areas of high and low prevalences within short distances. In an attempt to make this diagnostic tool more applicable in the field, a comparative study using serum and blood collected on filter paper was undertaken with 568 individuals living in 2 areas with different endemicity for brugian filariasis in south Sulawesi. In Mamuju district, where the microfilaria (mf) prevalence of the studied individuals was 18.4%, antifilarial IgG4 was present in 73.1% of the venepuncture samples and 72.5% of the filter paper samples, respectively. In Mangkutane district, where lymphatic filariasis is transmitted at a low level (mf rate 2.4%), antifilarial IgG4 was detected in 35.5% and 39.9% of similar samples, respectively. There was no significant difference in the IgG4 detection rate determined from venepuncture and filter paper samples from the same donors (P = 0.124), and the IgG4 values were highly correlated (p = 0.97, P < 0.001, n = 568). These results indicate that the filter paper technique for collection of blood samples is a suitable alternative to venepuncture for use in the IgG4 ELISA.