Development and characterization of a standardized ELISA including a reference serum on each plate to detect antibodies induced by experimental malaria vaccines

Abstract:Seasonal influenza viruses (IAV) are a common cause of acute respiratory illness worldwide and generate a significant socio-economic burden. IAV rapidly mutate, necessitating annual vaccine reformulation as traditional vaccines do not typically induce broad-spectrum immunity. In addition to seasonal infections, emerging pandemic influenza viruses present a continued threat to global public health. Pandemic influenza viruses have consistently higher attack rates and are typically associated with greater mortality as compared to seasonal strains. Ongoing strategies to improve vaccine efficacy typically focus on providing broad-spectrum immunity, and while both B and T cells can mediate heterosubtypic responses, typical vaccine development will augment either humoral or cellular immunity. However, multipronged approaches, targeting several antigens, may limit the generation of viral escape mutants. There are few vaccine platforms that can deliver multiple antigens and generate robust cellular and humoral immunity. In this work, we describe a novel vaccination strategy, tested pre-clinically in mice, for the delivery of novel bivalent viral-vectored vaccines. Here, we show this strategy elicits potent T cell responses toward highly conserved internal antigens, whilst simultaneously inducing high levels of antibodies towards hemagglutinin (HA). Importantly, these humoral responses generate longlived plasma cells and generate antibodies capable of neutralising variant HA-expressing pseudotyped lentiviruses. Significantly, these novel viral-vectored vaccines induce strong immune responses capable of conferring protection in a stringent influenza A virus challenge.Thus, this vaccination regimen induces lasting efficacy toward influenza. Importantly, the simultaneous delivery of dual antigens may alleviate the selective pressure thought to potentiate antigenic diversity in avian influenza viruses.

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“…Nunc Maxisorp® 96-well plates were coated with 0.1 g recombinant protein (H7HA protein was produced in-house as described (16) (14,17).…”

Section: Elispotmentioning

confidence: 99%

Novel Bivalent Viral-Vectored Vaccines Induce Potent Humoral and Cellular Immune Responses Conferring Protection against Stringent Influenza A Virus Challenge

Chinnakannan

Mullarkey

Ulaszewska

et al. 2017

The Journal of Immunology

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“…Enhancing the immunogenicities of these antigens to obtain a protective and sustained response in humans remains a formidable task that confronts malarial vaccine researchers today [3,4]. One strategy that we have used to increase immunogenicity was to chemically conjugate malarial antigens to a carrier protein [5][6][7][8][9].…”

Section: Introductionmentioning

confidence: 99%

“…A strong positive correlation between GIA activity and anti-AMA1 specific antibody levels was displayed. In addition, an adult phase I trial in the U.S. with AMA1-C1, a mixture of AMA1-FVO and AMA1-3D7 alleles formulated on Alhydrogel with or without the addition of CPG 7909, established that this formulation was safe and well-tolerated [1,4].…”

Section: Introductionmentioning

confidence: 99%

Addition of CpG ODN to recombinant Pseudomonas aeruginosa ExoProtein A conjugates of AMA1 and Pfs25 greatly increases the number of responders

Qian

Rausch

Muratova

et al. 2008

Vaccine

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Both the blood stage protein apical membrane antigen 1 (AMA1) and the 25 kDa sexual stage protein (Pfs25) of Plasmodium falciparum are two leading candidates in malarial vaccine development. We have previously demonstrated that conjugation of these malarial antigens to recombinant Pseudomonas aeruginosa ExoProtein A (rEPA) significantly increased the mean specific functional antibody responses in mice; however, some mice responded poorly and were unable to demonstrate a functional response. We hypothesized that the immunogenicities of these two malarial antigens could be further enhanced by inclusion of a CpG oligodeoxynucleotide in the formulation. Mice were immunized with either rEPA conjugated or unconjugated AMA1 and Pfs25 formulated on Alhydrogel with or without the addition of CPG 7909. Mice received the formulations on days 0 and 28, and mouse sera were collected on day 42. ELISA analyses on these sera showed that the addition of CPG 7909 to AMA1-rEPA and Pfs25-rEPA formulated on Alhydrogel induced significantly higher mean antibody titers than the formulations without CPG 7909, and led to a mixed Th1/Th2 response as demonstrated by the production of mouse IgG1 and IgG2a subclasses. The presence of CPG 7909 in the formulations of both conjugated antigens greatly increased the proportion of responders with antibody titers sufficient to inhibit blood-stage parasite growth in vitro or block transmission of sexual stage parasites to mosquitoes. The results obtained in this study indicate the potential use of a combination strategy to increase the number of responders to malarial antigens in humans.

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“…The SternVolmer with good linear relationships fi t under the narrow range of concentration. The four parameter logistic nonlinear regression model is commonly used for curvefi tting analysis in bioassays [27][28][29]. It has been described as follows:…”

Section: Determination Of Parathionmentioning

confidence: 99%

Development of a Probe Based on Quantum Dots Embedded with Molecularly Imprinted Polymers to Detect Parathion

Tang¹,

Li²

2016

Pol. J. Environ. Stud.

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Development and characterization of a standardized ELISA including a reference serum on each plate to detect antibodies induced by experimental malaria vaccines

Cited by 201 publications

References 17 publications

Novel Bivalent Viral-Vectored Vaccines Induce Potent Humoral and Cellular Immune Responses Conferring Protection against Stringent Influenza A Virus Challenge

Novel Bivalent Viral-Vectored Vaccines Induce Potent Humoral and Cellular Immune Responses Conferring Protection against Stringent Influenza A Virus Challenge

Addition of CpG ODN to recombinant Pseudomonas aeruginosa ExoProtein A conjugates of AMA1 and Pfs25 greatly increases the number of responders

Development of a Probe Based on Quantum Dots Embedded with Molecularly Imprinted Polymers to Detect Parathion

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