[PMC free article] [PubMed] [Google Scholar] 18. Avatrombopag been isolated from humans include (17), subsp. (49), subsp. (41), and (27). infections are associated with arthropod vector transmission; e.g., is usually transmitted by sand flies, by cat fleas, and by the human body louse. There is recent evidence of association with ticks, fleas, and flies (10, 14, 15, 46). Furthermore, several distinct strains have been isolated from various rodent (5, 7, 8, 22, 23, 29) and ruminant (19, 6, 13) species throughout the world. Because of the ubiquitous nature of and their association with arthropod vectors, this genus has been implicated as a potential causative agent in diverse disease manifestations seen in humans for which there have been no definitive clinical diagnoses. Our laboratory was interested in exploring whether the prevalence of residing in wild-mammal populations may Avatrombopag have had an association with unexplained human febrile illnesses in the North American Southwest, as there was some serological evidence suggesting such a link (M. Y. Kosoy et al., Abstr. Am. Soc. Rickettsiol. Bartonella Emerg. Pathog. Group 2001 Joint Conf., abstr. 108, 2001; F. Koster et al., Abstr. Am. Soc. Rickettsiol. Bartonella Emerg. Pathog. Group 2001 Joint Conf., abstr. 133, 2001). Moreover, a (in human infections and, as an initial step, to identify species-specific antigens that elicit antibody responses during contamination. The goal was to initiate a primary analysis of immunodominant antigens related to rodent-borne and to further develop a more detailed comparative analysis Avatrombopag of immunogenic proteins from human pathogens to define genus-specific antigens versus species-specific antigens. As a result of these aims, this report describes a novel immunodominant, surface-associated protein from subsp. encoded by one of the largest bacterial genes yet identified. Additionally, at least two more related genes are present upstream, forming a multigene family. The genes in this family are characterized by multiple internal repetitive regions that are conserved within and among the individual genes, which we have therefore termed (gene complex. Mice were infected with live strains isolated from rodent species to generate antibodies that could be used to clone and characterize genes encoding proteins associated with contamination. Strains na19103nm (isolated from [wood rat] in New Mexico), Pm15590co and Pm136co (isolated from [deer mouse] in Colorado), and Sb944nv (isolated from [California ground squirrel] in Nevada) were obtained from Michael Kosoy, Division of Vector-Borne Infectious Diseases, Centers for Disease Control and Prevention, Fort Collins, Colorado. subsp. subsp. were purchased from the American Type Culture Collection, Manassas, Va. strains were cultivated on Trypticase soy agar supplemented with 5% sheep blood. Plates were incubated at 37C in 5% CO2 until growth was visible, usually between 5 and 10 days. was incubated at 26C under ambient atmosphere. Antiserum was generated from mice infected with as follows. Female BALB/cJ mice (The Jackson Laboratory, Bar Harbor, Maine) were inoculated intraperitoneally with live strain Pm15590co resuspended in 0.5 ml of phosphate-buffered saline (PBS) following harvesting from a plate culture. A booster injection with live Pm15590co was given 5 weeks after the primary injection, followed by a third booster 6 weeks later with strain Sb944nv. The mice were bled 3 weeks later and provided a source of anti-polyclonal antiserum. Polyclonal antibodies against each of strains Sb944nv and Pm136co were generated in a similar fashion. The rodent-associated strains of were used in this initial experiment because there was an interest in identifying specific antigens related to these types of infections. The progression from initially obtaining an immunoreactive clone from a rodent strain genomic library to identifying the gene complex in Hpse subsp. is usually summarized in Fig. ?Fig.1.1. A positive expression clone, termed 6.1, was identified from the strain na19203nm genomic library using the mouse antiserum described in the previous paragraph. Clone 6.1 contained a 1.5-kb insert and expressed a recombinant protein product of 58 kDa.