Journal:

Article Title: Th1 Cytokines Facilitate CD8-T-Cell-Mediated Early Resistance to Infection with Mycobacterium tuberculosis in Old Mice

doi: 10.1128/IAI.01475-05

Figure Lengend Snippet: Th1 cytokine production in the lungs of old mice during infection with M. tuberculosis. Young and old mice were aerogenically infected with a low dose of M. tuberculosis Erdman. At specific times after infection the right middle lung lobe was removed, homogenized in Ultraspec, and frozen at −80°C. RNA was isolated and reverse transcribed, and cDNA for IL-12p40 (A) and IFN-γ (B) was amplified by real-time PCR. The data are means ± standard errors for the relative quantity of message from five mice at each time and are representative of three independent experiments. 18S was used as an endogenous normalizer, and naïve young mice were used as the calibrator. At the early times after infection (0 to 12 days) the total numbers of lung cells isolated from young and old mice were statistically indistinguishable (data not shown). (C and D) On day 12 postinfection aliquots of lung homogenate were analyzed to determine the levels of IL-12p40 (C) and IL-18 (D) protein by ELISA. The data are means ± standard errors for five mice per group and are representative of two independent experiments. An asterisk indicates that the P value is <0.05, as determined by Student's t test.

Article Snippet: M. tuberculosis Erdman (= ATCC 35801) was obtained from American Type Culture Collection (Manassas, VA) and grown in Proskauer-Beck liquid medium containing 0.05% Tween 80 to the mid-log phase.

Techniques: Infection, Isolation, Reverse Transcription, Amplification, Real-time Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Journal:

Article Title: Th1 Cytokines Facilitate CD8-T-Cell-Mediated Early Resistance to Infection with Mycobacterium tuberculosis in Old Mice

doi: 10.1128/IAI.01475-05

Figure Lengend Snippet: Early resistance of old mice to M. tuberculosis infection. Young and old mice were infected with a low dose of M. tuberculosis Erdman by aerosol, and bacterial burdens in the lungs were determined (A). The data are means ± standard errors for five mice per group and are representative of four independent experiments. BrdU was administered intraperitoneally 14 h before lung cells were harvested. (B and C) Single-cell suspensions from lungs were stained with fluorescently conjugated antibodies against CD3ɛ, CD8, and BrdU. (C) Representative dot plots for lung cells from young and old mice on day 15 postinfection. (B) Percentages were determined and used to calculate absolute numbers. The data are representative of two independent experiments. One asterisk indicates that the P value is <0.05 and three asterisks indicate that the P value is <0.0005, as determined by Student's t test.

Article Snippet: M. tuberculosis Erdman (= ATCC 35801) was obtained from American Type Culture Collection (Manassas, VA) and grown in Proskauer-Beck liquid medium containing 0.05% Tween 80 to the mid-log phase.

Techniques: Infection, Aerosol, Staining

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: Typical examples of MGC formation after 3 days of culture. Monocytes were stimulated with BCG (0.4/cell; a), T-SN (50%) (b), BCG–T-SN (c); or ConA-SN (50%) (d). Stimulation of cells in different fusion systems leads to very different fusion rates (in the examples shown, about 4% [a]), 10% [b], 30% [c], and 70% [d]). The inset in Fig. ​Fig.1c1c exemplifies that phagocytosis of whole cells by other cells might contribute to MGC formation. Giemsa staining; magnification, ×50 (×75 for inset).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Staining

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: Stimulation of human monocytes with mycobacteria and T-SN in combination leads to more than additive fusion rates compared to stimulation with one component alone. Monocytes were cultured for 3 days with viable BCG (0.4/cell), BCG-SN (50%), or T-SN (50%) alone or with viable BCG, heat-killed (h.k.) BCG (0.4/cell), or M. tuberculosis H37Ra (M.tb.; 1:64) in combination with T-SN. Results represent mean ± SEM (n = 4 to 18). Fusion rates induced by stimuli in combination were significantly different from those obtained with BCG or T-SN alone (P < 0.01).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: A relatively low BCG/cell ratio is optimal for MGC formation. Monocytes were cultured with indicated BCG numbers per cell and T-SN (50%) for 3 days. Results represent mean ± SEM (n = 4 to 10). ⧫, cell loss in 2 of 10 experiments; ⧫⧫, cell loss in 4 of 10 experiments.

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: MGC containing mycobacteria. Monocytes (1.2 × 105) were cultured with BCG (0.4/cell) and T-SN (50%) in Lab-Tek chamber slides. After 3 days, mycobacteria were stained with carbol fuchsin and nuclei of MGC were counterstained with malachite green. Magnification, ×100.

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture, Staining

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: MGC formation induced by BCG in combination with T-SN requires direct contact of monocytes and mycobacteria. Monocytes were stimulated with BCG (0.4/cell) and T-SN (50%) in cell culture plates containing transwell inserts. Monocytes and BCG were either cultured in the same compartment or separated by the semipermeable membrane of the insert. Results represent mean ± SEM (n = 10). The fusion rates obtained with these two culture conditions differed significantly (P < 0.01).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture, Membrane

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: Influence of CD18 MAb on MGC formation. Monocytes were cultured for 3 days with BCG (0.4/cell) and T-SN (50%) or ConA-SN (50%). CD18 MAb was added simultaneously in the concentrations indicated. Results represent mean ± SEM (n = 3). The asterisks indicate statistically significant differences between cultures with and without antibody (∗, P < 0.05; ∗∗, P < 0.01).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: Influence of MAb against IFN-γ on MGC formation. Monocytes were cultured for 3 days with BCG (0.4/cell) and T-SN (50%) or with ConA-SN (50%). Anti-IFN-γ MAb was added simultaneously in the concentrations indicated. Results represent mean ± SEM (n = 2 to 5). The asterisks indicate statistically significant differences between cultures with and without antibody (∗, P < 0.05; ∗∗, P < 0.01).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture

Journal:

Article Title: Generation of Multinucleated Giant Cells In Vitro by Culture of Human Monocytes with Mycobacterium bovis BCG in Combination with Cytokine-Containing Supernatants

doi:

Figure Lengend Snippet: Influence of MAb against TNF-α on MGC formation. Monocytes were cultured for 3 days with BCG (0.4/cell) and T-SN (50%) or ConA-SN (50%). Anti-TNF-α MAb was added simultaneously in the concentrations indicated. Results represent mean ± SEM (n = 4 to 5). The asterisks indicate statistically significant differences between cultures with and without antibody (∗, P < 0.05; ∗∗, P < 0.01).

Article Snippet: M. bovis bacillus Calmette-Guérin (BCG, strain Chicago, ATCC [American Type Culture Collection] 27289), BCG lysate (without membranes), and supernatant of BCG cultures (BCG-SN) were a kind gift from I. Flesch, Department of Immunology, University of Ulm, Ulm, Germany.

Techniques: Cell Culture