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Abstract Number: 1739

Immunomodulatory Properties of CD271+ and CD271- Synovial Mesenchymal Cells

Alicia Usategui1, Manuel J. Del Rey1, Regina Faré1, Gabriel Criado2, Vanessa Miranda1, Juan D. Cañete3 and Jose L. Pablos1, 1Servicio de Reumatología, Instituto de Investigación Hospital 12 de Octubre (I+12), Madrid, Spain, 2Grupo de Enfermedades Inflamatorias y Autoinmunes, Instituto de Investigación Hospital 12 de Octubre (I+12), Madrid, Spain, 3Arthritis Unit. Rheumatology Department, Hospital Clínic of Barcelona, Barcelona, Spain

Meeting: 2014 ACR/ARHP Annual Meeting

Keywords: Arthritis, Fibroblasts, immunoregulation, mesenchymal stem cells and synovial cells, synovial fluid

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Session Information

Title: T cell Biology in Rheumatoid Arthritis and Other Arthritis

Session Type: Abstract Submissions (ACR)

Background/Purpose

Mesenchymal stem cells (MSC) have been isolated from synovium and represent a fraction of synovial fibroblast (SF) cultures. CD271+ is considered a MSC surface marker that is also present in a small fraction of primary cultures of SF. Bone marrow or synovial derived CD271+ cells have shown increased chondrogenic potential but their immunosuppressive properties have not been analyzed. We have analyzed the immunosuppressive potential of CD271+ compared to CD271- primary SF cultures from human osteoarthritic synovial tissues.  

Methods

Osteoarthritic synovial membranes were obtained at knee joint replacement surgery (n=9). CD271+/- separation was carried out by magnetic sorting of passage 0 SF cultures and confirmed by flow cytometry. Multipotent differentiation capability of SF to adipocytes, chondroblasts and osteoclasts was studied using standard in vitrotissue culture-differentiating conditions and staining with oil red, alcian blue or alizarin red, respectively. Immunomodulatory properties of CD271+/- sorted cells were analyzed in co-cultures with T lymphocytes obtained from healthy donors (ratio 1:10), stimulated with anti-CD3/CD28 beads. T-cell proliferation was measured by CellVue dye dilution as detected by flow cytometry. T-cell cytokine production (IL-2, IL-10, IL-17 and IFN-γ) in supernatants was quantified by ELISA. Quantitative data were analyzed by Mann Whitney or one-sample t-test.

Results

Primary passage 0 OA SF cultures contained (11.27±7.25%) CD271+ cells. After sorting, both CD271+ and CD271- SF cultures contained multipotential MSC capacity to differentiate to adipocytes, chondroblasts and osteoclasts in specific differentiation media. Both types of cells significantly inhibited the proliferation of anti-CD3/CD28 stimulated T cells and there were no quantitative differences between CD271+ and CD271- cells (70.46±6.56% and 73.53±13.15% respectively compared to single T-cell parallel cultures where proliferation was set to 100%). Co-culture of SF and T lymphocytes induced a significant increase in IL-2, IL-10, IL-17 and IFN-γ production by T cells that was similar in CD271+ and CD271- cells.

Conclusion

These data confirm that SF cultures contain a CD271+ cell fraction. Both CD271+ and CD271- SF cultures showed multipotential and immunomodulatory properties previously described as characteristic of MSC. T-cell anti-proliferative effect and non-specific up-regulation of T-cell cytokine production were similar in CD271+ and CD271- SF.


Disclosure:

A. Usategui,
None;

M. J. Del Rey,
None;

R. Faré,
None;

G. Criado,
None;

V. Miranda,
None;

J. D. Cañete,
None;

J. L. Pablos,
None.

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