Donor selection for xenotransplantation: detection of Galalpha1-3Gal on different porcine organs.

The expression of the major porcine xenoantigens (Galalpha1-3Gal) in different tissues varies between species. The selection of suitable donors and the interpretation of studies which attempt to prevent hyperacute rejection are dependent on donor expression of Galalpha1-3Gal. Screening of large number of animals to find potential Galalpha1-3Gal negative donors requires a robust, tissue-based and practical method of assessing Galalpha1-3Gal expression. In this study, we have assessed the expression of Galalpha1-3Gal in a variety of pig organs using anti Galalpha1-3Gal antibody. Biopsies of heart, kidney, ear and tail were obtained from 20 outbred pigs. Biopsies were fixed in formalin and stained with a human anti Galalpha1-3Gal antibody obtained from pooled human AB serum passed down a Galalpha1-3Gal immunoadsorbent column. Tissue from all 4 organs from all 20 pigs expressed Galalpha13Gal. This study shows that detection of Galalpha1-3Gal on an ear or tail biopsy is a simple but very reliable method for assessing Galalpha1-3Gal expression on the heart and kidney and facilitates donor selection for xenotransplantation.

Anti-CD45 monoclonal antibody perfusion of human renal allografts prior to transplantation. A safety and immunohistological study. CD45 Study Group.

The perfusion of kidneys with anti-CD45 monoclonal antibodies prior to transplantation offers a means of targeting passenger antigen-presenting cells with the aim of reducing the subsequent incidence of rejection episodes. A safety study was performed in humans of such pretreatment in 40 unsensitized recipients of first cadaveric renal grafts, who were followed for 3 months after transplantation. A 50-ml solution containing 2 mg of each of the rat anti-CD45 mAbs YTH 24.5 and YTH 54.12 was injected into the allograft renal artery ex vivo and just before transplantation while the renal vein was kept clamped. No patients died, but 4 grafts were lost. Two were lost due to primary nonfunction, 1 was lost because of late renal artery thrombosis, and 1 was lost to rejection. There were no cases of renal vein thrombosis and 1 trivial renal artery stenosis, and only 2 patients produced human anti-rat antibodies. Between 63.5% and 100% (median 96.4%) of CD45+ cells in the postperfusion biopsies were coated with anti-CD45 as determined by double-immunolabeling. The number of patients experiencing rejection episodes was inversely associated with this "antibody uptake": 75% of the low uptake group (< 95%) had at least 1 rejection episode, compared with 22% of the high uptake group (> or = 95%) (P = 0.001). The complement components C3 and C5b-9 colocalized with perfused anti-CD45 in 32/33 (97.0%) and 11/33 (33.3%) of the biopsy specimens, respectively. We conclude that: (1) this technique appears free of adverse effects, (2) high antibody uptake within the kidney is associated with a lower incidence of rejection, and (3) the antibodies used fix and activate complement in vivo.

Pretreatment of renal transplants with anti-CD45 antibodies: optimization of perfusion technique.

Pig kidneys were perfused ex vivo with a monoclonal anti-pig CD45 antibody to determine optimum conditions for human kidney anti-CD45 perfusion, as such pretreatment may prolong renal allograft survival. Efficacy of perfused antibody uptake was assessed by immunohistochemical double-labelling of multiple renal biopsies. Antibody uptake by CD45+ cells was rapid, and maximal within one hour of perfusion. An antibody concentration of 40 micrograms/ml more effectively saturated available CD45 binding sites than lower concentrations. Whilst cortical perfusion was homogeneous (median uptake > 99%, minimum 90.0%) after 25, 50 or 100 ml perfusion with 40 micrograms/ml monoclonal antibody (mAb), the medulla was less evenly perfused with 25 ml (median uptake 82.6%) than with 50 or 100 ml (median uptake 98.5% and 98.7%, respectively). Prolongation of renal cold ischaemia of up to 46 hours prior to mAb perfusion did not adversely affect antibody binding. Clamping of the renal vein during and after mAb perfusion was associated with better cortical and medullary mAb uptake than if the vein was left unclamped. These data show that the vast majority of the kidney's CD45+ antigen-presenting cells can be bound by anti-CD45 mAb perfused ex vivo, if an adequate volume and concentration of mAb perfusate is employed and the renal vein is clamped during and after the process.

Pretreatment of kidney allografts with monoclonal antibodies to CD45: results of a multicentre study. CD45 Study Group.

The perfusion of human renal allografts with CD45-specific monoclonal antibodies (mAbs) may reduce their immunogenicity and the incidence of rejection. We performed a safety study in 40 patients receiving their first cadaveric renal transplant. Two milligrams each of the rat CD45 specific mAbs YTH 24.5 and YTH 54.12 were perfused into the grafts prior to transplantation. The patients were followed for 3 months. No patient died, and four grafts were lost, three to vascular causes not considered to be related to antibody perfusion and one to severe rejection. Two patients developed an anti-rat antibody response. Immunohistological double-labelling performed on cortical biopsies taken post-perfusion and prior to wound closure showed that at least 60% of the CD45+ cells were coated by perfused anti-CD45 ('antibody uptake'). Among the patients studied for uptake and episodes of rejection, the incidence of rejection was 75% in 12 patients whose antibody uptake was < 95% compared with 22% in 18 patients with antibody uptake > or = 95% (P = 0.01). We conclude that this treatment was free of adverse effects and that there is a correlation between the uptake of antibody by passenger leucocytes and reduction in acute rejection episodes.

Systemic lupus erythematosus complicated by panniculitis, optic atrophy, and hemiplegia: report of a case.

A case is presented of a patient with systemic lupus erythematosus who had three of its rarest complications--panniculitis, optic atrophy, and hemiplegia, the latter two occurring despite adequate corticosteroid therapy. There appears to be a missing link in this patient's immune mechanism, which faulted her complete response to corticosteroid therapy and led to the three complications.

Cutaneous Hodgkin's disease.

A case of specific cutaneous Hodgkin's disease, which is sparsely mentioned in the literature, is reported. The lesion was nodulo-ulcerative and corresponded to the Grosz-Hirschfield type. It was confirmed by biopsy and the pattern was similar to that of an excised supraclavicular node examined two years previously. The lesion responded to treatment with gentamicin topically, clindamycin orally and 500 r of cauterizing X-ray. There has been no recurrence to date and no other new cutaneous lesions. This report further confirms what Ferdinand von Hebra implied one hundred years ago--that a dermatologist should not only know skin diseases but also the cutaneous manifestations of visceral diseases.