Expression of major histocompatibility complex antigens and induction of human T-lymphocyte proliferation by astrocytes and macrophages from porcine fetal brain

Abstract

Porcine fetal brain cells are of potential use as donor cells for transplantation therapies of neurodegenerative diseases in humans. Our aim was to determine the immunestimulatory properties of astrocytes and macrophages from porcine fetal brain in vitro. By flow cytometry, freshly isolated porcine fetal brain cells were nonautofluorescent, while primary cultures of these cells, prepared to favor growth of astrocytes and macrophages/microglia, consisted of both an autofluorescent and a nonautofluorescent cell population. The cultured autofluorescent cells had qualities typical of macrophages: CD18 (beta(2) integrin subunit) expression, high granularity, and phagocytic activity. The cultured nonautofluorescent cells stained positive for the astrocyte marker glial fibrillary acidic protein and CD56 (NCAM isoform). While freshly isolated porcine fetal brain cells expressed very low levels of major histocompatibility complex (MHC) class I and no MHC class II antigens, primary culture of these cells resulted in upregulation of MHC class I antigens on astrocytes and macrophages and MHC class II antigens on a subpopulation of the macrophages. Single-cell suspensions prepared from the primary cultures were flow sorted into astrocyte and macrophage populations on the basis of cell granularity and autofluorescence or on the basis of CD56 expression. Pure suspensions (>98%) of astrocytes induced a low proliferative response in human T lymphocytes, as determined by [(3)H]thymidine incorporation after 4 days of coculture. A suspension of 91% macrophages was a strong inducer of human T-cell proliferation, even stronger than allogeneic mononuclear blood cells. For neural xenotransplantation, our findings suggest that depletion of macrophages from the donor-cell suspensions may enhance graft survival by reducing cell-mediated rejection.