Coccidioides spp. 5500 The tiny spores (2 × 4 μm) convert into multinucleate round cells (spherules) within the lungs of the host and undergo isotropic growth to produce large parasitic cells (60 to > 100 μm in diameter). The latter undergo an elaborate process of wall growth and cytoplasmic compartmentalization to form and release a multitude of endospores (4–10 μm in diameter). Each endospore grows and differentiates into a second-generation spherule, which is again able to yield an average of 200 to 300 endospores. Mature spherules most likely escape phagocytosis simply because they are too large to be ingested by neutrophils, macrophages, and dendritic cells (Hung et al., 2007). Coccidioidomycosis (California disease, Desert rheumatism, San Joaquin valley fever, and Valley fev Both clinical and experimental evidence have demonstrated that T-cell immunity is pivotal for defense against this respiratory disease. The ability of the host to elicit a strong delayed-type hypersensitivity response to the pathogen is essential (Tarcha et al., 2006). Coccidiomycosis is mainly a disease of immunocompromised in humans, but mice and monkeys have been used as animal models of disease (Ivey et al., 2003). Coccidioidomycosis is a human respiratory disease caused by inhalation of airborne spores produced by Coccidioides spp (Coccidioides immitis and C. posadasii). The pathogen is an anamorphic (asexual) ascomycetous fungus which resides in alkaline desert and semidesert soil in regions of the southwestern United States. The major areas where this disease is endemic include some of the most rapidly growing communities of Arizona and southern California. People who are frequently exposed to dust from the soil in these regions have a high chance of infection with Coccidioides. Recurrent epidemics of coccidioidomycosis appear to be correlated with climatic conditions. Periods of rain in the spring enhance growth of the soilborne saprobic phase of Coccidioides, which results in “blooms” of spores that are air dispersed during the late fall and winter seasons. Risk factors known to contribute to symptomatic coccidioidal infection include pregnancy (third trimester), immunosuppression, age (>65 years), and racial or geographic origin. Antifungal drug therapy for coccidioidomycosis (e.g., amphotericin B, fluconazole, or intraconazole) is typically continued for many months to years. Persons who contract coccidioidal meningitis require lifelong therapy. Coccidioides infection can be life threatening and, at the very least, is responsible for high medical costs incurred by patients as a result of hospitalization, clinic visits, lost wages, and long-term drug therapy (Tarcha et al., 2006). Baboon Papio cynocephalus 9556 Bank vole Clethrionomys glareolus 447135 Bear Ursus americanus 9643 Birds Passeroidea 175121 Brown Trout Salmo trutta 8032 Buffalo Bison bison 9901 Carnivores Vulpes 9625 Cat Felis catus 9685 Catfishes Siluriformes 7995 Cattle Bos taurus 9913 Chicken Gallus gallus 9031 Chimpanzee Pan troglodytes 9598 chinchillas Chinchillidae 10150 Copper Pheasant Syrmaticus soemmerringii 9067 Deer Cervus elaphus 9860 Deer mouse Peromyscus maniculatus 10042 Dog Canis familiaris 9615 Ducks Anas 8835 Ferret Mustela putorius furo 9669 Fish Hyperotreti 117565 Gerbil Gerbillina 10045 Goat Capra hircus 9925 Gray wolf Canis lupus 9612 Guinea pig Cavia porcellus 10141 Hamster Mesocricetus auratus 10036 Horse Equus caballus 9796 Human Homo sapiens 9606 Macaque Macaca fascicularis 9541 Mongolian Gerbil Meriones unguiculatus 10047 Monkey Platyrrhini 9479 Mouse Mus musculus 10090 None None Parrot Psittacidae 9224 Pig Sus scrofa 9823 Rabbit Oryctolagus cuniculus 9986 Rainbow trout Oncorhynchus mykiss 8022 Rat Rattus 10114 Raven Corvus corax 56781 sei whale Balaenoptera borealis 9768 Sheep Ovis aries 9940 Squirrel Spermophilus richardsonii 37591 Tree shrew Tupaiidae 9393 Trouts, salmons & chars Salmoninae 504568 Turkey Meleagris gallopavo 9103 Vole Microtus ochrogaster 79684 Water buffalo Bubalus bubalis 391902 C. immitis Subunit URE Protein Vaccine VO_0011463 Subunit vaccine Research Subcutaneous injection CpG ODN (Li et al., 2001) Subcutaneous injection Recombinant protein preparation BALB/c Mice each were immunized s.c. with recombinant protein (15 μg of rURE) or bovine serum albumin for control group (Li et al., 2001). The rURE-immune mice showed significantly lower counts of C. immitis in both lungs and spleens compared to control mice immunized with BSA plus CpG ODN (Li et al., 2001). Mice were challenged by the i.p. route at 14 days after the last protein immunization or 30 days after the last DNA immunization. The inoculum contained 100 viable arthroconidia in 100 μl of PBS (Li et al., 2001). C. posadasii DNA Vaccine encoding PRA Protein VO_0011461 DNA vaccine Research pVR1012 [Ref1392:Awasthi et al., 2005] Intramuscular injection (i.m.) Intramuscular injection (i.m.) DNA vaccine construction C57BL6 C57BL6 mice were anesthetized by i.m. injection of ketamine-xylazine (75 µg/g and 10 µg/g body weight, respectively). Transfected DCs (1–1.5 x 10^6 per 30 µl) were intranasally administered in both nares alternately. Comparative controls were Ag2/PRA plasmid DNA alone, nontransfected cells, and cells transfected with the vector plasmid alone. The immunization was performed twice at an interval of 1 wk (on days 0 and 7). The second immunization was given to boost the immune response (Awasthi et al., 2005). Upon necropsy after 10 days of infection, fungal burden in lung and spleen of immunized mice was significantly reduced as compared with the control animals. The lung tissue homogenates of immunized animals showed higher levels of IFN-gamma. Histologically, lung tissues of immunized mice were in better condition than the control mice (Awasthi et al., 2005). The immunized mice were infected with C. posadasii either i.p. or intranasally. Intranasal challenge was given on day 19 with 30 arthroconidia per 30 µl of saline, whereas the i.p. challenge was given on day 14 with 2500 arthroconidia per 500 µl of saline. The infected mice become symptomatic around day 9–10 and start dying on day 11. Therefore, the mice were sacrificed on 10th day of infection, and their lungs and spleens were collected (Awasthi et al., 2005). C. posadasii Subunit GEL1 Protein Vaccine VO_0011490 Subunit vaccine Research Intraperitoneal injection (i.p.) CpG ODN (Integrated DNA Technologies, Inc., Coralville, Iowa) (Delgado et al., 2003). Intraperitoneal injection (i.p.) Recombinant protein preparation BALB/c, C57BL/6 Mice were immunized subcutaneously (s.c.) with rGel1p plus adjuvant. The mice were then boosted by s.c. immunization 14 days later with the same amount of immunogen plus adjuvant (Delgado et al., 2003). rGel1p-immune mice infected with the pathogen showed a significant reduction in fungal burden and increased survival compared to nonimmune mice (Delgado et al., 2003). The animals were subsequently challenged with 100 viable arthroconidia by the i.p. route 2 weeks after the last immunization and then sacrificed 12 days later to determine the residual CFU in the lungs and spleen (Delgado et al., 2003). C. posadasii Subunit PMP1 Protein Vaccine VO_0011460 Subunit vaccine Research Intramuscular injection (i.m.) Monophosphoryl lipid A-stable emulsion (MPL-SE) adjuvant (Orsborn et al., 2006). Intramuscular injection (i.m.) Recombinant protein preparation C57BL/6 Mice were immunized subcutaneously with 1, 5, or 50 μg rPmp1 with monophosphoryl lipid A-stable emulsion (MPL-SE) adjuvant on day 0 and day 14. Control mice received adjuvant only (Orsborn et al., 2006). Vaccination with PMP1 elicited significant protection in mice and a reduction in colonization of organs by Coccidioides (Orsborn et al., 2006). Four weeks after boosting (day 42), the mice were infected intraperitoneally with 310 arthroconidia and sacrificed 2 weeks later (Orsborn et al., 2006). licensed Coccidioidomycosis human vaccine Generic Unknown VO_0012179 Inactivated or "killed" vaccine Licensed A generic representation of vaccines utilized to prevent coccidioidomycosis in humans, typically consisting of inactivated (killed) Coccidioides organisms to stimulate protective immunity without causing disease. These vaccines were historically developed and licensed for human use against Valley fever. GEL1 Coccidioides posadasii VO_0011330 14582416 CDD:304882 199306 ? glucanosyltransferase 8.01 15771.51 231 Glycosyl hydrolase family 1; cl23725 >AAK69492.1 glucanosyltransferase, partial [Coccidioides posadasii] SGGLVYEYPQEPSNYGLVQLGKGKPKELDDFKALAKAFKGTKNPSGDGGYNSTGGANPCPKKNAPNWDVD SESLPAIPEPAKKFMKDGPGEGPGLSGKGSQNAGTQSSGTATPGSGSVDPTSSAGAAAGLRPEFGIAPMM CAMLVVLSSMFGASFIFMV Protective antigen The GEL1 mRNA of C. posadasii was detected at the highest level during the endosporulation stage of the parasitic cycle, and the mature protein was immunolocalized to the surface of endospores. BALB/c or C57BL/6 mice were immunized subcutaneously with the bacterium-expressed recombinant protein (rGel1p) to evaluate its protective efficacy against a lethal challenge of C. posadasii by either the intraperitoneal or intranasal route. In both cases, rGel1p-immune mice infected with the pathogen showed a significant reduction in fungal burden and increased survival compared to nonimmune mice [Ref1389:Delgado et al., 2003]. pmp1 Coccidioides posadasii VO_0011332 78364922 CDD:239311 199306 ? peroxisomal matrix protein 5.21 16731.65 234 Peroxiredoxin (PRX) family, PRX5-like subfamily; members are similar to the human protein, PRX5, a homodimeric TRX peroxidase, widely expressed in tissues and found cellularly in mitochondria, peroxisomes and the cytosol. The cellular location of PRX5...; cd03013 >ABB42829.1 peroxisomal matrix protein [Coccidioides posadasii] MASLKAGDSFLSDVVFSYIPWTPDNKDIKACGMPQNYEASKLWADKKVVLFSLPGAFTPTCSASHLPGYI QKLPQLKEKGVDVVAVLAFNDAWVMSAWGKANGVTGDDILFLSDPEAKFSKSIGWNAGERTGRYAMIIDH GQVTYAEIEPGREVTVSGADAVISKL Protective antigen Recombinant Pmp1 was reactive with serum from individuals with both acute and protracted disease, and evoked protection in two murine models of infection with C. posadasii [Ref1391:Orsborn et al., 2006]. pra Coccidioides immitis VO_0011333 42742528 CDD:128986 CDD:282710 5501 ? proline-rich antigen 3.93 18554.97 254 eight cysteine-containing domain present in fungal extracellular membrane proteins; smart00747 >AAS45282.1 proline-rich antigen [Coccidioides immitis] MQFSHALIALVAAGLASAQLPDIPPCALNCFVEALGNDGCTRLTDFKCHCSKPELPGQITPCVEEACPLD ARISVSNIVVDQCSKAGVPIEIPPVDTTAAPEPSETAEPTAEPTEEPTAEPTAEPTAEPTHEPTEEPTAV PTGTGGGVPTGTGSFTVTGRPTASTPAEFPGAGSNVRASVGGIAAALLGLAAYL Protective antigen C57BL6 mice were immunized with syngeneic, bone marrow-derived DCs (JAWS II cells) transfected with a cDNA encoding the protective Coccidioides-Ag2/proline-rich Ag (pra). The immunized mice were lethally challenged with C. posadasii through either an i.p. or intranasal route. Upon necropsy after 10 days of infection, fungal burden in lung and spleen of immunized mice was significantly reduced as compared with the control animals. The lung tissue homogenates of immunized animals showed higher levels of IFN-gamma. Histologically, lung tissues of immunized mice were in better condition than the control mice [Ref1392:Awasthi et al., 2005]. URE Coccidioides immitis VO_0011331 73808050 CDD:294200 5501 ? urease 4.18 15939.32 219 Superfamily of metallo-dependent hydrolases (also called amidohydrolase superfamily) is a large group of proteins that show conservation in their 3-dimensional fold (TIM barrel) and in details of their active site. The vast majority of the members have a...; cl00281 >BAE20261.1 urease, partial [Coccidioides immitis] CGIGKAGNPDVMDGVTPNMIVGSSTDVIACEGKIVTAGGIDTHVHFICPQQVEEALASGVTTLLGGGTGP TEGTNATTCTPAPNQFKTMMQACDHLPINVGLTGKGNDSGLPSLRDQCRAGAAGLKVHEDWGATPAVIDT CLQVCDEFDIQCLIHTDTLNESGF Protective antigen Recombinant urease (rURE) of C. immitis was expressed in Escherichia coli and tested asa vaccine candidate in BALB/c mice. BALB/c mice immunized subcutaneously with rURE and subsequently challenged by the intraperitoneal (i.p.) route with a lethal inoculum of C. immitis arthroconidia demonstrated a significant reduction in the level of C. immitis infection compared to control animals [Ref1390:Li et al., 2001]. Awasthi et al., 2005 journal Awasthi S, Awasthi V, Magee DM, Coalson JJ 2005 175 6 3900-3906 Journal of immunology (Baltimore, Md. : 1950) Delgado et al., 2003 journal Delgado N, Xue J, Yu JJ, Hung CY, Cole GT 2003 71 6 3010-3019 Infection and immunity Hung et al., 2007 journal Hung CY, Xue J, Cole GT 2007 1111 225-235 Annals of the New York Academy of Sciences Ivey et al., 2003 journal Ivey FD, Magee DM, Woitaske MD, Johnston SA, Cox RA 2003 21 27-30 4359-4367 Vaccine Kirkland et al., 1998 journal Kirkland TN, Thomas PW, Finley F, Cole GT 1998 66 2 424-431 Infection and immunity Li et al., 2001 journal Li K, Yu JJ, Hung CY, Lehmann PF, Cole GT 2001 69 5 2878-2887 Infection and immunity Orsborn et al., 2006 journal Orsborn KI, Shubitz LF, Peng T, Kellner EM, Orbach MJ, Haynes PA, Galgiani JN 2006 74 3 1865-1872 Infection and immunity Tarcha et al., 2006a journal Tarcha EJ, Basrur V, Hung CY, Gardner MJ, Cole GT 2006 74 1 516-527 Infection and immunity Tarcha et al., 2006b journal Tarcha EJ, Basrur V, Hung CY, Gardner MJ, Cole GT 2006 74 10 5802-5813 Infection and immunity