mycobacterium abscessus nr 44261  (ATCC)


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    ATCC mycobacterium abscessus nr 44261
    Mycobacterium Abscessus Nr 44261, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mycobacterium abscessus nr 44261  (ATCC)


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    ATCC mycobacterium abscessus nr 44261
    Mycobacterium Abscessus Nr 44261, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    mav whole lysate wl  (ATCC)


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    ATCC mav whole lysate wl
    Previous exposure <t>to</t> <t>BCG</t> or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or <t>MAV</t> (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.
    Mav Whole Lysate Wl, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    1) Product Images from "BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity"

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    Journal: Frontiers in Immunology

    doi: 10.3389/fimmu.2019.00234

    Previous exposure to BCG or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or MAV (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.
    Figure Legend Snippet: Previous exposure to BCG or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or MAV (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.

    Techniques Used: Infection, Labeling, Flow Cytometry, Expressing, Software, In Vitro

    MAV and MAB cross-reactive immunity includes Th1 and Th17 responses. (A) Schematic of experiments conducted to measure MAV or MAB cross-reactive immunity. PBMC from BCG-vaccinated or latent TB infected individuals ( n = 6) were stimulated with BCG or rested in media for 7 days. Then, these expanded cells were co-cultured with MAV- or MAB-infected autologous monocytes at an E:T ratio of 10:1. On day 3 of co-culture, Th1, Th2, and Th17 responses were measured in co-culture supernatants using CBA. Fold changes for each cytokine was calculated by dividing the amount of cytokine produced following restimulation with MAV or MAB by the amount produced in medium-rested cultures. (B) Exposure of BCG-expanded T cells to MAV-infected macrophages increased IL-17, IFN-γ, and TNF-α by 156 ± 62, 11 ± 1.7, 10.3 ± 2.9 pg/ml (Mean ± SE), respectively. IL-10, IL-6, and IL-2 showed no marked changes with fold changes (mean ± SE) of 0.3 ± 0.1, 0.9 ± 0.2, 0.9 ± 0.2 pg/ml, respectively. (C) Similarly, exposure of BCG-expanded T cells to MAB-infected macrophages increased IL-17 and IFN-γ in by 7.2 ± 1.6, and 5.6 ± 2, mean fold ± SE. There were no marked changes in the levels of TNF-α, IL-10, IL-6, and IL-2 with fold changes (mean ± SE) of 1.2 ± 0.3, 0.2 ± 0.2, 0.6 ± 0.3, 0.8 ± 0.4 pg/ml, respectively.
    Figure Legend Snippet: MAV and MAB cross-reactive immunity includes Th1 and Th17 responses. (A) Schematic of experiments conducted to measure MAV or MAB cross-reactive immunity. PBMC from BCG-vaccinated or latent TB infected individuals ( n = 6) were stimulated with BCG or rested in media for 7 days. Then, these expanded cells were co-cultured with MAV- or MAB-infected autologous monocytes at an E:T ratio of 10:1. On day 3 of co-culture, Th1, Th2, and Th17 responses were measured in co-culture supernatants using CBA. Fold changes for each cytokine was calculated by dividing the amount of cytokine produced following restimulation with MAV or MAB by the amount produced in medium-rested cultures. (B) Exposure of BCG-expanded T cells to MAV-infected macrophages increased IL-17, IFN-γ, and TNF-α by 156 ± 62, 11 ± 1.7, 10.3 ± 2.9 pg/ml (Mean ± SE), respectively. IL-10, IL-6, and IL-2 showed no marked changes with fold changes (mean ± SE) of 0.3 ± 0.1, 0.9 ± 0.2, 0.9 ± 0.2 pg/ml, respectively. (C) Similarly, exposure of BCG-expanded T cells to MAB-infected macrophages increased IL-17 and IFN-γ in by 7.2 ± 1.6, and 5.6 ± 2, mean fold ± SE. There were no marked changes in the levels of TNF-α, IL-10, IL-6, and IL-2 with fold changes (mean ± SE) of 1.2 ± 0.3, 0.2 ± 0.2, 0.6 ± 0.3, 0.8 ± 0.4 pg/ml, respectively.

    Techniques Used: Infection, Cell Culture, Co-Culture Assay, Produced

    BCG-specific T cells cross-protect against MAV and MAB. Total PBMC or subsets of T cells purified after 7 days of optimal BCG stimulation were co-cultured with autologous macrophages infected with MAV or MAB (E:T of 10). Residual mycobacteria quantified 3 days after co-culture and percent inhibition calculated by dividing the number of residual mycobacteria in the presence BCG-stimulated PBMC by the number of residual mycobacteria in control cultures containing medium-rested PBMC. (A) BCG-expanded T cells inhibit intracellular MAV ( n = 8) and MAB ( n = 5) as potently as they inhibit intracellular BCG ( n = 8). BCG-expanded T cells inhibited intracellular MAV better than intracellular BCG (** p < 0.01, Mann-Whitney U test). (B) Pure CD4, CD8, and γδ T cells inhibited intracellular MAV, and the level of inhibition was similar to inhibition by total BCG-expanded PBMC. (C) Pure CD4, CD8, and γδ T cells inhibited intracellular MAB, and the level of inhibition is similar to the levels of inhibition mediated by total BCG-expanded PBMC.
    Figure Legend Snippet: BCG-specific T cells cross-protect against MAV and MAB. Total PBMC or subsets of T cells purified after 7 days of optimal BCG stimulation were co-cultured with autologous macrophages infected with MAV or MAB (E:T of 10). Residual mycobacteria quantified 3 days after co-culture and percent inhibition calculated by dividing the number of residual mycobacteria in the presence BCG-stimulated PBMC by the number of residual mycobacteria in control cultures containing medium-rested PBMC. (A) BCG-expanded T cells inhibit intracellular MAV ( n = 8) and MAB ( n = 5) as potently as they inhibit intracellular BCG ( n = 8). BCG-expanded T cells inhibited intracellular MAV better than intracellular BCG (** p < 0.01, Mann-Whitney U test). (B) Pure CD4, CD8, and γδ T cells inhibited intracellular MAV, and the level of inhibition was similar to inhibition by total BCG-expanded PBMC. (C) Pure CD4, CD8, and γδ T cells inhibited intracellular MAB, and the level of inhibition is similar to the levels of inhibition mediated by total BCG-expanded PBMC.

    Techniques Used: Purification, Cell Culture, Infection, Co-Culture Assay, Inhibition, MANN-WHITNEY

    BCG vaccination or Mtb infection of mice induces MAV and MAB reactive immunity. (A) Groups of C57BL/6 mice ( n = 4–5 per group) were vaccinated once or twice intranasally with BCG. Four weeks later mice were euthanized. Splenic cells were harvested from vaccinated and control mice. Cells were rested in medium or stimulated overnight with live BCG at MOI of 3, MAV at MOI of 3, and MAV-WL in IFN-γ ELISPOT assays. Shown are the means ± SE of IFN-γ SFC per million splenic cells. The number of IFN-γ SFC following stimulation with BCG, MAV, and MAV-WL were significantly higher in mice which received one or two BCG vaccinations compared to unvaccinated mice (* P < 0.05, Mann-Whitney U test). The number of mycobacteria-induced IFN-γ SFC were similar following one versus two BCG vaccination ( P > 0.05). (B) C57BL6 infected with aerosolized Mtb ( n = 5) were euthanized 4 weeks after infection. Splenic cells from uninfected and infected mice were used in IFN-γ ELISPOT assays. Mtb-infected mice had significantly more MAV and MAB cross-reactive IFN-γ SFC compared to uninfected mice (* P < 0.05, Mann-Whitney U test).
    Figure Legend Snippet: BCG vaccination or Mtb infection of mice induces MAV and MAB reactive immunity. (A) Groups of C57BL/6 mice ( n = 4–5 per group) were vaccinated once or twice intranasally with BCG. Four weeks later mice were euthanized. Splenic cells were harvested from vaccinated and control mice. Cells were rested in medium or stimulated overnight with live BCG at MOI of 3, MAV at MOI of 3, and MAV-WL in IFN-γ ELISPOT assays. Shown are the means ± SE of IFN-γ SFC per million splenic cells. The number of IFN-γ SFC following stimulation with BCG, MAV, and MAV-WL were significantly higher in mice which received one or two BCG vaccinations compared to unvaccinated mice (* P < 0.05, Mann-Whitney U test). The number of mycobacteria-induced IFN-γ SFC were similar following one versus two BCG vaccination ( P > 0.05). (B) C57BL6 infected with aerosolized Mtb ( n = 5) were euthanized 4 weeks after infection. Splenic cells from uninfected and infected mice were used in IFN-γ ELISPOT assays. Mtb-infected mice had significantly more MAV and MAB cross-reactive IFN-γ SFC compared to uninfected mice (* P < 0.05, Mann-Whitney U test).

    Techniques Used: Infection, Enzyme-linked Immunospot, MANN-WHITNEY

    BCG vaccination in humans induces MAV and MAB cross-reactive T cells. Paired pre-and post-vaccination PBMC from recently BCG vaccinated volunteers living in St. Louis, MO ( n = 5) were used. PBMC were labeled with CFSE and stimulated with BCG, MAB WL, or MAB WL. Medium rested PBMC were used as negative controls. On day 7, cells were restimulated with PMA/ionomycin for 2 h, viable cells were counted and cells were stained for surface and intracellular markers for flow cytometry study. (A–C) show the data for proliferating and IFN-γ producing T cells. (D–F) show the data for proliferating and GZM-A producing T cells. There were significantly higher absolute numbers (AN, per ml of cultures) of BCG-reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03, Wilcoxon Matched Pairs test), MAV WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), MAV WL reactive CFSE lo IFN-γ + CD8 + T cells ( P = 0.03), MAV WL reactive CFSE lo Granzyme A + CD8 + T cells ( P = 0.03), MAB WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), and MAB WL reactive CFSE lo Granzyme A + CD4 + T cells ( P = 0.03), indicating that BCG induces NTM cross-reactive immunity. * P < 0.05.
    Figure Legend Snippet: BCG vaccination in humans induces MAV and MAB cross-reactive T cells. Paired pre-and post-vaccination PBMC from recently BCG vaccinated volunteers living in St. Louis, MO ( n = 5) were used. PBMC were labeled with CFSE and stimulated with BCG, MAB WL, or MAB WL. Medium rested PBMC were used as negative controls. On day 7, cells were restimulated with PMA/ionomycin for 2 h, viable cells were counted and cells were stained for surface and intracellular markers for flow cytometry study. (A–C) show the data for proliferating and IFN-γ producing T cells. (D–F) show the data for proliferating and GZM-A producing T cells. There were significantly higher absolute numbers (AN, per ml of cultures) of BCG-reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03, Wilcoxon Matched Pairs test), MAV WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), MAV WL reactive CFSE lo IFN-γ + CD8 + T cells ( P = 0.03), MAV WL reactive CFSE lo Granzyme A + CD8 + T cells ( P = 0.03), MAB WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), and MAB WL reactive CFSE lo Granzyme A + CD4 + T cells ( P = 0.03), indicating that BCG induces NTM cross-reactive immunity. * P < 0.05.

    Techniques Used: Labeling, Staining, Flow Cytometry

    a mortivallis dsm 44261t  (ATCC)


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    ATCC a mortivallis dsm 44261t
    A Mortivallis Dsm 44261t, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a mortivallis dsm 44261t  (ATCC)


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    ATCC a mortivallis dsm 44261t
    A Mortivallis Dsm 44261t, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a mortivallis dsm 44261t  (ATCC)


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    ATCC a mortivallis dsm 44261t
    A Mortivallis Dsm 44261t, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    a mortivallis dsm 44261t  (ATCC)


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    ATCC a mortivallis dsm 44261t
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    dsm 44261  (ATCC)


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    ATCC dsm 44261
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    ATCC mycobacterium abscessus nr 44261
    Mycobacterium Abscessus Nr 44261, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC mav whole lysate wl
    Previous exposure <t>to</t> <t>BCG</t> or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or <t>MAV</t> (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.
    Mav Whole Lysate Wl, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC a mortivallis dsm 44261t
    Previous exposure <t>to</t> <t>BCG</t> or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or <t>MAV</t> (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.
    A Mortivallis Dsm 44261t, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    ATCC dsm 44261
    Previous exposure <t>to</t> <t>BCG</t> or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or <t>MAV</t> (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.
    Dsm 44261, supplied by ATCC, used in various techniques. Bioz Stars score: 91/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Previous exposure to BCG or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or MAV (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.

    Journal: Frontiers in Immunology

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    doi: 10.3389/fimmu.2019.00234

    Figure Lengend Snippet: Previous exposure to BCG or Mtb induces cross-reactive T cells against NTM. PBMC from BCG-vaccinated and/or latent TB infected individuals ( n = 6) were CFSE-labeled and stimulated with optimal concentrations of live BCG (Connaught) or MAV (ATCC 700898). On day 7, cells were restimulated with PMA/ionomycin and the total percentages of CFSE lo (proliferating) and IFN-γ producing T cells were determined by flow cytometry. (A) Flow cytometry plots of a single volunteer. Lymphocytes gated on the basis of forward and side scatter and then regated on CD3 + cells were analyzed for CFSE and IFN-γ expression by use of FlowJo software. The number in the upper left quadrant of each dot plot refers to the percentage of CFSE lo IFN-γ + CD3 + T cells detected after in vitro stimulation. (B) Composite data from 6 volunteers. Stimulation indices were calculated by dividing the absolute numbers of CFSE lo IFN-γ + CD3 + T cells in cultures containing BCG or MAV by the absolute numbers of CFSE lo /IFN-γ + CD3 + T cells in medium-rested cultures. Stimulation with MAV led to expansion of effector T cells comparable to the level obtained with BCG stimulation ( P > 0.05, Wilcoxon matched-pairs test). The bars show ranges and the lines show mean values. NS, not significant.

    Article Snippet: Connaught BCG, MAV (ATCC 700898), MAV-whole lysate (WL), MAB (NR-44261, BEI Resources) and MAB-WL were used for in vitro expansion of mycobacterium-specific T cells.

    Techniques: Infection, Labeling, Flow Cytometry, Expressing, Software, In Vitro

    MAV and MAB cross-reactive immunity includes Th1 and Th17 responses. (A) Schematic of experiments conducted to measure MAV or MAB cross-reactive immunity. PBMC from BCG-vaccinated or latent TB infected individuals ( n = 6) were stimulated with BCG or rested in media for 7 days. Then, these expanded cells were co-cultured with MAV- or MAB-infected autologous monocytes at an E:T ratio of 10:1. On day 3 of co-culture, Th1, Th2, and Th17 responses were measured in co-culture supernatants using CBA. Fold changes for each cytokine was calculated by dividing the amount of cytokine produced following restimulation with MAV or MAB by the amount produced in medium-rested cultures. (B) Exposure of BCG-expanded T cells to MAV-infected macrophages increased IL-17, IFN-γ, and TNF-α by 156 ± 62, 11 ± 1.7, 10.3 ± 2.9 pg/ml (Mean ± SE), respectively. IL-10, IL-6, and IL-2 showed no marked changes with fold changes (mean ± SE) of 0.3 ± 0.1, 0.9 ± 0.2, 0.9 ± 0.2 pg/ml, respectively. (C) Similarly, exposure of BCG-expanded T cells to MAB-infected macrophages increased IL-17 and IFN-γ in by 7.2 ± 1.6, and 5.6 ± 2, mean fold ± SE. There were no marked changes in the levels of TNF-α, IL-10, IL-6, and IL-2 with fold changes (mean ± SE) of 1.2 ± 0.3, 0.2 ± 0.2, 0.6 ± 0.3, 0.8 ± 0.4 pg/ml, respectively.

    Journal: Frontiers in Immunology

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    doi: 10.3389/fimmu.2019.00234

    Figure Lengend Snippet: MAV and MAB cross-reactive immunity includes Th1 and Th17 responses. (A) Schematic of experiments conducted to measure MAV or MAB cross-reactive immunity. PBMC from BCG-vaccinated or latent TB infected individuals ( n = 6) were stimulated with BCG or rested in media for 7 days. Then, these expanded cells were co-cultured with MAV- or MAB-infected autologous monocytes at an E:T ratio of 10:1. On day 3 of co-culture, Th1, Th2, and Th17 responses were measured in co-culture supernatants using CBA. Fold changes for each cytokine was calculated by dividing the amount of cytokine produced following restimulation with MAV or MAB by the amount produced in medium-rested cultures. (B) Exposure of BCG-expanded T cells to MAV-infected macrophages increased IL-17, IFN-γ, and TNF-α by 156 ± 62, 11 ± 1.7, 10.3 ± 2.9 pg/ml (Mean ± SE), respectively. IL-10, IL-6, and IL-2 showed no marked changes with fold changes (mean ± SE) of 0.3 ± 0.1, 0.9 ± 0.2, 0.9 ± 0.2 pg/ml, respectively. (C) Similarly, exposure of BCG-expanded T cells to MAB-infected macrophages increased IL-17 and IFN-γ in by 7.2 ± 1.6, and 5.6 ± 2, mean fold ± SE. There were no marked changes in the levels of TNF-α, IL-10, IL-6, and IL-2 with fold changes (mean ± SE) of 1.2 ± 0.3, 0.2 ± 0.2, 0.6 ± 0.3, 0.8 ± 0.4 pg/ml, respectively.

    Article Snippet: Connaught BCG, MAV (ATCC 700898), MAV-whole lysate (WL), MAB (NR-44261, BEI Resources) and MAB-WL were used for in vitro expansion of mycobacterium-specific T cells.

    Techniques: Infection, Cell Culture, Co-Culture Assay, Produced

    BCG-specific T cells cross-protect against MAV and MAB. Total PBMC or subsets of T cells purified after 7 days of optimal BCG stimulation were co-cultured with autologous macrophages infected with MAV or MAB (E:T of 10). Residual mycobacteria quantified 3 days after co-culture and percent inhibition calculated by dividing the number of residual mycobacteria in the presence BCG-stimulated PBMC by the number of residual mycobacteria in control cultures containing medium-rested PBMC. (A) BCG-expanded T cells inhibit intracellular MAV ( n = 8) and MAB ( n = 5) as potently as they inhibit intracellular BCG ( n = 8). BCG-expanded T cells inhibited intracellular MAV better than intracellular BCG (** p < 0.01, Mann-Whitney U test). (B) Pure CD4, CD8, and γδ T cells inhibited intracellular MAV, and the level of inhibition was similar to inhibition by total BCG-expanded PBMC. (C) Pure CD4, CD8, and γδ T cells inhibited intracellular MAB, and the level of inhibition is similar to the levels of inhibition mediated by total BCG-expanded PBMC.

    Journal: Frontiers in Immunology

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    doi: 10.3389/fimmu.2019.00234

    Figure Lengend Snippet: BCG-specific T cells cross-protect against MAV and MAB. Total PBMC or subsets of T cells purified after 7 days of optimal BCG stimulation were co-cultured with autologous macrophages infected with MAV or MAB (E:T of 10). Residual mycobacteria quantified 3 days after co-culture and percent inhibition calculated by dividing the number of residual mycobacteria in the presence BCG-stimulated PBMC by the number of residual mycobacteria in control cultures containing medium-rested PBMC. (A) BCG-expanded T cells inhibit intracellular MAV ( n = 8) and MAB ( n = 5) as potently as they inhibit intracellular BCG ( n = 8). BCG-expanded T cells inhibited intracellular MAV better than intracellular BCG (** p < 0.01, Mann-Whitney U test). (B) Pure CD4, CD8, and γδ T cells inhibited intracellular MAV, and the level of inhibition was similar to inhibition by total BCG-expanded PBMC. (C) Pure CD4, CD8, and γδ T cells inhibited intracellular MAB, and the level of inhibition is similar to the levels of inhibition mediated by total BCG-expanded PBMC.

    Article Snippet: Connaught BCG, MAV (ATCC 700898), MAV-whole lysate (WL), MAB (NR-44261, BEI Resources) and MAB-WL were used for in vitro expansion of mycobacterium-specific T cells.

    Techniques: Purification, Cell Culture, Infection, Co-Culture Assay, Inhibition, MANN-WHITNEY

    BCG vaccination or Mtb infection of mice induces MAV and MAB reactive immunity. (A) Groups of C57BL/6 mice ( n = 4–5 per group) were vaccinated once or twice intranasally with BCG. Four weeks later mice were euthanized. Splenic cells were harvested from vaccinated and control mice. Cells were rested in medium or stimulated overnight with live BCG at MOI of 3, MAV at MOI of 3, and MAV-WL in IFN-γ ELISPOT assays. Shown are the means ± SE of IFN-γ SFC per million splenic cells. The number of IFN-γ SFC following stimulation with BCG, MAV, and MAV-WL were significantly higher in mice which received one or two BCG vaccinations compared to unvaccinated mice (* P < 0.05, Mann-Whitney U test). The number of mycobacteria-induced IFN-γ SFC were similar following one versus two BCG vaccination ( P > 0.05). (B) C57BL6 infected with aerosolized Mtb ( n = 5) were euthanized 4 weeks after infection. Splenic cells from uninfected and infected mice were used in IFN-γ ELISPOT assays. Mtb-infected mice had significantly more MAV and MAB cross-reactive IFN-γ SFC compared to uninfected mice (* P < 0.05, Mann-Whitney U test).

    Journal: Frontiers in Immunology

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    doi: 10.3389/fimmu.2019.00234

    Figure Lengend Snippet: BCG vaccination or Mtb infection of mice induces MAV and MAB reactive immunity. (A) Groups of C57BL/6 mice ( n = 4–5 per group) were vaccinated once or twice intranasally with BCG. Four weeks later mice were euthanized. Splenic cells were harvested from vaccinated and control mice. Cells were rested in medium or stimulated overnight with live BCG at MOI of 3, MAV at MOI of 3, and MAV-WL in IFN-γ ELISPOT assays. Shown are the means ± SE of IFN-γ SFC per million splenic cells. The number of IFN-γ SFC following stimulation with BCG, MAV, and MAV-WL were significantly higher in mice which received one or two BCG vaccinations compared to unvaccinated mice (* P < 0.05, Mann-Whitney U test). The number of mycobacteria-induced IFN-γ SFC were similar following one versus two BCG vaccination ( P > 0.05). (B) C57BL6 infected with aerosolized Mtb ( n = 5) were euthanized 4 weeks after infection. Splenic cells from uninfected and infected mice were used in IFN-γ ELISPOT assays. Mtb-infected mice had significantly more MAV and MAB cross-reactive IFN-γ SFC compared to uninfected mice (* P < 0.05, Mann-Whitney U test).

    Article Snippet: Connaught BCG, MAV (ATCC 700898), MAV-whole lysate (WL), MAB (NR-44261, BEI Resources) and MAB-WL were used for in vitro expansion of mycobacterium-specific T cells.

    Techniques: Infection, Enzyme-linked Immunospot, MANN-WHITNEY

    BCG vaccination in humans induces MAV and MAB cross-reactive T cells. Paired pre-and post-vaccination PBMC from recently BCG vaccinated volunteers living in St. Louis, MO ( n = 5) were used. PBMC were labeled with CFSE and stimulated with BCG, MAB WL, or MAB WL. Medium rested PBMC were used as negative controls. On day 7, cells were restimulated with PMA/ionomycin for 2 h, viable cells were counted and cells were stained for surface and intracellular markers for flow cytometry study. (A–C) show the data for proliferating and IFN-γ producing T cells. (D–F) show the data for proliferating and GZM-A producing T cells. There were significantly higher absolute numbers (AN, per ml of cultures) of BCG-reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03, Wilcoxon Matched Pairs test), MAV WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), MAV WL reactive CFSE lo IFN-γ + CD8 + T cells ( P = 0.03), MAV WL reactive CFSE lo Granzyme A + CD8 + T cells ( P = 0.03), MAB WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), and MAB WL reactive CFSE lo Granzyme A + CD4 + T cells ( P = 0.03), indicating that BCG induces NTM cross-reactive immunity. * P < 0.05.

    Journal: Frontiers in Immunology

    Article Title: BCG Vaccination Induces M. avium and M. abscessus Cross-Protective Immunity

    doi: 10.3389/fimmu.2019.00234

    Figure Lengend Snippet: BCG vaccination in humans induces MAV and MAB cross-reactive T cells. Paired pre-and post-vaccination PBMC from recently BCG vaccinated volunteers living in St. Louis, MO ( n = 5) were used. PBMC were labeled with CFSE and stimulated with BCG, MAB WL, or MAB WL. Medium rested PBMC were used as negative controls. On day 7, cells were restimulated with PMA/ionomycin for 2 h, viable cells were counted and cells were stained for surface and intracellular markers for flow cytometry study. (A–C) show the data for proliferating and IFN-γ producing T cells. (D–F) show the data for proliferating and GZM-A producing T cells. There were significantly higher absolute numbers (AN, per ml of cultures) of BCG-reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03, Wilcoxon Matched Pairs test), MAV WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), MAV WL reactive CFSE lo IFN-γ + CD8 + T cells ( P = 0.03), MAV WL reactive CFSE lo Granzyme A + CD8 + T cells ( P = 0.03), MAB WL reactive CFSE lo IFN-γ + CD4 + T cells ( P = 0.03), and MAB WL reactive CFSE lo Granzyme A + CD4 + T cells ( P = 0.03), indicating that BCG induces NTM cross-reactive immunity. * P < 0.05.

    Article Snippet: Connaught BCG, MAV (ATCC 700898), MAV-whole lysate (WL), MAB (NR-44261, BEI Resources) and MAB-WL were used for in vitro expansion of mycobacterium-specific T cells.

    Techniques: Labeling, Staining, Flow Cytometry