995. A Murine Model of Klebsiella pneumoniae Gastrointestinal Colonization with Parenteral Vancomycin Administration

Abstract Background As a leading cause of nosocomial infections, Klebsiella pneumoniae poses a significant threat due to its propensity to acquire resistance to many classes of antibiotics, including carbapenems. Gastrointestinal (GI) colonization by K. pneumoniae is a risk factor for subsequent infection as well as transmission to other patients. To study this crucial step in pathogenesis, we developed a mouse model of K. pneumoniae GI colonization using a clinically relevant parenteral antibiotic regimen. Methods To improve the clinical relevance of our model, we elected to use intraperitoneal injections of vancomycin, one of the most highly utilized antibiotics in the United States. Results To optimize dosage in C57bl/6 mice, we injected 20mg/kg, 350mg/kg, or vehicle (PBS) for three days prior to gastric gavage with 108 colony forming units (CFU) of a low-resistance strain of K. pneumoniae. The mice who received 350mg/kg (a mouse equivalent of a human dose of 1g/day calculated through the FDA guidelines for estimating safe dosing) shed about 107 CFU/g of feces at Day 7 while those receiving the lower dose or vehicle shed 104 CFU/g. Next, we compared 3- or 5-day pre-treatments with vancomycin prior to inoculation with an ST258 (epidemic carbapenem-resistant) strain. At Day 7 post-inoculation, mice who received 5 days shed 1010 CFU/g feces while those who received vancomycin for 3 days or vehicle for 5 days (PBS) shed 106 or 104 CFU/g feces respectively. Thus, we chose 5 days of 350mg/kg vancomycin injection as our regimen for inducing robust GI colonization in mice. Finally, we tested the durability of colonization by following fecal shedding in mice up to Day 60 post-inoculation with a second ST258 strain. Shedding during the first 7 days occurs at about 1010 CFU/g feces, and from day 14 to day 60 fecal loads are stable around 107 CFU/g feces. Results are comparable between male and female mice. Conclusion In conclusion, we have developed a mouse model of robust, prolonged GI colonization with multiple strains of K. pneumoniae using controlled dosing of a clinically relevant antibiotic. This model may be used to study a key step in K. pneumoniae pathogenesis and infection prevention in the future. Disclosures All Authors: No reported disclosures


L, PCT, SS Comparison of Sepsis vs INSI
Conclusion. L is sub-optimal in discriminating sepsis from INSI. PCT with or without L was acceptable but not as robust as SS. SS alone or in any combination provided superior and significant discrimination between sepsis and INSI. Incorporation of SS into the clinical assessment process for suspected sepsis pts should be evaluated to determine the impact on early detection and Pt management.

A Murine Model of Klebsiella pneumoniae Gastrointestinal Colonization with Parenteral Vancomycin Administration
Bettina Cheung, BS 1 ; Marine Lebrun-Corbin, BS, MS 1 ; Alan R. Hauser, MD PhD 1 ; 1 Northwestern University, Chicago, Illinois Session: P-56. Microbial Pathogenesis Background. As a leading cause of nosocomial infections, Klebsiella pneumoniae poses a significant threat due to its propensity to acquire resistance to many classes of antibiotics, including carbapenems. Gastrointestinal (GI) colonization by K. pneumoniae is a risk factor for subsequent infection as well as transmission to other patients. To study this crucial step in pathogenesis, we developed a mouse model of K. pneumoniae GI colonization using a clinically relevant parenteral antibiotic regimen.
Methods. To improve the clinical relevance of our model, we elected to use intraperitoneal injections of vancomycin, one of the most highly utilized antibiotics in the United States.
Results. To optimize dosage in C57bl/6 mice, we injected 20mg/kg, 350mg/kg, or vehicle (PBS) for three days prior to gastric gavage with 10 8 colony forming units (CFU) of a low-resistance strain of K. pneumoniae. The mice who received 350mg/kg (a mouse equivalent of a human dose of 1g/day calculated through the FDA guidelines for estimating safe dosing) shed about 10 7 CFU/g of feces at Day 7 while those receiving the lower dose or vehicle shed 10 4 CFU/g. Next, we compared 3-or 5-day pre-treatments with vancomycin prior to inoculation with an ST258 (epidemic carbapenem-resistant) strain. At Day 7 post-inoculation, mice who received 5 days shed 10 10 CFU/g feces while those who received vancomycin for 3 days or vehicle for 5 days (PBS) shed 10 6 or 10 4 CFU/g feces respectively. Thus, we chose 5 days of 350mg/kg vancomycin injection as our regimen for inducing robust GI colonization in mice. Finally, we tested the durability of colonization by following fecal shedding in mice up to Day 60 post-inoculation with a second ST258 strain. Shedding during the first 7 days occurs at about 10 10 CFU/g feces, and from day 14 to day 60 fecal loads are stable around 10 7 CFU/g feces. Results are comparable between male and female mice.
Conclusion. In conclusion, we have developed a mouse model of robust, prolonged GI colonization with multiple strains of K. pneumoniae using controlled dosing of a clinically relevant antibiotic. This model may be used to study a key step in K. pneumoniae pathogenesis and infection prevention in the future. Background. Frequent plateletpheresis using the Time Accel leukoreduction system chamber may result in lymphopenia in healthy donors, with increased donation in the previous year associated with CD4+ T-cell count of less than 200 cells/µL. However, this finding has not been replicated and the clinical significance of plateletpheresis-associated lymphopenia remains unclear.

CD4+ T-Cell Lymphopenia Associated with Frequent Plateletpheresis in Healthy Donors
Methods. A prospective observational study of healthy plateletpheresis donors aged 18 or older who donated at least once in the previous 365 days was conducted at the Kraft Blood Center at Brigham and Women's Hospital/Dana Farber Cancer Institute, where the Time Accel system is used exclusively. Blood was drawn immediately before plateletpheresis or at least 2 weeks after the last donation to assess for total lymphocyte and CD4+ T-cell counts.