Abstract
Brucella abortus is a Gram negative facultative intracellular pathogen of cattle, and an important zoonosis in humans worldwide. Previous studies have shown that dendritic cells (DC) from humans and mice are highly permissive for Brucella survival and proliferation. Impairment of DC activation and maturation by Brucella infection has also been reported in these two species. The aim of this study was to characterize infection of bovine DC with B. abortus. Monocyte-derived DC (mdDC) were cultured from bovine peripheral blood mononuclear cells (PBMC) using the recombinant bovine cytokines IL-4 and GM-CSF. The resulting mdDC were DEC205+, MHC class IIhi. Approximately 70% of the cultured cells were DEC205+, MHC II+. MdDC were infected with B. abortus strain 2308 at an MOI of 1 and 100. Parallel infection experiments were performed in monocyte derived macrophages (mdM) isolated from the same subjects. Bacteria were successfully killed by mdDC by 24hours post infection (PI) with high and low dose of B. abortus, bacteria persisted in mdM infected with a high dose. Expression of IL-1b, IL-6, IL-10, IL-12p40, IFNγ, iNOS and TNFα in B. abortus infected and LPS stimulated mdDC at 6 and 24hours PI were evaluated using RT-qPCR. At 6hours PI all transcripts were increased over control cells and significantly less IL-10, IL-12p40, and IFNγ were expressed in mdDC infected with B. abortus compared to LPS stimulation. Evaluation of mdDC cultures by flow cytometry was performed. Flow cytometric analysis of infected and LPS stimulated mdDC 24hours PI showed expression of CD80 and CD86 was impaired in two of the three animals analyzed. MHC class II expression was equivocal between the groups. From these results we conclude that cultured bovine mdDC are not permissive for intracellular proliferation of B. abortus, and infected mdDC exhibit some signs of maturational and activational impairment.
| Original language | English (US) |
|---|---|
| Pages (from-to) | 255-261 |
| Number of pages | 7 |
| Journal | Veterinary Immunology and Immunopathology |
| Volume | 149 |
| Issue number | 3-4 |
| DOIs | |
| State | Published - Oct 15 2012 |
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Keywords
- Bovine dendritic cell
- Brucella abortus
- Dendritic cell maturation
ASJC Scopus subject areas
- Immunology
- veterinary(all)
Cite this
Characterization of Brucella abortus infection of bovine monocyte-derived dendritic cells. / Heller, Meera C; Watson, Johanna L; Blanchard, Myra; Jackson, K. A.; Stott, Jeffrey L; Tsolis, Renee M.
In: Veterinary Immunology and Immunopathology, Vol. 149, No. 3-4, 15.10.2012, p. 255-261.Research output: Contribution to journal › Article
}
TY - JOUR
T1 - Characterization of Brucella abortus infection of bovine monocyte-derived dendritic cells
AU - Heller, Meera C
AU - Watson, Johanna L
AU - Blanchard, Myra
AU - Jackson, K. A.
AU - Stott, Jeffrey L
AU - Tsolis, Renee M
PY - 2012/10/15
Y1 - 2012/10/15
N2 - Brucella abortus is a Gram negative facultative intracellular pathogen of cattle, and an important zoonosis in humans worldwide. Previous studies have shown that dendritic cells (DC) from humans and mice are highly permissive for Brucella survival and proliferation. Impairment of DC activation and maturation by Brucella infection has also been reported in these two species. The aim of this study was to characterize infection of bovine DC with B. abortus. Monocyte-derived DC (mdDC) were cultured from bovine peripheral blood mononuclear cells (PBMC) using the recombinant bovine cytokines IL-4 and GM-CSF. The resulting mdDC were DEC205+, MHC class IIhi. Approximately 70% of the cultured cells were DEC205+, MHC II+. MdDC were infected with B. abortus strain 2308 at an MOI of 1 and 100. Parallel infection experiments were performed in monocyte derived macrophages (mdM) isolated from the same subjects. Bacteria were successfully killed by mdDC by 24hours post infection (PI) with high and low dose of B. abortus, bacteria persisted in mdM infected with a high dose. Expression of IL-1b, IL-6, IL-10, IL-12p40, IFNγ, iNOS and TNFα in B. abortus infected and LPS stimulated mdDC at 6 and 24hours PI were evaluated using RT-qPCR. At 6hours PI all transcripts were increased over control cells and significantly less IL-10, IL-12p40, and IFNγ were expressed in mdDC infected with B. abortus compared to LPS stimulation. Evaluation of mdDC cultures by flow cytometry was performed. Flow cytometric analysis of infected and LPS stimulated mdDC 24hours PI showed expression of CD80 and CD86 was impaired in two of the three animals analyzed. MHC class II expression was equivocal between the groups. From these results we conclude that cultured bovine mdDC are not permissive for intracellular proliferation of B. abortus, and infected mdDC exhibit some signs of maturational and activational impairment.
AB - Brucella abortus is a Gram negative facultative intracellular pathogen of cattle, and an important zoonosis in humans worldwide. Previous studies have shown that dendritic cells (DC) from humans and mice are highly permissive for Brucella survival and proliferation. Impairment of DC activation and maturation by Brucella infection has also been reported in these two species. The aim of this study was to characterize infection of bovine DC with B. abortus. Monocyte-derived DC (mdDC) were cultured from bovine peripheral blood mononuclear cells (PBMC) using the recombinant bovine cytokines IL-4 and GM-CSF. The resulting mdDC were DEC205+, MHC class IIhi. Approximately 70% of the cultured cells were DEC205+, MHC II+. MdDC were infected with B. abortus strain 2308 at an MOI of 1 and 100. Parallel infection experiments were performed in monocyte derived macrophages (mdM) isolated from the same subjects. Bacteria were successfully killed by mdDC by 24hours post infection (PI) with high and low dose of B. abortus, bacteria persisted in mdM infected with a high dose. Expression of IL-1b, IL-6, IL-10, IL-12p40, IFNγ, iNOS and TNFα in B. abortus infected and LPS stimulated mdDC at 6 and 24hours PI were evaluated using RT-qPCR. At 6hours PI all transcripts were increased over control cells and significantly less IL-10, IL-12p40, and IFNγ were expressed in mdDC infected with B. abortus compared to LPS stimulation. Evaluation of mdDC cultures by flow cytometry was performed. Flow cytometric analysis of infected and LPS stimulated mdDC 24hours PI showed expression of CD80 and CD86 was impaired in two of the three animals analyzed. MHC class II expression was equivocal between the groups. From these results we conclude that cultured bovine mdDC are not permissive for intracellular proliferation of B. abortus, and infected mdDC exhibit some signs of maturational and activational impairment.
KW - Bovine dendritic cell
KW - Brucella abortus
KW - Dendritic cell maturation
UR - http://www.scopus.com/inward/record.url?scp=84866354379&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84866354379&partnerID=8YFLogxK
U2 - 10.1016/j.vetimm.2012.07.006
DO - 10.1016/j.vetimm.2012.07.006
M3 - Article
C2 - 22884262
AN - SCOPUS:84866354379
VL - 149
SP - 255
EP - 261
JO - Veterinary Immunology and Immunopathology
JF - Veterinary Immunology and Immunopathology
SN - 0165-2427
IS - 3-4
ER -