Upregulation of CC Chemokine Receptor 7 (CCR7) Enables Migration of Xenogeneic Human Adipose-Derived Mesenchymal Stem Cells to Rat Secondary Lymphoid Organs

Abstract

BackgroundCC chemokine receptor 7 (CCR7) expression is vital for cell migration to secondary lymphoid organs (SLOs). Our previous work showed that inducing CCR7 expression enabled syngeneic mesenchymal stem cells (MSCs) to migrate into SLOs, resulting in enhanced immunosuppressive performance in mice. Given that human adipose-derived stem cells (hASCs) are widely used in clinical therapy, we further investigated whether upregulation of CCR7 enables xenogeneic hASCs to migrate to rat SLOs.Material/MethodshASCs rarely express CCR7; therefore, hASCs were transfected with lentivirus encoding rat CCR7 (rCCR7) plus green fluorescence protein (GFP) or GFP alone. CCR7 mRNA and cell surface expression of rCCR7-hASCs and GFP-hASCs were examined by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM), respectively. The phenotype, differentiation, and proliferation capacity of each cell type was also determined. To examine migration, rCCR7-hASCs and GFP-hASCs were injected intravenously into Lewis rats, and the proportion of GFP-positive cells in the spleen and lymph nodes was determined with FCM.ResultsmRNA and cell surface protein expression of CCR7 was essentially undetectable in hASCs and GFP-ASCs; however, CCR7 was highly expressed in rCCR7-ASCs. rCCR7-hASCs, GFP-hASCs, and hASCs shared a similar immunophenotype, and maintained the ability of multilineage differentiation and proliferation. In addition, the average proportion of GFP-positive cells was significantly higher following transplantation of rCCR7-hASCs compared with GFP-hASCs (p<0.01).ConclusionsThese results suggest that upregulation of rat CCR7 expression does not change the phenotype, differentiation, or proliferation capacity of hASCs, but does enable efficient migration of hASCs to rat SLOs.

Chemokines and their receptors: predictors of the therapeutic potential of mesenchymal stromal cells

“…Specific chemokines have been shown attract immune cells expressing the cognate receptor to specific tissues: the expression of CCR1 in macrophages and neutrophils leads to kidney infiltration in renal ischemia–reperfusion injury [ 67 ]; CCR5 directs CD8 + T cells towards the brain [ 68 ]; CCR3, CCR4 and CCR10 are highly expressed by T cells in skin [ 69 , 70 ]; CXCR3 mediates T cell recruitment into the kidney [ 71 ]; and CXCR6 is highly expressed by liver-infiltrating CD8 + T cells [ 72 ]. Furthermore, CCR7 targets MSCs to secondary lymphoid organs [ 73 , 74 ], CXCR3-deficient MSCs fail to infiltrate into the nephritic kidney [ 75 ] and CXCR4 receptor overexpression in MSCs improves treatment of acute lung injury [ 76 ]. Thus, specific chemokine receptors could have an essential role in MSC homing into specific tissues.…”

Targeted Migration of Human Adipose-Derived Stem Cells to Secondary Lymphoid Organs Enhances Their Immunomodulatory Effect and Prolongs the Survival of Allografted Vascularized Composites

“…The preparation of hASCs has been described in previous study . Briefly, adipose tissue was washed and minced firstly, and then digested with hyaluronidase (0.05%, Hyclone, Logan/Utah, USA) and type I collagenase (0.1%, Hyclone) for 45 minutes at 37°C.…”

“…The preparation of hASCs has been described in previous study [34]. Briefly, adipose tissue was washed and minced firstly, and then digested with hyaluronidase (0.05%, Hyclone, Logan/Utah, USA) and type I collagenase (0.1%, Hyclone) for 45 minutes at 37 C. After centrifugation at 1,500 rpm for 10 minutes, the digested single cells were collected in low-glucose Dulbecco's modified Eagle's medium (Gibco, Waltham, MA, USA) supplemented with 100 U/ml penicillin, 100 U/ml streptomycin (Hyclone), and 10% fetal bovine serum (FBS, Gibco) in a humidified atmosphere of 5% CO 2 at 37 C. Adherent cells were passaged every 5-7 days.…”

Redirecting adult mesenchymal stromal cells to the brain: a new approach for treating CNS autoimmunity and neuroinflammation?

“…In support of this concept, Ad-MSCs genetically modified to express CCR7 homed more efficiently to SLOs where the CCR7 ligands CCL19 and CCL21 are constitutively expressed. 57 Chemokine receptors such as CXCR4 have also been induced by cytokine stimulation to promote the homing and engraftment of systemically administered human Ad-MSCs to the bone marrow. 58 Furthermore, modifications to pre-existing, albeit nonfunctional forms of surface proteins have also been tested by enzymatic modification and activation of CD44 glycoforms to promote tropism of MSCs to the bone marrow.…”

“…Modification of chemokine receptor and/or adhesion molecules to enhance migration of MSCs to the inflamed CNS is based on an existing premise that MSCs are amenable to manipulation and redirection to peripheral tissue sites. In support of this concept, Ad‐MSCs genetically modified to express CCR7 homed more efficiently to SLOs where the CCR7 ligands CCL19 and CCL21 are constitutively expressed . Chemokine receptors such as CXCR4 have also been induced by cytokine stimulation to promote the homing and engraftment of systemically administered human Ad‐MSCs to the bone marrow .…”