Induction of tolerance to heart allografts in rats using posttransplant total lymphoid irradiation and anti-T cell antibodies.

This study examined whether posttransplant anti-T cell monoclonal or polyclonal antibody therapy could provide a window of treatment to allow posttransplant total lymphoid irradiation (TLI) to induce tolerance. These experiments were conducted in a high responder strain combination of an ACI cardiac allograft into a Lewis rat. In this situation, treatment with antibody or posttransplant TLI alone is insufficient to induce tolerance, while similar treatments alone have been shown to induce tolerance in low responder strains. The affects of three anti-T cell therapies were compared: anti-CD4 mAb therapy, anti-CD3 mAb, and rabbit antithymocyte globulin (RATG). None of these antibody therapies alone prolonged graft survival indefinitely. Combining anti-CD4 therapy with posttransplant TLI markedly delayed rejection but failed to induce long-term graft survival. Tolerance could be induced by a combination of anti-pan T cell antibody (anti-CD3) and TLI, and, all grafts survived beyond 100 days. RATG failed to prevent graft rejection when used alone or in combination with TLI. However, posttransplant therapy with a combination of RATG, TLI, and single-donor blood transfusion resulted in graft survival beyond 100 days. Recipients bearing long-term donor grafts rejected third-party (PVG) grafts within 2 weeks. Low density donor bone marrow cells used instead of a blood transfusion did not facilitate tolerance. The results indicate that monoclonal or polyclonal anti-pan T cell antibodies, TLI, and a donor blood cell infusion function synergistically in facilitating tolerance to allografts in the posttransplant period.

Beta-blockade with bucindolol in heart failure caused by ischemic versus idiopathic dilated cardiomyopathy.

BACKGROUND: We investigated the effects of bucindolol, a nonselective, non-ISA beta-blocker with mild-vasodilatory properties, in patients with congestive heart failure from ischemic dilated cardiomyopathy (ISCDC, n = 27) and compared the results with those in subjects with heart failure from idiopathic dilated cardiomyopathy (IDC, n = 22). METHODS AND RESULTS: Patients were randomized in a double-blind fashion to receive 12 weeks' treatment with either bucindolol or placebo, with randomization stratified for IDC or ISCDC: Invasive (right heart catheterization) and noninvasive (echo, MUGA, central venous norepinephrine, exercise treadmill studies, and symptom scores) tests of heart failure severity were determined at baseline and end of the study. For all subjects (ISCDC plus IDC), relative to placebo treatment, bucindolol-treated patients had significant improvement in ejection fraction, left ventricular size and filling pressure, stroke work index, symptom score, and central venous norepinephrine. However, most of these differences could be attributed to improvement in the IDC subgroup, as the only parameter with a statistically significant degree of improvement in the bucindolol-treated ISCDC subgroup was left ventricular size. CONCLUSIONS: We conclude that beta-blockade may produce quantitatively different degrees of response in different kinds of heart muscle disease.

Lysophosphatidylcholine increases cytosolic calcium in ventricular myocytes by direct action on the sarcolemma.

Lysophosphatidylcholine (LPC) accumulates in myocardial tissues during ischemia, and has toxic effects which may contribute to the arrhythmias and relaxation abnormalities that occur during acute ischemia. These effects of LPC may be mediated in part by calcium overload. To test this hypothesis, spontaneously contracting cultured embryonic chick ventricular myocytes were superfused with various concentrations of LPC (10, 50 and 100 microM) while effects on contractile motion (video motion detector) and changes in free intracellular calcium ion concentration ([Ca2+]i indo-1 fluorescence) were determined. At concentrations greater than or equal to 10 microM, a dose-related, time-dependent effect occurred after exposure to LPC, consisting of the development of contracture and marked elevation of [Ca2+]i. LPC also produced a dose-related, time-dependent inhibition of K+ uptake, indicating there was inhibition of the Na(+)-K+ ATPase Na+ pump. However, the LPC-induced increase in [Ca2+]i was not due to Na+ overload caused by inhibition of the Na(+)-K+ ATPase Na+ pump because superfusion with a zero-Na+ solution did not prevent an increase in [Ca2+]i after LPC exposure; and the increase in [Ca2+]i after exposure to LPC occurred too rapidly to be accounted for by Na+ pump inhibition. Removal of extracellular Ca2+ prevented the rise in [Ca2+]i, after exposure to LPC but treatment with verapamil failed to inhibit the increase in [Ca2+]i induced by LPC. We conclude that LPC produces contracture due to an increase [Ca2+]i. These effects are seen at concentrations of 10 microM and greater, are not due to altered Na(+)-K+ ATPase Na+ pump or calcium channel function, and are probably related to the detergent properties of this amphiphile. There effects may account in part for myocardial dysfunction during ischemia in intact tissue.

Myocyte injury and contraction abnormalities produced by cytotoxic T lymphocytes.

BACKGROUND: The mechanisms by which ventricular function is altered during cardiac transplant rejection are not well understood. Therefore, an in vitro model system has been developed to facilitate investigation of lymphocyte-mediated myocyte injury. METHODS AND RESULTS: Splenic lymphoid cells were obtained from mice 8-10 days after placement of a vascularized abdominal cardiac allograft and were restimulated in vitro with irradiated donor-type splenocytes for 5 days. Cytotoxic effects of these allogenically stimulated lymphocytes on syngeneic and donor strain fetal cultured myocytes were determined by a 51Cr release assay at different lymphocyte to myocyte ratios. 51Cr release from donor strain myocytes was detectable within 1 hour of exposure, was maximal by 3-5 hours of coincubation with sensitized lymphocytes, and was allospecific. Cell injury manifest by 51Cr release was calcium dependent and was inhibited by pretreatment of lymphocytes with phorbol ester to deplete protein kinase C. Myocyte injury was also prevented by pretreatment of sensitized lymphocytes with anti-Thy 1.2 or anti-CD8 antibody plus complement but not by treatment with anti-CD4 antibody, indicating that CD8+ cytotoxic T cells are involved. Altered myocyte contractile motion preceded myocyte lysis (51Cr release), was characterized by an initial reversible decrease in amplitude of contraction, and was followed by rapid and irregular beating with eventual complete cessation of contraction. Contractile alterations induced by sensitized lymphocytes were inhibited by elimination of CD8+ cells. CONCLUSIONS: Myocyte injury can be produced by sensitized cytotoxic T lymphocytes in vitro and is calcium and protein kinase C dependent. The contractile abnormalities produced appear to be similar to those observed in cardiac transplant patients undergoing rejection, and thus this model system promises to allow investigation of the mechanisms involved.

Aseptic meningitis following cardiac transplantation: clinical characteristics and relationship to immunosuppressive regimen.

Neurologic disorders are uncommon but alarming complications of cardiac transplantation. Of 29 patients from the Utah Cardiac Transplant Program (UCTP) who had lumbar puncture because of change in neurologic function, or to assess fever of uncertain etiology, CSF pleocytosis was present in 14 patients, 4 of whom had an active infectious process involving the nervous system. In 10 other patients, CSF pleocytosis with negative cultures appeared following treatment with OKT3 monoclonal antibody. The most prominent clinical signs of this aseptic meningitis syndrome are fever and transient cognitive dysfunction.

Immunosuppression following cardiac transplantation.

Immunosuppression following cardiac transplantation can be divided into early rejection prophylaxis, chronic maintenance, and the treatment of established episodes of allograft rejection. Early rejection prophylaxis is the immunosuppressive protocol administered in the first few weeks following transplantation and consists of cyclosporine, azathioprine, and corticosteroids with or without the addition of specific anti-T cell agents such as antithymocyte globulins, antilymphoblast globulins, or the murine monoclonal anti-CD3 antibody (OKT3). Most programs now use triple therapy (cyclosporine, azathioprine, and prednisone) as chronic maintenance immunosuppression, although the feasibility of corticosteroid-free maintenance has been demonstrated. The treatment of acute allograft rejection involves optimization of cyclosporine and azathioprine doses along with the augmentation in corticosteroids and with or without the addition of a specific anti-T cell agent, depending on the histological grade hemodynamic consequences of the rejection episode. Further individualization of immunosuppressive therapy is likely to occur in the future.

Estrus, ovulation, and serum hormones in mares given prostaglandin F2 alpha, estradiol, and gonadotropin-releasing hormone.

A gonadotropin-releasing hormone (GnRH) was injected in mares given prostaglandin F2 alpha (PGF2 alpha) to induce luteolysis in an attempt to sunchronize ovulation. Pretreatment with estradiol-17 beta (E2-17 beta) was used to determine whether or not estradiol would enhance the release of luteinizing hormone (LH) after treatment with GnRH. Twelve mares were used in a balanced Latin square crossover design. Mares were injected with PGF2 alpha, treatment A; PGF2 alpha mgnRH, treatment B; or PGF2 alpha me2-17 beta mgnRH, treatment C. The interval +/- SEM from PGF2 alpha injection to estrus was 3.3 +/- 0.2, 3.2 +/- 0.3, and 2.3 +/- 0.1 days for treatments A, B, and C, respectively. The mean interval in days from injection to first ovulation averaged 1 day less (P less than 0.10) for treatments B (6.7 +/- 0.6) and C (6.2 +/- 0.3) than for treatment A (7.6 +/- 0.7). The furation of estrus and the interval from ovulation to end of estrus were similar among the three treatment groups. Mares treated with GnRH (treatments B and C) had a significant (P less than 0.01) two- to threefold increase in LH concentrations when compared with controls. Pretreatment with estradiol-17 beta did not appear to enhance LH release after GnRH was given, and the time of ovulation was not significantly changed by GnRH treatment.

Prolonged interovulatory interval after oestradiol treatment in mares.

This study was designed to test if oestradiol treatment would prevent or delay luteolysis in mares. Mares (5/group) received 0, 0.1, 1.0 or 10.0 mg oestradiol-17 beta daily from the day of ovulation until the next ovulation or for a maximum of 32 days. This treatment did not prevent luteolysis which occurred 15.8, 16.8, 15.8 and 17.3 days after the previous ovulation for the mares treated with 0, 0.1, 1.0 and 10.0 mg oestradiol respectively. Although oestradiol treatment failed to alter oestrous behaviour after luteolysis, daily treatment with 10.0 mg oestradiol prevented follicular growth and inhibited ovulation.