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Mycobacterium marinum: a potential immunotherapy for Mycobacterium tuberculosis infection.

Tian WW, Wang QQ, Liu WD, Shen JP, Wang HS - Drug Des Devel Ther (2013)

Bottom Line: Moreover, the M. marinum group expressed more interleukin (IL)-1B and IL-12p40 on day 3 (IL-1B: P = 0.003 and 0.004, respectively; IL-12p40: P = 0.001 and 0.011, respectively), a higher level of CXCL10 on day 1 (P = 0.006 and 0.026, respectively), and higher levels of chemokine (C-X-C motif) ligand (CXCL) 8 and chemokine (C motif) ligand (XCL) 1 on day 3 (CXCL8: P = 0.012 and 0.014, respectively; XCL1: P = 0.000 and 0.000, respectively).The M. marinum stimulated coculture group also secreted more tumor necrosis factor (TNF)-α, IL-1β, and IL-10 on day 1 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.000, respectively; IL-10: P = 0.002 and 0.019, respectively) and day 3 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.001, respectively; IL-10: P = 0.000 and 0.000, respectively).In addition, the colony-forming units (an index of viability) of M. marinum in the M. marinum stimulated coculture group was significantly less than that of BCG and H37Ra in their corresponding bacillus stimulated groups (P = 0.037 and 0.013, respectively).

View Article: PubMed Central - PubMed

Affiliation: Chinese Academy of Medical Sciences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanjing, Jiangsu, People's Republic of China.

ABSTRACT

Purpose: The aim of the present study was to investigate the immune response induced by Mycobacterium marinum infection in vitro and the potential of M. marinum as an immunotherapy for M. tuberculosis infection.

Methods: The potential human immune response to certain bacillus infections was investigated in an immune cell-bacillus coculture system in vitro. As a potential novel immunotherapy, M. marinum was studied and compared with two other bacilli, Bacillus Calmette-Guérin (BCG) and live attenuated M. tuberculosis. We examined the changes in both the bacilli and immune cells, especially the time course of the viability of mycobacteria in the coculture system and host immune responses including multinuclear giant cell formation by Wright-Giemsa modified staining, macrophage polarization by cell surface antigen expression, and cytokines/chemokine production by both mRNA expression and protein secretion.

Results: The M. marinum stimulated coculture group showed more expression of CD209, CD68, CD80, and CD86 than the BCG and M. tuberculosis (an attenuated strain, H37Ra) groups, although the differences were not statistically significant. Moreover, the M. marinum group expressed more interleukin (IL)-1B and IL-12p40 on day 3 (IL-1B: P = 0.003 and 0.004, respectively; IL-12p40: P = 0.001 and 0.011, respectively), a higher level of CXCL10 on day 1 (P = 0.006 and 0.026, respectively), and higher levels of chemokine (C-X-C motif) ligand (CXCL) 8 and chemokine (C motif) ligand (XCL) 1 on day 3 (CXCL8: P = 0.012 and 0.014, respectively; XCL1: P = 0.000 and 0.000, respectively). The M. marinum stimulated coculture group also secreted more tumor necrosis factor (TNF)-α, IL-1β, and IL-10 on day 1 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.000, respectively; IL-10: P = 0.002 and 0.019, respectively) and day 3 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.001, respectively; IL-10: P = 0.000 and 0.000, respectively). In addition, the colony-forming units (an index of viability) of M. marinum in the M. marinum stimulated coculture group was significantly less than that of BCG and H37Ra in their corresponding bacillus stimulated groups (P = 0.037 and 0.013, respectively).

Conclusion: Our results indicated that M. marinum could be a potentially safe and effective immunotherapy.

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Related in: MedlinePlus

The levels of cytokines secreted in the culture supernatant were detected by ELISA on day 1, 3, and 7.Notes: The group M+P, M+P+B, and M+B refer to the control group without bacilli, experimental groups, and control groups without peripheral blood mononuclear cells, respectively, while the bacilli B, M, and H refer to Bacillus Calmette-Guérin, M. marinum, and H37Ra, respectively.Abbreviations: IL, interleukin; TNF, tumor necrosis factor; ELISA, enzyme-linked immunosorbent assay.
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f4-dddt-7-669: The levels of cytokines secreted in the culture supernatant were detected by ELISA on day 1, 3, and 7.Notes: The group M+P, M+P+B, and M+B refer to the control group without bacilli, experimental groups, and control groups without peripheral blood mononuclear cells, respectively, while the bacilli B, M, and H refer to Bacillus Calmette-Guérin, M. marinum, and H37Ra, respectively.Abbreviations: IL, interleukin; TNF, tumor necrosis factor; ELISA, enzyme-linked immunosorbent assay.

Mentions: Cytokines were also detected by ELISA in the supernatants of various cell cultures on days 1, 3, and 7. In the M+P+B groups, TNF-α and IL-1β secretion peaked on day 1, IFN-γ was higher on days 3 and 7, whereas IL-10 showed higher levels on day 1 and 3 than on day 7. The culture supernatants from the M+P+B−M group contained higher levels of TNF-α, IL-1 β, and IL-10 than those from the M+P, M+B−M, M+P+B−B, and M+P+B−H groups on days 1 and 3 (day 1, TNF-α: P = 0.000, 0.001, 0.000, and 0.000, respectively; IL-1β: P = 0.000, 0.000, 0.000, and 0.000, respectively; IL-10: P = 0.001, 0.001, 0.002, and 0.019, respectively; day 3, TNF-α: P = 0.000, 0.001, 0.000, and 0.000, respectively; IL-1β: P = 0.000, 0.000, 0.000, and 0.001, respectively; IL-10: P = 0.000, 0.000, 0.000, and 0.000, respectively). On day 1, a higher level of IFN-γ was observed in the culture supernatant from the M+P+B−M group than those from the M+P and M+B−M groups (P = 0.012 and 0.039, respectively). No significant differences in IFN-γ secretion were found between the M+P+B−M and the M+P+B−H group on day 1 and 3. However, higher IFN-γ secretion was observed in the M+P+B−H group when compared with the M+P+B−B group (P = 0.048 and 0.012, respectively) (Table 4 and Figure 4). The amount of IL-12/23 was surprisingly low in the supernatants of all groups (IL-12 < 1.5 pg/mL, IL-23 < 4 pg/mL); nonetheless, IL-12/23 levels were slightly higher in the culture supernatant from the M+P+B−M group (data not shown). The IL-4 levels in the culture supernatants were below the detection level of ELISA (data not shown).


Mycobacterium marinum: a potential immunotherapy for Mycobacterium tuberculosis infection.

Tian WW, Wang QQ, Liu WD, Shen JP, Wang HS - Drug Des Devel Ther (2013)

The levels of cytokines secreted in the culture supernatant were detected by ELISA on day 1, 3, and 7.Notes: The group M+P, M+P+B, and M+B refer to the control group without bacilli, experimental groups, and control groups without peripheral blood mononuclear cells, respectively, while the bacilli B, M, and H refer to Bacillus Calmette-Guérin, M. marinum, and H37Ra, respectively.Abbreviations: IL, interleukin; TNF, tumor necrosis factor; ELISA, enzyme-linked immunosorbent assay.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3733875&req=5

f4-dddt-7-669: The levels of cytokines secreted in the culture supernatant were detected by ELISA on day 1, 3, and 7.Notes: The group M+P, M+P+B, and M+B refer to the control group without bacilli, experimental groups, and control groups without peripheral blood mononuclear cells, respectively, while the bacilli B, M, and H refer to Bacillus Calmette-Guérin, M. marinum, and H37Ra, respectively.Abbreviations: IL, interleukin; TNF, tumor necrosis factor; ELISA, enzyme-linked immunosorbent assay.
Mentions: Cytokines were also detected by ELISA in the supernatants of various cell cultures on days 1, 3, and 7. In the M+P+B groups, TNF-α and IL-1β secretion peaked on day 1, IFN-γ was higher on days 3 and 7, whereas IL-10 showed higher levels on day 1 and 3 than on day 7. The culture supernatants from the M+P+B−M group contained higher levels of TNF-α, IL-1 β, and IL-10 than those from the M+P, M+B−M, M+P+B−B, and M+P+B−H groups on days 1 and 3 (day 1, TNF-α: P = 0.000, 0.001, 0.000, and 0.000, respectively; IL-1β: P = 0.000, 0.000, 0.000, and 0.000, respectively; IL-10: P = 0.001, 0.001, 0.002, and 0.019, respectively; day 3, TNF-α: P = 0.000, 0.001, 0.000, and 0.000, respectively; IL-1β: P = 0.000, 0.000, 0.000, and 0.001, respectively; IL-10: P = 0.000, 0.000, 0.000, and 0.000, respectively). On day 1, a higher level of IFN-γ was observed in the culture supernatant from the M+P+B−M group than those from the M+P and M+B−M groups (P = 0.012 and 0.039, respectively). No significant differences in IFN-γ secretion were found between the M+P+B−M and the M+P+B−H group on day 1 and 3. However, higher IFN-γ secretion was observed in the M+P+B−H group when compared with the M+P+B−B group (P = 0.048 and 0.012, respectively) (Table 4 and Figure 4). The amount of IL-12/23 was surprisingly low in the supernatants of all groups (IL-12 < 1.5 pg/mL, IL-23 < 4 pg/mL); nonetheless, IL-12/23 levels were slightly higher in the culture supernatant from the M+P+B−M group (data not shown). The IL-4 levels in the culture supernatants were below the detection level of ELISA (data not shown).

Bottom Line: Moreover, the M. marinum group expressed more interleukin (IL)-1B and IL-12p40 on day 3 (IL-1B: P = 0.003 and 0.004, respectively; IL-12p40: P = 0.001 and 0.011, respectively), a higher level of CXCL10 on day 1 (P = 0.006 and 0.026, respectively), and higher levels of chemokine (C-X-C motif) ligand (CXCL) 8 and chemokine (C motif) ligand (XCL) 1 on day 3 (CXCL8: P = 0.012 and 0.014, respectively; XCL1: P = 0.000 and 0.000, respectively).The M. marinum stimulated coculture group also secreted more tumor necrosis factor (TNF)-α, IL-1β, and IL-10 on day 1 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.000, respectively; IL-10: P = 0.002 and 0.019, respectively) and day 3 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.001, respectively; IL-10: P = 0.000 and 0.000, respectively).In addition, the colony-forming units (an index of viability) of M. marinum in the M. marinum stimulated coculture group was significantly less than that of BCG and H37Ra in their corresponding bacillus stimulated groups (P = 0.037 and 0.013, respectively).

View Article: PubMed Central - PubMed

Affiliation: Chinese Academy of Medical Sciences and Peking Union Medical College, Jiangsu Key Laboratory of Molecular Biology for Skin Diseases and STIs, Nanjing, Jiangsu, People's Republic of China.

ABSTRACT

Purpose: The aim of the present study was to investigate the immune response induced by Mycobacterium marinum infection in vitro and the potential of M. marinum as an immunotherapy for M. tuberculosis infection.

Methods: The potential human immune response to certain bacillus infections was investigated in an immune cell-bacillus coculture system in vitro. As a potential novel immunotherapy, M. marinum was studied and compared with two other bacilli, Bacillus Calmette-Guérin (BCG) and live attenuated M. tuberculosis. We examined the changes in both the bacilli and immune cells, especially the time course of the viability of mycobacteria in the coculture system and host immune responses including multinuclear giant cell formation by Wright-Giemsa modified staining, macrophage polarization by cell surface antigen expression, and cytokines/chemokine production by both mRNA expression and protein secretion.

Results: The M. marinum stimulated coculture group showed more expression of CD209, CD68, CD80, and CD86 than the BCG and M. tuberculosis (an attenuated strain, H37Ra) groups, although the differences were not statistically significant. Moreover, the M. marinum group expressed more interleukin (IL)-1B and IL-12p40 on day 3 (IL-1B: P = 0.003 and 0.004, respectively; IL-12p40: P = 0.001 and 0.011, respectively), a higher level of CXCL10 on day 1 (P = 0.006 and 0.026, respectively), and higher levels of chemokine (C-X-C motif) ligand (CXCL) 8 and chemokine (C motif) ligand (XCL) 1 on day 3 (CXCL8: P = 0.012 and 0.014, respectively; XCL1: P = 0.000 and 0.000, respectively). The M. marinum stimulated coculture group also secreted more tumor necrosis factor (TNF)-α, IL-1β, and IL-10 on day 1 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.000, respectively; IL-10: P = 0.002 and 0.019, respectively) and day 3 (TNF-α: P = 0.000 and 0.000, respectively; IL-1β: P = 0.000 and 0.001, respectively; IL-10: P = 0.000 and 0.000, respectively). In addition, the colony-forming units (an index of viability) of M. marinum in the M. marinum stimulated coculture group was significantly less than that of BCG and H37Ra in their corresponding bacillus stimulated groups (P = 0.037 and 0.013, respectively).

Conclusion: Our results indicated that M. marinum could be a potentially safe and effective immunotherapy.

Show MeSH
Related in: MedlinePlus