Duthie M.S.,Protein Advances Incorporated |
Duthie M.S.,Infectious Disease Research Institute |
Hay M.N.,Protein Advances Incorporated |
Morales C.Z.,Protein Advances Incorporated |
And 10 more authors.
Clinical and Vaccine Immunology | Year: 2010
Despite the reduction in the number of leprosy cases registered worldwide as a result of the widespread use of multidrug therapy, the number of new cases detected each year remains stable in many countries. This indicates that Mycobacterium leprae, the causative agent of leprosy, is still being transmitted and that, without an earlier diagnosis, transmission will continue and infection will remain a health problem. The current means of diagnosis of leprosy is based on the appearance of clinical symptoms, which in many cases occur after significant and irreversible nerve damage has occurred. Our recent work identified several recombinant antigens that are specifically recognized by leprosy patients. The goal of the present study was to produce and validate the reactivity of a chimeric fusion protein that possesses the antibody binding properties of several of these proteins. The availability of such a chimeric fusion protein will simplify future test development and reduce production costs. We first identified the antibody binding regions within our top five antigen candidates by performing enzyme-linked immunosorbent assays with overlapping peptides representing the amino acid sequences of each protein. Having identified these regions, we generated a fusion construct of these components (protein advances diagnostic of leprosy [PADL]) and demonstrated that the PADL protein retains the antibody reactivity of the component antigens. PADL was able to complement a protein that we previously produced (the leprosy IDRI [Infectious Disease Research Institute] diagnostic 1 [LID-1] protein) to permit the improved diagnosis of multibacillary leprosy and that had a good ability to discriminate patients with multibacillary leprosy from control individuals. A serological diagnostic test consisting of these antigens could be applied within leprosy control programs to reduce transmission and to limit the appearance of leprosy-associated disabilities and stigmatizing deformities by directing treatment. Copyright © 2010, American Society for Microbiology. All Rights Reserved.
Wiley S.R.,Imdaptive Inc. |
Raman V.S.,Infectious Disease Research Institute |
Desbien A.,Infectious Disease Research Institute |
Bailor H.R.,Infectious Disease Research Institute |
And 7 more authors.
Science Translational Medicine | Year: 2011
Vaccination with an isolated antigen is frequently not sufficient to elicit a protective immune response. The addition of adjuvants to the antigen can increase the magnitude and breadth of the response generated, but quantification of this increase as a function of adjuvant has been intractable. We have directly determined the variation of the immunoglobulin G variable-chain repertoire of an entire organism as a function of vaccination. Using the well-established Plasmodium vivax antigen, PvRII, and massively parallel sequencing, we showed that the use of a Toll-like receptor (TLR) agonist in the vaccine formulation increased the diversity of the variable region sequences in comparison to the use of an oil-in-water emulsion adjuvant alone. Moreover, increased variable domain diversity in response to the use of TLR agonist - based adjuvants correlated with improved antigen neutralization. The use of TLR agonists also broadened the range of polymorphic variants against which these antibodies could be effective. In addition, a peptide microarray demonstrated that inclusion of adjuvants changed the profile of linear epitopes from PvRII that were recognized by serum from immunized animals. The results of these studies have broad implications for vaccine design - they may enable tailored adjuvants that elicit the broad spectrum of antibodies required to neutralize drifted and polymorphic pathogen strains as well as provide a method for rapid determination of correlates of adjuvant-induced humoral immunity.
Carter D.,Infectious Disease Research Institute |
Carter D.,Protein Advances Incorporated |
Reed S.G.,Infectious Disease Research Institute |
Reed S.G.,Immune Design
Current Opinion in HIV and AIDS | Year: 2010
Purpose of review: Recent clinical trial results have indicated that it may be possible for vaccines to induce protection against HIV. To build on this result, strategies should be designed to enhance duration, breadth, and magnitude of antibody production. Strategic formulation of agonists of the innate immune system and carriers that selectively present the target antigen yields a class of pharmaceuticals, named 'adjuvants', that greatly influence immunity resulting from vaccination. As researchers begin to focus not only on creating an immune response to an antigen, but also on the quality of that response, the role of adjuvants is becoming increasingly significant. This review is intended to give an overview of recent findings on how adjuvants model the immune response to antigens with a focus on the field of vaccines for HIV. Recent Findings: It is clear that innate and adaptive immunity are linked by communication channels that allow innate signals to influence the quality of adaptive responses as well as adaptive signals that temper innate responses. Adjuvants take advantage of this bridge to shape the immune response to antigens. In this review, we will discuss the different classes of adjuvants currently available; recent findings on the relationship between adjuvants and the type of immune profile generated; and the breadth of neutralizing antibodies as influenced by adjuvants. Summary: Because adjuvants influence the breadth of antibodies generated and the type of cells that proliferate in response to a vaccine this review is relevant for scientists clinicians involved in creating a new HIV vaccine. © 2010 Wolters Kluwer Health | Lippincott Williams & Wilkins.
Desbien A.L.,Infectious Disease Research Institute |
van Hoeven N.,Infectious Disease Research Institute |
Reed S.J.,Infectious Disease Research Institute |
Casey A.C.,Infectious Disease Research Institute |
And 6 more authors.
BioTechniques | Year: 2013
We have developed an influenza hemagglutinin protein microarray to assess humoral recognition of diverse influenza strains induced by vaccination and infection. Each array consists of controls and 127 hemagglutinin antigens from 60 viruses, spotted in replicates to generate a single array of 1296 spots. Six arrays are configured on a single slide, which in the following analysis was probed simultaneously with 2 isotype-specific fluorescent secondary antibodies yielding over 15,000 data points per slide. Here we report the use of this system to evaluate mouse, ferret, and human sera. The array allows simultaneous examination of the magnitude of antibody responses, the isotype of such antibodies, and the breadth of influenza strain recognition. We are advancing this technology as a platform for rapid, simple, high-throughput assessment of homologous and heterologous antibody responses to influenza disease and vaccination.