Journal: Science translational medicine

Article Title: Intestinal Commensal Bacteria Mediate Lung Mucosal Immunity and Promote Resistance of Newborn Mice to Infection

doi: 10.1126/scitranslmed.aaf9412

Figure Lengend Snippet: (A) Study design. (B) Intestinal commensal bacteria enumerated in postnatal day 4 (P4) newborn mice exposed to antibiotics (ABX) or no antibiotics (ABX-free) quantified using real-time PCR.. (C) Survival of ABX-free or ABX-exposed P4 mice or ABX-exposed newborn mice reconstituted with intestinal commensal bacteria and infected with S. pneumonia. (D) Survival of germ-free (GF) or conventionally housed (CNV) mice or age-matched GF mice reconstituted with intestinal commensal bacteria and infected with S. pneumoniae. (E) Survival of ABX-free or ABX-exposed P14 mice or ABX-exposed newborn mice reconstituted with intestinal commensal bacteria and infected with S. pneumoniae. (F) The amount of IL-22 in the bronchial lavage (BAL) fluid of P4 ABX-free or ABX-exposed mice or GF newborn mice or ABX-exposed or GF newborn mice reconstituted with intestinal commensal bacteria in early life. None of the newborn mice in this experimental group were inoculated with S. pneumoniae. (G) The amount of IL-22 in the BAL fluid of human newborns who were exposed to ABX or no ABX. (H) Survival of P4 ABX-free or ABX-exposed newborn mice treated with IL-22 intratracheally and infected with S. pneumoniae. (I) Survival of P4 ABX-free or ABX-exposed newborn mice treated with anti-IL22 antibody or isotype control antibody before reconstitution with intestinal commensal bacteria and infection with S. pneumoniae. Data are representative of three independent experiments. Results are shown as the mean ± s.e.m (Student’s t-test or ANOVA or Wilcoxon signed-rank test). *P ≤ 0.05; **P ≤ 0.01. Number of individual animals [n] are indicated.

Article Snippet: Murine neonatal pneumonia studies We grew S. pneumoniae serotype 19A (ATCC 700674) or E. coli serotype K1 ( 82 ) or C. albicans (37°C, 200 rpm) in tryptic soy (TS) broth to log-phase growth.

Techniques: Bacteria, Real-time Polymerase Chain Reaction, Infection, Control

Journal: Science translational medicine

Article Title: Intestinal Commensal Bacteria Mediate Lung Mucosal Immunity and Promote Resistance of Newborn Mice to Infection

doi: 10.1126/scitranslmed.aaf9412

Figure Lengend Snippet: (A) Representative flow cytometry plots of distinct subsets of IL-22+ cells in the lungs of postnatal day 4 (P4) newborn mice. Shown are relative frequencies of IL22+ T cells (CD45+CD3+) or neutrophils (CD45+Ly6G+) or lineage negative (CD3−CD8−CD11b−CD19−F4/80−CD161−Ly6G−F4/80−) lymphocytes in the lungs of P4 newborn mice. (B) The relative frequencies of distinct subsets of IL-22+ cells in the bronchial lavage (BAL) fluid of human newborn infants.. (C) Absolute numbers of IL22+ILC3 in the lungs of P4 wild-type (WT) or RorγtiDTR newborn mice treated with diphtheria toxin (DT) or no DT. (D) Survival of P4 WT or RorγtiDTR newborn mice treated with DT (ILC3-depleted) that received adoptive transfer of ILC3 and then were infected with S. pneumoniae. (E) The absolute number of IL-22+ILC3 in the lungs of ABX-free or ABX-exposed newborn mice at different time points after birth. (F) Representative flow cytometry plots and (G) absolute numbers of IL-22+ILC3 in the lungs of P4 GF or ABX-free or ABX-exposed or ABX-exposed newborn mice reconstituted with intestinal commensal bacteria in early life. (H) The absolute numbers of IL-22+ILC3 in the BAL fluid of human newborns exposed to ABX or no ABX. (I) ILC3 from P4 ABX-free newborn mice were labeled with carboxyflourosceinsuccimidylester (CFSE). ILC3 from age-matched P4 ABX-exposed or ABX-exposed newborn mice reconstituted with commensal bacteria were labeled with chloromethylbenozylaminotetramethylrhodamine (CMMTR). An equal number of CFSE- or CMMTR-labeled ILC3 were adoptively transferred into ABX-exposed newborn mice. Representative flow cytometry plots and absolute numbers of CFSE+ or CMMTR+ ILC3 in lung, spleen or small intestine were determined 12 h following adoptive transfer. (J) Relative capability of ILC3 from ABX-free or ABX-exposed or ABX-exposed newborn mice reconstituted with intestinal commensal bacteria in early life to traffic to the lungs (Homing index). Data and plots are representative of three independent experiments. Results are shown as the mean ± s.e.m (Student’s t-test or ANOVA or Wilcoxon signed-rank test). *P ≤ 0.05; **P ≤ 0.01. Number of individual animals [n] are indicated.

Article Snippet: Murine neonatal pneumonia studies We grew S. pneumoniae serotype 19A (ATCC 700674) or E. coli serotype K1 ( 82 ) or C. albicans (37°C, 200 rpm) in tryptic soy (TS) broth to log-phase growth.

Techniques: Flow Cytometry, Adoptive Transfer Assay, Infection, Bacteria, Labeling

Journal: Science translational medicine

Article Title: Intestinal Commensal Bacteria Mediate Lung Mucosal Immunity and Promote Resistance of Newborn Mice to Infection

doi: 10.1126/scitranslmed.aaf9412

Figure Lengend Snippet: (A) Representative flow cytometry histograms showing expression of C-C chemokine receptor (CCR) 4, 6, 7 and 9, C-X-C chemokine receptor (CXCR) 3 and 5 and C-C chemokine ligand (CCL) 20 by IL22+ILC3 innate lymphoid cells in the lung of postnatal day 4 (P4) ABX-free or ABX-exposed newborn mice. (B) An equal number of ILC3 from P4 WT or Ccr4−/− newborn mice were adoptively transferred into age-matched ABX-exposed newborn mice, and the ability of ILC3 to traffic to the lungs (homing index) was determined. (C) Survival of P4 WT or Ccr4−/− newborn mice that received adoptive transfer of WT ILC3 after infection with S. pneumoniae. (D) Representative flow cytometry plots and relative frequencies of distinct subsets of mononuclear phagocytes in the small intestine of P4 newborn mice. (E) The absolute numbers of IL22+ILC3 in the lungs of P4 WT or Zbtb46DTR newborn mice treated with DT (CD11b+CD103+ DC-depleted) that received adoptive transfer of CD11b+CD103+ DCs. (F) Survival of P4 WT or Zbtb46DTR newborn mice treated with DT (CD11b+CD103+ DC-depleted) that then received adoptive transfer of CD11b+CD103+ DCs, after infection with S. pneumoniae . (G) ILC3 isolated from lungs of P4 ABX-exposed mice were co-cultured with CD11b+CD103+ DCs isolated from age-matched ABX-exposed or ABX-free mice and examined for surface expression of various chemokine receptors. A representative flow cytometry plot is shown and (H) relative frequencies of IL-22+ILC3 cells expressing CCR4. (I) ILC3 isolated from lungs of P4 ABX-exposed mice were co-cultured either alone or with CD11b+CD103+ DCs isolated from age-matched ABX-exposed or ABX-free newborn mice. The ability of these ILC3 to migrate in vitro in response to a gradient of the chemokine ligand (CCL) 17 is shown. (J) The absolute numbers of IL22+ILC3 in the lungs of P4 Zbtb46DTR newborn mice either exposed to ABX or no ABX that were then treated with DT (CD11b+CD103+ DC-depleted) or no DT (no DC depletion) before they were reconstituted with commensal bacteria. Data and plots are representative of three independent experiments. Results are shown as the mean ± s.e.m (Student’s t-test or ANOVA or Wilcoxon signed-rank test,). *P ≤ 0.05; **P ≤ 0.01. Number of individual animals [n] are indicated.

Article Snippet: Murine neonatal pneumonia studies We grew S. pneumoniae serotype 19A (ATCC 700674) or E. coli serotype K1 ( 82 ) or C. albicans (37°C, 200 rpm) in tryptic soy (TS) broth to log-phase growth.

Techniques: Flow Cytometry, Expressing, Adoptive Transfer Assay, Infection, Isolation, Cell Culture, In Vitro, Bacteria

Journal:

Article Title: Nomenclature of Major Antimicrobial-Resistant Clones of Streptococcus pneumoniae Defined by the Pneumococcal Molecular Epidemiology Network

doi: 10.1128/JCM.39.7.2565-2571.2001

Figure Lengend Snippet: Properties of 16 pneumococcal clones a

Article Snippet: ATCC accession numbers for the 16 clones are given in Table . table ft1 table-wrap mode="anchored" t5 TABLE 1 caption a7 Clone Reference strain (ATCC accession no.) Serotype MIC (μg/ml) of: Reference(s) PEN CTX ERY CLI CHL TET SXT Spain 23F -1 SP264 (ATCC 700669) 23F 2 0.5 0.06 0.06 16 32 4/76 7, 32, 37 Spain 6B -2 GM17 (ATCC 700670) 6B 2 1 0.12 0.12 16 32 4/76 38 Spain 9V -3 TL7/1993 (ATCC 700671) 9V 2 1 0.12 0.12 2 ≤0.5 8/152 7, 29 Tennessee 23F -4 CS111 (ATCC 51916) 23F 0.12 32 32 0.12 2 ≤0.5 4/76 33, 49 Spain 14 -5 MS22 (ATCC 700902) 14 2 1 0.06 0.12 16 32 2/38 8 Hungary 19A -6 HUN663 (ATCC 700673) 19A 2 0.5 >32 >32 16 16 2/38 31, 38 South Africa 19A -7 17619 (ATCC 700674) 19A 0.5 0.5 0.25 0.25 2 ≤0.5 4/152 51 South Africa 6B -8 50803 (ATCC 700675) 6B 0.5 0.25 0.12 0.12 2 ≤0.5 1/19 51 England 14 -9 PN93/872/B (ATCC 700676) 14 0.03 0.03 32 0.12 2 ≤0.5 0.12/4.2 17 CSR 14 -10 87-029055 (ATCC 700677) 14 8 1 >32 >32 16 32 0.25/4.8 16, 19 CSR 19A -11 91-006571 (ATCC 700678) 19A 4 1 >32 >32 32 32 4/76 16, 19 Finland 6B -12 43362 Fi10 (ATCC 700903) 6B 1 >32 >32 4 32 8/152 48 South Africa 19A -13 51702 (ATCC 700904) 19A 8 2 >8 >8 32 8 8/152 51 Taiwan 19F -14 TW31 (ATCC 700905) 19F 2 1 4 0.12 2 16 4/76 47 Taiwan 23F -15 TW17 (ATCC 700906) 23F 1–2 1 >32 >32 2 16 0.5/9.5 47 Poland 23F -16 178 23F 8 8 >8 0.25 16 16 4/76 41 Open in a separate window a PEN, penicillin; CTX, cefotaxime; ERY, erythromycin; CLI, clindamycin; CHL, chloramphenicol; TET, tetracycline; SXT, trimethoprim-sulfamethoxazole; CSR, Czech Republic.

Techniques: Clone Assay

Journal: eLife

Article Title: Characterization of the kinetic cycle of an ABC transporter by single-molecule and cryo-EM analyses

doi: 10.7554/eLife.56451

Figure Lengend Snippet:

Article Snippet: Recombinant DNA reagent , pET15b-ACPs (from S. pneumoniae ) , Gift from Michael Johnson , Addgene Plasmid# 63687 , .

Techniques: Recombinant, Plasmid Preparation, Modification, Expressing, Labeling, Software

Journal:

Article Title: Performance of a Pneumolysin Enzyme-Linked Immunosorbent Assay for Diagnosis of Pneumococcal Infections ▿

doi: 10.1128/JCM.01030-07

Figure Lengend Snippet: PLY-ELISA data from lysates of different microorganisms

Article Snippet: Different studies have demonstrated that the Binax Now S. pneumoniae test lacks specificity to detect pneumococcal urinary antigen in children, since detectable pneumococcal C polysaccharide is present in the urine of healthy infants with upper respiratory tract colonization ( 8 , 9 , 17 ).

Techniques:

Journal:

Article Title: Performance of a Pneumolysin Enzyme-Linked Immunosorbent Assay for Diagnosis of Pneumococcal Infections ▿

doi: 10.1128/JCM.01030-07

Figure Lengend Snippet: Descriptive statistics for PLY-ELISA and Binax Now S. pneumoniae urinary antigen tests of the same urine samples

Article Snippet: Different studies have demonstrated that the Binax Now S. pneumoniae test lacks specificity to detect pneumococcal urinary antigen in children, since detectable pneumococcal C polysaccharide is present in the urine of healthy infants with upper respiratory tract colonization ( 8 , 9 , 17 ).

Techniques:

Journal:

Article Title: Performance of a Pneumolysin Enzyme-Linked Immunosorbent Assay for Diagnosis of Pneumococcal Infections ▿

doi: 10.1128/JCM.01030-07

Figure Lengend Snippet: Etiologic diagnose of patients with potential false-positive results by PLY-ELISA

Article Snippet: Different studies have demonstrated that the Binax Now S. pneumoniae test lacks specificity to detect pneumococcal urinary antigen in children, since detectable pneumococcal C polysaccharide is present in the urine of healthy infants with upper respiratory tract colonization ( 8 , 9 , 17 ).

Techniques: Biomarker Discovery, Infection

Journal: Clinical Microbiology and Infection

Article Title: The impact of 13-valent pneumococcal conjugate vaccination on virus-associated community-acquired pneumonia in elderly

doi: 10.1016/j.cmi.2017.10.006

Figure Lengend Snippet: (a) Flow-chart of ‘confirmed CAP’ study population ( n = 1653). $ No confirmed CAP = episodes not fulfilling the definition ‘confirmed CAP’, i.e. chest X-ray not consistent with pneumonia and/or less than two clinical symptoms. (b) Flow-chart of ‘suspected pneumonia’ study population ( n = 2917). *Symptom onset <14 days after vaccination. Abbreviations used: PCV13, 13-valent pneumococcal conjugate vaccine; CAP, community-acquired pneumonia; UAD, urinary antigen detection assay.

Article Snippet: Abbreviations used: CAP, community-acquired pneumonia; UAD, urinary antigen detection assay; Culture, either a sterile (e.g. blood) or non-sterile (e.g. sputum) sample available for culture; pneumococcal UAD, either conventional pneumococcal urinary antigen detection assay (i.e. Binax) or serotype-specific urinary antigen detection assay.

Techniques: Detection Assay

Journal: Clinical Microbiology and Infection

Article Title: The impact of 13-valent pneumococcal conjugate vaccination on virus-associated community-acquired pneumonia in elderly

doi: 10.1016/j.cmi.2017.10.006

Figure Lengend Snippet: Diagram of available diagnostic methods for individuals with confirmed CAP and a oropharyngeal swab available ( n = 1653). Abbreviations used: CAP, community-acquired pneumonia; UAD, urinary antigen detection assay; Culture, either a sterile (e.g. blood) or non-sterile (e.g. sputum) sample available for culture; pneumococcal UAD, either conventional pneumococcal urinary antigen detection assay (i.e. Binax) or serotype-specific urinary antigen detection assay. Please note that this diagram excludes 16 patients who did not have any of these diagnostic methods available (i.e. only an oropharyngeal swab for viral analyses).

Article Snippet: Abbreviations used: CAP, community-acquired pneumonia; UAD, urinary antigen detection assay; Culture, either a sterile (e.g. blood) or non-sterile (e.g. sputum) sample available for culture; pneumococcal UAD, either conventional pneumococcal urinary antigen detection assay (i.e. Binax) or serotype-specific urinary antigen detection assay.

Techniques: Diagnostic Assay, Detection Assay, Sterility