Increased IgG antibody responses to excretory/secretory antigens in neonates born from mothers infected with filarial nematodes.

The present investigation aimed to evaluate the extent to which maternal filarial infection influences IgG subclass immune responses in the cord blood of neonates. Prevalence of antigenaemia was detected using an Og4C3 assay. Filaria-specific IgG subclasses against excretory/secretory antigens were measured by ELISA. Transplacental transfer of circulating filarial antigen (CFA) was observed from 34.8% of CFA-positive mothers to their respective cord bloods. Filaria-specific IgG1, IgG2 and IgG4 responses were significantly higher among cord bloods of infected mothers compared to cord bloods of uninfected mothers. In contrast, the IgG3 response was significantly higher among cord bloods of uninfected mothers. The study shows that transplacental transfer of filarial antigens and filaria-specific IgG4 occurs more in mothers having high worm burdens, and transfer of filaria-specific IgG3 occurs more in the cord blood of uninfected mothers. The findings of the study provide evidence for the development of prenatal sensitization to filarial antigens in utero, and high filaria-specific IgG4 in cord blood may serve as a marker for in-utero sensitization.

Structural and thermal characterization of PHAs produced by Lysinibacillus sp. through submerged fermentation process.

In this investigation an attempt has been made to characterize and identify Lysinibacillus sp. 3HHX by 16S-rDNA sequencing. The bacterium exhibited occurrence of PHAs granules on an average 11±1 per cell of 1.0µm length and breadth 0.72µm, revealed from TEM studies. Under optimized condition, 4.006gm/L of PHAs was extracted using hypochlorite digestion and multi-solvent extraction process. PhaC gene of ∼540bp and higher PHA synthase activity was detected at 48h of cultivation. The extracted PHAs was structurally characterized by GC-MS and 1H NMR reported to be P(3HB-co-3HDD-co-3HTD) and amorphous in nature with 112°C melting point, -11.0°C glass transition point and 114.76°C decomposition temperature detected by DSC & TGA respectively. The C/O of biopolymer disc was 1:65 as revealed from C1s and O1s spectra of XPS, that was completely biodegradable within 30 days. This biopolymer was observed to be non-cytotoxic to NIH 3T3 mouse fibroblast cells. The report is of its kind in establishing the abilities of Lysinibacillus sp. 3HHX for non-growth associated PHA co-polymer production. Moreover the biocompatible and biodegradable nature of the biopolymer conferred to its substantial biomedical applications.

Hydrocele fluid: can it be used for immunodiagnosis of lymphatic filariasis?

BACKGROUND & OBJECTIVES: Diagnosis of lymphatic filariasis using serum has been established but the utility of hydrocele fluid for the purpose is not exactly known. Since, hydrocele is a chronic form of the disease manifestation in a variety of situations and often poses difficulty in diagnosing its origin, we have evaluated the usefulness usage of hydrocele fluid for diagnosis of filarial origin of hydrocele in this study. METHODS: Paired samples of serum and hydrocele fluid from 51 individuals with hydrocele, living in an endemic area of Wuchereria bancrofti were assessed. Circulating filarial antigen, filarial specific antibody and cytokine assay were performed in both serum and hydrocele fluid of patients. RESULTS: Og4C3 assay detected circulating filarial antigen (CFA) in serum and corresponding hydrocele fluids. The level of IgG, IFN-γ and IL-10 were found to be high in CFA-negative, while IgM and IgE were high in CFApositive hydrocele fluid and serum samples associated with hydrocele. On the other hand neither CFA-positive nor CFA-negative hydrocele fluid and serum samples associated with hydrocele showed any difference in IgG4 level. INTERPRETATION & CONCLUSION: This study showed that the filaria related antigens and antibodies found in serum can be detected with equal sensitivity in hydrocele fluid. Therefore, it can be used as an alternative to serum for immunodiagnosis of filariasis, and help monitoring the filarisis elimination programme.

Immuno-epidemiology of bancroftian filariasis: a 14-year follow-up study in Odisha, India.

Forty asymptomatic, circulating filarial antigen negative (CFA(-ve)) and ten asymptomatic, circulating filarial antigen positive (CFA(+ve)) individuals were followed up longitudinally over a period of 14 years at intervals of 7 years in order to investigate the immunological, parasitological and clinical changes that took place in an endemic area due to natural process. The clinical status, microfilaremia, circulating filarial antigenemia and immunological responses to filarial antigens (DSSd1 and Sd30) prepared from cattle filarial parasite Setaria digitata, were examined. The observations showed that 19 individuals had developed either antigenemia or filarial symptoms (acute filarial lymphangitis/hydrocele) from CFA(-ve) group. Three individuals had cleared antigenemia and one had developed microfilaremia from CFA(+ve) group after 7 years. Increased IgG and IgM and low IgG2 and IgG4 level responses along with high lymphocyte production were observed in CFA-negative individuals. This was in contrast to observations made in CFA(+ve) subjects. The results of the present study indicated that the changes taking place in the immunological, clinical and CFA status of individuals residing in filaria endemic regions developed different clinical manifestation with course of time.

In utero sensitization modulates IgG isotype, IFN-γ and IL-10 responses of neonates in bancroftian filariasis.

In utero exposure has been considered as a risk factor for filarial infection. To evaluate the influence of maternal infection on filarial-specific IgG subclass response in neonates and their correlation with plasma levels IL-10 and interferon-γ, 145 pairs of mothers and their respective cord bloods were examined. Transplacental transfer of circulating filarial antigen (CFA) was observed in 34·8% cord bloods from CFA positive mothers. Filarial-specific IgG1, IgG2 and IgG4 responses of cord bloods were found to be positively correlated with CFA of mothers. In contrast, IgG3 responses negatively correlated with CFA of mothers. The % of similarity of recognition pattern in the cord blood with maternal blood was high for IgG3 response than IgG4 in all three groups. An increased levels of IL-10 and decreased levels of interferon gamma (IFN-γ) were observed in cord blood of infected mothers. Interferon gamma was positively correlated with IgG3 and negatively correlated with IgG4 level. On the other hand, IL-10 was positively correlated with IgG4 and CFA, indicating that cytokines may play a role in modulating the immune responses in cord bloods of sensitized foetus. The findings of the study reveal that in utero tolerance or sensitization may influence the filarial-specific immunity to infection in neonates.

Bancroftian filariasis: circulating B-1 cells decreased in microfilaria carriers and correlate with immunoglobulin M levels.

B-1 cells play an important role in the outcome of infection in schistosomiasis, pneumonia and experimental filariasis. However, no information exists regarding status of B-1 cells in clinical manifestations of human filariasis. We investigated the levels of B-1 cells from the total B cells by flow cytometry. Significantly low levels of B-1 cells and IgM antibodies were detected against a wide variety of autoantigens in microfilariae carriers as compared to endemic controls and patients with chronic pathology. A positive correlation was found between IgM antibodies to actin and ss-DNA. Absorption of plasma with soluble actin, myosin and lipopolysaccharides (LPS) resulted in significant removal of antifilarial antibodies. Affinity-purified anti-ss-DNA antibodies were found to be reactive to filarial antigens and various autoantigens. Further, a positive correlation was found between polyreactive antibodies and B-1 cells in filarial-infected human subjects. After antifilarial treatment, levels of IgM antibodies to ss-DNA, actin, LPS and filarial antigen increased significantly indicating a role of polyreactive naturally occurring antibodies in filarial infection. Our findings add to the existing evidence that the B-cell defect in BALB.Xid mice account for susceptibility to murine filarial infection and indicate an important role for these antibodies in providing host protection against filarial infection.

Maternal filarial infection: association of anti-sheath antibody responses with plasma levels of IFN-γ and IL-10.

Maternal filarial infection influences the risk of acquiring infection and development of immunity in children. Here we have analysed the blood samples of 60 mothers (24 infected and 36 uninfected) and their corresponding cord bloods to assess the impact of maternal infection on the anti-sheath antibodies and cytokine production in neonates born from them. About 69·4% of non-infected mothers and their cord bloods showed the presence of anti-sheath antibodies, while only 16·6% of the cord bloods from infected mothers were positive for it. The IL-10 level was significantly high in cord bloods of infected mothers compared with non-infected mothers. At the same time the IL-10 level was also observed to be remarkably high in cord bloods of both infected and non-infected mothers negative for anti-sheath antibody. In contrast, IFN-γ levels were significantly high in cord bloods of non-infected mothers compared with infected mothers and the increment was prominent in cord bloods of both infected and non-infected mothers positive for anti-sheath antibody. The study reveals that the presence or absence of anti-sheath antibodies in association with cytokines skews the filarial specific immunity to either Th1 or Th2 responses in neonates. This may affect the natural history of filarial infection in early childhood.

Immunoprophylactic potential of filarial glutathione-s-transferase in lymphatic filariaisis.

OBJECTIVE: To elucidates the immunoprophylactic potential of glutathion-s-transferase (GST) from cattle filarial parasite Setaria digitata (S. digitata) against lymphatic filariasis. METHODS: GST was purified through affinity chromatography (SdGST) and chacterized by SDS-PAGE and Nano-LC MS/MS analysis. Antibody isotypes to SdGST were measured by ELISA. Antibody dependant cellular cytotoxicity (ADCC) was performed in vitro using sera from immunized animals and immune individuals. T-cell proliferation and cytokine response to SdGST in different groups of filariasis were measured. Immunoprophylactic potential of SdGST was evaluate in animal model. RESULTS: SdGST exhibited 30-fold enhancement of enzyme activity over crude parasitic extract. It was found to be 26 kDa by SDS-PAGE. Nano LC-MS/MS analysis followed by blast search showed 100% homology with Dirofilaria immitis (D. immitis) and only 43% with Homo sapiens (H. sapiens). Immunoblotting analysis showed putatively immune individuals carry significant level of antibodies to SdGST as compared with microfilaraemics. Immunized sera and sera endemic normal could neutralize the enzymatic activity of SdGST and inducing in vitro cytotoxicity of microfilariae. Peripheral blood mononuclear cells (PBMC) from endemic normals upon stimulation with SdGST showed a mixed type of Th1/Th2 response. SdGST immunization clear microfilariae from circulation in S. digitata implanted mastomys. CONCLUSIONS: The heterologous GST could be potentially developed as a vaccine candidate against lymphatic filarial parasite.

Lymphatic filariasis in children: age dependent prevalence in an area of India endemic for Wuchereria bancrofti infection.

Lymphatic filariasis has been considered as a disease of adults and most epidemiological surveys have excluded children. The prevalence of infection and clinical manifestations of the disease among children in the age group of 1-15 years was determined in a Wuchereria bancrofti endemic area. The 1383 children from the rural villages of a coastal district (Khurda), State of Orissa, India, were studied. The finger prick blood (50ìl) samples were collected between 20:30 and 23:30 hours for parasitological and immunological evaluation. At the same time clinical examination was also recorded. Circulating Filarial Antigen (CFA) status and antibody (IgG) to filarial antigen was also determined in the study population. The prevalence of asymptomatic microfilaraemic carriers (AS), acute disease (AC), hydrocele (Hyd) cases and cryptic infection (CFA +ve) were 9.9%, 14.6%, 3.8% and 17.1% respectively. It was observed that 45.4% of the children below 15 years of age were either infected or had clinical manifestations of the disease. IgG antibody positivity 75.4%, 84% and 95.8% were observed in 1-5 yr, 6-10 yr and 11-15 years age group respectively. The study suggested that asymptomatic infection and acute form of disease were common occurrence among the children and more than half of the children population were either infected or having clinical manifestations of the diseased by pre-adult stage (11-15 years of age) in the endemic area.