Detection of filarial antigen and antibody in serum and hydrocele fluid of 100 patients of hydrocele.

AIM: The present study was carried out to detect an association between isolated non-communicable hydrocoele and filariasis and to provide awareness to positive patients regarding sequel and advising methods for the reduction of morbidity. METHODS: Blood samples and hydrocoele fluids were used to detect filarial antigen and antibody by ICT Kit, Trop-bio kit and Sevafilachek Kit. These were followed by statistical evaluation by chi2 test. RESULTS: 14% of cases were positive for filarial antigen and antibody in hydrocoele patient serum, while 15% of cases were positive for filarial antigen and antibody in the serum of non-hydrocoele patients. CONCLUSION: Probability is less than 0.05, which is statistically significant.

Circulating filarial antigen in the hydrocele fluid from individuals living in a bancroftian filariasis area - Recife, Brazil: detected by the monoclonal antibody Og4C3-assay.

The purpose of this study was to examine the circulating filarial antigen (CFA) detected by the monoclonal antibody (mAb) Og4C3-ELISA in paired samples of serum and hydrocele fluid from 104 men with hydrocele, living in an endemic area of Wuchereria bancrofti. Nocturnal blood specimens were filtered and examined for microfilariae (MF) and ultrasound was used in order to identify the presence of adult worms (the filaria dance sign - FDS) in the lymphatic vessels of the scrotal area. Four groups were selected according to their parasitological status: group I - 71 MF- and FDS-; group II - 21 MF+ and FDS+; group III - 10 MF- and FDS+ and group IV- 2 MF+ and FDS-. CFA was identified simultaneously (fluid and serum) in 11 (15.5%), 21 (100%), 3 (30%), and 1 (50%) in groups I, II, III, and IV, respectively. In despite of high CFA+ level (antigen Og4C3) units/ml, the Geometrical Mean (GM) = 2696) in the sera of these 36/104 paired samples, when compared to the hydrocele fluid, (GM = 1079), showed a very good correlation between the CFA level in the serum and CFA level in the fluid (r = 0.731). CFA level in the serum of the 23 microfilaremics (groups II and IV) was extremely high (GM = 4189) and was correlated with MF density (r = 0.442). These findings report for the first time the potential alternative use of the hydrocele fluid to investigate CFA using the mAb Og4C3-ELISA.

Characterization of antibody responses to Wolbachia surface protein in humans with lymphatic filariasis.

Symbiotic Wolbachia organisms of filarial nematodes have received much attention as possible chemotherapy targets and disease-causing organisms. In order to further investigate the association between anti-Wolbachia immune responses and chronic filarial disease in humans, antibody responses to Wolbachia surface protein (WSP) were assayed in serum samples collected from 232 individuals living in Leogane, Haiti, an area where Wuchereria bancrofti infection is endemic, and from 67 North Americans with no history of lymphatic filariasis. As opposed to antifilarial antibody responses, which were largely influenced by the patient's infection status, the prevalence and levels of anti-WSP immunoglobulin G (IgG) antibodies among individuals with lymphedema or hydrocele were significantly greater than those in gender- and infection-matched individuals without disease. In at least one case, the anti-WSP IgG response was coincident with the onset of lymphedema development, and among anti-WSP-positive women with lymphedema, anti-WSP IgG levels were negatively correlated with the duration of lymphedema. The presence of anti-WSP IgG was also associated with the severity of inguinal adenopathy among men with hydrocele. In addition to the presence of anti-WSP antibodies among Haitians, 15 of 67 (22%) serum samples collected from individuals from North America, where filariasis is not endemic, were also positive for anti-WSP antibodies. In comparison to those from Haitians, anti-WSP antibodies from North Americans primarily recognized a distinct region of WSP located within the highly conserved second transmembrane domain. The results of this study demonstrate that anti-WSP antibody responses are associated with the presence of chronic filarial morbidity and not filarial infection status in humans and suggest that WSP should be further studied as a potential trigger for the development of filarial disease.

A four year follow up study of filaria specific IgE response in individuals with hydrocele.

BACKGROUND & OBJECTIVES: Hydrocele is the most common clinical manifestation in males infected with Wuchereria bancrofti. IgE response to a filarial allergen Sd30 was evaluated in hydrocele patients living in a W. bancrofti endemic region of Orissa, India. METHODS: IgE levels to Sd30, an antigenic fraction isolated from Setaria digitata, were determined by ELISA in sera from patients of filariasis (n = 93). IgG and IgM levels were also determined. RESULTS: It was observed that microfilaraemic patients with hydrocele (n = 16) exhibited considerably reduced (P < 0.01) IgE levels in comparison to hydrocele patients without microfilariae (n = 27) and to other filarial groups (elephantiasis, asymptomatic microfilaraemics). The reduced IgE response in microfilariae positive patients with hydrocele persisted even after a gap of four years in the majority of patients. INTERPRETATION & CONCLUSIONS: The results show low IgE levels in microfilaraemic patients with hydrocele in comparison to other groups in filaria endemic regions. The exact mechanism of this reduction in IgE is not known but these patients make a distinct group in filaria endemic regions and should be considered separately for immunological evaluation.

Clinical, parasitologic, and immunologic observations of patients with hydrocele and elephantiasis in an area with endemic lymphatic filariasis.

Hydrocele and elephantiasis, major clinical manifestations of bancroftian filariasis, are thought to share a common pathogenesis. The characteristics of 121 patients with hydrocele or elephantiasis in Leogane, Haiti, were compared: 39% of 57 men with hydrocele and 3% of 64 persons with lymphedema of the leg were microfilaria-positive (P < .001). Circulating filarial antigen, presumably from the adult worm, was detected in 15 (43%) microfilaria-negative men with hydrocele and 9 (15%) microfilaria-negative persons with leg edema (P = .004). Microfilaria-positive men had lower levels of filaria-specific IgG1 and hydroceles of significantly smaller volume and shorter duration than did microfilaria-negative men; hydrocele volume was inversely associated with microfilarial density (P = .001). In contrast, filarial antigen but not microfilariae was associated with filaria-specific IgG4 and decreased lymphocyte proliferation. Antigen status was not associated with severity of leg edema. In this filariasis-endemic area, men with hydrocele are more immunologically and parasitologically heterogeneous than are persons with elephantiasis.

Bancroftian filariasis in Kwale District of Kenya. II. Humoral immune responses to filarial antigens in selected individuals from an endemic community.

Humoral immune responses to filarial infection were investigated in 162 individuals, covering the full clinical and parasitological spectrum of bancroftian filariasis as seen in an endemic community of Kwale District, Kenya. Sera were tested for specific antifilarial antibodies (total immunoglobulins, IgM, IgGl, IgG4 and IgE) using ELISA. Most individuals showed an immunological response to the filarial antigen (prepared from adult Brugia pahangi). How these responses were related to the clinical manifestations, parasitological status and age was analysed by comparing the mean antibody levels among different categories of endemic individuals, and by comparing these to the levels in control groups from filariasis-free areas. IgM and IgE anti-filarial antibodies were detected with low specificity in unabsorbed sera. A higher specificity, clearly distinguishing the mean antibody levels in the endemic categories from those of control groups, was obtained for total specific immunoglobulins, and for IgE in sera absorbed with protein A. The most specific results were obtained for IgGl and IgG4; clear inter-category differences were observed for these classes. The mean level of specific IgG4 was significantly higher in microfilaraemic groups than in amicrofilaraemic groups, whereas the mean level of specific IgGl was significantly higher in amicrofilaraemic, symptomatic cases than in microfilaraemic, symptomatic cases. In most categories of endemic individuals, and for most antibody isotypes, the mean levels of specific antibodies tended to be higher (although not significantly) in the younger individuals than in the older individuals. Overall, the differences in the filarial antibody responses were more closely related to the presence or absence of microfilariae and to the age of the individuals than to the disease manifestations in this endemic population.

Diagnostic use of filarial antibody and antigen isolated from hydrocele fluid for human filariasis.

Paired samples of serum and hydrocele fluid of filarial patients associated with hydrocele were analysed for filarial antibody and antigen. Sera samples showed higher titers of filarial antibody and antigen compared to their corresponding hydrocele fluid samples. HFIgG isolated from hydrocele fluid was equally useful as FSIgG isolated from serum and detected filarial antigen in 23 out of 26 microfilaraemic sera, 7 out of 10 chronic sera and 3 out of 18 endemic normal sera by inhibition ELISA. Filarial antigen was isolated from hydrocele fluid. In inhibition ELISA antigen fraction, HFA-9 (20-25 kDa) isolated by sodium-dodecyl sulphate-polyacrylamide gel electrophoresis showed high reciprocal antigen titer of 2048. This active antigen fraction was evaluated for its diagnostic utility in comparison with Wuchereria bancrofti microfilariae excretory-secretory antigen (Wb mf ES Ag) in inhibition ELISA. Both antigen preparations detected filarial antigen in about 80% of microfilaraemic sera, 60% chronic sera and 20-30% of endemic normal sera. This study showed that antibody and antigen isolated from hydrocele fluid were equally sensitive as FSIgG and Wb mf ES Ag in the detection of filarial antigen by inhibition ELISA. Thus hydrocele fluid may be used as an alternative source for the isolation of antibody and antigen of immunodiagnostic importance.

Fractionation, characterization and diagnostic potential of filarial antigens isolated from hydrocoele fluid in bancroftian filariasis.

Filarial antigen was isolated from hydrocoele fluid and fractionated on Ultrogel AcA 44. Six protein fractions (HFA C1 to HFA C6) were chromatographically separated from the column. Of the 4 antigenic fractions (HFA C1, HFA C2, HFA C3 and HFA C5), HFA C3 was a major reactive fraction with filarial serum immunoglobulin G. Analysis of HFA C3 by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) yielded 18 bands of Mr 18,000-160,000. Two fractions, HFA C3-7 and HFA C3-9 of Mr 48,000-55,000 and 32,000-39,000 obtained by gel elution, were antigenic in an enzyme-linked immunosorbent assay (ELISA). SDS-PAGE immunoblotting confirmed the ELISA by identifying 3 immunoreactive bands of Mr 51,000, 38,000 and 35,000. The diagnostic potential of HFA C3-7 and HFA C3-9 was compared in serodiagnosis of active infections and diseased states. HFA C3-9 showed greater sensitivity in detection of filarial specific IgM antibody in cases with microfilaraemia. Physicochemical characterization indicated the protein nature of HFA C3-9.

Immune reactions to Wuchereria bancrofti infections in Tanzania. I. Serum-mediated adherence of leucocytes to microfilariae in vitro, using serum from different groups of patients.

Studies on serum-mediated adherence of leucocytes to microfilariae of Wuchereria bancrofti in vitro were carried out in an area of Tanzania endemic for filarial infections. Patients were divided into age-matched groups according to parasitological and clinical conditions. No adherence was seen with serum from microfilariae-positive patients (with or without clinical disease). A slightly, but not statistically significantly, higher prevalence of adherence-positive sera was found among hydrocele patients (27%) and lymphangitis/elephantiasis patients (33%) than among endemic control patients (20%). These three positive groups exhibited a marked difference in age distribution of prevalence of positivity, with very high prevalences found among young hydrocele patients (48%) and old lymphangitis/elephantiasis patients (63%). The hydrocele fluid from hydrocele patients with adherence-positive serum also mediated adherence.

Immune reactions to Wuchereria bancrofti infections in Tanzania. II. Characterization of the serum-mediated adherence reaction of leucocytes to microfilariae in vitro.

Investigations to characterize the in vitro reaction of serum-mediated leucocyte-adherence to microfilariae of Wuchereria bancrofti were carried out in Tanzania. The adherence reaction took place within one hour and at least two serum factors were involved: a heat-labile factor, present also in normal serum (probably complement), and a more heat-stable factor, present in positive serum only (probably an antibody). Neutrophils and eosinophils were involved in the reaction proportionately to Their numbers in the cell solutions used; the reaction killed significantly more microfilariae, than did adherence-negative tests. Microfilariae could be used for at least one week after their isolation from the donor, but a significant decrease in the percentage of reactive microfilariae occurred within 16 hours of isolation. A high degree of species specificity was observed when positive sera were tested against other nematodes. The involvement of complement and the speed with which the reaction took place is in contrast to the results of similar studies from India, and may indicate a difference in the parasite in the two continents.

Malignant mesothelioma of the tunica vaginalis testis: report of first case with preoperative diagnosis.

The cytologic, histologic and ultrastructural features of a malignant carcinomatous mesothelioma of the tunica vaginalis testis in a 30-year old patient are described. This is the first such case with preoperative diagnosis by cytologic examination of hydrocele fluid and the second with documented history of exposure to asbestos. The identity of the tumor was confirmed by a positive immunoperoxidase reaction directed against mesothelial cells. A review of the previously described ten cases is presented.