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Neuropilin-1 (NRP1) and its role in effector and regulatory CD4 T cell populations
E. J. Lee, MD; H. Nakayama, PhD; D. M. Briscoe, MD; H. B. Kim, MD; G. Demirci, MD, PhD
Department of Surgery and Department of Pediatrics, Division of Nephrology, Boston Childrenís Hospital, Harvard Medical School, Boston, MA

Background: Neuropilin-1 (NRP1) serves as a receptor for semaphorins and VEGF family members. Recently, NRP1 was shown to be highly expressed on CD4+Foxp3+ regulatory T cells (Treg) and impact the migration of Tregs into tumors. While NRP1 has been extensively studied in cancer models, its role in transplantation has not been investigated.

Methods: and

Results: 1) CD4+ T cells from wild type mouse spleen, thymus and peripheral lymph nodes were FACS stained for NRP1 and Treg specific markers. NRP1 was found on over 80% of all Tregs and its expression correlated with CD25, CTLA-4, GITR, Helios and OX40. T conventional (Tconv) cells expressed NRP1 at around 10%. 2) NaÔve Tconv were stimulated in vitro; CD25 was upregulated on all Tconv within 24 hours, while NRP1 levels only slightly increased over 96 hours. 3) Tconv were cultured in vitro under Treg inducing conditions for 5 days. Resulting CD4+Foxp3+CD25+ Treg expressed NRP1 levels of over 80%. 4.) An NRP1 expressing human glioblastoma cell line was treated with Sema3A, showing strong inhibition of Akt phosphorylation.

Conclusion:s: The expression of NRP1 on CD4+Foxp3+ T cells correlates with traditional Treg markers and confirms NRP1 as a Treg specific marker. Unlike CD25, NRP1 is not upregulated on CD4+ T effector cells. Contrary to earlier studies, we find strong NRP1 expression on induced Treg. Hence, NRP1 is a surface molecule that distinguishes Treg from effector T cells. Moreover, Sema3A-mediated inhibition of PI3K-Akt/mTOR signaling in NRP1 expressing cells suggests a functional role for NRP1 in regulatory immune responses.

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