After this period, surviving young infected mice regained weight. The number of estimated cases of infection (CDI) in people aged 65 years made up 57% of the total estimated CDI cases in 2011, but the same study found that 83% of total estimated deaths from CDI were from this same age group. In a separate review of the death registry from 2008, 92% of deaths from CDI occurred in people aged 65 years [4]. In a study of 336 patients with stool specimens positive for in Brigham and Women’s Hospital between 2004 and 2005, the odds ratio for severe disease was 3.35 for subjects aged 70 years [5]. Even when the analysis controlled for comorbidities by comparing patients with the same Charles comorbidity index, older patients did worse, although the difference was less notable when accounting KRT13 antibody for higher comorbidity scores [6]. There are a number of different possible etiologies for the higher susceptibility of elderly individuals to CDI. Immunosenescence, or the effect of aging on the immune system, may potentially affect the outcome of CDI. Aging is known to cause dysregulation of the innate immune system [7], as well as the adaptive PD173955 immune response, including T-cell activity [8C11]. Both the percentage and number of naive human B cells in blood decrease with age [12, 13]. There is also an age-related contraction of the B-cell repertoire with age that correlates with poor health status [14]. The most significant change with aging in humoral immunity is impairment in the ability to produce high-affinity immunoglobulins and undergo immunoglobulin class switching, which allows the production of high-affinity immunoglobulin G (IgG), immunoglobulin A (IgA), and immunoglobulin E (IgE) [15]. Serum IgA levels are elevated in elderly subjects as compared with younger controls, but these IgA antibodies are predominantly monomeric and are therefore not transported to the mucosal surface as secretory IgA [16]. These findings may be associated with the PD173955 decrease in specific antibody responses in humans vaccinated against tetanus toxoid, encephalitis viruses, organisms, toxins A and B have been shown to be an important part of host immune response against CDI, which can be affected by aging [17, 18]. Furthermore, intestinal dysbiosis is a key element in the pathogenesis of CDI. Previous studies analyzing bacterial populations in the gut indicated alterations of the gut microbiota with aging [19, 20]. These findings suggest that aging may affect the intestinal microbiota in ways that make patients more susceptible to CDI. Dissection of the underlying mechanisms of the increased PD173955 susceptibility to and worse outcome of CDI among aging persons is challenging because of the complexity of the immune response in aging hosts. Multiple interacting factors, such as comorbidities, polypharmacy, and healthcare exposure, in addition to biological factors, may contribute to the observed outcomes of infection in the elderly population. In this study, we aimed to simulate CDI in elderly individuals, using an aged C57/BL mouse model, and to investigate the biological reasons behind severe disease and poor outcomes with aging. METHODS Murine Model of Infection and Treatment The infection model is a modification of the protocol published by Chen et al [21]. This protocol has been approved by the Center for Comparative Medicine at the University of Virginia. Male C57BL/6 mice (Charles River), 2 and 18 months old, were used in all experiments. Mice were pretreated with an antibiotic cocktail (vancomycin [0.0045 mg/g], colistin [0.0042 mg/g], gentamicin [0.0035 mg/g], and metronidazole [0.0215 mg/g] in drinking water for 3 days, followed by clindamycin [32 mg/kg] intraperitoneally 1 day prior to infection) as previously described [22]. Food and water were allowed ad libitum. Although each mouse or treatment group was housed in a separate cage, all mice were housed in the same biosafety level 2 containment pod in the vivarium. Infection was performed with VPI 10463 (ATCC) in an inoculum of 104 or 105 administered by oral gavage. One set of.