Hum Vaccin 6:54C77

Hum Vaccin 6:54C77. effectively prevented parasites from developing within the mosquito vector in standard membrane feeding assays. Alga-produced Pfs25 in Prostratin combination with a human-compatible adjuvant composed of a TLR-4 agonist in a squaleneCoil-in-water emulsion is an attractive new vaccine candidate that merits head-to-head comparison with other modalities of vaccine production and administration. INTRODUCTION A vaccine to prevent gametocytemic humans from infecting mosquitoes by inducing antibodies against sexual stage antigens, termed a transmission-blocking vaccine (TBV), was first demonstrated experimentally as proof of principle using a chicken model of more than 30 years ago (1, 2). TBV development has been stated to be a key priority toward achieving the global goals of malaria control and elimination (3, 4), yet it lags behind other malaria vaccine development despite candidate proteins such as Pfs25 being deeply studied as a TBV. Pfs25, the first molecularly cloned sexual stage protein, was identified more than 25 years ago and remains a lead TBV candidate (5, 6). Parasite-produced Pfs25 contains 4 epidermal growth factor (EGF)-like domains and is not glycosylated, which makes it biotechnologically challenging to produce the properly folded, conformationally correct recombinant protein(s) required for the induction of effective transmission-blocking antibodies (7,C9). Heterologous expression systems used to produce Pfs25 as a recombinant subunit immunogen include (10), (11), (7, 12), baculovirus (13), (14), (wheat germ) extract (6, 15, 16), and most recently, the chloroplast of the microalga (17, 18). The microalga has previously been reported to produce nonglycosylated, immunogenic recombinant Pfs25 that induces transmission-blocking antibodies in combination with complete Freund’s adjuvant (17) and mucosal IgA antibodies when fused with cholera toxin beta subunit (CtxB) and administered orally (18). Production in algal chloroplasts is advantageous from an economic and biological perspective. The biotechnological production and scale-up of recombinant proteins in is remarkably inexpensive, requiring just light and simple chemical nutrients. Recombinant algae can be grown in transparent plastic bags as containment, and standardized scalable methods for processing the biological materials are available (19). The chloroplast lacks the machinery to glycosylate proteins, a distinct advantage for production of Pfs25, and contains nucleus-encoded chaperones that likely assist in folding complex EGF-like domains (20). A major issue that has arisen previously in designing a Pfs25-based vaccine is the need for a strong yet human-use-compatible adjuvant to overcome the inherent lack of antigenicity of Pfs25; in a phase I clinical trial, erythema nodosum associated with the Montanide ISA 51 oil-in-water adjuvant was observed in a small number of subjects, which led to cessation of that vaccine formulation (21). New adjuvants based on squaleneCoil-in-water emulsion in the context of Prostratin a nontoxic lipid A-like moiety, glucopyranosyl lipid A (GLA), have been developed in extensive animal/experimentation (22,C25) and used in human experimentation (26), and they have been demonstrated to have the desired effects of low reactogenicity, induction of high Prostratin levels of effector antibodies, and optimized affinity maturation. The purpose of the present study was to assess the ability of macrogamete and zygote Pfs25 were assessed in response to vaccination of mice with chloroplast-produced Pfs25 combined with alum, alum with GLA, squaleneCoil-in-water emulsion alone, and GLA plus squaleneCoil-in-water emulsion. Conventional and avidity enzyme-linked immunosorbent assay (ELISA), immunofluorescence assays, and standard membrane feeding assays (SMFAs) were used to measure the function of the vaccine-induced antibodies. This work is an important new LATS1 step along a promising pathway for transmission-blocking vaccine development for deployment in humans. MATERIALS AND METHODS Ethics statement. All procedures were approved by the University of CaliforniaSan Diego (UCSD) Institutional Animal Care and Use Committee under protocol number S09277, in compliance with the USDA Animal Welfare Act Prostratin (Public Law 89-544), the Health.