We applied an immunoscreening technique, in vivo-induced antigen technology (IVIAT), to identify immunogenic bacterial proteins expressed during human infection with Salmonella enterica serovar Typhi, the cause of typhoid fever. We were able to assign a functional classification to 25 of 35 proteins identified by IVIAT. Of these 25, the majority represent proteins with known or potential roles in the pathogenesis of S. enterica. These include proteins implicated in fimbrial structure and biogenesis, antimicrobial resistance, heavy metal transport, bacterial adhesion, and extracytoplasmic substrate trafficking as well as secreted hydrolases. The 10 remaining antigens represent proteins with unknown functions. Of the 35 identified antigens, four had no immunoreactivity when probed with control sera from individuals never exposed to serovar Typhi organisms; these four included PagC, TcfB, and two antigens of unknown function encoded by STY0860 and STY3683. PagC is a virulence factor known to be upregulated in vivo in S. enterica serovar Typhimurium infection of mice. TcfB is the major structural subunit of a fimbrial operon found in serovar Typhi with no homolog in serovar Typhimurium organisms. By examining differential immunoreactivities in acute-versus convalescent-phase human serum samples, we found specific anti-PagC and anti-TcfB immunoglobulin G responses in patients with serovar Typhi bacteremia. Serovar Typhi antigens identified by IVIAT warrant further evaluation for their contributions to pathogenesis, and they may have diagnostic, therapeutic, or preventive uses.In humans, infection with Salmonella enterica serovar Typhi causes a spectrum of illness that includes diarrhea, a selflimited febrile illness, and most significantly, typhoid fever, a systemic infection characterized by persistent fever, lymphadenopathy, hepatosplenomegaly, encephalopathy, and intestinal hemorrhage and perforation (16). Worldwide, approximately 20 million cases of typhoid fever resulting in 200,000 deaths occur annually (2). The increasing incidence of disease caused by multidrug-resistant serovar Typhi organisms underscores the importance of developing novel approaches to the diagnosis, treatment, and prevention of typhoid fever (22).Although many S. enterica serovars infect a broad range of animal hosts and cause gastroenteritis in humans, serovar Typhi is among the few serovars for which natural infection appears to be limited to human hosts. In humans, serovar Typhi organisms penetrate the gastrointestinal epithelial barrier and infect phagocytes within the lamina propria. However, unlike organisms from other broad-host-range serovars, serovar Typhi organisms are adapted for prolonged intracellular survival in human macrophages, allowing the bacteria to spread to reticuloendothelial organs, including the liver, spleen, and bone marrow (16).Because serovar Typhi organisms are human specific, there is no optimal animal model of serovar Typhi infection, and presumed factors that contribute to the pathogenesis and immunology of...