Interactions of neurotrophic factors GDNF and NT-3, but not BDNF, with the immune system following fetal spinal cord transplantation

Abstract

Glial cell line-derived neurotrophic factor (GDNF) is known to stimulate survival of dopaminergic and spinal cord motor neurons. However, little is known of the possible immune sequelae of GDNF exposure, or that of other putative trophic factors. To address these questions we utilized in oculo grafts of spinal cord, wherein we could induce different levels of immune responses via allogeneic vs. syngeneic combinations. Adult female Sprague-Dawley and Fisher rats were used as hosts for allogeneic and syngeneic grafts, respectively. Embryonic age 14–-15-day-old fetuses were taken from pregnant dams of each strain, and cervical spinal cords were removed and dissected. Pieces of the spinal cord were transplanted into the anterior chamber of the eye within each strain. At 5-day intervals, 0.5 μg of GDNF, brain-derived neurotrophic factor (BDNF), neurotrophin-3 (NT-3) or cytochrome c (CC) was injected into the anterior chamber of the eye and the sizes of the transplants were measured for the Sprague-Dawley rats. The same injections and measurements, but only for GDNF and CC, were carried out using Fisher rats. As expected, GDNF increased transplant survival and growth in both the Sprague-Dawley and Fisher animals. At day 41–42, all rats were sacrificed. Cameral graft appearance was evaluated by cresyl violet and immunohistochemically using antibodies against neurofilament (NF), calcitonin gene-related peptide (CGRP) and glial fibrillary acidic protein (GFAP). To monitor immune responses, the following monoclonal antibodies were used: OX38 against CD4, OX18 against MHC class I WHO), OX8 against CD8, OX6 against MHC class II (MHCII), OX42 against CD11b, R73 against α and β T cell receptor MR), and ED[. In the Sprague-Dawley grafts, significantly higher amounts of CD8 +, T lymphocyte + , MHCI + and MHCII + antigen-presenting cells (APC) were observed in GDNF-treated transplants. These markers were also increased in NT-3-treated groups. There were two types of OX-42 + cells, one was the ordinary ramified microglial cell, the other appeared to be a phagocytic cell, looking like the interstitial proliferating variety. Interestingly, the phagocytic OX-42 + cells had the same distribution as ED1 + and MHCII + cells. In contrast, there were few inununoreactive cells after GDNF treatment in the inbred Fisher animals, similar to the CC control group. These results suggest that GDNF and to some extent NT-3, can activate the immune system in allogeneic graft combinations, but that these trophic factors do not produce overt rejection, and do not per se induce immune responses.