Early prediction of acute rejection after inbred rat kidney transplantation using macrophage migration inhibition test

In order to formulate an early diagnostic method for acute rejection after kidney transplantation, macrophage migration inhibition test (MIT) was carried out with lapse of time after inbred rat kidney allotransplantation. The mean survival time of rat kidney allograft was found to be 7.07 +/- 1.34 days. On the other hand, in the group treated with rabbit anti-rat lymphocyte serum (ALS) the mean survival time was lengthened to 14.15 +/- 2.14 days (p less than 0.05). The corresponding antigen used for MIT was prepared with donor kidney by ultrasonication, and its protein concentration at 180 mug/ml was the most optimal as not to elicit non-specific inhibition of macrophages. In the control group, activity of macrophage inhibitory factor (MIF activity) turned positive 3 days after the transplantation, and it became strongly positive by 5 or 7 dyas at the period when rejection crisis appeared frequently. ALS-treated group showed a lower MIF activity than the control group (p less than 0.05) and on 7-12 dyas before rejection crisis appeared frequently, MIF activity became strongly positive. These findings suggest that this MIT is simple and will be proved to be useful in predicting the acute rejection as well as in controlling the immunosuppression.</p

Beneficial effect of EPC-K1 on the survival of warm ischemic damaged graft in rat cardiac transplantation.

A newly introduced compound, EPC-K1, represents a phosphate diester linkage of vitamin E and vitamin C. The effect of EPC-K1 on the reperfusion injury was evaluated in a heterotopic cardiac transplantation model using syngenic combination rats. Prior to the warm ischemia, 12mg EPC-K1/kg was administered intravenously to donor rats. After 15 min of warm ischemic time, hearts were harvested and perfused with 4 degrees C saline. After completion of the transplantation, recipient rats were also treated with intravenous 12 mg EPC-K1/kg, before reperfusion. Saline was used instead of EPC-K1 for both donors and recipients in the control group. On the 7th post-transplantation day, graft survival was 7 out of 8 in EPC-K1 group, versus 1 out of 9 in the control group (p &#60; 0.001). Thiobarbituric acid-reactive substance levels in the recipient serum, three hours after reperfusion, were significantly limited, in the group in which EPC-K1 was administered only to donors. But it was not possible to clarify whether the effect of EPC-K1 is primarily at the donor or recipient levels at this time. These results indicate that EPC-K1 may reduce reperfusion injury after cardiac transplantation. This beneficial effect may be mediated by the hydroxyl radical scavenging properties of EPC-K1.</p

Effect of serum fractions obtained from cancer patients by double filtration plasmapheresis combined with natural tumor necrosis factors and cyclophosphamide on murine pulmonary metastases.

We investigated the effects of fractionated sera obtained from cancer patients by double filtration plasmapheresis (DFPP) plus antitumor agents on murine pulmonary metastasis. Fractions of the sera, in combination with natural human tumor necrosis factors (nTNF) and cyclophosphamide (Cy), were systemically administered to Lewis lung carcinoma-bearing mice. When the second filtrate (a plasma fraction containing substances composed of smaller molecular weight compounds) combined with low-dose nTNF (1,000 U/kg) and Cy (250 micrograms/kg) was administered to the mice, the degree of metastasis was significantly suppressed compared with the control group (p less than 0.01). In contrast, the discarded fluid (a plasma fraction containing larger molecular weight compounds) combined with the same doses of nTNF and Cy caused little inhibition of metastasis. Also, the discarded fluid significantly suppressed natural killer activity compared with normal sera (p less than 0.01). The results suggested that DFPP combined with nTNF and Cy is an efficient procedure to remove immunosuppressive factors from the sera of cancer-bearing hosts, to enhance the host antitumor immunity, and to suppress tumor proliferation.</p

Effects of a single donor-specific blood transfusion on the survival of rat cardiac allografts.

It is now well recognized that pre-transplant donor-specific blood transfusion (DST) has a beneficial effect on the survival of allografts. To determine the optimal interval between DST and transplantation, and to analyze the mechanisms of this effect, the survival of cardiac allografts to rats which received a single DST was examined. The cardiac allograft survival was found to be prolonged when the DST was performed 1 to 6 weeks before grafting. In addition, recipient rat sera collected 1 to 6 weeks after a single DST showed significant inhibition of a mixed lymphocyte reaction (MLR). This MLR inhibition correlated with prolongation of survival of histoincompatible rat cardiac allografts. It thus appears that a single DST given from 1 to 6 weeks before transplantation has a beneficial effect on allograft survival and that MLR inhibition may be essential for inducing the effect of transfusion on organ transplantation.</p

Prolongation of Cardiac Allograft Survival in Rats by Treatment with Anti-Interleukin 2 Antiserum

Interleukin-2 (IL2) is the obligatory signal for both T cell mitogenesis and in vitro generation of alloreactive cytotoxic T lymphocytes (CTL). An investigation was made to determine whether an antibody directed against IL2 would suppress the rejection reaction of rat cardiac allografts. Rabbit anti-interleukin 2 (anti-IL2) antiserum was obtained by immunizing at 2 week intervals over a period of 8 weeks with 10(6) U of recombinant human IL2 along with complete Freund's adjuvant. The bioassay for inhibition of IL2 activity by anti-IL2 antiserum was carried out in conjunction with the IL2-dependent cytotoxic T cell (CTLL cell) assay. Cardiac allografts of F344 rats were heterotopically transplanted into ACI rats. Seven daily doses of 1 ml of anti-IL2 antiserum were administered intravenously following transplantation. IL2-driven [3H]thymidine incorporation in CTLL cells was significantly inhibited by rabbit anti-IL2 antiserum. Graft survival in the anti-IL2 serum-treated group was significantly prolonged in a dose-dependent fashion compared to control groups. In conclusion, these results indicate that rabbit anti-IL2 antiserum may prove to be of significant value as an immunosuppressive agent in clinical organ transplantation.</p

Effects of serum fractions obtained from cancer patients by double-filtration plasmapheresis on tumor growth and metastasis in mice.

We administered serum fractions obtained from cancer patients by double-filtration plasmapheresis (DFPP) to cancer-bearing mice to examine the effects on tumor growth and metastasis. Fraction 1 (whole plasma), fraction 2 (a plasma fraction containing substances with higher particle size), fraction 3 (a plasma fraction containing substances with smaller particle size) and saline were administered intravenously to cancer-bearing mice for 10 days following the inoculation of tumor cells. The tumor growth and metastasis in mice administered fraction 2 was far more rapid than that in the control mice. On the other hand, tumor growth in mice administered fraction 3 was significantly delayed compared with that in mice injected with fraction 2. These results suggest that factors in the higher particle-size fraction of cancer patients' sera promote the growth and the metastasis of tumors in mice, and that DFPP, which remove these factors, is an effective therapy against cancer.</p

Active Enhancement of Rat Cardiac Allografts by Donor-Specific B Lymphocytes

In an attempt to induce the active enhancement of the cardiac allografts, recipient rats were injected intravenously with 1 x 107 donor-specific spleen cells or an equal number of their T or B cell subpopulations on each of 7 days before transplantation. Mean survival times (MST) in the group pretreated with donor-specific spleen cells (MST 27.5 ± 7.8 days) and the group pretreated with donor-specific B cells (MST 37.5 ± 15.5 days) were significantly prolonged (p<0.01) compared with the untreated control (MST 11.6 ± 2.0 days) and the group pretreated with donor-specific T cells. To investigate the mechanisms of the beneficial effect of donor-specific B cells on rat cardiac allografts, inhibition assay of mixed lymphocyte reaction (MLR) were carried out by the addition of recipient rat sera or spleen cells harvested 7 days following the intravenous administration of donor-specific spleen cells or their T and B cell subpopulations. Recipient rat sera harvested 7 days following the intravenous administration of donor-specific B cells showed significant inhibition of MLR; this inhibition was correlated to the prolonged survival of histoincompatible rat cardiac allografts. In contrast, MLR suppressor cells could not be detected in any experimental group. Thus, donor-specific B cell given 7 days before transplantation may possibly have beneficial effect on rat cardiac allografts and MLR inhibition induced by the intravenous administration of donor-specific B cells may be essential for prolonged allograft survival

Retroperitoneal hematoma associated with femoral neuropathy: a complication under antiplatelets therapy.

We report a case of retroperitoneal hematoma presenting as femoral nerve pulsy on antiplatelet therapy. The patient, a 78-year-old man who had undergone antiplatelet treatment using ticlopidine, was admitted to our hospital with complaints of sudden-onset low abdominal and back pain. Computed tomography showed an iso-density mass in the right retroperitoneum within the psoas muscle. We made a diagnosis of retroperitoneal hematoma compressing the femoral nerve and performed an operation to remove the hematoma in order to decompress the femoral neuropathy. Postoperatively, the patient rapidly recovered from the femoral neuropathy. In the particular case in which no antagonist against the ticlopidine is available, surgical decompression could produce a good outcome.</p

Unresponsiveness of antidonor cytotoxic T cells in a long-term stable renal transplant recipient.

The antidonor immune response was examined in a one haplotype-mismatched renal transplant recipient with an allograft that had been well-functioning for more than 10 years. Although the relative response of the mixed lymphocyte reaction (MLR) was (45.8)% and the MLR responder cells stimulated by donor cells produced measurable amounts of interleukin-2 (IL-2) (11.6 U/ml), the cytotoxic T lymphocytes (CTL) could not be generated against donor cells, even with exogenous IL-2. These results indicate that antidonor CTL precursors were either deleted or inactivated in this recipient.</p

Nonicteric liver damage with a gamma-glutamyl transpeptidase level of 5,609 units/l in a renal-transplant recipient receiving azathioprine.

A 26-year-old male with renal allograft, who received immunosuppressive treatment with azathioprine, presented marked elevations of serum biliary tract enzymes, such as gamma-glutamyl transpeptidase (5,609 units/l) and alkaline phosphatase (60.5 Bessey-Lowry units), 14 months after transplantation. Two months later the patient became icteric; he died of respiratory failure 19 months after the renal allograft. Postmortem examination revealed intrahepatic cholestasis with minimal inflammatory cell infiltration, indicating drug hepatotoxicity.</p