Transplanted neural stem cells modulate regulatory T, γδ T cells and corresponding cytokines after intracerebral hemorrhage in rats

Lu Gao, Qin Lu, Li-Jie Huang, Lin-Hui Ruan, Jian-Jing Yang, Wei-Long Huang, Wei-Shan ZhuGe, Yong-Liang Zhang, Biao Fu, Kun-Lin Jin, Qi-Chuan ZhuGe, Lu Gao, Qin Lu, Li-Jie Huang, Lin-Hui Ruan, Jian-Jing Yang, Wei-Long Huang, Wei-Shan ZhuGe, Yong-Liang Zhang, Biao Fu, Kun-Lin Jin, Qi-Chuan ZhuGe

Abstract

The immune system, particularly T lymphocytes and cytokines, has been implicated in the progression of brain injury after intracerebral hemorrhage (ICH). Although studies have shown that transplanted neural stem cells (NSCs) protect the central nervous system (CNS) from inflammatory damage, their effects on subpopulations of T lymphocytes and their corresponding cytokines are largely unexplored. Here, rats were subjected to ICH and NSCs were intracerebrally injected at 3 h after ICH. The profiles of subpopulations of T cells in the brain and peripheral blood were analyzed by flow cytometry. We found that regulatory T (Treg) cells in the brain and peripheral blood were increased, but γδT cells (gamma delta T cells) were decreased, along with increased anti-inflammatory cytokines (IL-4, IL-10 and TGF-β) and decreased pro-inflammatory cytokines (IL-6, and IFN-γ), compared to the vehicle-treated control. Our data suggest that transplanted NSCs protect brain injury after ICH via modulation of Treg and γδT cell infiltration and anti- and pro-inflammatory cytokine release.

Figures

Figure 1.
Figure 1.
Total numbers of T lymphocyte cells in the brain and peripheral blood. Total numbers of T lymphocytes in the brain were significantly increased in intracerebral hemorrhage (ICH), vehicle-treated (ICH + V) and neural stem cells (NSCs)-treated groups (ICH + NSCs), compared to sham-operated group (Sham) (A); there was no difference of total numbers of T lymphocytes of each group in peripheral blood in ICH, vehicle-treated (ICH + V) and NSCs-treated groups (ICH + NSCs), compared to sham-operated group (Sham) (B), (n = 8 in each group). *p < 0.05 versus sham ICH.
Figure 2.
Figure 2.
Increased absolute numbers of γδT and Treg cells in the brain and peripheral blood after ICH. The absolute numbers of γδT and Treg cells were increased significantly in the brain (A) and peripheral blood; (B) after ICH. (n = 8 in each group) *p < 0.05 versus sham ICH.
Figure 3.
Figure 3.
Behavioral functional test before and after ICH. Transplanted NSCs improve functional recovery after ICH at day 3, 7 and 14. (n = 6 in each group) *p < 0.05 versus ICH and ICH + V.
Figure 4.
Figure 4.
Transplanted NSCs altered γδT and Treg cells in the brain and peripheral blood. Transplanted NSCs reduced the absolute numbers of γδT cells in the brain and peripheral blood (A–C); The percentage and the absolute numbers of Treg cells were significantly increased in the brain by transplantation of NSCs (D,E); An increased absolute Treg cell numbers in the peripheral blood after transplanted NSCs (F). (n = 8 in each group) *p < 0.05 versus ICH-vehicle.
Figure 5.
Figure 5.
Expressions of inflammatory cytokine in the cerebral and peripheral blood after transplantation of NSCs. ELASA (A,B) revealed the upregulations of IL-6 and IFN-γ in the haemorrhagic brains and peripheral blood in the groups transplanted witn NSCs, compared to the normal samples. In the peripheral blood, TGF-β and IL-10 were upregulated after ICH followed NSC transplantation. However, IL-4 was unchanged both in the brain and peripheral blood after ICH. In brain and peripheral blood samples, the groups treated with NSCs showed decreases of IL-6 and IFN-γ and increases of IL-4, IL-10 and TGF-β levels, compared with the ICH group. (n = 4 in each group) *p < 0.05 versus normal; **p < 0.05 versus ICH.

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