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Histopathological Analysis of Acinetobacter baumannii Lung Infection in a Mouse Model

SHIGERU TANSHO-NAGAKAWA
SHIGERU TANSHO-NAGAKAWA
, 
YOSHINORI SATO
YOSHINORI SATO
, 
TSUNEYUKI UBAGAI
TSUNEYUKI UBAGAI
, 
TAKANE KIKUCHI-UEDA
TAKANE KIKUCHI-UEDA
, 
GO KAMOSHIDA
GO KAMOSHIDA
, 
SATOSHI NISHIDA
SATOSHI NISHIDA
 y 
YASUO ONO
YASUO ONO
   | 20 dic 2021
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Article Category: original-paper
Publicado en línea: 20 dic 2021
Páginas: 469 - 477
Recibido: 25 ago 2021
Aceptado: 31 oct 2021
DOI: https://doi.org/10.33073/pjm-2021-044
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© 2021 Shigeru Tansho-Nagakawa et al., published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.

Fig. 1.

Survival rates of mice with a lung infection. Survival rates of CH3/He mice following intratracheal inoculation with ATCC 19606 (A), TK1090 (B), and PAO-1 (C) strains. Groups of mice (ATCC 19606, TK1090, and PAO-1; n = 7 or 8 for uninfected and infected mice, respectively) were intratracheally inoculated as indicated, and their clinical outcome was monitored daily for up to 14 days.
Survival rates of mice with a lung infection. Survival rates of CH3/He mice following intratracheal inoculation with ATCC 19606 (A), TK1090 (B), and PAO-1 (C) strains. Groups of mice (ATCC 19606, TK1090, and PAO-1; n = 7 or 8 for uninfected and infected mice, respectively) were intratracheally inoculated as indicated, and their clinical outcome was monitored daily for up to 14 days.

Fig. 2.

Bacterial densities in mice infected with Acinetobacter baumannii. A. baumannii bacterial load in the lungs and kidneys of female C3H/HeN mice (groups of 3 mice for ATCC 19606- and TK1090-infected mice, respectively) intratracheally inoculated with 107 CFU of ATCC 19606 (open) and TK109 (closed) strains. Bacterial load in the respective organs was determined by quantitative bacteriology at 1 day (A) and 14 days (B) post-infection. The data are presented as the mean + standard deviation (SD). The detection limits for bacterial load were 1.3 log10 CFU/organ for the lung and kidney. Asterisks indicate statistically significant differences in the number of bacteria (*p < 0.05; ATCC 19606 vs. TK1090; unpaired t-test).
Bacterial densities in mice infected with Acinetobacter baumannii. A. baumannii bacterial load in the lungs and kidneys of female C3H/HeN mice (groups of 3 mice for ATCC 19606- and TK1090-infected mice, respectively) intratracheally inoculated with 107 CFU of ATCC 19606 (open) and TK109 (closed) strains. Bacterial load in the respective organs was determined by quantitative bacteriology at 1 day (A) and 14 days (B) post-infection. The data are presented as the mean + standard deviation (SD). The detection limits for bacterial load were 1.3 log10 CFU/organ for the lung and kidney. Asterisks indicate statistically significant differences in the number of bacteria (*p < 0.05; ATCC 19606 vs. TK1090; unpaired t-test).

Fig. 3.

Histopathological analysis of the lung tissues from Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. The lung tissues from C3H/HeN mice infected with ATCC 19606 (A, B, C, and D), TK1090 (E, F, G, and H), and PAO-1 (I, J, K, and L) strains and uninfected mouse (M) are shown. The lung tissues from the mice infected with ATCC 19606 at 1 day (A), 2 days (B), 5 days (C), and 14 days (D) post-infection. The lung tissues from the mice infected with TK1090 at 1 day (E), 2 days (F), 5 days (G), and 14 days (H) post-infection. The lung tissues from the mice infected with PAO-1 at 1 day (I), 2 days (J), 5 days (K), and 14 days (L) post-infection. Photomicrograph images (magnification, 100×; hematoxylin and eosin staining). Squares represent high magnification (400×).
Histopathological analysis of the lung tissues from Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. The lung tissues from C3H/HeN mice infected with ATCC 19606 (A, B, C, and D), TK1090 (E, F, G, and H), and PAO-1 (I, J, K, and L) strains and uninfected mouse (M) are shown. The lung tissues from the mice infected with ATCC 19606 at 1 day (A), 2 days (B), 5 days (C), and 14 days (D) post-infection. The lung tissues from the mice infected with TK1090 at 1 day (E), 2 days (F), 5 days (G), and 14 days (H) post-infection. The lung tissues from the mice infected with PAO-1 at 1 day (I), 2 days (J), 5 days (K), and 14 days (L) post-infection. Photomicrograph images (magnification, 100×; hematoxylin and eosin staining). Squares represent high magnification (400×).

Fig. 4.

Infiltration of neutrophils and macrophages in the lung tissues of Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. The lung tissues from C3H/HeN mice infected with ATCC19606 (A, B, C, and D), TK1090 (E, F, G, and H), and PAO-1 (I, J, K, and L) strains and uninfected mice (M and N) are shown. The lung tissues from the mice infected with ATCC 19606 at 1 day (A and B) and 5 days (C and D) post-infection. The lung tissues from the mice infected with TK1090 at 1 day (E and F) and 5 days (G and H) post-infection. The lung tissues from the mice infected with PAO-1 at 1 day (I and J) and 5 days (K and L) post-infection. Neutrophils and macrophages were detected by immunostaining with anti-Ly-6G antibody (A, C, E, G, I, and K) and anti-CD68 antibody (B, D, F, H, J, and L) using diaminobenzidine (DAB) method and restained with hematoxylin, respectively. Photomicrograph images (magnification, 100×).
Infiltration of neutrophils and macrophages in the lung tissues of Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. The lung tissues from C3H/HeN mice infected with ATCC19606 (A, B, C, and D), TK1090 (E, F, G, and H), and PAO-1 (I, J, K, and L) strains and uninfected mice (M and N) are shown. The lung tissues from the mice infected with ATCC 19606 at 1 day (A and B) and 5 days (C and D) post-infection. The lung tissues from the mice infected with TK1090 at 1 day (E and F) and 5 days (G and H) post-infection. The lung tissues from the mice infected with PAO-1 at 1 day (I and J) and 5 days (K and L) post-infection. Neutrophils and macrophages were detected by immunostaining with anti-Ly-6G antibody (A, C, E, G, I, and K) and anti-CD68 antibody (B, D, F, H, J, and L) using diaminobenzidine (DAB) method and restained with hematoxylin, respectively. Photomicrograph images (magnification, 100×).

Fig. 5.

Frequency of neutrophils and macrophages accumulated in the lung tissues of Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. Infiltrated neutrophils and macrophages in the lung tissues of the infected mice were detected by monoclonal antibodies using diaminobenzidine (DAB) method and restained with hematoxylin, and were measured by the HALO image analysis system for counting of cell numbers. The frequencies of neutrophils (A) and macrophages (B) in the mice post-infection are shown. The frequencies of infected mice on day 2 and day 5 are shown as open and closed bars, respectively. Symbols indicate statistically significant differences (*p < 0.05, C3H/HeN mice infected with A. baumannii strains ATCC 19606 or TK1090 at 2 days post-infection vs. C3H/HeN mice infected with P. aeruginosa PAO-1 at 2 days post-infection, unpaired t-test; †p < 0.05, C3H/HeN mice infected with A. baumannii strains ATCC 19606 or TK1090 at 5 days post-infection vs. C3H/HeN mice infected with P. aeruginosa PAO-1 at 5 days post-infection, unpaired t-test).
Frequency of neutrophils and macrophages accumulated in the lung tissues of Acinetobacter baumannii- and Pseudomonas aeruginosa-infected mice. Infiltrated neutrophils and macrophages in the lung tissues of the infected mice were detected by monoclonal antibodies using diaminobenzidine (DAB) method and restained with hematoxylin, and were measured by the HALO image analysis system for counting of cell numbers. The frequencies of neutrophils (A) and macrophages (B) in the mice post-infection are shown. The frequencies of infected mice on day 2 and day 5 are shown as open and closed bars, respectively. Symbols indicate statistically significant differences (*p < 0.05, C3H/HeN mice infected with A. baumannii strains ATCC 19606 or TK1090 at 2 days post-infection vs. C3H/HeN mice infected with P. aeruginosa PAO-1 at 2 days post-infection, unpaired t-test; †p < 0.05, C3H/HeN mice infected with A. baumannii strains ATCC 19606 or TK1090 at 5 days post-infection vs. C3H/HeN mice infected with P. aeruginosa PAO-1 at 5 days post-infection, unpaired t-test).
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