Recombinant hemagglutinin proteins formulated in a novel PELC/CpG adjuvant for H7N9 subunit vaccine development

Novel vaccination strategies against Mycobacterium tuberculosis (MTB) are urgently needed. The use of recombinant MTB antigens as subunit vaccines is a promising approach, but requires adjuvants that activate antigen-presenting cells (APCs) for elicitation of protective immunity. The mycobacterial cord factor Trehalose-6,6-dimycolate (TDM) and its synthetic analogue Trehalose-6,6-dibehenate (TDB) are effective adjuvants in combination with MTB subunit vaccine candidates in mice. However, it is unknown which signaling pathways they engage in APCs and how these pathways are coupled to the adaptive immune response. Here, we demonstrate that these glycolipids activate macrophages and dendritic cells (DCs) via Syk–Card9–Bcl10–Malt1 signaling to induce a specific innate activation program distinct from the response to Toll-like receptor (TLR) ligands. APC activation by TDB and TDM was independent of the C-type lectin receptor Dectin-1, but required the immunoreceptor tyrosine-based activation motif–bearing adaptor protein Fc receptor γ chain (FcRγ). In vivo, TDB and TDM adjuvant activity induced robust combined T helper (Th)-1 and Th-17 T cell responses to a MTB subunit vaccine and partial protection against MTB challenge in a Card9-dependent manner. These data provide a molecular basis for the immunostimulatory activity of TDB and TDM and identify the Syk–Card9 pathway as a rational target for vaccine development against tuberculosis.

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2838. Safety and Immunogenicity of a gp120-CD4 Chimeric Subunit Vaccine: A Phase 1a Randomized Controlled Trial

Open Forum Infectious Diseases ◽

10.1093/ofid/ofz359.143 ◽

2019 ◽

Vol 6 (Supplement_2) ◽

pp. S65-S66

Author(s):

Joel V Chua ◽

Charles Davis ◽

Amy Nelson ◽

Ka Wing J Lam ◽

Lydiah Mutumbi ◽

...

Keyword(s):

Subunit Vaccine ◽

Vaccine Development ◽

Controlled Trial ◽

Chimeric Protein ◽

Controlled Study ◽

Control Group ◽

Single Chain ◽

Double Blind ◽

Humoral Responses ◽

Hiv 1

Abstract Background A primary challenge for HIV vaccine development is to raise antiviral antibodies capable of recognizing highly variable viral antigens. The full-length single chain (FLSC) gp120-CD4 chimeric protein was designed to present a highly conserved CD4-induced HIV-1 envelope structure that evokes cross-reactive humoral responses (Figure 1). IHV01 is an FLSC subunit vaccine formulated in alum adjuvant. The safety and immunogenicity of IHV01 was evaluated in this first-in-human phase 1a trial. Methods This randomized, double-blind placebo-controlled study involved three dose-escalating cohorts (75 µg, 150 µg, and 300 µg doses). Eligible participants were HIV-1 uninfected healthy volunteers aged 18 to 45 years. Participants in each cohort were block randomized in groups of four in a 3:1 ratio to receive either vaccine or placebo. Intramuscular injections were given on weeks 0, 4, 8, and 24. Participants were followed for an additional 24 weeks after the last immunization. Crossreactive antibody binding titers against diverse HIV envelopes and antigens and specific CD4i epitopes on gp120 were assessed. Results Sixty-five volunteers were enrolled—49 vaccine and 16 placebo. Majority (81%) of vaccinations with IHV01 produced no localized or systemic reactions; no different from the control group. The overall incidence of adverse events (AEs) was not significantly different between groups. Majority (89%) of vaccine-related AEs were mild in severity. The most common vaccine-related AEs were injection site pain (31%), pruritus (10%), and headache (10%). There were no vaccine-related serious AE, discontinuation due to AE, or intercurrent HIV infection. By the final vaccination, all subjects in all cohorts had developed antibodies against IHV01; all placebo recipients were negative. The antibodies induced by IHV01 reacted with envelope antigens from diverse HIV-1 strains (Figure 2). Conclusion IHV01 vaccine was safe, well tolerated, and immunogenic in all doses tested. The vaccine raised broadly reactive humoral responses against multiple gp120 domains, transition state structures, and CD4i epitopes. Disclosures All Authors: No reported Disclosures.

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Vaccination with Recombinant Cryptococcus Proteins in Glucan Particles Protects Mice against Cryptococcosis in a Manner Dependent upon Mouse Strain and Cryptococcal Species

mBio ◽

10.1128/mbio.01872-17 ◽

2017 ◽

Vol 8 (6) ◽

Cited By ~ 27

Author(s):

Charles A. Specht ◽

Chrono K. Lee ◽

Haibin Huang ◽

Maureen M. Hester ◽

Jianhua Liu ◽

...

Keyword(s):

Cryptococcus Neoformans ◽

Mouse Strain ◽

Subunit Vaccine ◽

Vaccine Development ◽

Inflammatory Responses ◽

Cryptococcus Gattii ◽

Candidate Vaccine ◽

Effective Vaccine ◽

Mouse Strains ◽

Vaccine Antigens

ABSTRACT Development of a vaccine to protect against cryptococcosis is a priority given the enormous global burden of disease in at-risk individuals. Using glucan particles (GPs) as a delivery system, we previously demonstrated that mice vaccinated with crude Cryptococcus -derived alkaline extracts were protected against lethal challenge with Cryptococcus neoformans and Cryptococcus gattii . The goal of the present study was to identify protective protein antigens that could be used in a subunit vaccine. Using biased and unbiased approaches, six candidate antigens (Cda1, Cda2, Cda3, Fpd1, MP88, and Sod1) were selected, recombinantly expressed in Escherichia coli , purified, and loaded into GPs. Three mouse strains (C57BL/6, BALB/c, and DR4) were then vaccinated with the antigen-laden GPs, following which they received a pulmonary challenge with virulent C. neoformans and C. gattii strains. Four candidate vaccines (GP-Cda1, GP-Cda2, GP-Cda3, and GP-Sod1) afforded a significant survival advantage in at least one mouse model; some vaccine combinations provided added protection over that seen with either antigen alone. Vaccine-mediated protection against C. neoformans did not necessarily predict protection against C. gattii . Vaccinated mice developed pulmonary inflammatory responses that effectively contained the infection; many surviving mice developed sterilizing immunity. Predicted T helper cell epitopes differed between mouse strains and in the degree to which they matched epitopes predicted in humans. Thus, we have discovered cryptococcal proteins that make promising candidate vaccine antigens. Protection varied depending on the mouse strain and cryptococcal species, suggesting that a successful human subunit vaccine will need to contain multiple antigens, including ones that are species specific. IMPORTANCE The encapsulated fungi Cryptococcus neoformans and Cryptococcus gattii are responsible for nearly 200,000 deaths annually, mostly in immunocompromised individuals. An effective vaccine could substantially reduce the burden of cryptococcosis. However, a major gap in cryptococcal vaccine development has been the discovery of protective antigens to use in vaccines. Here, six cryptococcal proteins with potential as vaccine antigens were expressed recombinantly and purified. Mice were then vaccinated with glucan particle preparations containing each antigen. Of the six candidate vaccines, four protected mice from a lethal cryptococcal challenge. However, the degree of protection varied as a function of mouse strain and cryptococcal species. These preclinical studies identify cryptococcal proteins that could serve as candidate vaccine antigens and provide a proof of principle regarding the feasibility of protein antigen-based vaccines to protect against cryptococcosis.

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Improving immunogenicity of influenza virus H7N9 recombinant hemagglutinin for vaccine development

Vaccine ◽

10.1016/j.vaccine.2018.09.034 ◽

2019 ◽

Vol 37 (13) ◽

pp. 1897-1903 ◽

Cited By ~ 1

Author(s):

Ting-Hui-Lin ◽

Min-Yuan Chia ◽

Chun-Yang Lin ◽

Yi-Qi Yeh ◽

U-Ser Jeng ◽

...

Keyword(s):

Influenza Virus ◽

Vaccine Development ◽

Recombinant Hemagglutinin

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Antigens ofPlasmodiwn falciparumblood stages with clinical interest cloned and expressed inE. coli

Parasitology ◽

10.1017/s0031182000085735 ◽

1986 ◽

Vol 92 (S1) ◽

pp. S119-S137 ◽

Cited By ~ 4

Author(s):

J. Scaife ◽

N. Bone ◽

M. Goman ◽

R. Hall ◽

I. A. Hope ◽

...

Keyword(s):

Subunit Vaccine ◽

Malaria Parasite ◽

Recombinant Dna ◽

Vaccine Development ◽

Human Malaria Parasite ◽

A Subunit ◽

Clinical Interest ◽

Recombinant Dna Techniques ◽

Species Specific ◽

Potential Use

The human malaria parasite,Plasmodium falciparum, is currently being actively studied by molecular biologists. It is hoped that the use of recombinant DNA techniques in this area will give new insights into the biology of the organism and, at the same time, provide new approaches to diagnosis and vaccine development.Our own studies employ the blood stages of the parasite and cover three main areas: enzymes of importance in parasite metabolism; antigens of potential use in a subunit vaccine; and repetitive DNA as a probe able to distinguish genetically different isolates ofP. falciparumand as a species-specific diagnostic tool in human and mosquito infections.

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Expression and Solubilization of Insect Cell-Based Rabies Virus Glycoprotein and Assessment of Its Immunogenicity and Protective Efficacy in Mice

Clinical and Vaccine Immunology ◽

10.1128/cvi.05258-11 ◽

2011 ◽

Vol 18 (10) ◽

pp. 1673-1679 ◽

Cited By ~ 15

Author(s):

R. Ramya ◽

B. Mohana Subramanian ◽

V. Sivakumar ◽

R. L. Senthilkumar ◽

K. R. S. Sambasiva Rao ◽

...

Keyword(s):

Rabies Virus ◽

Insect Cell ◽

Neutralizing Antibodies ◽

Subunit Vaccine ◽

Vaccine Development ◽

Protective Efficacy ◽

Vector System ◽

Soluble Form ◽

Rabies Virus Glycoprotein ◽

Virus Glycoprotein

ABSTRACTRabies is a fatal zoonotic disease of serious public health and economic significance worldwide. The rabies virus glycoprotein (RVG) has been the major target for subunit vaccine development, since it harbors domains responsible for induction of virus-neutralizing antibodies, infectivity, and neurovirulence. The glycoprotein (G) was cloned using the baculovirus expression vector system (BEVS) and expressed inSpodoptera frugiperda(Sf-9) cells. In order to obtain a soluble form of G suitable for experimentation in mice, 18 different combinations of buffers and detergents were evaluated for their ability to solubilize the insect cell membrane-associated G. The combination that involved 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulfonate (CHAPS) detergent in lysis buffer 1, formulated with Tris, NaCl, 10% dimethyl sulfoxide (DMSO), and EDTA, gave the highest yield of soluble G, as evidenced by the experimental data. Subsequently, several other parameters, such as the concentration of CHAPS and the duration and temperature of the treatment for the effective solubilization of G, were optimized. The CHAPS detergent, buffered at a concentration of 0.4% to 0.7% (wt/vol) at room temperature (23 to 25°C) for 30 min to 1 h using buffer 1, containing 10% DMSO, resulted in consistently high yields. The G solubilized using CHAPS detergent was found to be immunogenic when tested in mice, as evidenced by high virus-neutralizing antibody titers in sera and 100% protection upon virulent intracerebral challenge with the challenge virus standard (CVS) strain of rabies virus. The results of the mice study indicated that G solubilized with CHAPS detergent retained the immunologically relevant domains in the native conformation, thereby paving the way for producing a cell-free and efficacious subunit vaccine.

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Advances and challenges in malaria vaccine development

Expert Reviews in Molecular Medicine ◽

10.1017/s1462399409001318 ◽

2009 ◽

Vol 11 ◽

Cited By ~ 15

Author(s):

Ruobing Wang ◽

Joseph D. Smith ◽

Stefan H.I. Kappe

Keyword(s):

Life Cycle ◽

Subunit Vaccine ◽

Malaria Vaccine ◽

Vaccine Development ◽

Control Strategies ◽

Complex Life Cycle ◽

Mosquito Vector ◽

Specific Stage ◽

Specific Species ◽

Induced Immunity

Malaria remains one of the most devastating infectious diseases that threaten humankind. Human malaria is caused by five different species ofPlasmodiumparasites, each transmitted by the bite of femaleAnophelesmosquitoes. Plasmodia are eukaryotic protozoans with more than 5000 genes and a complex life cycle that takes place in the mosquito vector and the human host. The life cycle can be divided into pre-erythrocytic stages, erythrocytic stages and mosquito stages. Malaria vaccine research and development faces formidable obstacles because many vaccine candidates will probably only be effective in a specific species at a specific stage. In addition,Plasmodiumactively subverts and escapes immune responses, possibly foiling vaccine-induced immunity. Although early successful vaccinations with irradiated, live-attenuated malaria parasites suggested that a vaccine is possible, until recently, most efforts have focused on subunit vaccine approaches. Blood-stage vaccines remain a primary research focus, but real progress is evident in the development of a partially efficacious recombinant pre-erythrocytic subunit vaccine and a live-attenuated sporozoite vaccine. It is unlikely that partially effective vaccines will eliminate malaria; however, they might prove useful in combination with existing control strategies. Elimination of malaria will probably ultimately depend on the development of highly effective vaccines.

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Cloning, expression and purification of the outer membrane protein28 of Salmonella enterica serovar Typhimurium for subunit vaccine development - a short communication

Veterinarski arhiv ◽

10.24099/vet.arhiv.170526 ◽

2018 ◽

Vol 88 (4) ◽

pp. 559-568

Author(s):

Mamta Pandey ◽

◽

Mumtesh K. Saxena ◽

Anjani Saxena ◽

Richa Jha ◽

...

Keyword(s):

Outer Membrane ◽

Salmonella Enterica ◽

Subunit Vaccine ◽

Short Communication ◽

Salmonella Enterica Serovar Typhimurium ◽

Vaccine Development ◽

Expression And Purification ◽

Serovar Typhimurium

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Mycoplasma suis Alpha-Enolase Subunit Vaccine Induces an Immune Response in Experimental Animals

Vaccines ◽

10.3390/vaccines9121506 ◽

2021 ◽

Vol 9 (12) ◽

pp. 1506

Author(s):

Shujiang Xue ◽

Kangseok Seo ◽

Miaosen Yang ◽

Chengdu Cui ◽

Meng Yang ◽

...

Keyword(s):

Subunit Vaccine ◽

Vaccine Development ◽

Protective Efficacy ◽

Immunological Response ◽

Proliferation Index ◽

Protein Subunit ◽

Cd8 Cells ◽

Mycoplasma Suis ◽

T Lymphocyte Proliferation ◽

Ifn Γ

Recombinant protein technology has emerged as an excellent option for vaccine development. However, prior to our study, the immune induction ability of recombinant Mycoplasma suis alpha-enolase (rMseno) in animals remained unclear. The purpose of this study was to develop a rMseno protein subunit vaccine and to determine its ability to elicit an immunological response. To accomplish this, we cloned the gene into pET-15b, expressed it in BL21 cells, and purified it. Following the establishment of immunity, the immunogenicity and potential for protection of rMseno were evaluated in mice and piglets. The results demonstrate that anti-M. suis serum recognized the pure rMseno protein in both mice and piglets as evidenced by high levels of specific anti-rMseno antibodies, significantly increased levels of IFN-γ and IL-4 cytokines, and significantly increased T lymphocyte proliferation index. Piglets also had significantly increased levels of specific IgG1, IgG2a, CD4+, and CD8+ cells. The rMseno findings demonstrated a robust immunological response in mice and piglets, affording partial clinical protective efficacy in piglets.

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