The Wuchereria bancrofti orthologue of Brugia malayi SXP1 and the diagnosis of bancroftian filariasis

Rao, K. V. Narasinga; Eswaran, M. Arthanari; Ravi, Varatharajalu; Gnanasekhar, B; Narayanan, R. B.; Kaliraj, Perumal; Jayaraman, Kunthala; Marson, Aimee L.; Raghavan, Nithyakalyani; Scott, Alan L.

doi:10.1016/s0166-6851(99)00231-5

Cited by 82 publications

(91 citation statements)

References 22 publications

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“…The full length ALT-2 gene from Brugia malayi (BmALT-2) was cloned into the prokaryotic expression vector pRSET (Invitrogen). Cloning of the W. bancrofti SXP-1 (WbSXP-1) gene has been previously described (32). Recombinant clones were transformed into the E. coli strain BL21 (DE3) pLysS for production of recombinant protein.…”

Section: Methodsmentioning

confidence: 99%

“…Levels of ALT-2 specific antibodies in sera from human subjects (Table 1) were studied by ELISA as previously described (32). Briefly, polyvinyl microtiter plates were coated with rBmALT-2 protein at a concentration of 10 µg/ml at 4 C and blocked with 3% bovine serum albumin in 0.01 M PBS for 2 hr at 37 C. Sera diluted (1:200) were incubated in microtiter plates at 37 C for 2 hr.…”

Section: Methodsmentioning

confidence: 99%

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The Larval Specific Lymphatic Filarial ALT‐2: Induction of Protection Using Protein or DNA Vaccination

Ramachandran

Kumar

Rami

et al. 2004

Microbiology and Immunology

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Genes from the infective stage of lymphatic filarial parasites expressed at the time of host invasion have been identified as potential vaccine candidates. By screening an L3 cDNA library with sera from uninfected longstanding residents of an area endemic for onchocerciasis, so‐called “endemic normals” (EN), we have cloned and characterized one such gene termed the abundant larval transcript two (ALT‐2). The stage specificity of ALT‐2 gene transcription and protein synthesis was confirmed by PCR using gene‐specific primers, and by western blot analysis of protein extracts from various stages of the parasite life cycle using specific antisera. Significant differences in antibody response to the recombinant ALT‐2 were observed in endemic populations with differing clinical manifestations of lymphatic filariasis with an antibody response present in sera from 18 of 25 (72%) EN subjects compared to 9 of 25 (36%) with subclinical microfilaracmia (MF) and 14 of 25 (52%) of those with chronic lymphatic obstruction (CP) (P=0.01 for comparison of EN to CP or to MF). This differential responsiveness suggests that the protective immunity postulated to account for their uninfected status might be associated with a response to this protein. When the utility of ALT‐2 as a vaccine candidate was tested in a murine model using either recombinant protein or a DNA vaccine construct, statistically significant protection was observed when compared to a control filarial gene product expressed across all stages of the parasite lifecycle (SXP‐1; P=0.02 for protein and P=0.01 for the DNA vaccine) or compared to adjuvant alone. This level of protection indicates that this vaccine is a promising candidate for further development.

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Section: Methodsmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

The Larval Specific Lymphatic Filarial ALT‐2: Induction of Protection Using Protein or DNA Vaccination

Ramachandran

Kumar

Rami

et al. 2004

Microbiology and Immunology

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“…which is required for SXP/RAL-2 is specially expressed in the subventral pharyngeal glands few nematodes and the protein is most likely secreted. 18 The similarity of WbL 2 to other proteins from the SXP/RAL-2 family, such as from Ancylostoma caninum (Ac-16), Ascaris suum (As14), Acanthocheilonema viteae (Av-RAL-2), Setaria digitata (Sd-SXP-1), Brugia malayi (Bm-SXP-1), and Onchocerca volvulus (Ov17 and P36991) suggests that WbL 2 might share some biological properties with these (apparently) nematode-specific proteins. 19 While little is known of the functions of these proteins, some of them were identified as targets for nematode vaccines due to the protection they conferred upon immunization.…”

Section: Effect On the Splenocytes Proliferation And Cytokine Analysismentioning

confidence: 99%

Role of recombinant SXP/RAL-2 family protein Wuchereria bancrofti L2 (rWbL2) as vaccine candidate in lymphatic filariasis in mastomys

et al. 2016

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“…Circulating microfilariae in the blood samples were identified as described previously (Gnanasekar et al 2004) and the circulating filarial antigens were detected using a commercially available Og4C3 kit (Lalitha et al 1998) and a WbSXP-based ELISA (Rao et al 2000). Individuals with no circulating antigen or microfilariae were considered as EN, whereas, subjects with circulating microfilariae and/or circulating antigen as detected by Og4C3/WbSXP ELISA were considered as MF.…”

Section: Collection Of Sera Samplesmentioning

confidence: 99%

“…These two parasites are closely related and show significant antigenic overlap that the antibodies generated against W. bancrofti in infected or immune individuals show signifiant cross-reactivity with B. malayi antigens (Lalitha et al 2002;Rao et al 2000;Rathaur et al 2003). In the endemic areas, some individuals are naturally immune to the disease (Endemic Normal, EN); whereas, some carry the infection and show acute symptoms (Microfilaremics, MF) and others exhibit morphological evidence of lymph edema in the dependable parts as the infection becomes chronic (Chronic Pathology, CP).…”

Section: Introductionmentioning

confidence: 99%

Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

2005

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Iterative screening of a phage display cDNA expression library of the third-stage larvae (L3) of Brugia malayi with sera from putatively immune individuals (endemic normal, EN) identified a novel clone with insert showing significant homology to Onchocerca volvulus novel immunogenic protein-3 (Ov-NIP3) gene and Caenorhabditis elegans NIP3-like protein and hence the gene was designated Brugia malayi NIP3-like protein (BmNIP3). EST database analysis showed that ESTs from several gastrointestinal nematodes such as Ancylostoma caninum, Teladorsagia circumcincta, Haemonchus contortus and Strongyloides stercoralis has BmNIP3 homologues, but the gene has not been described from these parasites. Sequence analyses showed that BmNIP3 has three potential mucin-type O-glycosylation sites and several serine/threonine phosphorylation sites. As expected, BmNIP3 protein isolated from the parasite was serine/threonine phosphorylated. Further analyses showed that BmNIP3 is differentially transcribed, with highest level of expression present in the larval (L3 and L4) stages. Mice immunized with rBmNIP3 developed strong antibody responses predominantly of the IgG1 and IgG2a subtype. A similar analyses of the sera samples from EN individuals showed that they also carry high levels of IgG1 and IgG2 antibodies against BmNIP3, whereas, chronically infected patients carry largely IgG3 antibodies and MF individuals carry high levels of IgG1 antibodies against BmNIP3. This study thus describes a novel protein from B. malayi that appears to be highly immunogenic in both humans and mice.

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The Wuchereria bancrofti orthologue of Brugia malayi SXP1 and the diagnosis of bancroftian filariasis

Cited by 82 publications

References 22 publications

The Larval Specific Lymphatic Filarial ALT‐2: Induction of Protection Using Protein or DNA Vaccination

The Larval Specific Lymphatic Filarial ALT‐2: Induction of Protection Using Protein or DNA Vaccination

Role of recombinant SXP/RAL-2 family protein Wuchereria bancrofti L2 (rWbL2) as vaccine candidate in lymphatic filariasis in mastomys

Cloning and characterization of a novel immunogenic protein 3 (NIP3) from Brugia malayi by immuno screening of a phage-display cDNA expression library

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