Serum IgM/IgG binding to, and complement-dependent cytotoxicity (CDC) of, PBMCs and RBCs from (a) wild-type (WT), (b) 1,3-galactosyltransferase gene-knockout (GTKO), (c) GTKO/beta-1,4-N-acety1 galactosaminyltransferase 2-knockout (GTKO/4GalNT2KO), (d) GTKO/cytidine monophosphate-N-acetylneuraminic acid hydroxylase-knockout (GTKO/CMAHKO), and (e) GTKO/4GalNT2KO/CMAHKO/hCD55 (TKO/hCD55) pigs were measured by flow cytometry

Serum IgM/IgG binding to, and complement-dependent cytotoxicity (CDC) of, PBMCs and RBCs from (a) wild-type (WT), (b) 1,3-galactosyltransferase gene-knockout (GTKO), (c) GTKO/beta-1,4-N-acety1 galactosaminyltransferase 2-knockout (GTKO/4GalNT2KO), (d) GTKO/cytidine monophosphate-N-acetylneuraminic acid hydroxylase-knockout (GTKO/CMAHKO), and (e) GTKO/4GalNT2KO/CMAHKO/hCD55 (TKO/hCD55) pigs were measured by flow cytometry. acid hydroxylase-knockout (GTKO/CMAHKO), and (e) GTKO/4GalNT2KO/CMAHKO/hCD55 (TKO/hCD55) pigs were measured by flow cytometry. We obtained the following results: (i) Serum IgM/IgG binding and CDC in Volunteers were significantly greater to WT, GTKO, and GTKO/4GalNT2KO PBMCs or RBCs than to GTKO/CMAHKO and TKO/hCD55 cells; (ii) ESRD, DBD, and Allo-TCMR serum antibody binding and CDC to WT pig PBMCs were significantly greater than to GTKO, GTKO/4GalNT2KO, GTKO/CMAHKO, and TKO/hCD55 cells; (iii) antibody binding to GTKO/CMAHKO pig cells was significantly lower in hemodialysis than peritoneal dialysis patients. (iv) Two of twenty allotransplantation recipients serum IgG binding to GTKO pig PBMCs increased on POD14 compared with Pre, but IgG binding to GTKO pig RBCs did not; (v) In all sera, the lowest antibody binding and CDC were to GTKO/CMAHKO and TKO/CD55 pig cells. We conclude (i) CMAHKO in the pig may be critical to the success of clinical pig kidney xenotransplantation, and may be the most important after GTKO, at least c-met-IN-1 in Chinese patients; (ii) subjects with ESRD, or who are immunosuppressed after kidney allotransplantation, and DBD, have lower levels of antibody binding and CDC to genetically-engineered pig cells than do volunteers; (iii) TKO pigs with selected human protective transgenes, e.g., CD55, are likely to prove to be the optimal sources of kidneys for clinical xenotransplantation. studies have indicated that HLA-sensitized patients will not be at greater risk of rejecting a pig organ than HLA-non-sensitized patients (9C14), but other studies indicate that HLA-sensitized patients have a greater risk of rejecting a pig organ (15C17), and so it would be prudent not to select HLA-sensitized patients for the first clinical trials of pig kidney transplantation (18). Subjects with brain-death (DBD subjects) are a frequent source of organs for transplantation, and transplantation of a pig organ into a brain-dead human has recently been carried out (19). However, brain death is associated with dysfunction of the cardiovascular, pulmonary, endocrine, thermoregulation, renal, hematologic and inflammatory systems (20C23). If DBD subjects are used as in preclinical models of pig renal xenotransplantation, there is concern that these pathophysiological consequences may affect the xenograft, e.g., by activation of T and B lymphocytes, release of cytokines, etc. (19). The aims of the present study were (i) to measure serum anti-pig antibodies in healthy human volunteers, patients with ESRD pre- and post-renal allotransplantation, DBD subjects, and patients with renal allografts who were currently experiencing acute T cell-mediated rejection (Allo-TCMR), and (ii) to provide further data to help select pigs with the optimal genotype for clinical renal xenotransplantation. Materials and Methods Human Sera Blood was drawn from (i) healthy volunteers (Volunteers, n=20; ABO blood types A n=6; B n=6; AB n=3; O n=5), (ii) patients with ESRD (n=20), Rabbit Polyclonal to ZNF682 pre-renal transplantation (Pre) and on Day 1 (POD 1) and Day 14 (POD 14) after renal allotransplantation, (iii) brain-dead organ donors (DBD, n=20) and (iv) patients with renal allografts who were currently experiencing episodes of acute T cell-mediated rejection (Allo-TCMR, n=20) ( Table?1 ). Sera were obtained from de-identified remnant/discarded clinical laboratory samples. Sera from Volunteers were obtained from the Second Affiliated Hospital of Hainan?Medical?University, and all experimental protocols were approved by the ethics committee of the Second Affiliated Hospital of Hainan Medical University. All procedures involving humans were performed in accordance with the relevant guidelines and regulations, and had no c-met-IN-1 adverse effects on the subjects. Pigs Blood was obtained from wild-type (WT, i.e., genetically-data indicating that TKO pig organs will prove to be a major advance over GTKO organs for transplantation into humans (5, 7, 35) which is consistent with our conclusions from the present study. Healthy Human Volunteers vs Other Groups A second major observation made in this research was that (i) sufferers with ESRD, brain-dead donors, and immunosuppressed sufferers with kidney allografts generally acquired lower degrees of anti-pig antibodies than healthful individual volunteers considerably, except in regards to WT pig cells. The tendencies in CDC were comparable to those of IgG and IgM. Furthermore, the ABO c-met-IN-1 bloodstream kind of the donor.