Circulating fibrosis markers, eosinophil cationic protein and eosinophil protein X in patients with Wuchereria bancrofti infection: association with clinical status.

We measured the concentrations of several circulating fibrosis markers (type I collagen I, type III procollagen, hyaluronan) and eosinophil granule proteins (ECP and EPX) in lymphatic filariosis patients to investigate their relationship with clinical, parasitological and immunological data. This study was conducted in Polynesian patients with various stages of the disease (acute lymphangitis, chyluria, hydrocoele, elephantiasis), a closely related microbial lymphangitis and endemic controls. We observed modifications of the different markers in this pathology. Serum type I collagen and PIIINP were decreased. Serum hyaluronan, linked to perilymphatic granulomatous inflammation, was significantly increased in acute lymphangitis and elephantiasis patients. Serum ECP was also increased, at the limit of significance in our sample, in elephantiasis patients. These two last markers, already validated in another helminth disease, schistosomiasis, have potential interest in terms of follow-up of morbidity in these parasitic diseases.

Soluble cellular adhesion molecules, selectins, VEGF and endothelin-1 in patients with Wuchereria bancrofti infection and association with clinical status.

Lymphatic filariasis, a mosquito-transmitted disease commonly known as Bancroftian filariasis, is characterized by debilitating pathology linked to the progression of lymphoedema to a chronic state of elephantiasis. We performed longitudinal measurements of endothelial adhesion and angiogenic molecules in 63 Polynesian patients living in an hyperendemic focus of Wuchereria bancrofti. Decreased serum concentrations of soluble (s-) L selectin (CD62L) were noticed in sera of of patients with chronic conditions (hydrocele and elephantiasis). Chyluria was associated with increased vascular endothelial growth factor (VEGF) levels, whereas elephantiasis presented a high endothelin-1 (ET-1) profile. By contrast, increased serum concentrations of soluble intercellular (sICAM-1, CD54), but not of vascular cell (sVCAM-1, CD106), adhesion molecules were observed in sera of patients with bacterial lymphangitis used as controls. These trends are consistent with the increased permeability of vascular structures, a major clinical feature observed in acute lymphatic pathology (of bacterial or filarial origin), and of fundamental differences in the pathogenesis of hydrocele and elephantiasis. Using markers correlated with the clinical status (high ET-1 and VEGF levels for elephantiasis and chyluria, respectively; low CD62L levels for hydrocoele and elephantiasis) it should be possible to monitor disease progression in lymphatic filariasis.

Development and standardization of a rapid, PCR-based method for the detection of Wuchereria bancrofti in mosquitoes, for xenomonitoring the human prevalence of bancroftian filariasis.

PCR has recently been studied as a promising tool for monitoring the progress of efforts to eliminate lymphatic filariasis. PCR can be used to test concurrently at least 30 pools, with as many as 40 mosquitoes in each pool, for the presence of filarial larvae. The SspI PCR assay for the detection of Wuchereria bancrofti DNA in pools of mosquitoes has been used since 1994 in a variety of laboratories worldwide. During that time, the original assay has been modified in these different laboratories and no standardized assay currently exists. In an effort to standardize and improve the assay, a meeting was held on 15-16 November 2001, at Emory University in Atlanta, with representatives from most of the laboratories currently using the assay. The first round of testing was designed to test the four most promising methods for DNA extraction from pools of mosquitoes. Two of the four methods stood out as clearly the best and these will be now optimised and evaluated in two further rounds of testing.

The impact of 34 years of massive DEC chemotherapy on Wuchereria bancrofti infection and transmission: the Maupiti cohort.

Semi-annual mass DEC chemotherapy combined with vector control at the beginning of the programme, has been administered on the remote island of Maupiti (French Polynesia) since 1955 (except two periods in 1960-67 and 1970-74). The results of two surveys in 1985 and 1989, reporting 0% microfilaraemia, led to the hope that the eradication of lymphatic filariasis had been achieved. We combined parasitological criteria (microfilaraemia by membrane filtration), immunological (antigenaemia and serum levels of specific IgG antibodies) and molecular (PCR-based evaluation of infection in mosquitoes) techniques and found only good control of the parasite: We found residual microfilaraemia in 0.4% of the sample (mean level in carriers: 101.2 mf/ml), antigenaemia in 4.6% (mean level in positive persons: 714.4 units/ml) and specific IgG in 21.6% (including in one very young child). In addition, an infection rate of 1.4% was calculated in the Aedes polynesiensis vector population. These data, obtained in 1997 just before a hurricane, were partially confirmed in 1999 (0.1% of infection rate in the vector). Together with the possibility of some resistance to DEC, various epidemiological factors critical for the eradication of lymphatic filariasis are discussed.

Role of streptococcal infection in the acute pathology of lymphatic filariasis.

Growing evidence suggest that secondary bacterial, mainly streptococcal, infections contribute significantly to recurrent episodes of acute adenolymphangitis (ADL) of filarial origin. We examined the role of group A streptococci in the progression of lymphedema in Polynesian patients with filariasis-related ADL (22 cases) or chronic pathology (10 cases), or with erysipela (10 patients) and, as controls, in 20 healthy adults. Antistreptolysin O (ASLO) and anti-streptodornase B (ASDB) titers were systematically determined in parallel to parasitological and biochemical tests. ASLO and ASDB assays were positive in 100% of erysipela, 75% of filarial ADL as compared to 50% of chronic pathology and 39% of healthy controls. Interestingly, by opposition to ASLO titers which were not significantly different between the four groups, ASDB titers were higher in ADL (p = 0.019) and erysipela (p = 0.002) than in controls. These results support the hypothesis that recurrent streptococcal infections may have an important role in the pathogenesis of ADL in lymphatic filariasis.

Filarial antibody responses in Wuchereria bancrofti transmission area are related to parasitological but not clinical status.

In Wuchereria bancrofti transmission areas, three groups of individuals have been identified, according to the presence or absence of microfilariae or adult worm derived molecules in the blood compartment. These groups likely reflect individuals with different permissivity/resistance to the complete development of W. bancrofti cycle. The profile of filarial-specific immunoglobulins was analysed in W. bancrofti-exposed individuals in French Polynesia, according to the presence or absence of microfilariae (Mf) and adult worms, measured by Og4C3 circulating antigen. Individuals harbouring adult worms, have higher filarial-specific IgG4 but lower IgG3 and IgE levels, than adult worm-free individuals, independently of the presence of Mf. Low filarial-specific IgG1 and IgG2 levels were associated with the presence of Mf but independent of the presence/absence of adult worms. The filarial antibody responses were associated with the parasitological status of individuals but not with clinical symptoms such as hydroceles or limb lymphangitis or elephantiasis. The reduction of filarial-specific immunoglobulin levels was higher after treatment with diethylcarbamazine, than ivermectin, which likely reflects the better effect of the former on W. bancrofti adult worms. However, reduction of antibody levels was also observed in Mf-and adult worm-negative individuals. This could be due to the overall reduction of W. bancrofti transmission in the island where this study took place.

Assessment of immunochromatographic test for rapid lymphatic filariasis diagnosis.

Two rapid immunodiagnostic tests (ICT Filariasis test), developed for the quick diagnosis of Wuchereria bancrofti infection, have been validated in laboratory and field situation. The aim of this study was to assess the performance and usefulness of this antigen capture assay as a diagnostic method in three foci of lymphatic filariasis, located in the South Pacific (Society archipelago, French Polynesia), with different levels of endemicity. A sample of 1,595 patients was tested with this assay in parallel with a reference Og4C3 antigen capture assay and microfilariae detection. A second-generation ICT test, available for whole blood analysis, was also tested in parallel with the first generation test, developed for serum analysis, on a sample of 50 reference cases. The correspondence between the results obtained with the two rapid tests was excellent, without any influence of rheumatoid factors, but the sensitivity was in both cases slightly inferior to the one obtained with the ELISA reference test. This seems particularly true in epidemiological situation where a high proportion of amicrofilaraemic, adult worm carriers are observed.

The immunodominant Brugia malayi paramyosin as a marker of current infection with Wuchereria bancrofti adult worms.

The full-length cDNA sequence encoding Brugia malayi L3 paramyosin has been isolated by immunoscreening a cDNA library with a mouse antiserum raised against Wuchereria bancrofti L3 infective larvae. A recombinant truncated form of paramyosin was expressed as a glutathione S-transferase fusion protein and used to evaluate humoral responses of adults from a W. bancrofti-endemic area in French Polynesia according to their parasitological status. Immunoglobulin G4 (IgG4) preferentially bound to paramyosin in W. bancrofti-parasitized individuals, in contrast to unparasitized individuals, who harbored neither microfilaria nor Og4C3 adult worm circulating antigen. Reduction of the anti-paramyosin IgG4 titer following combined chemotherapy with diethylcarbamazine and ivermectin was significantly correlated with a reduction in the adult worm burden. This indicates that the presence of paramyosin-reactive IgG4 is associated with the presence of parasites and that reduction can be used as an immunological marker for W. bancrofti clearance.

Reduction of Wuchereria bancrofti adult worm circulating antigen after annual treatments of diethylcarbamazine combined with ivermectin in French Polynesia.

Circulating filarial antigen (CFA), determined with Og4C3 ELISA, is a marker of Wuchereria bancrofti adult worm infection. The reduction of CFA over 2 years was determined in 185 microfilaremic and 111 amicrofilaremic but CFA+ adults given an annual dose of either diethylcarbamazine (DEC) or ivermectin or the two combined. Reduction of CFA level was good with DEC but weak with ivermectin and followed the same pattern in amicrofilaremic and microfilaremic groups. Combinations and DEC alone had a similar impact on CFA level. CFA clearance was observed in amicrofilaremic but not in microfilaremic persons in all DEC-containing treatments. However, the highest clearance rate was observed in persons treated with DEC at 6 mg/kg combined with ivermectin. Continuous reduction of CFA level after repeated treatments shows that elimination of W. bancrofti infection, monitored by CFA clearance, might be achieved within a few years with annual treatments of DEC combined with ivermectin.

A universally applicable internal standard for PCR detection of Wuchereria bancrofti in biological samples.

A PCR-based assay have been previously described to detect Wuchereria bancrofti in mosquitoes and in human blood samples. However, the efficiency of PCR amplification may vary between samples depending on the presence of PCR inhibitors, leading sometimes to false negative results. To overcome this drawback, an internal standard plasmid (pWB11) was constructed. It can be added to each PCR reaction for coamplification along with the target W. bancrofti DNA (Sspl DNA repeat) using the same pair of primers. PCR products from W. bancrofti DNA or from pWB11 are 34 bp different in size and can be visualized either on agarose gel or by DNA ELISA using two different oligonucleotides probes.

A polymerase chain reaction assay for the detection of Wuchereria bancrofti in blood samples from French Polynesia.

A polymerase chain reaction (PCR) assay based on a highly repeated deoxyribonucleic acid (DNA) sequence found in Wuchereria bancrofti (the SspI repeat) has been developed to address the shortcomings of traditional diagnostic methods. In this field study in a W. bancrofti endemic region of French Polynesia, 373 human blood samples were collected and 100 microL of blood were screened by the SspI PCR assay and 1 microL by membrane filtration. The SspI PCR assay detected 99 of 113 blood samples in which microfilariae had been detected by filtration (sensitivity of 88%) with a specificity of 100%. All the samples missed by the SspI PCR assay had less than 8 microfilariae per mL of blood. To evaluate the efficacy of screening larger blood samples by PCR, both 100 microL and 500 microL samples from 50 patients with very low-level microfilaraemia were screened by the SspI PCR assay; the sensitivity increased from 60% to 84% when using the larger volume of blood. Finally, an enzyme-linked immunosorbent assay-based version of the SspI PCR assay was used to screen blood from 12 patients following treatment with diethylcarbamazine, ivermectin, or both. These results showed that the PCR assay closely paralleled the presence or absence of microfilariae in the blood and that no increase in the DNA level was seen immediately following drug treatment.

Wuchereria bancrofti filariasis in French Polynesia: age-specific patterns of microfilaremia, circulating antigen, and specific IgG and IgG4 responses according to transmission level.

The age-specific patterns of microfilaremia, Og4C3 antigenemia, anti-Brugia malayi IgG and IgG4 were assessed in 3 villages of low, medium and high transmission level for Wuchereria bancrofti filariasis. The prevalence rates for each of the 4 markers were clearly age dependent and their patterns strongly associated with the transmission level. The antigenemia prevalence rate was consistently higher than the microfilaremia prevalence rate, in all age groups. The prevalences of anti-B. malayi IgG and IgG4 responses were very similar and much higher than those of microfilaremia or antigenemia. Antibody responses reached the plateau at an earlier age and at a higher prevalence with increased intensity of transmission. For all the markers, the prevalence rates were significantly higher in males than in females.

Low positive predictive value of anti-Brugia malayi IgG and IgG4 serology for the diagnosis of Wuchereria bancrofti.

Enzyme-linked immunosorbent assays (ELISAs) for anti-Brugia malayi immunoglobulin (Ig) G and IgG4 were evaluated on sera from 1561 subjects in French Polynesia for the serodiagnosis of Wuchereria bancrofti filariasis, compared with the test for Onchocerca gibsoni circulating antigen (Og4C3) as a 'gold standard'. The sensitivity of the ELISA-IgG and ELISA-IgG4 assays was 90.8% and 94.5%, and the specificity was 45.9% and 50.7%. The positive predictive values were 41% and 45% respectively for an antigen prevalence rate of 30%. Thus antibody prevalences exceeded by two-fold the antigen prevalence, which itself exceeded by two-fold the prevalence of microfilaraemia.

Og4C3 circulating antigen, anti-Brugia malayi IgG and IgG4 titers in Wuchereria bancrofti infected patients, according to their parasitological status.

This study involved 221 microfilaremic (Mf+), 302 amicrofilaremic (Mf-) antigen positive (AG+) and 1454 Mf-antigen negative (AG-) individuals living in endemic villages. Whatever the group considered, antigen and antibody titers were widely distributed. Og4C3 antigen, detected both in Mf- and Mf+ patients, was significantly higher in Mf+ patients. The Mf parasitological status did not significantly influence the antifilarial antibodies levels in the infected AG+ individuals, although IgG4 was more discriminant. In the supposedly uninfected individuals (Mf-AG-), anti-filarial IgG and IgG4 could be detected in a large proportion of the group. Og4C3 circulating antigen test was confirmed to be a good marker of active Wuchereria bancrofti infection.

Og4C3 circulating antigen: a marker of infection and adult worm burden in Wuchereria bancrofti filariasis.

Og4C3 circulating filarial antigen was detected in the sera of 94.5% (259/274) of microfilaremic patients, 32% (239/751) of persons with presumption of filariasis, and 23% (11/48) of chronic filariasis patients. The antigen level was correlated with the microfilariae (Mf) density and patient age (P < .01). It remained stable in patients treated with microfilaricidal drugs. Og4C3 antigen, undetectable in Mf culture media, was demonstrated to be a rare somatic Mf antigen. It appears to be an excreted or secreted antigen from adult filaria. It could be used as a marker of infection and an indicator of adult worm burden.

Low predictive value of PGL-I serology for the early diagnosis of leprosy in family contacts: results of a 10-year prospective field study in French Polynesia.

In 1983, a cohort study to follow up the family contacts of leprosy cases was implemented in French Polynesia to assess the usefulness and applicability of phenolic glycolipid-I (PGL-I) serology in a leprosy control program. A total of 1201 contacts (666 females, 535 males) have been included in the study. The IgM anti-PGL-I seroprevalence determined on the initial sera was 17%. It was significantly higher among females than males (20% vs 15%, p = 0.02). From 1983 to 1992, 4 out of 204 (2%) anti-PGL-I seropositive contacts developed the disease (1 indeterminate, 1 BT, 1 BL, 1 LL) compared with 10 out of 997 (1%) seronegative contacts (4 indeterminate, 3 BT, 1 BB, 2 TT). Of these 10 patients, only 3 (2 indeterminate, 1 BT) converted to seropositivity when leprosy was diagnosed. The risk of developing leprosy was not significantly higher among seropositive than among seronegative groups (2% vs 1%, p = 0.2). A PGL-I circulating antigen test performed on 216 selected sera at entry into the trial showed a higher antigen prevalence when the antibody level was higher. PGL-I antigen was detectable in 5 of 12 patients tested prior to diagnosis (1 LL, 1 BL, 3 indeterminate). The median time to externalize the disease was not significantly different among antibody-positive and -negative contacts (17 vs 25 months, p = 0.3). The relative risk of developing leprosy for contact individuals was 30.8 times that of noncontacts, and 15% of the total new cases detected between 1983 and 1992 emerged from the study population.(ABSTRACT TRUNCATED AT 250 WORDS)