OBJECTIVES AND DESIGN: At the present time there are two waiting list for patients with vascular prosthetic infection indicated for arterial transplantation in the Czech Republic. The inclusion of each patient for cold-stored or cryopreserved arterial transplantation is the preference of indicating surgeon. In this experimental work we studied the immunogenicity of rat aortal allografts treated by our new clinical cryopreservation/slow thawing protocol. MATERIAL AND METHODS: Brown-Norway (BN) (N = 6, 203-217 g) or Lewis (LEW) (N = 6, 248-254 g) abdominal aortal grafts treated in accordance with our new clinical cryopreservation/slow thawing protocol were orthotopically transplanted to Lewis recipients (N = 12, 191-245 g). Aortal wall histology and infiltration by recipient immune cells, as well as donor specific anti MHC class I and II antibodies in recipient serum were studied in both isografts and allografts on day 30 postransplant. Core data of cryopreserved allografts were compared to our previous data of cold-stored aortal allografts treated in accordance with our clinical cold-storage protocol. RESULTS: Cryopreserved allografts showed regular morphology of aortal wall with clear differentiation of all three basic anatomical layers on day 30 postransplant. Intimal layer showed no hyperplasia, luminal surface was covered by endothelial cells. No statistical difference was observed in tunica media thickness between isografts and allografts. The medial layer showed no necrosis, shrinkage or immunoglobuline G deposition in any experimental group. The adventitial infiltration by immune cells was significantly higher (P<0.05) in allografts. Cryopreserved allografts showed significant lower activation of both cell- and antibody mediated immunity compared to historical data of cold-stored allografts. CONCLUSION: Aortal wall histology of rat allografts treated by our new standardized clinical cryopreservation/slow thawing protocol was comparable to that of the cryopreserved isografts on day 30 posttranspant. The immunogenicity of cryopreserved aortal allografts was significantly lower compared to that of cold-stored aortal allografts.
- MeSH
- alografty fyziologie MeSH
- aorta transplantace MeSH
- arterie transplantace MeSH
- homologní transplantace metody MeSH
- kryoprezervace metody normy MeSH
- krysa rodu rattus MeSH
- modely u zvířat MeSH
- potkani inbrední BN MeSH
- potkani inbrední LEW MeSH
- rejekce štěpu imunologie MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- Geografické názvy
- Česká republika MeSH
OBJECTIVES AND DESIGN: The aim of our study was to simulate in rats all aspects and techniques used in our new clinical program of cryopreserved alloarterial transplantation and investigate the influence of two immunosuppressive protocols with tacrolimus on acute rejection of these allografts. MATERIALS AND METHODS: Cryopreserved abdominal aortic grafts were transplanted between Brown-Norway and Lewis rats. Tacrolimus (0.2 mg/kg daily) was administered from day 1 to day 30 (TAC1) or from day 7 to day 30 (TAC7), respectively. No immunosuppressed isogeneic (ISO) and allogeneic (ALO) rats combination served as control. Aortal wall infiltration by immunocompetent cells (MHC II+ cells of recipient origin) was studied on day 30 after transplantation. Flow cytometry was used for the analysis of day 30 sera for the presence of donor specific anti-MHC class I and II antibodies. RESULTS: The aortal allografts in both immunosuppressed groups showed regular morphology of aortal wall with no depositions of immunoglobulin G on day 30. The adventitial infiltration of non-immunosuppressed aortal allografts by MHC class II positive cells of recipient origin was significantly higher (ALO 20.7±6.7 cells, P<0.001) compared to both immunosuppressed groups (TAC1 5.9±5.5 cells, TAC7 6.1±5.1 cells). Day 30 sera from the allogeneic non-immunosuppressed animals decreased significantly the binding of fluorescence-labelled MHC class I (46.9±19.4%) and class II (65.8±11.9%) antibody to donors spleen cells compared with day 30 sera from both immunosuppressed groups (TAC1, anti-MHC class I 102.4±4.2%, p < 0.001, anti-MHC class II 102.6±6.0%), (TAC7, anti-MHC class I 79.9±3.3%, p < 0.001, anti-MHC class II 80.9±2.7%). CONCLUSION: Both immunosuppressed protocols with tacrolimus (administration from day 1 or from day 7 following transplantation) were able to suppress acute cell- and antibody-mediated rejection of cryopreserved abdominal aortic allografts processed in accordance with our new standardized clinical protocol.
- MeSH
- aorta fyziologie transplantace MeSH
- cévní protézy * MeSH
- homologní transplantace MeSH
- imunosupresiva aplikace a dávkování MeSH
- imunosupresivní léčba MeSH
- kryoprezervace * MeSH
- krysa rodu rattus MeSH
- potkani inbrední BN MeSH
- potkani inbrední LEW MeSH
- přežívání štěpu MeSH
- průtoková cytometrie MeSH
- rejekce štěpu imunologie MeSH
- rozvrh dávkování léků MeSH
- takrolimus aplikace a dávkování MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Geografické názvy
- Česká republika MeSH
BACKGROUND: The aim of our experimental work was to assess morphological changes of arterial wall that arise during different thawing protocols of a cryopreserved human aortic root allograft (CHARA) arterial wall. METHODS: The experiment was performed on CHARAs. Two thawing protocols were tested: 1, CHARAs were thawed at a room temperature at +23°C; 2, CHARAs were placed directly into a water bath at +37°C. MICROSCOPIC SAMPLES PREPARATION: After fixation, all samples were washed in distilled water for 5 min, and dehydrated in a graded ethanol series (70, 85, 95, and 100%) for 5 min at each level. The tissue samples were then immersed in 100% hexamethyldisilazane for 10 minutes and air dried in an exhaust hood at room temperature. Processed samples were mounted on stainless steel stubs, coated with gold. RESULTS: Thawing protocol 1: All 6 (100%) samples showed loss of the endothelium and damage to the subendothelial layers with randomly dispersed circular defects and micro-fractures without smooth muscle cells contractions in the tunica media. Thawing protocol 2: All 6 (100%) samples showed loss of endothelium from the luminal surface, longitudinal corrugations in the direction of blood flow caused by smooth muscle cells contractions in the tunica media with frequent fractures in the subendothelial layer. CONCLUSION: All the samples thawed at the room temperature showed smaller structural damage to the CHARA arterial wall with no smooth muscle cell contraction in tunica media when compared to the samples thawed in a water bath.
- MeSH
- alografty * patologie MeSH
- aorta patologie transplantace MeSH
- aortální chlopeň patologie transplantace MeSH
- chirurgická náhrada chlopně škodlivé účinky metody MeSH
- dárci tkání MeSH
- dospělí MeSH
- homologní transplantace škodlivé účinky metody MeSH
- kryoprezervace metody MeSH
- lidé středního věku MeSH
- lidé MeSH
- srdeční chlopně umělé škodlivé účinky MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Arterial allografts are used as vascular conduits in the treatment of prosthetic graft infection. Immunosuppression decreases their rupture risk rate. However, immunosuppression can be unprofitable in florid infection. Previously, we confirmed inhibition of cell-mediated destruction of rat aortic grafts by delayed use of tacrolimus. In this work, we studied the influence of this protocol on the antibody-mediated rejection. MATERIAL AND METHODS: Flow cytometry was used for the retrospective analysis of day 0, 14, and 30 sera obtained from Lewis rat recipients of isogeneic fresh infrarenal aortic grafts (group A) or Brown-Norway rat aortic grafts (group B,C,D) for the presence of donor-specific anti-MHC class I and II antibodies. Tacrolimus in daily dose of 0.2 mg/kg was administered from day 1 to day 30 (group C) or from day 7 to day 30 (group D). RESULTS: Inhibition of fluorescence-labeled anti-BN MHC class I and MHC class II antibodies binding to BN-splenocytes was observed only by day 14 and day 30 sera of allogeneic non-immunosuppressed Lewis rats (group B). The day 30 sera significantly decreased anti-MHC I (42±3%) and anti-MHC II antibody binding (56±3%) compared to day 0 (76±9%, p=0.005 and 79±5%, p=0.003, respectively). Deposition of immunoglobulins G into the tunica media was observed only in non-immunosuppressed aortic allografts on day 30. CONCLUSIONS: Fresh aortic allografts induce donor-specific anti-MHC class I and anti-MHC class II antibody production. Delayed administration of tacrolimus completely suppressed antibody production and antibody-mediated destruction of aortic allografts.
- MeSH
- aorta transplantace MeSH
- B-lymfocyty imunologie MeSH
- CD8-pozitivní T-lymfocyty imunologie MeSH
- homologní transplantace MeSH
- imunosupresiva farmakologie MeSH
- isoprotilátky imunologie MeSH
- krysa rodu rattus MeSH
- MHC antigeny I. třídy imunologie MeSH
- MHC antigeny II. třídy imunologie MeSH
- potkani inbrední BN MeSH
- potkani inbrední LEW MeSH
- rejekce štěpu farmakoterapie imunologie MeSH
- slezina cytologie imunologie MeSH
- takrolimus farmakologie MeSH
- transplantace cév metody MeSH
- vztah mezi dávkou a účinkem léčiva MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
Arterial allografts are used now-a-days as a modality in the treatment of vascular prosthesis infections. Prolonged administration of immunosuppressive drugs seemed to be essential for long-time patency rates of alloarterial vascular reconstructions. Nevertheless, the use of immunosuppressives if there exist an acute infection is controversial. The experimental work described herein studied effects of a delayed low-dose FK 506 administration on the development of acute rejection changes 30 days after aortal transplantation in rats. The response of the recipient's immune system to aortal wall antigens of the donor in the field of no immunosuppression resulted in an intimal proliferation and its infiltration by immunocompetent cells of the recipient, necrosis of medial smooth muscle cells, including deposition of immunoglobulins, and a massive adventitial infiltration of CD4 and CD8 positive cells. On the other hand, all the principal histological signs of rejection listed above were suppressed by FK 506 administration, no matter whether the immunosuppressive was administered on day 0 or day 7 after the transplantation.
- MeSH
- aorta patologie transplantace MeSH
- CD4-pozitivní T-lymfocyty cytologie MeSH
- CD8-pozitivní T-lymfocyty cytologie MeSH
- financování organizované MeSH
- imunohistochemie MeSH
- imunosupresiva aplikace a dávkování MeSH
- krysa rodu rattus MeSH
- potkani inbrední BN MeSH
- potkani inbrední LEW MeSH
- rejekce štěpu farmakoterapie patologie MeSH
- takrolimus aplikace a dávkování MeSH
- tunica intima patologie účinky léků MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- mužské pohlaví MeSH
- zvířata MeSH
- MeSH
- aorta metabolismus transplantace MeSH
- apolipoproteiny E nedostatek MeSH
- arterioskleróza patofyziologie MeSH
- finanční podpora výzkumu jako téma MeSH
- inbrední kmeny myší MeSH
- lidé MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- kongresy MeSH
- MeSH
- aorta transplantace MeSH
- aortální aneurysma chirurgie MeSH
- chirurgie operační metody MeSH
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- pooperační komplikace MeSH
- senioři MeSH
- Check Tag
- dospělí MeSH
- lidé středního věku MeSH
- lidé MeSH
- mladiství MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- MeSH
- antigeny CD4 imunologie MeSH
- aorta patologie transplantace MeSH
- cévní endotel patologie účinky léků MeSH
- cyklosporiny terapeutické užití MeSH
- kombinovaná farmakoterapie MeSH
- krysa rodu rattus MeSH
- monoklonální protilátky terapeutické užití MeSH
- rejekce štěpu MeSH
- zvířata MeSH
- Check Tag
- krysa rodu rattus MeSH
- zvířata MeSH
- MeSH
- aorta transplantace MeSH
- cévní protézy MeSH
- lidé středního věku MeSH
- Check Tag
- lidé středního věku MeSH
- mužské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
