Estudio sobre quimerismo reverso y el efecto de la movilización de células madre de médula ósea sobre la expresión de citoquinas en un modelo experimental de trasplante de tejidos compuestos / Study on reverse chimerism and the effect of mobilization of bone marrow stem cells on the expression of cytokines in an experimental model of composite tissue transplantation

Introducción y Objetivo: Los trasplantes de tejidos compuestos sufren rechazo crónico modulado entre otros factores por citoquinas. El quimerismo reverso o quimerismo del aloinjerto se define como la repoblación del tejido trasplantado por células circulantes del receptor. Plerixafor produce la movilización de células madre de médula ósea CD34+ hacia la sangre periférica. El objetivo del estudio fue conocer los mecanismos moleculares que intervienen en el rechazo crónico y el quimerismo reverso tras la administración de plerixafor. Material y Método: Realizamos 16 trasplantes osteomusculares heterotópicos de pata posterior entre ratas Brown-Norway hembra y Wistar Lewis macho bajo inmunosupresión subterapéutica con tacrólimus. Establecimos 2 grupos de estudio según la administración postoperatoria de plerixafor. Transcurridas 9 semanas estudiamos la expresión de citoquinas y el infiltrado leucocitario en distintas localizaciones musculares, así como el grado de rechazo crónico y porcentaje de quimerismo reverso en diferentes tejidos del aloinjerto. Resultados: Encontramos diferencias estadísticas en la expresión de factor estimulante de colonias granulocíticas e interleucina 12 a nivel de los tercios medio y distal del aloinjerto, y de interleucina 6 a nivel del tercio medio del aloinjerto. La intensidad del infiltrado leucocitario fue mayor en el grupo que no recibió plerixafor. Ambos grupos desarrollaron rechazo crónico y pudimos observar la aparición de quimerismo reverso. Sin embargo no observamos diferencias significativas en el infiltrado leucocitario, el rechazo crónico ni el quimerismo reverso. Conclusiones: La movilización de células madre de médula ósea CD34+ se asoció con una menor expresión de factor estimulante de colonias granulocíticas, interleucina 6 e interleucina 12. Estos hallazgos contribuyen a elucidar los mecanismos moleculares que podrían conducir a la creación de quimeras en el aloinjerto

Background and Objective: Vascularized composite allotransplantation suffer chronic rejection modulated by cytokines. Reverse chimerism or allograft chimerism is defined as the repopulation of the transplanted tissue by circulating cells of the recipient. Plerixafor mobilizes CD34+ bone marrow stem cells to the peripheral blood. The aim of the study was to know the molecular mechanisms involved in chronic rejection and reverse chimerism after plerixafor administration. Methods: Sixteen heterotopic osteomuscular hindlimb transplants were performed between female Brown-Norway rats as donors and male Wistar Lewis rats as recipients under subtherapeutic immunosuppression with tacrolimus. Two groups were established according to the postoperative administration of plerixafor. After 9 weeks, expression of cytokines and leukocyte infiltration were studied in different muscle locations, as well as the degree of chronic rejection and percentage of reverse chimerism in different tissues of the allograft. Results: Statistical differences were found in granulocyte colony stimulating factor and interleukin 12 expression at middle and distal allograft thirds, and interleukin 6 expression at middle allograft third. The intensity of leukocyte infiltration was greater in the group that did not receive plerixafor. Both groups developed chronic rejection and the appearance of reverse chimerism could be observed. However, no significant differences were observed in leukocyte infiltration, chronic rejection or reverse chimerism. Conclusions: The mobilization of CD34+ bone marrow stem cells was associated with a lower expression of granulocytic colony stimulating factor, interleukin 6 and interleukin 12. These findings contribute to elucidate the molecular mechanisms that could lead to the creation of chimeras in the allograft

Pilot study: use of contrast-enhanced ultrasonography in feline renal transplant recipients.

Objectives The study aims were to evaluate the feasibility of contrast-enhanced ultrasonography in feline renal transplant recipients in the post-transplantation period and to report findings in a case with presumptive delayed allograft ischemia. Methods Cats were imaged postoperatively using contrast harmonic ultrasonography after a bolus injection of a microbubble contrast medium. Time/mean pixel intensity curves were generated for cortical and medullary regions of interest in the renal allograft in each cat. Arrival time, time to peak, wash-in slope, wash-out slope, mean transit time and renal blood flow were calculated for each area. Results Within the renal cortices of cats without ureteral obstruction 1 day post-transplantation, arrival time was 2.0-6.3 s, time to peak was 3.6-30.1 s, wash-in rate was 2.45-41.14 mean pixel intensity (MPI)/s, wash-out rate was -2.01 to -0.47 MPI/s and blood flow was 6.1-106.5 MPI/s. Ratio mean transit time was 0.29-1.29. Typical cortical and medullary perfusion patterns were observed in these cats. In one cat with delayed graft ischemia followed by presumptive acute transplant rejection, dynamic and heterogeneous cortical and medullary perfusion was demonstrated. Decreases in cortical blood flow were paralleled by elevated serum creatinine. Conclusions and relevance Contrast-enhanced ultrasonography can be used in feline renal transplant recipients and provides both qualitative and quantitative data regarding renal allograft perfusion.

Corneal grafting for the treatment of full-thickness corneal defects in dogs: a review of 50 cases.

OBJECTIVE: To describe corneal grafting for the treatment of full-thickness corneal defects in dogs and to determine its effectiveness in preserving vision. METHODS: A review of the medical records of dogs that underwent corneal grafting following corneal perforations (≥3 mm) at the VTH-UAB from 2002 to 2012 was carried out. RESULTS: Fifty dogs of different breed, age and gender were included. Brachycephalic breeds were overrepresented (37/50;74%). All cases were unilateral, with euryblepharon being the most common concurrent ocular abnormality (20/50;40%). Full-thickness penetrating keratoplasties (FTPK) were performed in 21/50 eyes (42%) and lamellar keratoplasties (LK) in 29/50 eyes (58%). Frozen grafts (FroG) were used in 43/50 eyes (86%) and fresh homologous grafts (FreHoG) in 7/50 (14%). Of the former group, 26 were homologous (FroHoG:60%) and 17 heterologous (FroHeG:40%). A combination of topical medication (antibiotics, corticosteroids, cycloplegics, and 0.2% cyclosporine A) and systemic mycophenolate mofetil was administered. Median follow-up time was 200 days. Postsurgical complications included wound dehiscence (6/50;12%) and glaucoma (4/50;8%). Clinical signs of graft rejection were diagnosed as follows: FroHoG (13/26;50%), FroHeG (11/17;65%), FreHoG (4/7;57%), FTPK (12/21;57%), and LK (16/29;55%). Medical treatment successfully controlled graft rejection in 11/28 eyes (39%). Good anatomical outcome was achieved in 86% (43/50), of which 95% (41/43) were visual at last examination, with moderate opacification to complete transparency of the graft present in 48.2%. CONCLUSIONS: Corneal grafting is an effective surgical treatment for full-thickness corneal defects in dogs. If graft rejection is present, additional medical or surgical therapy may be necessary, achieving a highly satisfactory visual outcome.

Liver transplantation in the mouse: Insights into liver immunobiology, tissue injury, and allograft tolerance.

The surgically demanding mouse orthotopic liver transplant model was first described in 1991. It has proved to be a powerful research tool for the investigation of liver biology, tissue injury, the regulation of alloimmunity and tolerance induction, and the pathogenesis of specific liver diseases. Liver transplantation in mice has unique advantages over transplantation of the liver in larger species, such as the rat or pig, because the mouse genome is well characterized and there is much greater availability of both genetically modified animals and research reagents. Liver transplant experiments using various transgenic or gene knockout mice have provided valuable mechanistic insights into the immunobiology and pathobiology of the liver and the regulation of graft rejection and tolerance over the past 25 years. The molecular pathways identified in the regulation of tissue injury and promotion of liver transplant tolerance provide new potential targets for therapeutic intervention to control adverse inflammatory responses/immune-mediated events in the hepatic environment and systemically. In conclusion, orthotopic liver transplantation in the mouse is a valuable model for gaining improved insights into liver biology, immunopathology, and allograft tolerance that may result in therapeutic innovation in the liver and in the treatment of other diseases.

Canine DLA-79 gene: an improved typing method, identification of new alleles and its role in graft rejection and graft-versus-host disease.

Developing a preclinical canine model that predicts outcomes for hematopoietic cell transplantation in humans requires a model that mimics the degree of matching between human donor and recipient major histocompatibility complex (MHC) genes. The polymorphic class I and class II genes in mammals are typically located in a single chromosome as part of the MHC complex. However, a divergent class I gene in dogs, designated dog leukocyte antigen-79 (DLA-79), is located on chromosome 18 while other MHC genes are on chromosome 12. This gene is not taken into account while DLA matching for transplantation. Though divergent, this gene shares significant similarity in sequence and exon-intron architecture with other class I genes, and is transcribed. Little is known about the polymorphisms of DLA-79 and their potential role in transplantation. This study was aimed at exploring the reason for high rate of rejection seen in DLA-matched dogs given reduced intensity conditioning, in particular, the possibility that DLA-79 allele mismatches may be the cause. We found that about 82% of 407 dogs typed were homozygous for a single, reference allele. Owing to the high prevalence of a single allele, 87 of the 108 dogs (∼80%) transplanted were matched for DLA-79 with their donor. In conclusion, we have developed an efficient method to type alleles of a divergent MHC gene in dogs and identified two new alleles. We did not find any statistical correlation between DLA-79 allele disparity and graft rejection or graft-versus-host disease, among our transplant dogs.

Oral immunosuppressive medication for growing pigs in transplantation studies.

Immunosuppressive (IS) medication is needed to avoid graft rejection in porcine transplantation models. An ideal IS therapy should have no side-effects, but increased susceptibility to infections, disturbed intestinal microflora and toxic effects on organs and tissues are commonly reported. The aim of the present study was to design an IS protocol with tacrolimus and mycophenolic acid to be used for maintenance therapy in the post-transplant period. An eligible whole blood trough value for tacrolimus was 5-15 µg/L. Conventional specific pathogen-free pigs were fitted with an indwelling catheter under general anaesthesia, and after the acclimatization period three groups were formed: group A (n= 4) received 0.15 mg/kg body weight (BW) twice daily tacrolimus and 500 mg twice daily mycophenolic acid; group B (n= 4) received 0.3 mg/kg BW twice daily tacrolimus and 500 mg twice daily mycophenolic acid; group C (n= 2) did not receive any medication. Daily clinical examinations and analyses of blood concentrations of tacrolimus and glucose were performed. Total and differential white blood cell counts, enzyme activities, bilirubin and electrolyte concentrations were measured every fourth day. At the end of the experiment, the pigs were killed with an overdose of pentobarbital intravenously and a necropsy was performed immediately. All animals seemed to tolerate the IS treatment well. No alterations in their clinical state of health were observed throughout the study and daily weight gain was similar for the three groups. The necropsy did not reveal any pathological findings related to medication. The study showed that 0.25 mg/kg BW twice daily tacrolimus and 500 mg twice daily mycophenolic acid would be an appropriate maintenance dosage for conventional pigs.

Outcome after renal transplantation in 26 dogs.

OBJECTIVES: To evaluate clinical outcome in dogs after renal transplantation and determine predictors of outcome. STUDY DESIGN: Retrospective case series. ANIMALS: Dogs (n = 26) that had renal allograft transplantation. METHODS: Medical records (1994-2006) of 26 consecutive cases of dogs that had kidney transplantation were reviewed. History, signalment, pre- and postoperative clinicopathologic and monitoring variables, postoperative complications, immunosuppressive therapy, and survival were recorded. RESULTS: Median survival was 24 days (range, 0.5 to 4014 days) with a probability of survival to 15 days of 50% and the 100-day survival probability was 36%. Cause of death was attributed to thromboembolic disease in 8 dogs, infection in 6 dogs, and rejection in 1 dog. The only factor significantly associated with an increased likelihood of death was increasing age at time of surgery (P = .024). CONCLUSIONS: Canine renal transplantation in clinical patients is associated with a high morbidity and mortality and increasing recipient age has a negative association with outcome. Thromboembolic complications are a major cause of death in the immediate postoperative period and effective anticoagulation protocols may greatly improve survival in the future.

Effect of cyclosporine, dexamethasone, and human CTLA4-Ig on production of cytokines in lymphocytes of clinically normal cats and cats undergoing renal transplantation.

OBJECTIVE: To evaluate effects of cyclosporine, dexamethasone, and the immunosuppressive agent human CTLA4-Ig on cytokine production by feline lymphocytes in vitro and to assess patterns of cytokine production for 5 immunosuppressed renal transplant recipient cats. ANIMALS: 21 clinically normal cats and 5 immunosupressed renal transplant recipient cats. PROCEDURES: Peripheral blood mononuclear cells were isolated from clinically normal cats and stimulated with concanavalin A (Con A; 10 µg/mL) alone or Con A with cyclosporine (0.05 µg/mL), dexamethasone (1 × 10(-7)M), a combination of cyclosporine-dexamethasone, or human CTLA4-Ig (10 g/mL). Cells from transplant recipients were stimulated with Con A alone. An ELISA was performed to measure production of interferon (IFN)-γ, granulocyte macrophage-colony stimulating factor (GM-CSF), interleukin (IL)-2, IL-4, and IL-10. Proliferation of CD4+ and CD8+T cells from immunosuppressed cats were also evaluated. Pairwise comparisons were performed via a Wilcoxon signed rank test or Wilcoxon rank sum test. RESULTS: Cyclosporine, dexamethasone, cyclosporine-dexamethasone combination, and CTLA4-Ig caused a significant decrease in IL-2, IFN-γ, and GM-CSF production. Cyclosporine and cyclosporine-dexamethasone, but not human CTLA4-Ig, caused a significant decrease in IL-10 production. High basal concentrations of IL-2 and IL-10 were identified in transplant recipients, and IL-10 was significantly increased in stimulated cultures. In immunosuppressed cats, there was a decrease in frequency of responders and proliferative capacity of CD4+ and CD8+T cells. CONCLUSIONS AND CLINICAL RELEVANCE: CTLA4-Ig successfully inhibited proinflammatory cytokines while sparing cytokines critical for allograft tolerance. These data may be useful for developing better strategies to prevent rejection while sparing other immune functions.

Effects of fenbendazole on routine immune response parameters of BALB/c mice.

Fenbendazole (FBZ) is an anthelmintic drug widely used to treat and prevent pinworm outbreaks in laboratory rodents. Although data in nonrodent species indicate possible effects of fenbendazole on the bone marrow and lymphocyte proliferation and function, little has been reported regarding possible effects on the rodent immune system. The purpose of the current study was to determine the effects of a therapeutic regimen of FBZ on immune parameters in BALB/c mice. Both 9-wk on-off and 5-wk continuous medicated feed protocols were assessed. No significant differences between normal and FBZ diet treated mice were observed in the following parameters: complete blood count, blood chemistry, quantitation of major T and B cell markers in spleen, quantitation of T cell markers in the thymus, spleen cell proliferation to T and B cell mitogens, bone marrow colony-forming cell assays, skin graft rejection, and primary and secondary humoral immune responses. These data indicate that FBZ treatment does not affect many standard broad measures of immune function.

Gingival overgrowth in dogs associated with clinically relevant cyclosporine blood levels: observations in a canine renal transplantation model.

OBJECTIVE: To investigate the development of gingival hyperplasia in dogs after renal transplantation and administration of microemulsified cyclosporine A (MCsA). STUDY DESIGN: Experimental study. ANIMALS: Healthy adult mongrel dogs (n=5). METHODS: As part of study on renal transplantation, dogs administered MCsA (20 mg/kg/day), azathioprine, and prednisolone to prevent graft rejection were monitored for development of gingival changes. Prednisolone was discontinued after 3 months. MCsA dose was adjusted to maintain whole blood trough concentration of 400-700 ng/mL. Gingival change was evaluated by weekly examination and photodocumentation, and gingival biopsy for histopathology was performed at 28 weeks. RESULTS: One dog was lost because of acute graft rejection. Gingival hyperplasia developed in 3 of 4 dogs. The earliest gingival changes occurred in the interdental papillae at 20 weeks after transplantation. On histopathology, the underlying connective tissue was thickened and contained increase numbers of fibroblasts and inflammatory infiltrates. CONCLUSIONS: Long-term immunosuppression with an MCsA-based treatment likely induces substantial gingival hyperplasia when therapeutic, immunosuppressive blood levels of MCsA were maintained for 32 weeks. CLINICAL RELEVANCE: MCsA is used for immune-mediated diseases and preventing rejection after transplant in dogs. MCsA blood levels, and gingival hyperplasia should be monitored by routine examination of the interdental papilla in dogs administered MCsA for long periods.

Corneal transplantation for inflammatory keratopathies in the horse: visual outcome in 206 cases (1993-2007).

OBJECTIVE: To evaluate the visual outcome of three techniques of corneal transplantation surgery in treating severe inflammatory keratopathies in the horse. DESIGN: Retrospective medical records study. ANIMALS STUDIED: Medical records of 206 horses that received corneal transplantation surgery at the University of Florida Veterinary Medical Center from 1993 to 2007 were reviewed. PROCEDURE: Data collected from the medical records included signalment, types of ocular lesions, type of transplant surgery performed, length of follow-up, complications, and visual outcomes. RESULTS: Full thickness penetrating keratoplasty (PK) was performed in 86 horses for melting ulcers, iris prolapse/descemetoceles, and medically nonresponsive full thickness stromal abscesses (SA). Posterior lamellar keratoplasty (PLK) and deep lamellar endothelial keratoplasty (DLEK) are split thickness penetrating keratoplasties that were utilized for medically nonresponsive deep stromal abscesses (DSA) in 54 and 66 eyes, respectively. The most common postoperative surgical complication was graft rejection and varying degrees of graft opacification. Wound dehiscence and aqueous humor leakage was also a common postoperative problem. A positive visual outcome was achieved for PK, PLK, and DLEK in 77.9%, 98.1%, and 89.4%, respectively. CONCLUSIONS: Corneal transplantation is a tectonically viable surgery in the horse with an overall success rate of 88.5% in maintaining vision when treating vascularized and infected corneal disease in the horse.

Ultrasonographic determination of resistive index and graft size for evaluating clinical feline renal allografts.

Ultrasonographic examination is a commonly employed technique for postoperative renal allograft evaluation after transplantation. Allograft size and resistive index (RI) are two objective ultrasonographic measures that may help establish a diagnosis and direct postoperative management for grafts with suboptimal function but their diagnostic efficacy has not been evaluated in clinical veterinary patients. Results of 69 feline renal transplant ultrasonographic examinations and RI determinations were studied. Based on clinical parameters at the time of the ultrasonographic examination, patients were grouped into six clinical/functional categories including evaluations of clinically normal grafts, delayed graft function, ureteral obstruction, uroabdomen, graft thrombosis, and rejection. RI, graft size (length, cross-sectional area, and volume), cyclosporine A whole blood trough concentration, Doppler blood pressure, creatinine concentration, and days from transplantation were compared between these categories and associations with each other were examined. RI was of little value in differentiating among the clinical categories with the exception of graft thrombosis. Graft volume and time from transplantation were significantly greater in grafts with signs of rejection and ureteral obstruction compared to clinically normal ultrasound examinations. Graft volume, cross-sectional area and length were generally associated. Cyclosporine A blood concentrations was associated with RI in both the pooled data and in the delayed graft function category. These results indicate RI should be used only as part of a larger clinical picture and in light of other factors including cyclosporine A concentration and the timing of the study relative to the implantation surgery for the diagnosis of postoperative transplantation complications. Graft volume may provide a more sensitive, albeit, nonspecific, indicator of allograft dysfunction.

Survival, complications, and analysis of risk factors after renal transplantation in cats.

OBJECTIVE: To report survival, complications, and analyze risk factors for survival after renal transplantation (RTr) and cyclosporine-A based immunosuppression in cats. STUDY DESIGN: Historical cohort. ANIMALS: Cats (n=60). METHODS: Data were obtained from medical records of cats that had RTr. Influence of various perioperative factors on survival and complications was evaluated. Occurrence of postoperative hypertension (HT), seizures, infection, acute allograft rejection (AR), congestive heart failure (CHF), and delayed graft function (DGF) was evaluated. RESULTS: Survival to discharge after RTr was 77.5%. Estimated median overall survival time was 613 days; 6 month and 3 year overall survival proportions were 65% and 40%, respectively. Age, weight, and blood pressure influenced overall survival. Increased preoperative creatinine concentration, blood urea nitrogen, postoperative creatinine concentration, left ventricular wall thickness, and reduced creatinine reduction ratio influenced survival until discharge. HT was identified in 9/30 (30%) cats; however, no risk factors were identified, nor was HT related to seizures. AR was identified in 8/62 (13%) grafts. Infection, predominantly bacterial, developed in 22/60 (37%) cats. CHF occurred in 7/60 (12%) cats before discharge. Cats experiencing CHF were younger, had an increased incidence of heart murmurs, and poor initial graft function. DGF was identified in 5 cats and seizures in 2 cats. CONCLUSIONS: RTr affords cats with CRF long survival times. Older cats and cats with severe azotemia, HT, and cardiovascular disease may have increased mortality after RTr. Complications after RTr were common. CLINICAL RELEVANCE: Clinicians should be aware of these risk factors when recommending feline RTr.

Aspectos clínicos e radiográficos do pericárdio bovino como substituto do ligamento cruzado cranial de cães / Clinical and radiographic aspects of the bovine pericardium as a substitute of the canine cranial cruciate ligament

Avaliaram-se os aspectos clínicos e radiográficos do enxerto de pericárdio bovino, preservado em glicerina, como substituto do ligamento cruzado cranial. Quinze cães machos, sem raça definida, pesando entre 17,4 e 31,6 kg, foram submetidos à ruptura experimental do ligamento cruzado cranial e à substituição por pericárdio, via videoartroscopia. Os cães foram divididos em três grupos de cinco e avaliados aos 30, 90 e 120 dias. O membro operado foi imobilizado por duas semanas e procederam-se avaliações clínicas semanais. Radiografias foram feitas mensalmente e foram realizadas colheita de líquido sinovial nos tempos descritos. Clinicamente, os cães mostraram claudicação acentuada a moderada, hipotrofia muscular no membro operado e acentuado deslocamento cranial da tíbia em relação ao fêmur. As alterações degenerativas foram observadas nas radiografias. Observou-se instabilidade acentuada em todos os animais. O líquido sinovial tinha características de inflamação. Concluiu-se que o enxerto rompeu precocemente, provocou reação inflamatória persistente e fenômenos de rejeição, não sendo, portanto, recomendado para substituição do ligamento cruzado cranial de cães

The clinical and radiographic aspects of the bovine pericardium preserved in glicerin, were evaluated as a substitute for canine cranial cruciate ligament. Fifteen male mongrel dogs weighing between 17.4 and 31.6kg had the ligament experimentally ruptured and the stifle joint stabilized by an arthroscopical technique with bovine pericardium as a graft. The dogs were divided into three groups of five animals each. They were evaluated at 30, 90 and 120 days. The operated limb was imobillized for two weeks and clinical examination was performed weekly. Radiographs were taken monthly and the sinovial fluid was collect at 30, 90 and 120 days. Clinically, dogs presented high to moderate lameness, muscle hipotrophy in the operated limb and accentuated cranial drawer movement. Degenerative disease was detected in radiography. All dogs showed total rupture of the graft. Sinovial fluid analysis showed characteristics of inflammation. It can be concluded that pericardium graft failed prematurely, incited persistent inflammatory reaction and rejection phenomena. Thus, it cannot be recommended as a xenograft for cranial cruciate ligament replacement

Use of capecitabine to prevent acute renal allograft rejection in dog erythrocyte antigen-mismatched mongrel dogs.

OBJECTIVE: To assess efficacy and toxicity of a capecitabine (CAP)-based regimen for preventing rejection of renal allografts in dog erythrocyte antigen (DEA)-mismatched mongrel dogs. STUDY DESIGN: Prospective, pilot study. ANIMALS: Eight healthy, unrelated, DEA mismatched, adult mongrel dogs. METHODS: All dogs received CAP, starting at 50 mg/m2 PO b.i.d. 4 days preoperatively, increasing to 200 mg/m2 PO b.i.d. by the day of surgery. All dogs received cyclosporine-A (CsA) and prednisolone starting 2 days preoperatively. Standard heterotopic renal transplantation with native nephrectomy was performed. After 90 days, surviving dogs were euthanatized and histopathologic examination was performed. RESULTS: Two of 8 dogs developed acute neurotoxicity leading to death or euthanasia within 5 days of surgery. For the 6 remaining dogs, there were no statistically significant changes in complete blood count or serum biochemical values. No opportunistic infections developed during the study period. Five of 6 dogs had no to minimal evidence of graft rejection. Two of 6 dogs developed superficial and pigmentary keratitis. Significant histopathologic findings in all dogs included mild lymphoplasmacytic gastroenteritis, steroid hepatopathy, and corneal epithelial thinning. One dog had moderate interstitial nephritis and pyelitis. CONCLUSIONS: In this experimental model, a CAP-CsA-prednisolone immunosuppressive regimen was effective in preventing rejection of allografts in DEA-mismatched dogs. Severe, unpredictable neurotoxicity and variable ocular toxicity significantly limit clinical applications at this time. CLINICAL RELEVANCE: A CAP-CsA-prednisolone protocol is an effective, oral immunosuppressive regimen for prevention of allograft rejection in DEA-mismatched mongrel dogs. For clinical application, identification of patients susceptible to toxic side effects would be necessary.

Results of clinical renal transplantation in 15 dogs using triple drug immunosuppressive therapy.

OBJECTIVE: To evaluate outcome of renal transplantation in dogs administered cyclosporine, azathioprine, and prednisolone immunosuppression. STUDY DESIGN: Prospective clinical study. ANIMALS: Fifteen dogs with chronic renal failure. RESULTS: Nine dogs died within 1 month of surgery; 5 died from complications associated with generalized thromboembolism. Three dogs survived for 6-25 months. Three dogs alive at the time of this report have survived 22-48 months; however, all 3 dogs have had bacterial infections that responded to antibiotic therapy. There was no biochemical evidence of acute allograft rejection in any dog. Perioperative use of enoxaparin may have prevented thromboembolism in 5 dogs. CONCLUSIONS: Triple drug immunosuppressive therapy used in this study prevented acute renal allograft rejection in 6 dogs that survived >4 weeks; however, immunosuppression was excessive, resulting in an unacceptable frequency of infection and other drug-related complications. Perioperative anticoagulation therapy seem to be warranted. CLINICAL RELEVANCE: Survival time and quality of life for this group of dogs was poor; however, there was no evidence of acute rejection in the dogs surviving >4 weeks. This protocol should only be used if the degree of immunosuppression is reduced, and early evidence of allograft rejection is monitored by renal biopsy or markers of lymphocyte activation.

Renal allograft histopathology in dog leukocyte antigen mismatched dogs after renal transplantation.

OBJECTIVE: To evaluate allograft histopathology in dog leukocyte antigen (DLA)-mismatched dogs undergoing renal transplantation, with transient immunosuppression. STUDY DESIGN: Prospective study. ANIMALS: Ten healthy adult mongrel dogs. METHODS: Reciprocal renal transplantation and bilateral nephrectomy were performed. Immune conditioning consisted of nonmyeloablative (200 cGy), total body irradiation (TBI), bone marrow transplantation (BMT; 7 dogs), cyclosporine (CSA; 15 mg/kg every 12 hours), mycophenolate mofetil (MMF; 10 mg/kg every 12 hours) and intermittent prednisone (1 mg/kg every 12-24 hours). Biopsies were collected at transplantation, during full immunosuppression (44-90 days), and once medications were reduced or discontinued (228-580 days). Biopsies were evaluated for interstitial, tubular, vascular, and glomerular lesions. Blood urea nitrogen, creatinine, serum CSA concentrations, and clinical score were determined at each biopsy. RESULTS: Seven dogs survived >200 days (mean, 380 days). Transient CSA toxicity was suspected in 6 dogs. Lymphocytic, plasmacytic interstitial inflammation, and tubulitis progressed when immunosuppressive medications were decreased. All 7 dogs had histologic lesions consistent with some degree of allograft rejection at study end. CONCLUSION: Nonmyeloablative TBI, BMT, and short-term immunosuppression with CSA, MMF, and prednisone allowed renal allograft function and dog survival for >200 days. It appears unlikely that total drug withdrawal will be possible in unrelated DLA-mismatched dogs using this protocol. CLINICAL RELEVANCE: Transient immunosuppression with MMF, CSA, and prednisone along with BMT and nonmyeloablative TBI may make kidney transplantation a clinical reality for treatment of kidney failure in dogs. Initiating both MMF and CSA at lower dosages may potentially eliminate early renal allograft injury.

Use of capecitabine after renal allograft transplantation in dog erythrocyte antigen-matched dogs.

OBJECTIVES: To investigate the use of a capecitabine (CAP)-based regimen after renal transplantation in dogs. STUDY DESIGN: Prospective, pilot study. ANIMALS: Healthy, unrelated, dog erythrocyte antigen (DEA)-matched, adult beagles. METHOD: Standard heterotopic renal transplantation with native nephrectomy was performed in 7 dogs. Dogs received oral, twice daily, CAP (250 mg/m2), cyclosporine-A (CsA) (4 mg/kg), ketoconazole (5 mg/kg), and prednisolone (0.25 mg/kg). After 90 days the surviving dogs were euthanatized and complete necropsy was performed. RESULTS: Seven transplants were performed. All dogs survived surgery. Six dogs had acute neurotoxicity, which resulted in death or euthanasia of 2 dogs within 2 days of surgery. In the remaining dogs, toxicity resolved rapidly with cessation of drug administration. Thereafter, modification of the regimen minimized toxicity. The 5 remaining dogs survived to study end; 4 dogs had no evidence of graft rejection. Necropsy examination was mostly unremarkable in all dogs. There were no major changes in CBC or biochemical values, except for a significant increase in serum calcium. CONCLUSIONS: CAP appeared well tolerated in most dogs. Toxicity occurred but abated with modification of the drug regimen. Efficacy for postoperative immunosuppression cannot be determined by this study, although results are promising. CLINICAL RELEVANCE: CAP-CsA-prednisolone is an effective, oral immunosuppressive regimen for prevention of acute allograft rejection in DEA-matched beagles. Further studies on dose, toxicity, and efficacy compared with current immunosuppressive regimens are needed before use in clinical practice.

Evaluation of the prevalence of infections in cats after renal transplantation: 169 cases (1987-2003).

OBJECTIVE: To determine the prevalence of infections developing postoperatively, document the contribution of infection to increased risk of death, and identify risk factors associated with the development of infectious complications in cats after renal transplantation. DESIGN: Retrospective study. ANIMALS: 169 cats that received renal allograft transplants. PROCEDURES: Medical records of cats receiving renal transplants at the University of California from January 1987 through December 2003 were reviewed. RESULTS: 47 infections developed in 43 of 169 cats. Bacterial infections were most common (25/47 cats), followed by viral (13/47), fungal (6/47), and protozoal (3/47) infections. The median duration from transplant surgery to development of infection was 2.5 months. Infection was the second most common cause of death after acute rejection of the transplant, accounting for 14% of deaths overall. Cats with concurrent diabetes mellitus had a significantly increased risk of developing an infection after renal transplantation. Sex, increasing age, concurrent neoplasia, and previous treatment for transplant rejection were not associated with development of infection. CONCLUSIONS AND CLINICAL RELEVANCE: Infection was a common complication and an important cause of death or euthanasia in cats after renal transplantation. Development of diabetes mellitus after transplantation significantly increased the risk of infection.

Evaluation of oxidative stress markers for the early diagnosis of allograft rejection in feline renal allotransplant recipients with normal renal function.

The purpose of this study was to identify oxidative damage to renal allografts during graft rejection by evaluating changes in oxidative markers and plasma lactate levels in feline renal allotransplant recipients. Heterotopic renal allotransplantations were performed between 8 adult feline cross-matched donors. Following 14 d of immunosuppression, the drugs were discontinued to allow allograft rejection. Baseline and serial postoperative evaluations of serum creatinine, plasma lactate, plasma thiobarbituate reactive substances (TBARS), plasma creatol, urine creatol, and renal sonographic cross-sectional area were performed. When sonographic evaluation revealed the absence of blood flow to the allograft, the rejected kidney was nephrectomized and evaluated histopathologically. Allograft rejection occurred in all cats by day 26. A significant elevation in body temperature occurred during the rejection period. No significant change was observed between any of the time periods for plasma TBARS, creatol, or urine creatol. There was a significant decrease in plasma lactate levels throughout the study. Markers of oxidative stress from venous blood did not reflect renal allograft rejection in cats with a normally functioning native kidney. Renal allograft rejection may be associated with significant increases in body temperature and warrants further investigation.