title: Porcine neural xenografts in the immunocompetent rat: immune response following grafting of expanded neural precursor cells.
abstract: Intracerebral neural xenografts elicit a host immune response that results in their rapid rejection. This forms a key barrier to the therapeutic use of xenogeneic tissue transplantation for conditions such as Parkinson's disease. The current study sought to provide insight into the cellular components of donor cell suspensions that are important in stimulating the host rejection response and thereby to suggest rational manipulations of xenogeneic donor tissue that might ultimately enhance its clinical utility. The neural stem cell mitogens, epidermal growth factor and fibroblast growth factor-2, have been used to isolate and expand populations of primordial neural precursor cells from the embryonic pig brain. The immune response elicited by these cells on transplantation into the non-immunosuppressed rat has been fully characterised. In the first experiments, expanded neural precursors were grafted into the hemi-parkinsonian, non-immunosuppressed Sprague-Dawley rat and graft status and host response examined 10, 21, 35 and 60 days post-transplantation. While equivalent primary tissue grafts were completely eliminated at 35 days, grafts of expanded neural precursors with healthy neurofilament-positive projections were present at all time-points, and two large grafts remained even at 60 days. Some grafts appeared to elicit minimal host immune responses at the time-points they were examined, although most did appear to be undergoing a rejection process since a co-ordinated response involving host cytotoxic T-lymphocytes, microglia/macrophages, immunoglobulin M and complement could be demonstrated to varying degrees. Subsequent experiments went on to demonstrate further that expanded precursor populations and primary tissue suspensions differed in their immunogenic profile. Firstly, when primary tissue was injected intraperitoneally into immunocompetent rats a vigorous primary humoral response was generated. No such response was detected following injection of expanded neural precursors. Secondly, flow cytometric analysis revealed small but significant levels of class II porcine major histocompatibility complex expression in primary cell suspensions but no such expression in expanded precursor populations.The results of this study therefore demonstrate that the immunogenicity of porcine neural cell suspensions used for intracerebral grafting is reduced when neural stem cell mitogens are used to expand precursor cells. The implications of these findings in the development of novel xenogeneic cellular therapies for neurodegenerative conditions such as Parkinson's disease are discussed.
authors.affiliation: Cambridge Centre for Brain Repair, University of Cambridge, Forvie Site, Robinson Way, Cambridge CB2 2PY, UK.
authors.forename: R J
authors.initials: RJ
authors.lastname: Armstrong
authors.forename: T P
authors.initials: TP
authors.lastname: Harrower
authors.forename: C B
authors.initials: CB
authors.lastname: Hurelbrink
authors.forename: M
authors.initials: M
authors.lastname: McLaughin
authors.forename: E L
authors.initials: EL
authors.lastname: Ratcliffe
authors.forename: P
authors.initials: P
authors.lastname: Tyers
authors.forename: A
authors.initials: A
authors.lastname: Richards
authors.forename: S B
authors.initials: SB
authors.lastname: Dunnett
authors.forename: A E
authors.initials: AE
authors.lastname: Rosser
authors.forename: R A
authors.initials: RA
authors.lastname: Barker
mesh_terms: D000818:Animals; D000906:Antibodies / Q000097:blood / Q000276:immunology; D015478:Antigens, Heterophile / Q000097:blood / Q000276:immunology; D015415:Biomarkers / Q000097:blood; D016380:Brain Tissue Transplantation / Q000009:adverse effects* / Q000379:methods; D002455:Cell Division / Q000187:drug effects / Q000276:immunology; D002540:Cerebral Cortex / Q000166:cytology / Q000276:immunology / Q000637:transplantation; D005260:Female; D005333:Fetus; D005434:Flow Cytometry; D006084:Graft Rejection / Q000276:immunology* / Q000378:metabolism / Q000503:physiopathology; D006085:Graft Survival / Q000276:immunology; D007121:Immunocompetence / Q000276:immunology*; D008934:Mitogens / Q000494:pharmacology; D017072:Neostriatum / Q000276:immunology / Q000503:physiopathology / Q000601:surgery*; D009474:Neurons / Q000166:cytology / Q000276:immunology / Q000637:transplantation; D020734:Parkinsonian Disorders / Q000378:metabolism / Q000503:physiopathology / Q000601:surgery*; D051381:Rats; D017207:Rats, Sprague-Dawley; D033581:Stem Cell Transplantation; D013234:Stem Cells / Q000166:cytology / Q000276:immunology; D013552:Swine; D014183:Transplantation, Heterologous / Q000009:adverse effects* / Q000379:methods
chemical_list: D000906:Antibodies; D015478:Antigens, Heterophile; D015415:Biomarkers; D008934:Mitogens