Vaccine Research and Development for the Prevention of Filarial Nematode Infections

Grieve, Robert B.; Wisnewski, Nancy; Frank, Glenn; Tripp, Cynthia A.

doi:10.1007/978-1-4615-1823-5_33

Cited by 21 publications

(15 citation statements)

References 159 publications

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“…Further, the vaccine has been subject to animal trials in a number of systems, including murine and jird models, each resulting in significant and similar levels of protection, ranging from 25 to 76% (20).…”

Section: Discussionmentioning

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: Discussionmentioning

confidence: 99%

“…Vaccination with larval specific antigens, such as ALT-2 should avoid this issue; this gene product is exclusively expressed in the L3 stage of the parasite, and available evidence indicates that the immune responses to the L3 or L4 stages are nonpathogenic (20).…”

Section: Discussionmentioning

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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“…These drugs pose problems due to their undesirable side effects. Thus, there is a need to develop a suitable agent such as vaccine for the successful elimination of human lymphatic filariasis [8][9][10][11]. According to various research reports, Brugia malayi abundant larval transcript-2 (BmALT-2) is the most abundant of the L3-expressed, stage-specific novel proteins [12][13].…”

Section: Introductionmentioning

confidence: 99%

Cloning, expression and characterization of Brugia malayi abundant larval protein transcript-2 (BmALT-2) expressed in Pichia pastoris

Paul

Saha

Selvaraja

et al. 2016

Biotechnology & Biotechnological Equipment

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Besides mass drug administration, successful elimination of human lymphatic filariasis, a mosquitoborne tropical parasitic disease, requires developing other suitable prophylactic agents such as vaccines. The Brugia malayi abundant larval transcript-2 (BmALT-2) protein has been identified as one possible candidate. So far, it (E-ALT-2) has been expressed as a 24-kDa non-glycosylated protein in Escherichia coli. Easy and low-cost downstream purification of secreted BmALT-2 in Pichia pastoris may be a vital option to inexpensive large-scale production. This study focused on expression and molecular characterization of BmALT-2 (P-ALT-2) in P. pastoris. Sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis showed that some P. pastoris colonies produced 27 and 24 kDa bands and few colonies produced a 24-kDa band. Preliminary studies confirmed glycosylation of 27 kDa P-ALT-2. The ratio of glycosylated and non-glycosylated P-ALT-2 was 20:80-70:30 in various colonies. The maximum yield of glycosylated and non-glycosylated P-ALT-2 was measured as 7.99 § 1.12 and 9.18 § 1.35 mg L ¡1 compared to 6.5 § 1.2 mg L ¡1 of E-ALT-2 in shake flasks. Their overall expression was about 25% and 35% higher than that in E. coli. The glycosylated P-ALT-2 exhibited about 15% higher immunoreactivity with human endemic normal sera. The enhanced secreted production by P. pastoris may lead to cost-effective large-scale production of BmALT-2.

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“…Therefore, it is imperative to identify the key enzymes and biochemical pathways that are pivotal to the parasites' survival in the host's hostile environment, including their oxidative stresses and immune responses. These enzymes should provide excellent biochemical targets for developing eective chemotherapies and vaccines (Grieve et al 1995;Lazdins and Kron 1999). One such enzyme is glutathione S-transferase (GST; EC 2.5.1.18; Brophy and Pritchard 1994), which is involved in xenobiotic metabolism, intracellular binding, and biosynthesis of endogenous substrates such as prostaglandins and leukotrienes (Boyer 1989;Armstrong 1991;van Bladeen and van Ommen 1991).…”

Section: Introductionmentioning

confidence: 99%

Identification and localization of glutathione S-transferase as a potential target enzyme in Brugia species

Rao

Salinas

Mehta

et al. 2000

Parasitology Research

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Brugia filarial nematodes are pathogenic lymphatic-dwelling parasites that, like other helminths, may modify the host's defense mechanisms by a major detoxification process involving glutathione-binding proteins such as glutathione S-transferases (GSTs). In the present study, soluble extracts of third-stage larvae, adult male and female worms, microfilariae of either B. pahangi or B. malayi or the adult worm excretory-secretory products of B. malayi were used to determine GST activity. These extracts and affinity-purified fractions of B. pahangi adult worms had a specific enzymatic activity when 1-chloro-2,4-dinitrobenzene was used as a substrate. The observance of this enzyme in all life cycle stages of Brugia spp. demonstrates its ubiquitous nature. Lavage of intraperitoneally infected jirds, but not that of uninfected jirds, also showed increased enzymatic activity, suggesting that GST is secreted in vivo. Soluble proteins of both Brugia spp. were strongly recognized by antibodies in sera from rabbits immunized with affinity-purified native GST of Onchocerca volvulus. Immunohistochemical studies localized these proteins in adult worms, demonstrating cross-reactivity between the GST of these two filarial nematodes. The effect of this enzyme on the motility and viability of adult worms, microfilariae, and larvae was tested in vitro using a battery of known GST inhibitors. Of all those tested, ethacrynic acid, N-ethylmalemide, 4-nitropyridine-oxide, or 1-chloro-2,4-dinitrobenzene at micromolar concentrations reduced the viability and motility of microfilariae, third-stage larvae, and adult worms. These results suggest that Brugia GSTs are major metabolic enzymes and may play an important role in the parasite's survival.

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Vaccine Research and Development for the Prevention of Filarial Nematode Infections

Cited by 21 publications

References 159 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

Cloning, expression and characterization of Brugia malayi abundant larval protein transcript-2 (BmALT-2) expressed in Pichia pastoris

Identification and localization of glutathione S-transferase as a potential target enzyme in Brugia species

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