OBJECTIVETo observe the survival of hand allograft under the state of immunosuppression and the pathological changes of rejection in the recovery process.

METHODSThe biopsies of the skin, nerve, muscle, tendon and bone tissue of hand allografts during different stages from 1 day to 7 months after operation were observed using routine histological technique.

RESULTSNo significant changes due to rejection in skin, nerve, muscle and bone tissue were observed. But different degrees of weak rejective changes were found on the wall of blood vessels; in the muscle and nerve the reactions were markedly stronger than those found in skin tissues.

CONCLUSIONSThe rejection in deep tissues should be monitored in controlling the rejection of hand allograft.

Adult ; Biopsy ; Graft Rejection ; pathology ; Hand Transplantation ; Humans ; Immunosuppression ; Male ; Skin ; immunology ; pathology ; Transplantation, Homologous

OBJECTIVETo investigate the pathological and immunological changes of renal grafts in recipients experiencing graft rejection.

METHODSThe clinicopathologic data of 56 renal needle biopsy samples obtained from renal transplant recipients were analyzed retrospectively. The specimens were classified histopathologically according to the Banff 2009 classification system and analyzed by immunohistochemical labeling and immunofluorescence.

RESULTSIn the 56 recipients, 1 (1.79%) experienced hyperacute rejection, 8 (14.29%) had suspected acute rejection, 12 (21.43%) developed acute T-cell rejection, 6 (10.71%) had acute antibody-mediated rejection, 2 (3.57%) had acute T-cell rejection with acute antibody-mediated rejection, 12 (21.43%) had chronic active T cell-mediated rejection, 2 (3.57%) had chronic active antibody-mediated rejection, 2 (3.57%) had chronic active T cell-mediated rejection with antibody-mediated rejection, 8 (14.29%) had non-specific interstitial fibrosis and tubular atrophy, and 3 (5.36%) had normal graft function. The expression levels of immune markers CD3, CD4, CD8, CD20, GrB and perforin differed with the types of T cell-mediated graft rejection, and the positivity and expression levels of these markers tended to increased with the severity of graft rejection. The expression of C4d was positive in all cases with antibody-mediated graft rejection.

CONCLUSIONSThe pathological characteristics of the renal biopsy specimens and expression levels of the immune markers allow timely and accurate evaluation of graft rejection type to provide a reliable pathological and etiological basis for clinical treatment and prognostic assessment.

Adolescent ; Adult ; Aged ; Female ; Graft Rejection ; immunology ; pathology ; Graft Survival ; Humans ; Kidney ; immunology ; pathology ; Kidney Transplantation ; adverse effects ; Male ; Middle Aged ; Retrospective Studies ; Young Adult

An orthotopic penetrating keratoplasty model was developed in the rat. An oversized (0.5 mm) graft was used and 8 interrupted sutures were applied. These sutures were not removed. Eleven grafts out of 13 were rejected by the 3rd week in the disparate group (Brown Norway rat to Lewis rat transplantation group), which was characterized by edema, opacity, and neovascularization. All grafts remained clear in the syngeneic group (Lewis rat to Lewis rat transplantation group). Immunohistochemical examination was performed. This model seems to be a reliable and reproducible one to evaluate rejection reaction in corneal transplantation.
Animals ; Female ; Graft Rejection ; Keratoplasty, Penetrating/immunology/*methods/pathology ; Lymphocyte Subsets/immunology/pathology ; Macrophages/immunology/pathology ; Rats ; Rats, Inbred BN ; Rats, Inbred Lew ; Transplantation, Homologous ; Transplantation, Isogeneic

OBJECTIVETo gain insight into the role of dendritic cells in graft rejection following penetrating keratoplasty by investigating their distribution in rat cornea.

METHODSOrthotopical corneal transplantation was performed and immunohistochemical staining of the whole-mount cornea and the spleen tissue specimen employed to determine the distribution of the dendritic cells in the cornea.

RESULTSGraft rejection occurred in all rats following the transplantation. No OX-62(+) dendritic cells were found in normal cornea but they were present in the epithelium of the cornea graft with allograft rejection.

CONCLUSIONOX-62(+) dendritic cells presenting in the rejected cornea may be related to acute graft rejection after penetrating keratoplasty.

Animals ; Cornea ; immunology ; pathology ; surgery ; Dendritic Cells ; immunology ; physiology ; Female ; Graft Rejection ; etiology ; immunology ; physiopathology ; Keratoplasty, Penetrating ; adverse effects ; methods ; Male ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar

OBJECTIVETo investigate the pathologic monitoring of intestinal graft rejection in auxiliary en-bloc liver-small bowel transplantation in pigs.

METHODSFifty outbred long-white pigs were randomized into three groups, and the auxiliary composite liver-small bowel allotransplantations were undertaken in 10 pigs in group A and group B while segment small bowel allotransplantations were undertaken in 10 pigs in group C. Group A and C were not treated with immunosuppressive drugs while group B was treated with cyclosporine A and methylprednisolone. The postoperative intestinal graft rejections were monitored by biopsy through the jejunostomy or ileuostomy on 1, 3, 5, 7, 14, 21 and 30 days after operation. Through routine management, the specimens were directly examined via optical and electronic microscope respectively.

RESULTSAs shown from pathological data, the median initial time of postoperative rejection in group A was 8 days (ranged from 7 to 12), later than that in group C (5 days:ranged from 3 to 5), P<0.05). On the 7th day postoperatively, the rejection scores in group A was 1.11+/-0.20, lower than that in group C(2.56+/-0.18, P<0.05), but higher than that in group B(0.20+/-0.13, P<0.05). Ultrastructure also showed more severe intestinal graft rejection in intestinal transplantation than that in combined transplantation. The median survival time was 9 days(ranged from 7 to 25) in group A and 12 days(ranged from 7 to 20) in group C, while all the pigs in group B lived longer than 30 days.

CONCLUSIONThe pathological assessment through the jejunostomy or ileuostomy biopsy is a convenient method to monitor the postoperative graft rejections in intestinal related transplantation.

Animals ; Female ; Graft Rejection ; prevention & control ; Graft Survival ; Intestine, Small ; pathology ; transplantation ; ultrastructure ; Liver Transplantation ; adverse effects ; immunology ; Male ; Swine ; Transplantation, Homologous

In this study antigen-independent factor in the pathogenesis of chronic rejection of organ transplants was examined. Kidney isografts and allografts were transplanted orthotopically into bilaterally nephroectomized rat recipients and studied functionally, morphologically and immunohistologically, at serial intervals up to 52 weeks after transplantation. Allograft recipients developed progressive proteinuria after 12 weeks, with gradual renal failure ultimately leading to death. At the same time, morphological changes, including progressive arteriosclerosis and glomerulosclerosis, tubular atrophy and interstitial fibrosis, developed. Immunohistologically, macrophages infiltrated glomeruli during this period and cytokines became unregulated. Our results showed that antigen-independent functional and morphological changes occurred in long-term kidney isografts and mimicked those appearing much earlier in allografts that reject chronically. Initial injury and extent of functioning renal mass is suggested to be important factor for such late changes.
Animals ; Graft Rejection ; etiology ; immunology ; pathology ; Graft Survival ; physiology ; Kidney ; immunology ; pathology ; Kidney Transplantation ; immunology ; methods ; pathology ; Proteinuria ; etiology ; Rats ; Rats, Inbred Strains ; Time Factors ; Transplantation, Homologous ; Transplantation, Isogeneic

OBJECTIVETo investigate the mechanism of immune hyporesponsiveness induced by donor-antigen- unloaded recipient-derived immature dendritic cell (imDC) of liver grafts in rats.

METHODSForty Sprague-Dawley rats (donor) and forty male Wistar rats (recipient) were randomly divided into 4 groups: control, cyclosporine A (CsA), mature DC (mDC), and imDC groups respectively, with 10 donor rats and 10 recipient rats in each group. Recipient rats in CsA group were treated with 10 mg•kg⁻¹•d⁻¹ CsA starting day 2 after the transplantation. Recipients in the mDC or imDC groups were given Wistar rat derived mDCs (1 × 10⁶/rat) or imDCs (1 × 10⁶/rat) via dorsal vein of the penis respectively 1 day before the transplantation. In each group, 5 recipients were kept for determination of survival time and the other 5 rats were executed at day 10 after transplantation. Blood samples were collected for the measurement of serum alanine aminotransferase (ALT), total bilirubin (TBIL), interleukin 2 (IL-2), interferon gamma (IFN-γ), IL-4, and IL-10 levels. Liver tissue was harvested for HE staining and acute rejection evaluation. Expression levels of Fas-L/Fas in the grafts were detected by immunohistochemical staining; and Western blot was used to detect the expression level of Scurfin.

RESULTSThe survival time of CsA and imDC groups was significantly longer than that of control and mDC groups (all P < 0.05). The levels of serum ALT and TBIL in the control group (2072.20 ± 217.93 IU/L and 147.42 ± 22.02 µmol/L) and mDC group (2117.00 ± 285.13 IU/L and 141.58 ± 20.82 µmol/L) were significantly higher than those in the CsA group (59.68 ± 13.48 IU/L and 15.40 ± 2.13 µmol/L) or imDC group (50.80 ± 9.63 IU/L and 14.44 ± 3.49 µmol/L) (all P < 0.05). In the CsA and imDC groups, the levels of IL-2 (22.52 ± 3.75 pg/mL and 22.12 ± 3.90 pg/mL) and IFN-γ (309.20 ± 25.19 pg/mL and 321.00 ± 21.64 pg/mL) were significantly lower, but the levels of IL-4 (297.60 ± 25.07 pg/mL and 277.00 ± 22.47 pg/mL) and IL-10 (1226.00 ± 140.49 pg/mL and 1423.00 ± 106.39 pg/mL) were higher than those of the control (IL-2: 147.78 ± 12.80 pg/mL, IFN-γ: 1758.60 ± 106.22 pg/mL, IL-4: 17.40 ± 4.77 pg/mL, IL-10: 81.00 ± 9.47 pg/mL) and mDC groups (IL-2: 142.34 ± 9.29 pg/mL, IFN-γ: 1835.00 ± 82.63 pg/mL, IL-4: 15.60 ± 3.96 pg/mL, IL-10: 68.80 ± 11.23 pg/mL) (all P < 0.01). The expression level of Scurfin protein on CD4+ CD25+ T cells of the imDC group (1.34 ± 0.29) was significantly higher than that in the control (0.72 ± 0.13), CsA (0.37 ± 0.11), and mDC groups (0.78 ± 0.17) (all P < 0.05).

CONCLUSIONDonor-antigen-unloaded recipient-derived imDC is an effective treatment in inducing immune hyporesponsiveness through induction of T cell apoptosis, shift in Thl/Th2 balance, and proliferation of regulatory T cell.

Animals ; Antigens ; immunology ; Cytokines ; immunology ; Dendritic Cells ; cytology ; immunology ; transplantation ; Fas Ligand Protein ; immunology ; Forkhead Transcription Factors ; metabolism ; Graft Rejection ; immunology ; Graft Survival ; immunology ; Humans ; Immunity ; physiology ; Liver ; cytology ; immunology ; pathology ; Liver Transplantation ; immunology ; Male ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Rats, Wistar ; fas Receptor ; immunology

OBJECTIVETo study the roles of granzyme B and perforin in diagnosing acute rejection after liver transplantation, and the relationship between their activity index (AI) and Banff's histological grading criteria.

METHODSLiver biopsies were processed as for routine surgical specimens and labeled with granzyme B and perforin monoclonal antibodies. The number of positive cells/mm(2) was determined as activity index (AI) by IPP image analysis software. Histologic findings were used as the "gold standard" in diagnosing acute rejection.

RESULTSOf 41 liver biopsy samples studied, acute rejection was noted in 21 cases, the remaining 20 cases showed no evidence of rejection. The AI of granzyme B and perforin in the acute rejection group was significantly higher than that in the non-acute rejection group (< 0.001). In the acute rejection group, the AI in moderate to severe acute rejection was higher than that in mild to indeterminate acute rejection (< 0.001). Compared with the "golden" histologic criteria, the sensitivity, specificity, positive predictive value, negative predictive value and accuracy of granzyme B in diagnosing acute rejection were 90.0%, 95.2%, 94.7%, 90.9% and 92.7% respectively. The values of these parameters for perforin were also above 80%.

CONCLUSIONSGranzyme B and perforin are key markers of activated immune cells in acute rejection and highly expressed during acute liver rejection episodes. As ancillary investigations, these parameters demonstrated high sensitivity and specificity in diagnosing acute rejection in allograft post-transplant liver biopsies.

Biomarkers ; Biopsy ; Graft Rejection ; diagnosis ; metabolism ; Granzymes ; metabolism ; Humans ; Liver ; metabolism ; pathology ; Liver Transplantation ; immunology ; Membrane Glycoproteins ; metabolism ; Perforin ; Pore Forming Cytotoxic Proteins ; metabolism ; Sensitivity and Specificity

OBJECTIVETo evaluate the association of major histocompatibility complex class I chain related gene A (MICA) antibodies with acute rejection (AR), chronic rejection (CR) and renal function after renal transplantation.

METHODSSerum MICA antibodies were detected with ELISA before and after transplantation with also examinations of panel reactive antibodies (PRA), serum creatinine, urine, graft ultrasound, lymphocyte subsets and the pathology of graft biopsy. The study was carried out in two parts to monitor MICA antibodies in acute and chronic rejections after renal transplantation.

RESULTSIn the first part of the study 18 of the 41 recipients experienced episodes of acute rejection, and the incidence rate was markedly higher in MICA(+) group than in MICA(-) group (P<0.05). Compared with the recipients with stable renal functions, the patients with acute graft rejection showed a significantly higher positivity rate of MICA antibodies. Postoperative MICA antibody monitoring showed that MICA antibody level increased gradually 2-3 days after the occurrence of acute rejection; anti-rejection treatment lowered serum creatinine to a normal level but MICA antibodies remained positive. In the second part, 21 of 40 patients had chronic graft rejection and showed significantly higher positivity rate of MICA than the patients with stable renal functions (P<0.05). In patients with chronic rejections, the serum creatinine levels were significantly higher in MICA(+) than in MICA(-) cases (P<0.05). Graft biopsy of all MICA(+) cases showed C4d deposition.

CONCLUSIONThe status of MICA antibodies can predict the occurrence and treatment outcomes of acute rejection, and also as one of the major causes of chronic graft rejection, they affect the long-term survival of the renal grafts.

Adolescent ; Adult ; Antibodies ; blood ; immunology ; Complement C4b ; metabolism ; Creatinine ; blood ; Follow-Up Studies ; Graft Rejection ; blood ; immunology ; pathology ; HLA Antigens ; immunology ; Histocompatibility Antigens Class I ; immunology ; Humans ; Kidney ; metabolism ; physiopathology ; Kidney Transplantation ; Peptide Fragments ; metabolism ; Young Adult

The aim of this study was to evaluate whether the Th17 and Treg cell infiltration into allograft tissue is associated with the severity of allograft dysfunction and tissue injury in acute T cell-mediated rejection (ATCMR). Seventy-one allograft tissues with biopsy-proven ATCMR were included. The biopsy specimens were immunostained for FOXP3 and IL-17. The allograft function was assessed at biopsy by measuring serum creatinine (Scr) concentration, and by applying the modified diet in renal disease (MDRD) formula, which provides the estimated glomerular filtration rate (eGFR). The severity of allograft tissue injury was assessed by calculating tissue injury scores using the Banff classification. The average numbers of infiltrating Treg and Th17 cells were 11.6 +/- 12.2 cells/mm2 and 5.6 +/- 8.0 cells/mm2, respectively. The average Treg/Th17 ratio was 5.6 +/- 8.2. The Treg/Th17 ratio was significantly associated with allograft function (Scr and MDRD eGFR) and with the severity of interstitial injury and tubular injury (P < 0.05, all parameters). In separate analyses of the number of infiltrating Treg and Th17 cells, Th17 cell infiltration was significantly associated with allograft function and the severity of tissue injury. By contrast, Treg cell infiltration was not significantly associated with allograft dysfunction or the severity of tissue injury. The results of this study show that higher infiltration of Th17 cell compared with Treg cell is significantly associated with the severity of allograft dysfunction and tissue injury.
Acute Disease ; Creatinine/metabolism ; Forkhead Transcription Factors/metabolism ; Graft Rejection/*etiology/pathology ; Humans ; Immunoenzyme Techniques ; Interleukin-17/*metabolism ; Kidney Transplantation/*adverse effects ; Retrospective Studies ; T-Lymphocytes, Regulatory/*immunology/pathology ; Th17 Cells/*immunology/pathology ; Transplantation, Homologous