Cyclosporine A Group Research Articles

Conversion from cyclosporine A to mycophenolate mofetil protects recipient kidney and prevents intimal hyperplasia in rat aortic allografts

Background Recent studies have demonstrated that complete conversion from cyclosporine A (CsA) to mycophenolate mofetil (MMF) prolongs graft survival in patients undergoing clinical organ transplantation. We investigated the effects of conversion from CsA to MMF on recipient kidneys and transplant arteriosclerosis in a rat aortic allograft model as a high responder combination. Methods DA (MHC haplotype, RT1 a) rat abdominal aortic grafts were orthotopically transplanted into Lewis (RT1 l) rats. The recipients were divided into four oral treatment groups: (1) vehicle group, (2) CsA group (15 mg/kg/day), (3) CsA/MMF40 group (conversion from CsA 15 mg/kg/day to MMF 40 mg/kg/day on day 14), and (4) CsA/MMF20 group (conversion from CsA 15 mg/kg/day to MMF 20 mg/kg/day on day 14). On day 28 after transplantation, the rats were sacrificed and the hematoserological parameters were analyzed. The grafted aortas and recipient kidneys also were evaluated histologically and immunohistochemically. Results The CsA group developed serological renal dysfunction, arteriolar hyalinosis, and apoptosis in the recipient kidneys, whereas the CsA/MMF40 and CsA/MMF20 groups did not. In the vehicle group, we observed remarkable intimal hyperplasia and marked inflammatory cell infiltration including macrophages and T cells. In the CsA group, intimal hyperplasia was evident without infiltration of macrophages or T cells. In the CsA/MMF40 and CsA/MMF20 groups, intimal hyperplasia was abrogated, while adventitial infiltration of and adhesion to the endothelium by macrophages and T cells occurred. Conclusions Conversion from CsA to MMF protected recipient kidneys and prevented transplant arteriosclerosis. However, insufficient immunosuppression by MMF might reactivate immune cells. This conversion therapy has preventive potential in transplant patients with CsA-associated nephrotoxicity and transplant arteriosclerosis.

TWO IMMUNOSUPPRESSIVE PROTOCOLS FOR RENAL PATIENTS TRANSPLANTED FROM MARGINAL DONORS ACCORDING TO AN “OLD FOR OLD” ALLOCATION.

P729 Aims: When transplanting kidneys from marginal donors (MDs) with an “old for old” allocation, immunosuppression should be tailored either on the recipient or on the donor‘s characteristics, because elderly recipients are more fragile and marginal kidneys are more susceptible to nephrotoxic injuries. A calcineurin-inhibitors (CIN) free strategy during the induction phase may be a good solution but one cannot deny the existence of a greater risk for acute rejection. As for the maintenance phase tacrolimus (TAC) increases the diabetes occurence in comparison with Cyclosporine A (CyA), but the latter, in spite of the association with mycophenolate mofetil (MMF) seems to increase the acute rejection rate, particularly when tapering steroids. Aim of the study: on the basis of these informations (from literature and from personal experience) we decided to compare the short-term effects (6 months) of the two drugs used in 135 recipients of single/dual grafts from MDs with a protocol CIN free in the induction phase (basiliximab+steroids+MMF). Methods: MDs all ≥ 50 years old, renal biopsy before grafting, TAC (n=90 patients) or CyA (n=45 patients) added when serum Creatinine (sCr) ≤ 2.5 mg/dl. TAC and CyA groups were homogeneus for all parameters (donors gender and age, recipients gender) with the exception of cold ischemia time (longer in CyA group) and recipient’s age (older in TAC group). The groups were compared as for survival (Kaplan Meier estimates log rank for significance) and adverse events. Results: TAC and CyA patients behaved similarly as for delayed graft function (DGF; TAC 47%, CyA 37%) and acute rejection rate (total: TAC 14%, CyA 20%; during the CIN–free phase: TAC 8%, CyA 11%), sCr (TAC 1.6 ± 0.6, CyA 1.6 ± 0.5 mg/dL), steroid dose (TAC 3.3 ± 1.4, CyA 5 ± 6 mg/day). De novo insulin therapy was mostly required in TAC group as it was expected (18.2% vs 4.7%). Diabetes rate after 6 months, again, was higher in TAC group but statistically significance versus CyA group was not reached (11.4% vs 2.4% p=NS), perhaps because of small numbers. No statistically significant difference as for graft and patient survival was noted, in spite of the fact that TAC patients exihibit better results. Conclusions: When adopting a CIN free protocol for grafts from MDs with an “old for old” allocation, acute rejection rate is low in the induction phase and also afterwards, when either TAC or CyA are added. Graft and patient survivals are good with both drugs. TAC is associated with a statistically greater number of post transplantation diabetes but it allows an easier and safer tapering of steroids. Also by means of the steroid dose reduction, insulin is no more required in 50% of the patients at 6 months. In a CyA regimen, diabetes rate is very low and acute rejection rate is slightly, but not significatly, increased.

CYCLOSPORINE AND LOW-DOSE KETOCONAZOLE IN RENAL TRANSPLANT RECIPIENTS: A SINGLE-CENTER EXPERIENCE

The high cost of cyclosporine A (CsA) is an impediment for low-income patients. Previous studies have used ketoconazole at doses between 200 and 400 mg/day to lower CsA consumption. Ketoconazole and CsA were administered prospectively to renal transplant patients. Patients treated historically with CsA were used as a reference group. At different intervals posttransplant, clinical and laboratory data were recorded. The reference group (n=14) was treated with CsA from 1992 to 1997 and the CsA plus ketoconazole group (n=17) from 1998 to 2002. Follow-up was 76+/-22 and 29+/-14 months, respectively. CsA doses throughout the study were 4.0+/-1.3 and 1.6+/-0.6 mg/kg/day (a 60% reduction, P =0.00). Trough levels of CsA were 194+/-87 and 193+/-69 ng/mL, respectively. The ketoconazole dose was 54+/-17 mg/day. The monthly cost of CsA was reduced by 60%, including the cost of ketoconazole. CsA with ketoconazole resulted in a substantial dose and cost reduction that proved safe and effective.

Vitamin E attenuates biochemical and morphological features associated with development of chronic pancreatitis

The objective was to investigate the effects of vitamin E on collagen deposition induced by Cyclosporin A (CsA) administration in rats with caerulein (Cr) pancreatitis. CsA transforms the fully regenerative, self-limited form of Cr pancreatitis into a chroniclike disease in conjunction with increased transforming growth factor (TGF)-beta and myofibroblast proliferation. Vitamin E inhibits TGF-beta release in mesangial cells and reduces CsA cytotoxicity. Wistar rats received CsA daily (20 mg/kg), and CR pancreatitis was induced on days 1 and 8 (Cr + CsA group). In a separate group, vitamin E (600 mg.kg(-1).day(-1)) was administered starting 4 days before CsA. Three other groups received either vehicle, CsA, or Cr alone. Thiobarbituric acid-reactive substance (TBARS), 8-isoprostanes, and hyaluronic acid were measured in plasma obtained on the day the animals were killed (day 15). Pancreases were weighed and processed for light microscopy to assess connective tissue and myofibroblast number. Pancreatic homogenates were also assayed for collagen (hydroxyproline) and TBARS content. TBARS, 8-isoprostane, and TGF-beta were elevated in CsA and Cr + CsA rats. Vitamin E treatment greatly decreased these parameters. Vitamin E also decreased the fall in pancreatic weight observed in Cr + CsA pancreas. Pancreatic hydroxyproline and plasma hyaluronic acid were increased in Cr + CsA rats but were effectively reduced by vitamin E. Morphology showed improvement in fibrosis score and a decreased number of myofibroblasts in vitamin E-treated rats. Vitamin E reduces oxidative stress and collagen deposition during the development of experimental chronic pancreatitis. Adjuvant antioxidants may be of value in the treatment of chronic pancreatitis.

The effect of Cyclosporine A on survival time in salicylate-poisoned rats

Salicylate (SAL) produces mitochondrial membrane permeability transition (MPT) with resultant oxidative phosphorylation uncoupling. Cyclosporine A (CSA) inhibits SAL-induced MPT. This study determined if CSA pretreatment prolonged survival time in SAL-poisoned rats. Twenty-nine rats were randomized to receive pre-treatment with either 30 mg/kg CSA or equal volume of control diluent intraperitoneally (i.p.). Four hours later, all rats received 1700 mg/kg sodium salicylate i.p. Survival time, whole blood CSA ([CSA]), and serum sodium ([Na]), glucose and SAL ([SAL]) concentrations were determined. The results showed median survival time for controls was 18 min (95% CI 14–22 min) and for CSA animals was 14 min (95% CI 13–15 min). Univariate and multivariate analyses and Cox proportional hazard regression revealed CSA treatment was associated with higher [SAL], which was associated with shortened survival times. The CSA group also demonstrated shorter survival times for a given [SAL]. In conclusion, CSA pre-treatment shortened survival in SAL-poisoned rats.

A Randomized, Double-Blind, Multicenter, Controlled Clinical Trial of Cyclosporine Plus Chloroquine vs. Cyclosporine Plus Placebo in Early-Onset Rheumatoid Arthritis

Background Our objective was to assess the efficacy and safety of cyclosporine-A (CsA) plus chloroquine (Clq) in early-onset rheumatoid arthritis (RA) compared to CsA plus placebo. Methods We conducted a prospective, 12-month follow-up, multicenter, double-blind, placebo-controlled study of CsA (2.5–5 mg/kg/day[d]) plus Clq (150 mg/d) vs. CsA plus placebo in active RA of <2 years of evolution. Results A total of 149 patients were included; 111 patients (74.4%) completed the 12-month follow-up period. Evaluation at 6 and 12 months showed improvement for all clinical disease parameters. In both groups there was a decrease in tender joint count, swollen joint count, pain, assessment of efficacy by both investigator and patient, functional assessment, and morning stiffness, all differences statistically significant. With an intention-to-treat analysis, there was 64% in the CsA plus Clq group (CsA/Clq) and 63% in the CsA plus placebo group (CsA/Plac) at 12 months in the American College of Rheumatology (ACR)-20 criteria of improvement. Response rate for ACR-50 was 48 and 47%, and for ACR-70 it was 29% in both groups; the difference was not statistically significant between study groups. Gastrointestinal complaints were common in both groups. Four patients in CsA/Clq group and five patients in CsA/placebo group increased creatinine levels; two patients in each group discontinued treatment due to this reason. Conclusions There was no advantage to adding chloroquine to cyclosporine in patients with RA.

Gene Expression of Extracellular Matrix Proteoglycans in Human Cyclosporin‐Induced Gingival Overgrowth

Gingival overgrowth is one of the side effects associated with the systemic use of cyclosporin A (CsA). In vitro studies on the extracellular matrix of gingival tissues have demonstrated an altered composition, particularly an accumulation of proteoglycans and collagen. We investigated the gene expression of extracellular matrix proteoglycans in CsA-induced gingival tissue alterations. mRNA expression of the proteoglycans perlecan, decorin, biglycan, and versican was analyzed by reverse transcription polymerase chain reaction (RT-PCR) in gingival samples obtained from 12 individuals, six with CsA-induced gingival overgrowth (CsA group) and six with a normal gingiva (control group). The RT-PCR products were subjected to 1% agarose gel electrophoresis containing ethidium bromide and analyzed qualitatively and semiquantitatively by densitometry. Density values were normalized by determining the expression of the housekeeping gene beta-actin in the same sample. Groups were compared by the Student's t test. Perlecan expression showed a marked increase (54%) in the CsA group compared to the control group (P < 0.01), while no significant differences were observed for the other proteoglycans. CsA-induced gingival overgrowth seems to be associated with increased expression of perlecan, a typical basement membrane proteoglycan, but not decorin, biglycan, or versican.

Effects of cessation of immunosuppression on skeleton reconstructed by vascularized bone allograft in rats

In the present study, we investigated the effects of cessation of immunosuppression on skeleton reconstructed by vascularized allogenic bone transplantation in a rat tibio–fibula graft model. Twelve-week-old male 25 Dark Agouti rats with the major histocompatibility antigen (MHC) RT1 a were used as donors and age-matched male 25 Lewis rats with MHC RT1 l were used as recipients. Among them, 20 rats were randomly allocated to 8-week cyclosporine A (CsA) followed by 8-week CsA vehicle group or continuous 16-week CsA group. The remaining 5 rats received CsA for 8 weeks followed by no further treatment for next 40 weeks (long-term observation group). In the CsA followed by vehicle group as well as the continuous CsA group, the structure of the reconstructed bones was maintained, though the transplanted bones in former group were found to be partly non-vital. The CsA followed by vehicle group had higher bone mineral density of the transplanted bones and stronger strength of the reconstructed bones than the continuous CsA group. In the long-term observation group, the structure of the reconstructed bones was still maintained and the transplanted bones were almost vital. These results suggest that long-term strong immunosuppression may not be necessary for successful reconstruction of large bone defect by vascularized bone allograft.

Cyclosporine A effectively inhibits graft-versus-host disease during development of Epstein-Barr virus-infected human B cell lymphoma in SCID mouse.

We previously constructed human peripheral blood lymphocyte (hu-PBL)/severe combined immunodeficiency mouse (SCID) chimeras and induced human B-cell lymphomas associated with Epstein-Barr virus (EBV) in SCID mice. However, a number of SCID mice died of graft-versus-host disease (GVHD) during the early experimental course. The aim of this study was to test the efficacy of cyclosporine A (CSA) for prevention of GVHD and to define how CSA inhibits the occurrence of GVHD and the production of soluble interleukin (IL) 2 receptor (sIL-2R) in hu-PBL/SCID mice. No mouse died in the active EBV infection group with CSA administration, while 17 mice in three groups without CSA administration died of GVHD. Mortalities in these three groups were 55.56% (5/9), 30.43% (7/23), and 27.78% (5/18), and the medium life span was 17 days. Over the first 33 days after hu-PBL transplantation, serum level of human sIL-2R in hu-PBL/SCID chimeras was stable in the active EBV infection plus CSA group, while sIL-2R concentration gradually increased in the sera of mice with active EBV infection without CSA administration and peaked at 22 days. Thirty-two mice developed tumors among the 43 surviving SCID mice. There was no significant difference of tumor incidence between the active EBV infection groups with CSA and without CSA administration (P > 0.05). From their morphological and immunohistochemical features, as well as detection of human Alu-sequence and EBV in tumor cells, these EBV-induced tumors were identified as human B-cell lymphomas. Thus, CSA can strikingly inhibit GVHD in hu-PBL/SCID chimeras, and should therefore be effective to establish a stable SCID mouse model of human lymphoma associated with EBV. Treatment with CSA had no effect on the tumor incidence in hu-PBL/SCID chimeras after active EBV infection. Accordingly, serum level of sIL-2R is a valuable indicator of GVHD occurrence in hu-PBL/SCID chimeras.

Hypertrophy of vascularized bone isograft in rats treated with cyclosporine A.

The aim of this study was to investigate the effects of cyclosporine A (CsA) on vascularized tibio-fibula isograft between 12-week-old male Lewis rats. After transplantation, 45 rats were randomly allocated to one of the following 7 treatment groups: (1) 4-week vehicle (n = 5), (2) 4-week CsA (n = 5), (3) 8-week vehicle (n = 10), (4) 8-week CsA (n = 10), (5) 4-week CsA followed by 4-week vehicle (n = 5), (6) 16-week vehicle (n = 5), or (7) 4-week CsA followed by 12-week vehicle (n = 5). In soft X-ray and micro-computed tomography examination, hypertrophic change of the grafted bones was apparent in the 4- and 8-week CsA groups. Mineral apposition rate and bone formation rate of the grafted bones in the 4-week CsA group were markedly higher than those in the 4-week vehicle group. In the 4- and 8-week CsA groups, however, bone mineral density (BMD) of the grafted bones was lower and strength of the reconstructed bones was weaker than the 4- and 8-week vehicle groups. Urinary deoxypyridinoline (DPD) level was higher in the 4- and 8-week CsA groups than in the 4- and 8-week vehicle groups. The group of 4-week CsA followed by 4-week vehicle had a level of urinary DPD equal to the 8-week vehicle group, but their BMD of the grafted bones was lower and strength of the reconstructed bones was weaker than the 8-week vehicle group. By contrast, the group of 4-week CsA followed by 12-week vehicle had BMD of the grafted bones and strength of the reconstructed bones similar to the 16-week vehicle group. These findings demonstrate that short-term CsA treatment induces hypertrophic change of vascularized bone graft with high-turnover bone loss, and strength of the reconstructed bone is gradually restored after the cessation of CsA treatment.

Influence of cyclosporin A therapy on bone healing around titanium implants: a histometric and biomechanic study in rabbits.

Immunosuppressive agents may induce severe changes on bone metabolism. The purpose of the present study was to evaluate the influence of the administration of cyclosporin A (CsA) on the bone tissue around titanium implants. Eighteen New Zealand rabbits were randomly divided into 2 groups of 9 each. The test group (CsA) received daily subcutaneous injection of CsA (10 mg/kg body weight) and the control group (CTL) received saline solution by the same administration route. Three days after therapy began, 2 implants (7.0 mm long and 3.75 mm in diameter) were inserted bilaterally at the region of the tibial methaphysis. After 4, 8, and 12 weeks the animals were sacrificed and biomechanical tests and histometrical procedures, consisting of the determination of the percentages of bone-implant contact and bone area within the limits of the implant threads, were performed. Intergroup analysis showed that the removal torque and the percentage of bone contact with the implant surface for CsA group were significantly lower than those of the CTL group at 12 weeks (28.5 and 39.2 N cm, P = 0.01; 7.76% and 18.52%, P = 0.02, respectively). The data from the present study suggest that long-term administration of cyclosporin A may negatively influence bone healing around dental implants.

Isosorbide-5-mononitrate treatment prevents cyclosporin A-induced platelet hyperactivation and the underlying nitric oxide–cyclic guanosine-3′,5′-monophosphate disturbances

Introduction: The clinical use of cyclosporin A (CsA) is commonly associated with the development of hypertension and increased risk of thromboembolic events. Decreased endothelium-dependent relaxation and increased platelet activation seems to be involved on those side effects, but the underlying mechanisms are not yet elucidated. The present study aimed to evaluate the CsA effect on the platelet NO–cyclic guanosine-3′,5′-monophosphate (cGMP) pathway and the putative benefits of concomitant isosorbide-5-mononitrate (IS-5-MN) administration on CsA-induced hypertension and on platelet hyperactivation. Materials and Methods: Blood pressures, platelet NO synthase activity and cGMP content, intracellular free calcium concentration ([Ca 2+] i) and whole blood platelet aggregation were assessed in three rat groups orally treated, during 7 weeks, with the following diets: orange juice (control group), 5 mg/kg/day of CsA (CsA group) and 150 mg/kg/day, b.i.d., of IS-5-MN for 2 weeks and IS-5-MN plus 5 mg/kg/day of CsA for 7 weeks (IS-5-MN+CsA group). Results: IS-5-MN treatment has prevented hypertension development obtained in the solely CsA-treated rats. CsA treatment has inhibited NOS activity, which was reverted by the concomitant IS-5-MN and CsA administration. On the contrary, platelets from CsA-treated rats had cGMP content increased when compared with the control rats. The variation obtained when ISMN was present was less predominant. Therefore, the organic nitrate treatment has prevented platelet hyperactivation, namely, by decreasing thrombin-evoked [Ca 2+] i and collagen-evoked platelet aggregation, when compared with the solely CsA-treated group. The preventive effect of IS-5-MN was reinforced by electron microscopy studies of platelet activation. Conclusions: By increasing [Ca 2+] i and aggregation, CsA induces platelet hyperactivation and simultaneously increases cGMP content, which might represent a compensatory inhibitory mechanism. The concomitant IS-5-MN treatment prevents the above-mentioned platelet hyperreactivity and tends to normalize the NO–cGMP pathway as well as the development of hypertension.

FTY720, an immunosuppressant that alters lymphocyte trafficking, abrogates chronic rejection in combination with cyclosporine A

Chronic rejection remains the leading cause of failure after transplantation (Tx). FTY720, a new immunosuppressant altering lymphocyte trafficking, is effective against acute rejection, but its activity against chronic rejection is not known. A valid model of chronic rejection was produced. Heart transplantation (HTx) was performed using fully mismatched RA (RT1p) and PVG (RT1c) rats. Administration of donor-specific blood transfusion 12 days before HTx prolongs graft survival, but features of chronic rejection including intimal hyperplasia and vascular obliteration (VO) develop with time only in allogeneic Tx. This is therefore a valid model of chronic rejection. VO was assessed on post-Tx day 90 in six groups differing according to the maintenance immunosuppressive regimen administered. group 1, donor-specific blood transfusion only and no other treatment; group 2, FTY720 (0.3 mg/kg/day orally) for 90 days; group 3, cyclosporine A (CsA) (1 mg/kg/day orally) for 90 days; group 4, combined administration of FTY720 and CsA for 90 days; group 5, transient administration of combined FTY720 and CsA for 7 days; and group 6, syngeneic HTx (RA to RA). Graft infiltrate, endothelial immunoglobulin (Ig) G deposition, and complement binding were also examined on post-Tx day 90. In control group 1, severe VO was observed, compared with syngeneic HTx (group 6). Monotherapy with FTY720 (group 2) or with CsA (group 3) significantly but partially reduced VO. On the contrary, combined administration of FTY720 and CsA (group 4) abrogated VO. A 1-week treatment with combined FTY720 and CsA (group 5) reduced VO but only partially. In group 1, arteriosclerosis was accompanied by graft infiltrate, endothelial IgG deposition, and complement binding. In groups 2, 3, and 5, graft infiltrating scores were partially decreased compared with group 1 but remained higher than in syngeneic controls; endothelial IgG deposition and complement binding were still present. In group 4, continuous administration of combined FTY720 and CsA reduced graft infiltrate to the level of syngeneic control and abrogated both endothelial IgG deposition and complement binding. Maintenance treatment with either FTY720 or CsA monotherapy partially prevents chronic rejection; short-term treatment with combined FTY720 and CsA reduces chronic rejection only partially; and continuous treatment with combined FTY720 and CsA abrogates chronic rejection, and this is accompanied by dramatic reduction of graft infiltrating cells, endothelial IgG deposition, and complement binding. Prevention of chronic rejection by maintenance treatment with FTY720 and CsA represents indirect evidence that normal lymphocyte trafficking and function are mandatory for development of chronic rejection.

Cyclosporine induces progressive attenuation of baroreceptor heart rate response and cumulative pressor response in conscious unrestrained rats.

Cyclosporine A (CsA) use is associated with hypertension and reduced baroreceptor sensitivity (BRS), but the underlying mechanisms remain unresolved. In this study, we investigated whether CsA attenuation of BRS is 1) dependent on treatment regimen, and 2) causative of the pressor response. Furthermore, we investigated whether a reduction in plasma testosterone contributes to BRS attenuation caused by short-term CsA administration. The effects of the clinically used CsA formulation (15 mg/kg/day i.v. for 5 days) on mean arterial pressure (MAP), heart rate, BRS, and body weight were investigated in conscious rats. CsA caused reproducible pressor responses (15.1 +/- 3.0 mm Hg) starting after the first dose and continuing through the 5 days of the study. BRS and baseline MAP were inversely related in the CsA group because of a progressive reduction in BRS, which started on day 2 and reached approximately 50% of baseline on day 5 and a cumulative elevation in MAP. The inverse BRS and MAP responses required daily administration of CsA because neither response was evident throughout the 5-day observation period after a single dose of CsA. Plasma testosterone levels were similar in all groups, whereas the body weight decreased approximately 10% in the CsA group on day 5. These findings suggest 1) CsA attenuation of BRS is relatively rapid and cumulative; 2) the attenuation of BRS may contribute to the delayed, but not to the acute, pressor elicited by CsA; and 3) the cumulative reduction in BRS caused by short-term (5-day) CsA treatment is not testosterone-related.

Immunotolerance of liver allotransplantation induced by intrathymic inoculation of donor soluble liver specific antigen.

To study the effects of liver specific antigen (LSA) on the immunoreaction of liver allotransplantation and its significance. Orthotopic liver transplantation was used in this study. Group I: syngeneic control (Wistar-to-Wistar); Group II: acute rejection (SD-to-Wistar). Group III: acute rejection treated by intramuscular injection of cyclosporine A (CsA) (SD-to-Wistar+CsA). Group IV: Intrathymic inoculation of SD rat LSA one week before transplantation (LSA+SD-to-Wistar). The common situation and survival time, rejection grades, NF-kappaB activity of splenocytes and intragraft cytokine gene expression were observed to analyze the acute rejection severity and immune state of animals. The common situation of Wistar-to-Wistar group was very good after the transplantation and no signs of rejection were found. Recipients of SD-to-Wistar group lost body weight progressively. All died within 9 to 13 days after transplantation with the median survival time of 10.7+/-0.51 days. It was an optimal control for acute rejection. The common situation of SD-to-Wistar+CsA group was bad during CsA medication but only with mild rejection. As for LSA+SD-to-Wistar group, 5 of 6 recipients survived for a long time and common situation was remarkably better than that of SD-to-Wistar group and SD-to-Wistar+CsA group. Its rejection grades were significantly lower than that of SD-to-Wistar group (P=0.026). Furthermore, no significant discrepancies of rejection were found between SD-to-Wistar group and LSA+SD-to-Wistar group at day7 and day12 (P=0.067). NF-kappaB activity, IFN-gamma and IL-2mRNA expression were significantly inhibited in LSA+SD-to-Wistar group compared with that of SD-to-Wistar group (P<0.05). LSA is an important transplantation antigen which involves in the immunorejection of liver transplantation directly. We reported for the first time that intrathymic inoculation of LSA can induce immnotolerance of liver allotransplantation and grafts can survive for a long time thereby, thus leading to a novel way to liver transplantation immunotolerance.

Gene expression in cardiac and lung xenografts

Objectives: We have previously shown that lung xenografts could undergo “accommodation” in the same fashion as cardiac xenografts treated with cobra venom factor (CVF) and cyclosporin A (CsA). In this study, we asked if the gene expression profile of accommodated cardiac and lung xenografts was universal or specific to each organ. Methods: We used hamster-to-rat cardiac and lung xenotransplantation (XT) models. Cardiac XTs were treated with CVF CsA and compared to untreated controls. Lung XTs were treated with either CVF CsA or FK506 cyclophosphamide (Cp) and compared to untreated controls. We analyzed graft survival, protein expression of protective genes, histological and immunohistological findings and used microarray technology to identify other genes potentially involved in accommodation. Results: Rejection occurred rapidly in untreated rats. CVF CsA or FK506 Cp treatment significantly influenced graft survival. 8 of 12 heart transplants treated with CVF CsA survived at least 21 days. 7 of 16 in the CVF CsA group and 5 of 12 in the FK506 Cp group survived 21 days after lung transplantation. As expected, we observed significant protein expression of bcl-2, bcl-XL and HO-1 in cardiac and lung xenografts treated with CVF CsA, compared with untreated donor hearts. The overall pattern of gene expression in lung xenografts was distinct from that seen in accommodated cardiac xenografts. However, 38 genes were expressed in both organs after CVF CsA treatment, including insulin-like growth factor binding protein-6 and interferon regulatory factor6. Also, twelve genes, including interleukin-11and interferon regulatory factor-6, were expressed in accommodated cardiac xenografts and both groups of lung xenografts. Microarray results were validated with western blotting. Conclusion: After induction of accommodation, distinct genetic changes are seen in lungs compared to hearts, with only a few genes regulated in parallel.

Effects of Cyclosporin A, FK506, and 3-Deazaadenosine on Acute Graft-versus-host Disease and Survival in Allogeneic Murine Hematopoietic Stem Cell Transplantation

Background: We investigated the effect of donor marrow T cell depletion, administration of FK506, cyclosporin A (CSA), and 3-deazaadenosine (DZA) on graft versus host disease (GVHD) after allogeneic murine hematopoietic stem cell transplantation (HSCT). Methods: We used 4 to 6 week old Balb/c (H-2, recipient), and C3H/He (H-2, donor) mice. Total body irradiated recipients received 1×10 bone marrow cells (BM) and 0.5×10 splenocytes of donor under FK506 (36 mg/kg/day), CSA (5 mg/kg/day, 20 mg/kg/day), and DZA (45 mg/kg/day), which were injected intraperitoneally from day 1 to day 14 daily and then three times a week for another 2 weeks. To prevent the GVHD, irradiated Balb/c mice were transplanted with 1×10 rotor-off (R/O) cells of donor BM. The severity of GVHD was assessed daily by clinical scoring method. Results: All experimental groups were well grafted after HSCT. Mice in experimental group showed higher GVHD score and more rapid progression of GVHD than the mice with R/O cells (R/O group) (p<0.01). There were relatively low GVHD scores and slow progressions in FK506 and low dose CSAgroups than high dose CSA group (p<0.01). The survival was better in FK506 group than low dose CSA group. All mice treated with CSA died within 12 days after HSCT. The GVHD score in DZA group was low and slow in comparison with control group (p<0.05), but severity and progression were similar with low dose CSA group (p=0.11). All mice without immunosuppressive treatment died within 8 days, but all survived in R/O group (p< 0.01). Survival in low dose CSA group was longer than in control group (p<0.05), but in high dose CSA group, survival was similar to control group. The survival benefit in DZA group was similar with low dose CSA group. FK506 group has the best survival benefit than other groups (p<0.01), comparable with R/O group (p=0.18), although probability of survival was 60%. Conclusion: We developed lethal GVHD model after Effects of Cyclosporin A, FK506, 3-Deazaadenosine on Acute GVHD 151

Long-term results of a prospective randomized study comparing two immunosuppressive regimens, one with and one without CsA, in low-risk renal transplant recipients

Abstract Due to the nephrotoxicity of cyclosporin A (CsA), its benefit on long-term graft survival remains controversial, especially in low-risk patients. Here we report the 12-year results of a calcineurin-inhibitor-free regimen. One hundred and seventeen low-risk kidney recipients were prospectively randomized to maintenance therapy with either a combination of azathioprine and prednisone (group NoCsA, n= 58), or with cyclosporine, azathioprine, and prednisone (group CsA, n= 59). Both groups received induction therapy with anti-lymphocyte globulins (ALG). Twelve-year patient survival was 75% and 82.5% in the CsA and NoCsA groups, respectively [P= not significant (NS)]. Twelve-year graft survival was 59% and 56% (P= NS) in the CsA and NoCsA groups, respectively (NS). Transplant rejection rates were similar in both groups. Mean serum creatinine levels after 10 years were 161 and 136 μmol/I in the CsA and NoCsA groups, respectively. Rejection-free patients of the CsA group had poorer renal function (168 μmol/I) than those of the NoCsA group (121 μmol/I; P= 0.0060). We concluded that a 12-year graft survival of 56% and a graft half-life of 15 years can be achieved without the primary use of a calcineurin inhibitor in low-risk patients receiving ALG. Patients treated with CsA had poorer graft function at 12 years.

Actions of Immunosuppressor Drugs on the Development of an Experimental Ovarian Tumor1

Immunosuppression has been related to the incidence of tumor apparition, including endocrine tumors. The intrasplenic ovarian tumor (luteoma) is a typical benign endocrine tumor that develops under high gonadotropin stimulation and, from the immunological perspective, is located in a critical organ involved in immune response. To establish if immunosuppression could alter the development of this experimental tumor, the effects of cyclosporin A (CsA) and dexamethasone (Dex) were evaluated. After surgery, tumor-bearing and sham animals were kept without treatment for 4 weeks; thereafter, they were distributed into CsA (25 mg/kg), Dex (0.1 mg/kg), or vehicle (75:25 castor oil:ethanol) groups and were injected on alternate days for 50 days. Body weight was evaluated weekly. Animals were sacrificed after a jugular vein blood sample was obtained. Thymi were weighed. Tumors were measured and placed in formaline for histological studies. Serum luteinizing hormone (LH), follicle-stimulating hormone (FSH), prolactin (PRL), and estradiol were measured by radioimmunoassay. Hematological parameters were determined. CsA induced a significant decrease in survival rates both in tumor-bearing and sham animals (P < 0.01). Dex significantly impaired weight increase in both groups of animals. CsA induced a significant weight loss in sham animals, not observed in tumor-bearing animals. Dex induced thymus weight loss in both groups, whereas CsA induced thymus weight loss only in sham animals. Only Dex induced a decrease in lymphocyte number in both groups. CsA induced an increase in monocyte number only in sham animals. Treatments did not alter LH, FSH, or estradiol, whereas PRL was increased by CsA only in sham rats. Neither Dex nor CsA induced any significant variations in tumor volume, nor did they alter tumor histology. In addition, no visible metastases or alterations in other organs were observed. We conclude that, though immunological parameters were altered by the treatments, immunosuppressor drugs did not condition tumor development. In addition, tumors secrete one or more factor/s that counteract CsA effect.

Cyclosporine withdrawal and mycophenolate mofetil treatment effects on the progression of chronic cyclosporine nephrotoxicity

Recent clinical trials of mycophenolate mofetil (MMF) in chronic allograft nephropathy (CAN) demonstrated that the dose of cyclosporine A (CsA) is one of the critical factors in determining graft function in CAN, but the effect of MMF on chronic CsA nephropathy is undetermined. We undertook this study to evaluate the effect of MMF on CsA-induced nephrotoxicity in an animal model of chronic CsA nephropathy. In the first experiment, Sprague-Dawley rats on a low-salt diet were treated with CsA (7.5 mg/kg per day) for 10 weeks, or were treated with CsA for five weeks and then MMF (20 mg/kg per day) was administered five weeks later. In the second experiment, rats were treated with CsA for five weeks, and CsA was then withdrawn for five weeks with or without MMF treatment. Renal function, histologic parameters (tubulointerstitial fibrosis, arteriolopathy, ED-1-positive cells, renin-positive glomeruli, TUNEL-positive cells) and the expression of osteopontin and transforming growth factor (TGF)-beta1 mRNA expressions were compared for different treatment groups. CsA-treated rats showed decreased renal function and increased histologic parameters compared with the vehicle (VH)-treated rats. The addition of MMF did not improve these parameters compared with the CsA-treated rats. With CsA withdrawal, renal function and histologic parameters were significantly improved compared with the CsA-treated rats, and MMF treatment after CsA withdrawal further improved the histologic parameters. At the molecular level, the addition of MMF did not decrease the expression of osteopontin and transforming growth factor-beta1 (TGF-beta1) mRNAs, which were increased in the CsA-treated rat kidney. With CsA withdrawal, the expression of both mRNAs was significantly decreased compared with the CsA group, and a further decrease was observed with MMF treatment after CsA was withdrawn. The combined treatment of CsA and MMF does not prevent the development of chronic CsA nephrotoxicity, but MMF treatment after CsA withdrawal does improve chronic CsA nephrotoxicity. This finding provides a rationale for MMF treatment in chronic CsA nephrotoxicity.