IgG subclass responses to proinflammatory fraction of Brugia malayi in human filariasis.

Background & objectives: Earlier we demonstrated that immunization with F6, a proinflammatory molecular fraction isolated from the human filarial parasite Brugia malayi, protected the host and eliminated the infection in Mastomys coucha by a Th1/Th2 response including IgG2a antibody response. Whether F6 molecules become accessible to human host during natural course of infection and elicit similar response is not known. The present study was undertaken to determine the profile of IgG subclasses specifically reactive to F6 in different categories of bancroftian filariasis cases to infer any relationship between the levels of a particular F6-specific IgG subclass and the infection or disease status. Methods: Serum samples of normal individuals from filariasis non-endemic regions of India like Jammu & Kashmir, Uttarakhand, and Chandigarh [(NEN-W; n=10), healthy subjects from USA (NEN-U; n=10) and three categories of bancroftian filariasis cases from endemic areas: endemic normals (EN; n=10) with no symptoms and no microfilariae, asymptomatic microfilaremics (ASM; n=10) and chronic symptomatic amicrofilaremics (CL; n=10) were assayed for F6-specific IgG1, IgG2, IgG3 and IgG4 by ELISA using SDS-PAGE-isolated F6 fraction of B. malayi adult worms. Results: Significantly high levels of F6-specific IgG1, IgG2 and IgG3 were found in CL (P<0.001) and EN (P<0.01-0.001) bancroftian filariasis cases compared to NEN-U. Significant levels of F6-specific IgG1 (P<0.01) and IgG2 (P<0.01) but not IgG3 were found in ASM cases compared to NEN-U. The most abundant was IgG2 which when compared to NEN-U, was significantly high in CL (P<0.001) and EN cases (P<0.001), followed by ASM (P<0.01). F6-specific IgG4 response in EN, ASM and CL subjects was not significantly different from the levels of NEN-U. Among the non-endemic normals, the NEN-W subjects showed significant reactivity with IgG2 (P<0.001) but not with IgG1, IgG3 and IgG4 as compared to NEN-U subjects. IgG subclass levels were different in different categories. Interpretation & conclusions: The high levels of F6 reactive IgG1, IgG2 and IgG3 in endemic normals and chronic symptomatic bancroftian patients, and IgG1 and IgG2 in asymptomatic microfilaraemics, suggest that F6 molecules of parasite are accessible in these subjects for IgG subclass-specific immune response and IgG2 may be related to pathogenesis. Studies using individual F6 molecules will be done to identify the molecule(s) involved in infection and protective immunity.

Antifilarial activity of Caesalpinia bonducella against experimental filarial infections.

BACKGROUND & OBJECTIVE: Lymphatic filariasis is a disabling disease that continues to cripple population in tropical countries. Currently available antifilarial drugs are not able to control the disease. Therefore, a better antifilarial is urgently required for proper management of the disease. We undertook this study to assess the antifilarial activity of Caesalpinia bonducella-seed kernel against rodent filarial parasite in experimental model. METHODS: Microfilaraemic cotton rats and Mastomys coucha harbouring Litomosoides sigmodontis and Brugia malayi respectively, were treated with crude extract or fractions of the seed kernel C. bonducella through oral route for 5 consecutive days. Microfilaricidal, macrofilaricidal and female worm sterilizing efficacy was assessed. RESULTS: Crude extract showed gradual fall in microfilariae (mf) count in L. sigmodontis-cotton rat model from day 8 post-treatment attaining more than 95 per cent fall by the end of observation period. It also exhibited 96 per cent macrofilaricidal and 100 per cent female sterilizing efficacy. The butanol fraction F018 caused 73.7 per cent reduction in mf count and 82.5 per cent mortality in adult worms with 100 per cent female sterilization. The aqueous fraction F019 exerted more than 90 per cent microfilaricidal activity and 100 per cent worm sterilization. Two chromatographic fractions, F024 and F025 of hexane soluble fraction exhibited 64 and 95 per cent macrofilaricidal activity, respectively. Both the fractions caused gradual fall in microfilaraemia and 100 per cent worm sterilization. In B. malayi-M. coucha model F025 showed gradual reduction in microfilaraemia and caused 80 per cent sterilization of female parasites INTERPRETATION & CONCLUSION: In conclusion, C. bonducella- seed kernel extract and fractions showed microfilaricidal, macrofilaricidal and female-sterilizing efficacy against L. sigmodontis and microfilaricidal and female-sterilizing efficacy against B. malayi in animal models, indicating the potential of this plant in providing a lead for new antifilarial drug development.

Transmission of filarial infection through blood transfusion.

Microfilariae can be transmitted by blood transfusion and they may be circulated in the recipient's blood but they do not develop into adult worms. Mortality associated with transfusion associated filarial infection is not documented but it may give rise to morbidity in transfusion recipients in terms of allergic reaction. The present study was carried out to investigate the association of post transfusion reactions and filarial infections in an endemic area. About 11,752 transfusion recipients were followed up and in 15 months period, 47 (0.4%) post transfusion reactions (PTR) were reported. Routine investigations for post transfusion reaction were carried out in all 47 patients and their respective blood donor. Moreover, blood culture, microfilaria detection by concentration technique, filarial antibody and antigen detection (both by ELISA) were done in all subjects. Out of 47 patients showing post transfusion reaction, 29 (61.7%) patients developed allergic reaction. Eighteen (38.3%) patients having allergic reaction did not have previous history of blood transfusion and 14 (29.8%) of them received transfusion from blood donors who was either positive for microfilaria, filarial antigen or antibody. Microfilaremia was demonstrated in 4 (8.5%) patients and 5 (10.6%) blood donors. Microfilaria was concurrently present in 2 patients and their respective donors. Filarial antibody was detected in 27 (56.5%) patients and 26 (55.3%) blood donors but microfilaria was detected in 3 (6.4%) and 4 (8.5%) subjects, respectively. Antigen detection test correlated with microfileraemic state of subjects. The result shows that transfusion associated filarial infection may be a probable cause for transfusion-associated morbidity in endemic areas. In 14 (29.8%) patients having allergic reactions, the probable cause was transfusion-associated filarial infection. Filarial antigen detection test was found to be more useful in detecting infections. Blood donors with active history of filarial infection should be deferred from donating blood. Filarial antigen detection test may be employed as screening test for blood donors, if possible.

Time lag between case registration and commencement of treatment in a leprosy control unit.

Analysis of client-based data as a part of computerised management information system in a Government leprosy control unit in Tamil Nadu reveals that there was delay in initiating treatment of leprosy patients. The mean and standard deviation of the period of delay for cases registered before, within 6 months and after 6 months of start of MDT in the Unit were 6.80 +/- 6.40, 1.97 +/- 3.60 and 0.90 +/- 2.21 months respectively. Further, the delay was longer in PB, female and child cases. Giving priority to therapy for backlog cases and an effective monitoring system with specific indicator for time lag in starting treatment is indicated.

A computerized information system for evaluation of NLEP through monthly progress reports.

A computerized system for monitoring district-wise operational performance and epidemiological progress using existing regular and special monthly reports of the National Leprosy Eradication Programme (NLEP) is presented. The same system, with some minor modifications could be used for programme assessment at the Leprosy Control Unit level also. The advantage of the system is the speed with which it can generate output in the form of comparative tables and graphs for different regions for use by programme managers for making overall assessments in time and for sending feedback reports to workers at various levels, for self-assessment and for taking timely corrective action. The system presented provides immediate and easy access to the stored and/or processed information (indicators etc.,) at any time. The system has been pilot-tested using monthly reports from eighteen districts of Tamil Nadu.

Evaluation of filarial skin test with Brugia malayi larval antigen in the field.

To obtain comparable and reproducible results by filaria skin test with B. malayi larval antigen, standard procedure has been evolved. Antigen protein of 2 micrograms per test, injected intracutaneously, was found optimum for positive skin reaction. The reaction ratio based on increase of wheal area by 2 times or more was found to be statistically significant for interpreting positive results. However, in larger field trials, the simpler measurement of increase of wheal diameter by 1 1/2 times or more was found to be equally reliable. Patients treated with diethylcarbamazine citrate, antihistaminics and anti-inflammatory drugs are likely to be unresponsive to filarial antigen and will yield incorrect information.

Comparative analysis of surface components of adult, micro-filariae and infective larvae of Brugia malayi.

A comparative analysis of surface proteins of adult, microfilariae and infective larvae of Brugia malayi, the human filarial parasite, has been carried out using IODOGEN (1,3,4,6-tetrachloro-3,alpha 6 alpha-diphenyl-glycoluril) and lactoperoxidase methods. SDS-polyacrylamide gel electrophoretic and autoradiographic analyses revealed the presence of 9 proteins (15-200 kDa) in adults, while microfilariae and infective larvae showed 8 and 6 proteins (15-120 kDa), respectively. The pattern of proteins radiolabelled by IODOGEN method was very similar to that of proteins labelled by the lactoperoxidase method. Since these proteins are released by the protease treatment of whole parasites, they are likely to be present on the surface of the parasite.