Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 1. HSV co-infection induces persistent chlamydial morphology. Cultures of HeLa (A, B, C and F) or HEC-1B (D and E) cells were C. trachomatis-infected or co-infected with C. trachomatis and HSV-2 strain 333 (A, B, D–F) or HSV-1 strain KOS (C) at 10 moi (A–E) or 1 moi (F). Cells were harvested for TEM analyses. Black arrows on electron micrographs indicate EBs and white arrows indicate RBs.

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection

Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 2. HSV co-infection decreases chlamydial infectivity. Cultures of HeLa or HEC-1B cells were mock, C. trachomatis, or HSV singly infected and C. trachomatis/HSV co-infected. Cells were harvested for EB titration assays. A. HeLa cells were co-infected with HSV-2 strain 333 at 10 moi. B. HeLa cells were co-infected with HSV-1 strain KOS at 10 moi. C. HEC-1B cells were co-infected with HSV-2 strain 333 at 10 moi. D. HeLa cells were co-infected with HSV-2 strain 333 at, 0.1, 1, or 10 moi. EB titres are expressed as ifu ml-1 sample SEM; n = 3. Asterisks (*) indicate titres that are significantly different (by T-test) compared with those from C. trachomatis singly infected cells (P < 0.05). The data shown are representative of three independent experiments.

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection, Titration

Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 3. Purified HSV-2 co-infection decreases chlamydial infectivity. Cultures of HeLa cells were infected with C. trachomatis, or co-infected with chlamydia and HSV-2 crude stock, purified stock, mock crude stock, or mock purified stock at 10 moi or the equivalent volume of mock stock. Cells were harvested for EB titration analyses. EB titres are expressed as ifu ml-1 sample SEM; n = 3. Asterisks (*) indicate titres that are significantly different (by T-test) compared with those from C. trachomatis singly infected cells (P < 0.05). The data shown are representative of three independent experiments.

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection, Titration

Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 4. Co-infection with HSV-2 in the presence of cyclohexamide induces chlamydial persistence. Cultures of HeLa cells were infected with C. trachomatis. At 23 h post chlamydial infection the cells were refed with medium containing either diluent (ddH2O) (A–C) or 1 mg ml-1

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection

Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 5. Co-infection with HSV-2UV induces chlamydial persistence. Cultures of HeLa cells were C. trachomatis-infected followed by infection with replication-competent HSV-2 at 10 moi or an equal amount of HSV-2UV. Cells were harvested for TEM (A–C), C. trachomatis titre analyses (D) and total DNA isolation (E) at 0 or 20 h post HSV infection. HSV-2 viral stocks were UV-inactivated by exposure to 2.5 J cm-2 UV irradiation at 4°C; plaque assays demonstrate that HSV-2UV stocks contain no replication-competent virions (data not shown). EBs on electron micrographs are indicated by black arrows; RBs by white arrows. EB titres are expressed as ifu ml-1 sample SEM; n = 3. Asterisks (*) indicate titres that are significantly different (by T-test) compared with those from C. trachomatis singly infected cells (P < 0.05). The data shown are representative of three independent experiments.

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection, DNA Extraction, Irradiation

Journal: Cellular microbiology

Article Title: An early event in the herpes simplex virus type-2 replication cycle is sufficient to induce Chlamydia trachomatis persistence.

doi: 10.1111/j.1462-5822.2006.00823.x

Figure Lengend Snippet: Fig. 6. Co-infection does not alter accumulation of chlamydial DNA. Total DNA was purified and subjected to PCR analyses using host genome (GAPDH) and chlamydial genome (16S rRNA gene) specific DNA oligonucleotides. Amplimers were electrophoresed and quantified as described. The data in A were quantified after conversion from Macintosh to personal computer format whereas the data in B were quantified using the Macintosh format, thus causing differences in the intensity scales. Chlamydial DNA quantities were normalized to host DNA as ascertained by PCR with human GAPDH-specific primers (data not shown). These data are representative of three independent experiments and are averages of three biological replicates. The average integrated intensity for each amplimer was plotted SEM. A. HeLa cells were co-infected with HSV-2/C. trachomatis. Total cellular DNA was collected at 0 (T0) and 20 h (T20) post HSV-2 infection. B. HeLa cells were C. trachomatis-infected, or co-infected with HSV-2, HSV-2UV, or HSV-2 in the presence of cyclohexamide. Total cellular DNA was collected at 20 h post HSV infection. The asterisk (*) indicates that the value obtained at 20 h post HSV co-infection was significantly different (by t-test) from that at 0 h post HSV-2 co-infection (P < 0.05).

Article Snippet: Fluorescent and transmission electron microscopy Fluorescence analyses were performed as described previously (Deka et al., 2006) except FITC-conjugated monoclonal antibodies generated against C. trachomatis MOMP (Pathfinder C. trachomatis monoclonal antibody 30702, Bio-Rad) were used to stain chlamydial inclusions.

Techniques: Infection

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Murine Endometrial Organoids to Model Chlamydia Infection

doi: 10.3389/fcimb.2020.00416

Figure Lengend Snippet: Infection of murine EMOs with C. trachomatis . (A) Cartoon diagram depicting the method of infecting EMOs with C. trachomatis by mechanical fragmentation. (B) Confocal micrographs of an EMO infected with mCherry-expressing C. trachomatis (red, left panel, mCherry). Infected EMOs were fixed at 32 hpi and immunostained with anti- C. trachomatis MOMP (green, middle panel, MOMP). The merge, including a brightfield image, is shown on the right. Higher magnification of the indicated boxed area is shown in the inset of each panel. A single plane across the middle of the inclusion is shown. Scale bar: 40 μm. (C) Confocal micrographs of an EMO infected with mCherry-expressing C. trachomatis (red, middle panels, mCherry) fixed at 32 hpi and stained with SYTOX TM Green (green, left panels, DNA) and Phalloidin (red, middle panels, F-Actin). The merge is shown on the right. A single plane across the middle of the EMO and inclusions is shown. Top panels: whole EMO. Scale bar: 40 μm. Bottom panels: higher magnification of indicated boxed area. Inclusions are indicated by arrow heads. Scale bar: 20 μm.

Article Snippet: The following primary antibodies and dyes were used: goat polyclonal anti- Chlamydia major outer membrane protein (MOMP; 1:200, Virostat) and mouse monoclonal anti-FLAG (1:1,000, Sigma).

Techniques: Infection, Expressing, Staining