Immune cells subpopulations in cerebrospinal fluid and peripheral blood of patients with Aneurysmal Subarachnoid Hemorrhage

Abstract

Background: There is growing evidence supporting the role of inflammation in aneurysmal subarachnoid hemorrhage (aSAH) pathophysiology and it is of great interest to elucidate which immune mechanisms are involved.

Methods: 12 aSAH patients and 28 healthy controls were enrolled prospectively. We assessed leukocytes subpopulations and their activation status by flow cytometry in cerebrospinal fluid (CSF) and peripheral blood (PB) of SAH patients at the same time and in PB of controls.

Results: Monocytes and neutrophils were activated in CSF of aSAH patients. The percentage of CD14(++)CD16(+) monocytes were higher in CSF than in PB of aSAH patients, and were also increased in PB of aSAH patients compared with controls. An enhanced expression of CD69 was shown in CSF neutrophils compared with PB in aSAH patients. PB of aSAH patients showed lower percentage of total lymphocytes compared with controls PB. Additionally, lymphocytes were activated in CSF and PB of aSAH patients. CD4(+) and CD8(+) T cells had a decreased expression on CD3 and higher levels of CD69 in CSF compared with PB in aSAH patients. Moreover, PB CD4(+) and CD8(+) T cells of aSAH patients were activated compared with controls. Additionally, CD28 expression was decreased on CSF T lymphocytes.

Conclusions: Our data suggest an important recruitment of leukocytes to the site of injury in aSAH as well as an increased activation at this level. Overall, these results indicate that aSAH probably stimulates both the innate and adaptive immune responses.

Keywords: Aneurysmal subarachnoid hemorrhage; Cerebrospinal fluid; Flow cytometry; Neuroinflammation; Systemic inflammation.

Figure 1

Monocytes and neutrophils. A) Percentages of total monocytes and CD14⁺⁺CD16+ monocytes in CSF and PB of aSAH patients and in PB of controls. Data shown are median ± standard error. B) CD69 expression on neutrophils in CSF and PB of aSAH patients and in PB of controls. Data shown are median of CD69 mean fluorescence intensity (MFI) on neutrophils ± standard error.

Figure 2

Lymphocytes subsets. Percentages of CD4⁺ T cells, CD8⁺ T cells, NK cells and B cells lymphocytes of total lymphocytes in CSF and PB of aSAH patients and in PB of controls. Data shown are median ± standard error.

Figure 3

Activation pattern in CD4⁺ and CD8⁺ T cells. A) CD3 mean fluorescence intensity (MFI) on CD4⁺ T cells in PB and CSF of aSAH patients. Lines indicate samples from the same patient on the same time. B) CD3 MFI on CD8⁺ T cells in PB and CSF of aSAH patients. Lines indicate samples from the same patient on the same time. C) Ratio of CD4⁺ CD69⁺ T cells/CD4⁺ T cells and CD8⁺ CD69⁺ T cells/CD8⁺ T cells. Data shown are median ± standard error.

Figure 4

CD28 expression on CD4⁺ and CD8⁺ T cells. A) CD28 mean fluorescence intensity (MFI) on CD4⁺T cells of PB and CSF of aSAH patients. Lines indicate samples from the same patient on the same time. B) CD28 MFI on CD8⁺T cells on PB and CSF of aSAH patients. Lines indicate samples from the same patient on the same time.