Evaluation of the Proline-Rich Antigen of
            <i>Coccidioides immitis</i>
            as a Vaccine Candidate in Mice

Kirkland, Theo N.; Finley, Fred; Orsborn, Kris I.; Galgiani, John N.

doi:10.1128/iai.66.8.3519-3522.1998

Cited by 62 publications

(23 citation statements)

References 29 publications

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“…2a). This kind of discrepancy is very often observed in proline-rich proteins expressed in E. coli, due to presumably increased rigidity caused by high proline content, leading to slower migration than the same molecular weight protein [38][39][40][41]. Abaecin (3.9 kDa, 34 amino acids) contains 10 prolines in total of 34 amino acids, accounting for 34.9%, so it is assumed that the high content of proline residues affected the migration of all three fusion proteins (136 amino acids in length for each construct).…”

Section: Evaluation Of Expression Of His-tagged Sumo Fused Abaecin Inmentioning

confidence: 99%

A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag

Kim

Song

et al. 2019

BMC Biotechnol

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Background Despite the growing demand for antimicrobial peptides (AMPs) for clinical use as an alternative approach against antibiotic-resistant bacteria, the manufacture of AMPs relies on expensive, small-scale chemical methods. The small ubiquitin-related modifier (SUMO) tag is industrially practical for increasing the yield of recombinant proteins by increasing solubility and preventing degradation in expression systems. Results A new vector system, pKSEC1, was designed to produce AMPs, which can work in prokaryotic systems such as Escherichia coli and plant chloroplasts. 6xHis was tagged to SUMO for purification of SUMO-fused AMPs. Abaecin, a 34-aa-long antimicrobial peptide from honeybees, was expressed in a fusion form to 6xHis-SUMO in a new vector system to evaluate the prokaryotic expression platform of the antimicrobial peptides. The fusion sequences were codon-optimized in three different combinations and expressed in E. coli . The combination of the native SUMO sequence with codon-optimized abaecin showed the highest expression level among the three combinations, and most of the expressed fusion proteins were detected in soluble fractions. Cleavage of the SUMO tag by sumoase produced a 29-aa-long abaecin derivative with a C-terminal deletion. However, this abaecin derivative still retained the binding sequence for its target protein, DnaK. Antibacterial activity of the 29-aa long abaecin was tested against Bacillus subtilis alone or in combination with cecropin B. The combined treatment of the abaecin derivative and cecropin B showed bacteriolytic activity 2 to 3 times greater than that of abaecin alone. Conclusions Using a SUMO-tag with an appropriate codon-optimization strategy could be an approach for the production of antimicrobial peptides in E.coli without affecting the viability of the host cell. Electronic supplementary material The online version of this article (10.1186/s12896-019-0506-x) contains supplementary material, which is available to authorized users.

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Section: Evaluation Of Expression Of His-tagged Sumo Fused Abaecin Inmentioning

confidence: 99%

A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag

Kim

Song

et al. 2019

BMC Biotechnol

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“…68 With advances in laboratory technology, vaccine preparations for prevention of coccidioidomycosis have focused on subcellular and more recently to specifically cloned antigens, as well as improved ways to present the antigens, the use of engineered chimeric proteins and different adjuvants, many of which have shown some protective activity. 34,56,[69][70][71][72][73][74][75][76][77][78][79][80][81][82][83] These studies have primarily used mouse models of infection and have been performed by pretreatment of the mice with the vaccine preparation, usually multiple doses given subcutaneously, intramuscularly, or intraperitoneally, and challenged by intraperitoneal or pulmonary route 2 to 4 weeks later. Evaluation of protection is then based on prolonged survival and in some studies on the number of CFUs of Coccidioides remaining in the target organs (e.g., lungs).…”

Section: Vaccine Studiesmentioning

confidence: 99%

Experimental Animal Models of Coccidioidomycosis

Clemons¹,

Capilla²,

Stevens³

2007

Annals of the New York Academy of Sciences

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Experimental models of coccidioidomycosis performed using various laboratory animals have been, and remain, a critical component of elucidation and understanding of the pathogenesis and host resistance to infection with Coccidioides spp., as well as to development of more efficacious antifungal therapies. The general availability of genetically defined strains, immunological reagents, ease of handling, and costs all contribute to the use of mice as the primary laboratory animal species for models of this disease. Five types of murine models are studied and include primary pulmonary disease, intraperitoneal with dissemination, intravenous infection emulating systemic disease, and intracranial or intrathecal infection emulating meningeal disease. Each of these models has been used to examine various aspects of host resistance, pathogenesis, or antifungal therapy. Other rodent species, such as rat, have been used much less frequently. A rabbit model of meningeal disease, established by intracisternal infection, has proven to model human meningitis well. This model is useful in studies of host response, as well as in therapy studies. A variety of other animal species including dogs, primates, and guinea pigs have been used to study host response and vaccine efficacy. However, cost and increased needs of animal care and husbandry are limitations that influence the use of the larger animal species.

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“…The use of Th1 stimulators has demonstrated an increase in the protective effect of vaccines in animal models. For example, unmethylated CpG oligonucleotides in combination with A. fumigatus allergens [361], or antigens from C. immitis [362] stimulated Th1 response increasing resistance. However, the use of CpG increased the susceptibility of mice to challenge with C. albicans [363].…”

Section: Vaccinesmentioning

confidence: 99%

Animal models: an important tool in mycology

2007

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Animal models of fungal infections are, and will remain, a key tool in the advancement of the medical mycology. Many different types of animal models of fungal infection have been developed, with murine models the most frequently used, for studies of pathogenesis, virulence, immunology, diagnosis, and therapy. The ability to control numerous variables in performing the model allows us to mimic human disease states and quantitatively monitor the course of the disease. However, no single model can answer all questions and different animal species or different routes of infection can show somewhat different results. Thus, the choice of which animal model to use must be made carefully, addressing issues of the type of human disease to mimic, the parameters to follow and collection of the appropriate data to answer those questions being asked. This review addresses a variety of uses for animal models in medical mycology. It focuses on the most clinically important diseases affecting humans and cites various examples of the different types of studies that have been performed. Overall, animal models of fungal infection will continue to be valuable tools in addressing questions concerning fungal infections and contribute to our deeper understanding of how these infections occur, progress and can be controlled and eliminated.

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Evaluation of the Proline-Rich Antigen of Coccidioides immitis as a Vaccine Candidate in Mice

Cited by 62 publications

References 29 publications

A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag

A new prokaryotic expression vector for the expression of antimicrobial peptide abaecin using SUMO fusion tag

Experimental Animal Models of Coccidioidomycosis

Animal models: an important tool in mycology

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