Immunopathology of an hDAF transgenic pig model liver xenotransplant into a primate.

BACKGROUND: hDAF transgenic pigs do not display the inherent hyperacute rejection reactions of pig-to-primate xenotransplants. The purpose of this study was to determine the immunopathologic phenomena following an hDAF transgenic pig hepatic orthotopic xenotransplant into a baboon. METHODS: Donor animals were unmodified pigs (n=4) and hDAF transgenic pigs (n=2). Recipient animals were baboons (Papio anubis). Liver biopsies were immunostained using monoclonal antibodies to C3, C5b-9, IgG, IgM, CD2, CD4, CD8, CD68, CD20, Bric 216, CD31, and fibrin, and polyclonal antibody to C4. RESULTS: hDAF transgenic grafts showed IgG, IgM, and C4 endothelial deposits. However, no fibrin, C3, or C5b9 deposits were observed after reperfusion. hDAF xenografts displayed CD31 staining in the portal spaces, perilobular areas, and at hepatic sinuisoidal levels. The baboon that lived for 4 days displayed either CD4 or CD8 T-cells periportal infiltrate. CONCLUSIONS: Future studies will seek to determine the physiologic role of CD31 hepatic sinusoidal expression in transgenic xenotransplants, and will also study the role of T-cell infiltrates in xenograft rejection.

Human anti-porcine gammadelta T-cell xenoreactivity is inhibited by human FasL expression on porcine endothelial cells.

BACKGROUND: The role of gammadelta T cells during an immune response is still elusive and has been proposed to play a first line of defense along with other cells of the innate immune system, such as macrophages and natural killer cells, before alphabeta T-cell activation occurs. Innate cellular immune response plays a major role in xenograft rejection. We investigated the response of human gammadelta T cells to unmodified and human FasL (hFasL)-expressing xenogenic porcine endothelial cells. METHODS: A 51Cr release assay was used to study the xenoreactivity of human gammadelta T-cell clones against porcine endothelial cells. Stable transfectants of porcine endothelial cells expressing hFasL were established and analyzed for their effectiveness in controlling this response. RESULTS: Of the gammadelta T-cell clones tested, 38.9% were cytotoxic for porcine endothelial target cells. This cytotoxic response of human gammadelta T-cell clones was significantly inhibited by a monoclonal antibody against human CD3. Incubation of gammadelta T-cell clones with concanamycin A, an inhibitor of the perforin/granzyme B pathway, caused inhibition of lysis of porcine endothelial cells. Inhibition was not observed upon incubation with either anti-FasL or anti-tumor necrosis factor-alpha monoclonal antibodies. Expression of hFasL on porcine endothelial cells significantly reduced lysis by human gammadelta T cells. CONCLUSION: These results imply that human gammadelta T cells may represent an important obstacle to xenotransplantation. Specific strategies targeted at this subset of T cells could be important in controlling innate cellular response to xenografts and facilitate graft survival.

Changes in hypothalamic paraventricular nucleus catecholaminergic activity after acute and chronic morphine administration.

The participation of hypothalamic noradrenaline in the expression of neuroendocrine signs of morphine withdrawal has been proposed. The present study in rats examined: (1) the relationships between corticosterone secretion and the possible modifications in noradrenaline and dopamine content and turnover in the hypothalamic paraventricular nucleus after acute and chronic morphine administration; (2) the changes in cyclic adenosine monophosphate (cAMP) levels in the paraventricular nucleus after the same treatments. The results showed that acute morphine injection in control rats increased corticosterone release, 3-methoxy-4-hydroxyphenylethylene glycol (MHPG) production, and noradrenaline turnover. Dopamine turnover in the paraventricular nucleus was decreased and the cAMP levels remained unchanged. In chronic morphine-treated rats, there was no elevation in noradrenaline turnover or in corticosterone secretion, indicating that tolerance developed to the acute effects of the opioid. Correspondingly, no alterations in dopamine turnover were observed when chronic morphine-treated rats were compared with control rats acutely injected with morphine. cAMP levels in the paraventricular nucleus were unchanged during the tolerant state. The results raise the possibility that noradrenergic afferents play a significant role in the alterations of paraventricular nucleus function and pituitary-adrenal axis activity in response to acute and chronic morphine and suggest that these modifications are not mediated through adenylate cyclase activation. The present data provide further support for the idea of adaptive changes in noradrenergic neurons projecting to the paraventricular nucleus during chronic morphine exposure.