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Coxiella burnetii dormancy in a fatal ten-year multisystem dysfunctional illness: case report.

Sukocheva OA, Manavis J, Kok TW, Turra M, Izzo A, Blumbergs P, Marmion BP - BMC Infect. Dis. (2016)

Bottom Line: During life, extensive clinical and laboratory investigations from different disciplinary stand points failed to deliver a definitive identification of a cause.PCR analysis (COM1/IS1111 genes) confirmed the presence of C.b.The possible mechanisms and molecular adaptations for this alternative C.b. life style are discussed.

View Article: PubMed Central - PubMed

Affiliation: Q Fever Research Group (1993-2009), Hanson Institute, Adelaide, South Australia.

ABSTRACT

Background: In a previous study of a Q fever outbreak in Birmingham, our group identified a non-infective complex of Coxiella burnetii (C.b.) antigens able to survive in the host and provoked aberrant humoral and cell-mediated immunity responses. The study led to recognition of a possible pathogenic link between C.b. infection and subsequent long-term post Q fever fatigue syndrome (QFS). This report presents an unusually severe case of C.b. antigen and DNA detection in post-mortem specimens from a patient with QFS.

Case presentation: We report a 19-year old female patient who became ill with an acute unexplained febrile encephalitis-like illness, followed by increasingly severe multisystem dysfunction and death 10 years later. During life, extensive clinical and laboratory investigations from different disciplinary stand points failed to deliver a definitive identification of a cause. Given the history of susceptibility to infection from birth, acute fever and the diagnosis of "post viral syndrome", tests for infective agents were done starting with C.b. and Legionella pneumophila. The patient had previously visited farms a number of times. Comprehensive neuropathological assessment at the time of autopsy had not revealed gross or microscopic abnormalities. The aim was to extend detailed studies with the post-mortem samples and identify possible factors driving severe disturbance of homeostasis and organ dysfunction exhibited by the course of the patient's ten-year illness. Immunohistochemistry for C.b. antigen and PCR for DNA were tested on paraffin embedded blocks of autopsy tissues from brain, spleen, liver, lymph nodes (LN), bone marrow (BM), heart and lung. Standard H&E staining of brain sections was unrevealing. Immuno-staining analysis for astrocyte cytoskeleton proteins using glial fibrillary acidic protein (GFAP) antibodies showed a reactive morphology. Coxiella antigens were demonstrated in GFAP immuno-positive grey and white matter astrocytes, spleen, liver, heart, BM and LN. PCR analysis (COM1/IS1111 genes) confirmed the presence of C.b. DNA in heart, lung, spleen, liver & LN, but not in brain or BM.

Conclusion: The study revealed the persistence of C. b. cell components in various organs, including astrocytes of the brain, in a post-infection QFS. The possible mechanisms and molecular adaptations for this alternative C.b. life style are discussed.

No MeSH data available.


Related in: MedlinePlus

Patient BI and control brain samples from occipital cortex grey matter and occipital subcortical white matter were analysed using confocal microscopy for presence of C.b. antigens and co-stained for GFAP. Tissues were stained with anti-GFAP primary antibodies/FITC (green)-conjugated secondary antibodies; and co-stained with anti-C.b. primary antibodies/PE-conjugated (red) secondary antibodies as described in Material and Methods. Hoechst (blue) was used to stain cell nuclei. Magnification was set at 400X. a BI’s occipital subcortical white matter; b control patient’s occipital subcortical white matter; c BI’s occipital cortex grey matter; d control patient’s occipital cortex grey matter
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Fig2: Patient BI and control brain samples from occipital cortex grey matter and occipital subcortical white matter were analysed using confocal microscopy for presence of C.b. antigens and co-stained for GFAP. Tissues were stained with anti-GFAP primary antibodies/FITC (green)-conjugated secondary antibodies; and co-stained with anti-C.b. primary antibodies/PE-conjugated (red) secondary antibodies as described in Material and Methods. Hoechst (blue) was used to stain cell nuclei. Magnification was set at 400X. a BI’s occipital subcortical white matter; b control patient’s occipital subcortical white matter; c BI’s occipital cortex grey matter; d control patient’s occipital cortex grey matter

Mentions: Astrocytes in the occipital cortex and subcortical white matter from patient BI and control brain tissue were stained using GFAP antiserum with green-fluorescent (FITC) secondary antibodies, co-stained with Texas-red labelled monoclonal C.b. Phase 1 antiserum [2, 3] and viewed with confocal microscopy (Fig. 2). Astrocytes in the white and grey matter from patient BI (Fig. 2a and c, white and grey matter respectively) and control brain tissue (Fig. 2b and d) stained with anti-GFAP (green). Specific C.b. stained red and showed co-localization (yellow) with GFAP (green) protein (Fig. 2a). Staining with C.b. antisera revealed C.b. antigen complex in patient’s astrocytes, supporting the possible involvement of Q fever infection in the illness. Note that only a minority of astrocytes contained the C.b. Phase 1 antigen complex. C.b. antigens were also detected in cells from hippocampal white matter using dual staining with anti-GFAP and confocal microscopy (data not shown). White and grey matter staining for C.b. was negative in a control (uninfected) brain. Microglia, the innate immune cells mediating inflammatory responses in the CNS [10–12] were identified using the specific Iba-1 marker [13, 14]. Double immune-staining of the Iba-1 reactive cells with C.b. antisera was negative (data not shown).Fig. 2


Coxiella burnetii dormancy in a fatal ten-year multisystem dysfunctional illness: case report.

Sukocheva OA, Manavis J, Kok TW, Turra M, Izzo A, Blumbergs P, Marmion BP - BMC Infect. Dis. (2016)

Patient BI and control brain samples from occipital cortex grey matter and occipital subcortical white matter were analysed using confocal microscopy for presence of C.b. antigens and co-stained for GFAP. Tissues were stained with anti-GFAP primary antibodies/FITC (green)-conjugated secondary antibodies; and co-stained with anti-C.b. primary antibodies/PE-conjugated (red) secondary antibodies as described in Material and Methods. Hoechst (blue) was used to stain cell nuclei. Magnification was set at 400X. a BI’s occipital subcortical white matter; b control patient’s occipital subcortical white matter; c BI’s occipital cortex grey matter; d control patient’s occipital cortex grey matter
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4835832&req=5

Fig2: Patient BI and control brain samples from occipital cortex grey matter and occipital subcortical white matter were analysed using confocal microscopy for presence of C.b. antigens and co-stained for GFAP. Tissues were stained with anti-GFAP primary antibodies/FITC (green)-conjugated secondary antibodies; and co-stained with anti-C.b. primary antibodies/PE-conjugated (red) secondary antibodies as described in Material and Methods. Hoechst (blue) was used to stain cell nuclei. Magnification was set at 400X. a BI’s occipital subcortical white matter; b control patient’s occipital subcortical white matter; c BI’s occipital cortex grey matter; d control patient’s occipital cortex grey matter
Mentions: Astrocytes in the occipital cortex and subcortical white matter from patient BI and control brain tissue were stained using GFAP antiserum with green-fluorescent (FITC) secondary antibodies, co-stained with Texas-red labelled monoclonal C.b. Phase 1 antiserum [2, 3] and viewed with confocal microscopy (Fig. 2). Astrocytes in the white and grey matter from patient BI (Fig. 2a and c, white and grey matter respectively) and control brain tissue (Fig. 2b and d) stained with anti-GFAP (green). Specific C.b. stained red and showed co-localization (yellow) with GFAP (green) protein (Fig. 2a). Staining with C.b. antisera revealed C.b. antigen complex in patient’s astrocytes, supporting the possible involvement of Q fever infection in the illness. Note that only a minority of astrocytes contained the C.b. Phase 1 antigen complex. C.b. antigens were also detected in cells from hippocampal white matter using dual staining with anti-GFAP and confocal microscopy (data not shown). White and grey matter staining for C.b. was negative in a control (uninfected) brain. Microglia, the innate immune cells mediating inflammatory responses in the CNS [10–12] were identified using the specific Iba-1 marker [13, 14]. Double immune-staining of the Iba-1 reactive cells with C.b. antisera was negative (data not shown).Fig. 2

Bottom Line: During life, extensive clinical and laboratory investigations from different disciplinary stand points failed to deliver a definitive identification of a cause.PCR analysis (COM1/IS1111 genes) confirmed the presence of C.b.The possible mechanisms and molecular adaptations for this alternative C.b. life style are discussed.

View Article: PubMed Central - PubMed

Affiliation: Q Fever Research Group (1993-2009), Hanson Institute, Adelaide, South Australia.

ABSTRACT

Background: In a previous study of a Q fever outbreak in Birmingham, our group identified a non-infective complex of Coxiella burnetii (C.b.) antigens able to survive in the host and provoked aberrant humoral and cell-mediated immunity responses. The study led to recognition of a possible pathogenic link between C.b. infection and subsequent long-term post Q fever fatigue syndrome (QFS). This report presents an unusually severe case of C.b. antigen and DNA detection in post-mortem specimens from a patient with QFS.

Case presentation: We report a 19-year old female patient who became ill with an acute unexplained febrile encephalitis-like illness, followed by increasingly severe multisystem dysfunction and death 10 years later. During life, extensive clinical and laboratory investigations from different disciplinary stand points failed to deliver a definitive identification of a cause. Given the history of susceptibility to infection from birth, acute fever and the diagnosis of "post viral syndrome", tests for infective agents were done starting with C.b. and Legionella pneumophila. The patient had previously visited farms a number of times. Comprehensive neuropathological assessment at the time of autopsy had not revealed gross or microscopic abnormalities. The aim was to extend detailed studies with the post-mortem samples and identify possible factors driving severe disturbance of homeostasis and organ dysfunction exhibited by the course of the patient's ten-year illness. Immunohistochemistry for C.b. antigen and PCR for DNA were tested on paraffin embedded blocks of autopsy tissues from brain, spleen, liver, lymph nodes (LN), bone marrow (BM), heart and lung. Standard H&E staining of brain sections was unrevealing. Immuno-staining analysis for astrocyte cytoskeleton proteins using glial fibrillary acidic protein (GFAP) antibodies showed a reactive morphology. Coxiella antigens were demonstrated in GFAP immuno-positive grey and white matter astrocytes, spleen, liver, heart, BM and LN. PCR analysis (COM1/IS1111 genes) confirmed the presence of C.b. DNA in heart, lung, spleen, liver & LN, but not in brain or BM.

Conclusion: The study revealed the persistence of C. b. cell components in various organs, including astrocytes of the brain, in a post-infection QFS. The possible mechanisms and molecular adaptations for this alternative C.b. life style are discussed.

No MeSH data available.


Related in: MedlinePlus