Article Figures & Data
Figures
- FIG 1
Role of NAC species in protection against lethal polymicrobial IAI. Mice (n = 10/group) were injected i.p. with 3.5 × 107 CFU of C. dubliniensis (Cd) C. glabrata (Cg), C. tropicalis (Ct), or C. krusei (Ck) alone (standard inocula) or in combination with 8 × 107 CFU of S. aureus (Sa) (standard inocula) as a primary challenge, and then rechallenged with C. albicans/S. aureus after 14 days (A) or injected i.p. with C. dubliniensis/S. aureus as the primary challenge and rechallenged with C. albicans (Ca), C. tropicalis, or C. krusei in combination with S. aureus after 14 days (standard inocula) (B). Animals receiving no primary challenge served as the positive (lethal) control. Mice were monitored for 10 days post-rechallenge. Data are representative of 2 separate experiments. *, P < 0.05; **, P < 0.0001 (significantly different from control by log rank Mantel-Cox test).
- FIG 2
Limits of C. dubliniensis-mediated protection against lethal polymicrobial IAI. Mice (n = 10/group) were injected i.p. with different permutations of viable and nonviable C. dubliniensis and S. aureus as the primary challenge followed by rechallenge with C. albicans/S. aureus (standard inocula). Animals receiving no primary challenge served as the positive (lethal) control. Mice were monitored for 10 days post-rechallenge. Data are representative of 3 separate experiments. *, significantly different from control (P < 0.05) by log rank Mantel-Cox test.
- FIG 3
C. dubliniensis induces long term protection against polymicrobial IAI. Mice (n = 10/group) were injected i.p. with C. dubliniensis and S. aureus as the primary challenge 14, 30, and 60 days prior to rechallenge with C. albicans / S. aureus (standard inocula). Animals receiving no primary challenge served as the positive (lethal) control. Mice were monitored for 10 days post-rechallenge. *, significantly different from control (P < 0.05) by log rank Mantel-Cox test.
- FIG 4
Role of T and B cells in C. dubliniensis-induced protection. RAG mice (deficient in T and B cells) (n = 10) and the background congenic strain, C57BL/6J mice (n = 10) were given the primary challenge of C. dubliniensis and S. aureus 30 days or 14 days prior to rechallenge with C. albicans/S. aureus (standard inocula). Animals receiving no primary challenge served as the positive (lethal) controls. Mice were monitored for 10 days post-rechallenge. *, significantly different from control (P < 0.05) by log rank Mantel-Cox test.
- FIG 5
Role of macrophages in C. dubliniensis-induced protection. Mice (n = 10/group) previously given the primary challenge of C. dubliniensis/S. aureus (14 days prior) were injected i.p. with liposome-encapsulated clodronate (which results in ~90% depletion of resident peritoneal macrophages), liposomes only, or PBS 1 day prior to rechallenge with C. albicans/S. aureus. Animals receiving no primary challenge also served as the positive (lethal) controls. Mice were monitored for 10 days post-rechallenge. *, significantly different from control (P < 0.02) by log rank Mantel-Cox test.
- FIG 6
Presence of PMNLs in C. albicans/S. aureus rechallenged, protected animals. Mice (n = 10/group) were given the primary challenge of C. dubliniensis/S. aureus and rechallenged with C. albicans/S. aureus 14 days later. Control mice (n = 10) received no primary challenge. (A) H&E-stained smears of PMNLs from peritoneal lavage fluid collected 4 h after rechallenge. The illustration is representative of several individual mice evaluated. (B) Flow cytometry analysis results of PMNLs from peritoneal lavage fluid prior to rechallenge through 96 h post-rechallenge with C. albicans/S. aureus. Percentages indicate proportions of Gr-1hi PMNLs present in the total cell population. MFI of Gr-1hi PMNLs within the encircled areas are shown in red. The illustration is representative of results for several individual mice evaluated. (C) Microbial burden (C. albicans and S. aureus) in peritoneal lavage fluid of mice 4 h post-rechallenge through 10 days post-rechallenge with C. albicans/S. aureus in those that remained alive. Data are cumulative for all animals from each group. Mφ, macrophage(s).
- FIG 7
PMNL depletion abrogates protection. Mice (n = 10/group) given the primary challenge of C. dubliniensis/S. aureus were injected i.p. with 200 µg anti-Gr-1 (Ly6G/C) antibodies to deplete PMNLs or isotype control antibodies 48 h prior to and 2 h after rechallenge with C. albicans/S. aureus. Antibodies were given every 2 days to the remaining live animals for the duration of the study. Mice were monitored for 10 days post-rechallenge. *, significantly different from control (P = 0.02) by log rank Mantel-Cox test.
Tables
- TABLE 1
NAC species (with or without S. aureus coinfection) primary challenge survival
Primary challenge % survivalc after
primary challengeMTDd (days) NAC speciesa S. aureusb C. dubliniensis − 100 NA C. glabrata − 100 NA C. krusei − 100 NA C. tropicalis − 100 NA C. dubliniensis + 90 3 C. glabrata + 100 NA C. krusei + 50 4 C. tropicalis + 40 6 - TABLE 2
Limits of C. albicans-mediated protection
Primary challenge % survival after: C. albicansa S. aureusb Primary
challengecRechallenge with
C. albicans/S. aureus− − NA 25 Live C. albicans − 40 (9) 50 Killed C. albicans − 100 0 − Live S. aureus 100 25 − Killed C. albicans 100 20 Live C. albicans Killed S. aureus 60 (5) 15 Killed C. albicans Live S. aureus 100 90 Killed C. albicans Killed S. aureus 100 0 ↵a Inoculum of 1.75 × 107 live or killed C. albicans was injected i.p.
↵b Inoculum of 8 × 107 live or killed S. aureus was injected i.p.
↵c Results are cumulative from 6 studies with a 14-day observation period. NA, not applicable. Values in parentheses indicate the median time to death of mice that succumbed to infection.
FIG S1
Limits of C. albicans-mediated protection. Mice (n = 10/group) were injected i.p. with the standard 2.5× inocula of viable Ca (1.75 × 107), a 1× inocula (7 × 106), or a 0.14× inocula of 1 × 106 as the primary challenge followed by re-challenge with Ca/Sa (standard inocula) after 14 days. Animals receiving no primary challenge served as the positive (lethal) control. Mice were monitored for 10 days post re-challenge. Data are representative of 4 separate experiments. (A) Survival of animals in the 14-day primary challenge. (B) Survival of animals after re-challenge with Ca/Sa. Download FIG S1, PDF file, 0.1 MB.
Copyright © 2018 Lilly et al.
This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.
FIG S2
Confirmation of macrophage and PMNL depletion. (A) Cd/Sa mice (n = 10/group) were injected i.p. with liposome-encapsulated clodronate one day prior to re-challenge with Ca/Sa to deplete macrophages. Empty liposomes or PBS alone were administered to control animals. Peritoneal lavage fluid was analyzed by flow cytometry to confirm depletion with red arrows indicating F4/80+ cells (macrophages). (B) Mice (n = 10/group) given the primary challenge of Cd/Sa were injected i.p. with 200 μg anti-Gr-1 (Ly6G/C) antibodies to deplete PMNLs or isotype control antibodies 48 h prior to and 2 h after re-challenge with Ca/Sa. Peritoneal lavage fluid was analyzed by flow cytometry to confirm depletion just prior to re-challenge with Ca/Sa. PMNLs are shown within the red encircled areas. Download FIG S2, PDF file, 0.1 MB.
Copyright © 2018 Lilly et al.
This content is distributed under the terms of the Creative Commons Attribution 4.0 International license.
