Endogenous memory CD8 T cells are activated within cardiac allografts without mediating rejection.

Endogenous memory CD8 T cells infiltrate MHC-mismatched cardiac allografts within 12-24 h posttransplant in mice and are activated to proliferate and produce IFN-γ. To more accurately assess the graft injury directly imposed by these endogenous memory CD8 T cells, we took advantage of the ability of anti-LFA-1 mAb given to allograft recipients on days 3 and 4 posttransplant to inhibit the generation of primary effector T cells. When compared to grafts from IgG-treated recipients on day 7 posttransplant, allografts from anti-LFA-1 mAb-treated recipients had increased numbers of CD8 T cells but these grafts had marked decreases in expression levels of mRNA encoding effector mediators associated with graft injury and decreases in donor-reactive CD8 T cells producing IFN-γ. Despite this decreased activity within the allograft, CD8 T cells in allografts from recipients treated with anti-LFA-1 mAb continued to proliferate up to day 7 posttransplant and did not upregulate expression of the exhaustion marker LAG-3 but did have decreased expression of ICOS. These results indicate that endogenous memory CD8 T cells infiltrate and proliferate in cardiac allografts in mice but do not express sufficient levels of functions to mediate overt graft injury and acute rejection.

Association of a single nucleotide polymorphism in the SH2D1A intronic region with systemic lupus erythematosus.

SH2D1A, also known as signaling lymphocytic activation molecule (SLAM)-associated protein (SAP), is an adaptor protein. Recently, it was reported that SAP deficient mice were protected from systemic lupus erythematosus (SLE). In this study, we postulated SH2D1A gene to be a candidate susceptibility gene for SLE and analyzed its association with SLE. A case-control association study was conducted on 5 tag single nucleotide polymorphisms (SNPs) in SH2D1A region in 506 Japanese female SLE patients and 330 healthy female controls. The luciferase assay was performed to determine the functional role of the SNP associated with SLE. One SNP in the intron 2, rs2049995, showed association with SLE (p=0.0110, odds ratio (OR) 1.97, 95% confidence interval (CI) 1.16-3.34, under the dominant model). The association of rs2049995 seemed to be stronger in the subset with the age of onset less than 20 years (p=0.0067, OR 2.65, 95% CI 1.28-5.46). Functional evaluation of rs2049995 showed that reporter gene activity was increased 1.9-fold for the susceptible allele compared with the resistant allele. An intronic SNP of SH2D1A is associated with SLE.

Clinical outcomes of recipients with aplastic bone disease after renal transplantation.

OBJECTIVES: We expect that if chronic renal failure (CRF) is improved after renal transplantation (RTx), dialysis osteopathy bone lesions would also recover to normal. Nevertheless, it is controversial whether bone lesions really improve after RTx. In this study, we evaluated whether pathological dialysis osteopathy improved after RTx. MATERIALS AND METHODS: The 84 patients who had undergone living related RTx had also undergone a bone biopsy (Bx) since January 2004, including 13 (16.0%) with a diagnosis of aplastic osteopathy. They included 7 men and 6 women. The average hemodialysis (HD) period was 40.3 months. The immunosuppression was tacrolimus (FK); mycophenalate mofetil (MMF) and steroid. We examined Ca, P, intact-PTH (i-PTH), metabolic bone markers, and bone density (DXA) before and 1 year after RTx. In addition, a Bx was performed after having osteal labeling twice before Bx. In addition 2 cases (15.3%) also underwent a Bx after RTx. RESULTS: All cases survive with well functioning renal grafts. The mean levels of Ca and P before RTx were 8.7 mg/mL and 6.6 mg/dL, respectively. The mean i-PTH level was 137.8 pg/mL before RTx and of alkaline phosphatase (ALP) was 202.1 U/L before RTx. The total density and % age match of DXA before RTx averaged 398.7 mg/ccm and 96.7%, respectively. The mean bone volume fraction (BV/TV: Bone Volume/Tissue Volume) before RTx was 17.2%. The mean osteoid volume (OV/TV) before RTx was 2.7%. The mean fibrosis volume (Fb.V/TV) before RTx was 0%. The mean bone formation rate (BFR/BV) before RTx was 2.1 %/y. Two cases were also pathologically diagnosed as renal osteodystrophy at 1 year after RTx: 1 case was mixed type, and another was osteomalacia, which was accompanied by a lumbar compression fracture (Fx) during the clinical course. CONCLUSIONS: Bone metabolism in patients with aplastic ROD histologically improved at 1 year after RTx, presumably due to good renal transplant function. However, it is unknown whether both hypophosphatemia and decrease of FGF-23 improved bone However, patients with aplastic ROD were not completely normalized histologically at 1 year after RTx.

The SNP c.1326T>G in the non-SMC condensin I complex, subunit G (NCAPG) gene encoding a p.Ile442Met variant is associated with an increase in body frame size at puberty in cattle.

Recently, we had located a bovine carcass weight QTL, CW-2, to a 591-kb interval on BTA6 and have identified the SNP c.1326T>G in the NCAPG (non-SMC condensin I complex, subunit G) gene that leads to the amino acid change p.Ile442Met in the NCAPG protein, which is a candidate causative variation. Here, we examined the association of the NCAPG:c.1326T>G locus with linear skeletal measurements of growth-associated traits during adolescence, which is a period of intensive growth, using two historically and geographically distant cattle populations: 792 Japanese Black steers and 161 F(2) bulls of an experimental cross from Charolais and German Holstein. In both populations, the SNP NCAPG:c.1326T>G was associated with each component of body frame size: height, length and width at puberty. The associations of CW-2 with height- and length-associated traits were observed at an earlier growth period compared to the associations with thickness- and width-associated traits, indicating that the primary effect of the CW-2 QTL may possibly be exerted on skeletal growth. The significant associations of the NCAPG:c.1326T>G locus with growth-associated skeletal measurements are similar to the effects of the syntenic region on human chromosome 4 that are associated with adult height in humans, supporting the hypothesis that CW-2 is analogous to the human locus and pointing to a conserved growth-associated locus or chromosomal region present in both species.

LFA-1 antagonism inhibits early infiltration of endogenous memory CD8 T cells into cardiac allografts and donor-reactive T cell priming.

Alloreactive memory T cells are present in virtually all transplant recipients due to prior sensitization or heterologous immunity and mediate injury undermining graft outcome. In mouse models, endogenous memory CD8 T cells infiltrate MHC-mismatched cardiac allografts and produce IFN-γ in response to donor class I MHC within 24 h posttransplant. The current studies analyzed the efficacy of anti-LFA-1 mAb to inhibit early CD8 T cell cardiac allograft infiltration and activation. Anti-LFA-1 mAb given to C57BL/6 6 (H-2(b)) recipients of A/J (H-2(a)) heart grafts on days -1 and 0 completely inhibited CD8 T cell allograft infiltration, markedly decreased neutrophil infiltration and significantly reduced intragraft expression levels of IFN-γ-induced genes. Donor-specific T cells producing IFN-γ were at low/undetectable numbers in spleens of anti-LFA-1 mAb treated recipients until day 21. These effects combined to promote substantial prolongation (from day 8 to 27) in allograft survival. Delaying anti-LFA-1 mAb treatment until days 3 and 4 posttransplant did not inhibit early memory CD8 T cell infiltration and proliferation within the allograft. These data indicate that peritransplant anti-LFA-1 mAb inhibits early donor-reactive memory CD8 T cell allograft infiltration and inflammation suggesting an effective strategy to attenuate the negative effects of heterologous immunity in transplant recipients.

Acute antibody-mediated rejection in living ABO-incompatible kidney transplantation: long-term impact and risk factors.

The impact of acute antibody-mediated rejection (AAMR) on the long-term outcome on ABO-incompatible (ABOI) kidney transplantation is not well understood. We retrospectively analyzed the long-term impact of AAMR and risk factors for AAMR in 57 consecutive recipients performed between 1999 and 2004. Nineteen patients (33%) who developed AAMR within 3 months posttransplantation constituted of the AMR group. The graft survival rate was significantly lower in the AMR group (AMR vs. non-AMR, respectively; 5 years: 84% vs. 95%; 8 years: 45% vs. 95%; p = 0.009). The prevalence of transplant glomerulopathy at 1 year posttransplantation was significantly higher in the AMR group (AMR 64% vs. non-AMR 3%, p < 0.001). Multivariate analysis demonstrated that anti-blood group IgG antibody titers of 1:32 at the time of transplantation (OR, 9.52; p = 0.041) and donor-specific anti-HLA antibodies (DSHA) detected by Luminex single bead method (OR, 5.68; p = 0.015) were independent risk factors for AAMR regardless of baseline anti-blood group IgG antibody titers. Our results indicate that AAMR has a heavy impact on the long-term outcome and preoperative DSHA appears to have a more significant association with poor graft outcomes than anti-blood group antibodies, even in ABOI kidney transplantation.

Continent orthotopic ileal neobladder after kidney transplantation in a patient with urothelial cell carcinoma associated with Chinese herb nephropathy.

A 58-year-old man underwent kidney transplantation on November 14, 2002 for end-stage kidney disease after Chinese herb nephropathy. Immunosuppressive therapy was maintained with tacrolimus, mycophenolate mofetil, and methylpredonisolone. He was diagnosed with right ureteral cancer and underwent right nephroureterectomy on December 13, 2003. Then, he underwent left nephroureterectomy for left ureteral cancer on March 5, 2004. Subsequently, he was diagnosed with multiple bladder cancers and carcinoma in situ. On August 31, he underwent radical cystectomy with an orthotopic ileal neobladder (Studer's method). The postoperative course was uneventful. After 3 years follow-up, this patient shows no evidence of recurrence and his serum creatinine level is stable (1.7 mg/dL). The continence is maintained during both day and night; he voids without intermittent self-catheterization. We suggest that an orthotopic ileal neobladder is a safe method of urinary diversion after cystectomy in kidney transplant recipients.

Analysis of renal transplant protocol biopsies in ABO-incompatible kidney transplantation.

Numerous studies have shown that protocol biopsies have predictive power. We retrospectively examined the histologic findings and C4d staining in 89 protocol biopsies from 48 ABO-incompatible (ABO-I) transplant recipients, and compared the results with those of 250 controls from 133 ABO-compatible (ABO-C) transplant recipients given equivalent maintenance immunosuppression. Others have shown that subclinical rejection (borderline and grade I) in ABO-C grafts decreased gradually after transplantation. In our study, however, subclinical rejection in the ABO-I grafts was detected in 10%, 14% and 28% at 1, 3 and 6-12 months, respectively. At 6-12 months, mild tubular atrophy was more common in the ABO-C grafts whereas the incidence of transplant glomerulopathy did not differ between the two groups (ABO-C: 7%; ABO-I: 15%; p = 0.57). In the ABO-I transplants, risk factors for transplant glomerulopathy in univariate analysis were positive panel reactivity (relative risk, 45.0; p < 0.01) and a prior history of antibody-mediated rejection (relative risk, 17.9; p = 0.01). Furthermore, C4d deposition in the peritubular capillaries was detected in 94%, with diffuse staining in 66%. This deposition, however, was not linked to antibody-mediated rejection. We conclude that, in the ABO-I kidney transplantation setting, detection of C4d alone in protocol biopsies might not have any diagnostic or therapeutic relevance.

Evaluation of immunosuppressive regimens in ABO-incompatible living kidney transplantation--single center analysis.

Several protocols allow the successful ABO incompatible living-related kidney transplantation (ABO-ILKT), yet no single method has emerged as the best. We have made several substantial changes to our ABO-ILKT protocol over the past decade and a half and have attempted to determine whether the changes in immunosuppressive agents have resulted in a better outcome. We used methylprednisolone (MP), cyclosporine (CsA), azathioprine (AZ), antilymphocyte globulin (ALG) and deoxyspergualine (DSG) in the 105 cases of ABO-ILKT (group 1) between 1989 and 1999, and MP, tacrolimus (FK506), mycophenolate mofetil (MMF) in the 117 cases of ABO-ILKT (group 2) between 2000 and 2004. We compared the patient and graft survival rates as well as the incidence rate of acute rejection in these two eras, when different regimens were used. There were significant differences in the 1- and 5-year graft survival rates between groups 1 and 2 (1-year: 78% in group 1 vs. 94% in group 2; 5-year: 73% in group 1 vs. 90% in group 2, p = 0.008). Also, a higher incidence rate of acute rejection was significantly observed in group 1 (50/105, 48%) than in group 2 (18/117, 15%) (p < 0.001). We conclude that the FK/MMF combination regimen provides excellent graft survival results in ABO-ILKT.

Analysis of cause of death with a functioning graft: a single-center experience.

INTRODUCTION: After the introduction of new immunosuppressants, such as tacrolimus and mycophenolate mofetil, we have achieved excellent results for kidney transplantation with a low acute rejection rate. Currently, nonimmunological factors are considered to be the main cause of graft loss for long-term transplant patients. In this study, we analyzed the cause of death with a functioning graft. PATIENTS AND METHODS: We performed 1375 cases of living kidney transplantation (LKT) and 219 cases of cadaveric kidney transplantation (CKT) between January 1983 and December 2002. Of these patients, 86 LKT patients and 19 CKT patients died with a functioning graft. RESULTS: The mean duration of graft function was 4.8 +/- 4.5 years. The incidence of the causes of death were: infection, 24%; stroke, 17%; cardiovascular disease, 16%; malignant disease, 15%; hepatic failure, 11%; gastric ulcer, 4%; and accident/suicide 2%. Five- and 10-year graft survivals for LKT were 80.2 and 62.0%, respectively. The corresponding values for patients (with the exception of the patients who died with a functioning graft) was 83.0% and 66.1%, respectively. The 5- and 10-year graft survival rates for cadaveric kidney transplants were 70.8% and 48.9%, respectively. The corresponding values for patients (with the exception of the patients who died with a functioning graft) were 75.3% and 52.6%, respectively. CONCLUSION: To prevent death with a functioning graft, management of vascular disorders such as stroke and cardiovascular disease, malignant disease, and infectious disease is crucial for kidney transplant patients.

Microscopic examination of the urine helps to confirm the recovery of acute tubular necrosis after cadaveric renal transplantation: a case report.

INTRODUCTION: Delayed graft function due to acute tubular necrosis (ATN) is frequently seen in kidney transplants from non-heart-beating donors. However, only a biopsy can be used to assess the severity of ATN. Therefore, we studied the validity of microscopic findings in tubular epithelial cells (TECs) from urine as a means to monitor ATN. MATERIALS AND METHODS: The first voided urine in the morning was examined for the appearance and nuclear cytoplasmic (N/C) ratio of the TECs, using a murine staining with URO-3 monoclonal antibody to detect proximal tubular cells (PTCs). CASE: A 58-year-old man underwent cadaveric kidney transplantation in January, 2003 using tacrolimus, mycophenolate mofetil, and prednisone following basiliximab induction therapy. His graft did not function immediately; needle biopsy was performed on day 17. The pathological findings showed severe ATN without evidence of acute rejection. A large quantity of TECs was seen in his urine between days 7 and 14. After day 28, TECs with a large N/C ratio and that were URO-3 antibody-positive were detected. Urine volume increased gradually and hemodialysis was not necessary after day 36. CONCLUSION: The presence of URO-3-positive TECs with large N/C ratios suggests the reconstruction of PTCs. Therefore, it may be useful to monitor TEC findings to assess the severity ATN after cadaveric kidney transplantation.

Impact of HLA-identity on results of ABO-incompatible living kidney transplantation.

INTRODUCTION: Due to the continuing shortage of cadaveric donors in Japan, ABO-incompatible living kidney transplantation is being performed. Our previous studies showed that the long-term graft survival in ABO-incompatible living kidney transplantation was comparable to that in ABO-compatible living kidney transplantation. However, the impact on HLA-identity on the results of ABO-incompatible living donor kidney transplantation had not been investigated previously. MATERIALS AND METHODS: One hundred twenty-seven recipients underwent ABO-incompatible living kidney transplantation between January 1989 and December 2000. Five were grafted from HLA-identical sibling donors group (I). The remaining 122 recipients received grafts from an HLA-nonidentical donor (group N). Both groups were similar in terms of recipient age, donor age, warm ischemic time, and total ischemic time. Three or four sessions of plasmapheresis were performed prior to transplantation. Cyclosporine or tacrolimus, methylprednisolone, and azathioprine or mycophenolate mofetil were used for immunosuppression. Splenectomy was done at the time of kidney transplantation in all patients. RESULTS: Graft loss was seen in one of the five HLA-identical recipients due to chronic rejection. Five- and 10-year graft survival rates were 80.0% I vs 72.0% N, and 80.0% I vs 54.2% N, respectively. The incidence of acute rejection in the HLA-identical recipients was lower than that in the HLA-nonidentical recipients (20% I vs 67.2% N). In conclusion, long term graft survival among ABO-incompatible kidney transplants from HLA-identical sibling donors was much better than that from HLA-nonidentical sibling donors.

Impact of positive PRA on the results of ABO-incompatible kidney transplantation.

INTRODUCTION: Due to the continuing shortage of cadaveric donors in Japan, ABO-incompatible living kidney transplantation (LKT) is being performed. It is well known that highly sensitized patients with positive panel reactive antibodies (PRA) often present with acute rejection. Therefore, we examined the impact of a positive PRA on the results of ABO-incompatible LKT. MATERIALS AND METHODS: One hundred seventy-seven recipients underwent ABO-incompatible LKT between January 1989 and March 2003. Of these patients, 37 who had been examined for PRA before transplantation were included in this study. There were 25 men and 12 women of mean age 37.3 years. Plasmapheresis was performed to remove anti-ABO antibodies before transplantation. During the induction phase, methylprednisolone, azathioprine, or mycophenolate mofetil and cyclosporine or tacrolimus were used for immunosuppression. Splenectomy was performed at the time of kidney transplantation in all patients. PRA was measured using FlowPRA by flow cytometer. RESULTS: Eight of the 37 patients had a positive PRA before transplantation (class I, 5; class II, 1; class I and class II, 2). The incidence of acute rejection was 37.9% in the patients with a negative PRA and 37.5% in patients with a positive PRA. One patient with a negative PRA and one patient with a positive PRA lost grafts due to acute rejection. CONCLUSIONS: Positive PRA may not increase the incidence of acute rejection in ABO-incompatible LKT because plasmapheresis and splenectomy are performed to eliminate anti-ABO antibody.

Excellent outcome of ABO-incompatible living kidney transplantation under pretransplantation immunosuppression with tacrolimus, mycophenolate mofetil, and steroid.

INTRODUCTION: ABO-incompatible living kidney transplantation (LKT) has been performed to widen the indications for kidney transplantation. Since 2001, using a 7-day period of pretransplantation immunosuppression with tacrolimus (FK) plus mycophenolate mofetil (MMF) plus methylprednisolone (MP), we have observed a marked reduction in acute humoral/vascular rejection without any serious complications. PATIENTS AND METHODS: Forty-five adult patients underwent ABO-incompatible LKT at our institute between January 2000 and September 2002. There were 20 men and 25 women of mean age 33 years. Plasmapheresis was performed to remove anti-AB antibodies prior to kidney transplantation. In 2000, 13 patients were treated with FK plus MMF plus MP without 7-day pretransplantation immunosuppression (group 1). Since January 2001, we have administered FK (0.1 mg/kg/d) plus MMF (1-2 g/d) plus MP (125 mg/d) concomitantly with plasmapheresis starting from 7 days before transplantation in 32 patients (group 2). Splenectomy was performed at the time of kidney transplantation in all patients. RESULTS: Patient survival rate was 100% in both treatment groups. Graft survival rate was 92% and 97% in groups 1 and 2, respectively. One patient in group 1 lost the graft due to severe pancreatitis and 1 patient in group 2, due to severe humoral rejection. The incidence of acute rejection was 56% and 19% in group 1 and group 2, respectively. No patient experienced any lethal infectious complication. CONCLUSION: Pretransplantation immunosuppression for 7 days using FK, MMF, and MP in ABO-incompatible LKT provides an excellent outcome without severe infectious complications.

Systemic lupus erythematosus and thrombotic thrombocytopenic purpura: a case report and literature review.

We describe a patient with systemic lupus erythematosus (SLE) who developed severe and acute thrombotic thrombocytopenic purpura (TTP). Detection of the fragmentation of peripheral red blood cells (RBC) helped the early diagnosis of TTP and the patient was rescued by extensive plasma exchange started promptly after the diagnosis. Because manifestations of TTP are similar to those in SLE, it is sometimes difficult to make an accurate diagnosis of TTP in SLE patients. We emphasise here the significance of the early diagnosis of TTP by the observation of fragmented RBC and the intensive therapy, including plasma exchange, for this very severe condition.

Incidence of peripheral blood eosinophilia and the threshold eosinophile count for indicating hypereosinophilia-associated diseases.

BACKGROUND: A definite threshold of the peripheral blood eosinophile count that indicates the presence of hypereosinophilia-associated diseases has not yet been determined. METHODS: The threshold eosinophile count at which cases of hypereosinophilia-associated diseases (n = 25) can be differentiated from those of bronchial asthma (n = 101) was determined. Then, the incidences of eosinophile counts greater than 1.0 x 10(9)/l or the threshold level, were studied by analysis of 43,805 samples sent to the laboratory, and the diseases associated with the increased counts were determined. RESULTS: The eosinophile count in cases of hypereosinophilia-associated diseases and in those of bronchial asthma were 10.967 +/- 1.680 x 10(9)/l and 0.574 +/- 0.045 x 10(9)/l, respectively (P < 0.001); the threshold was 2.052 x 10(9)/l. The percentages of samples with an eosinophile count of more than 1.0 x 10(9)/l and 2.052 x 10(9)/l were 0.6% and 0.1%, respectively; the latter comprised of 41 samples from 24 patients including eight with hypereosinophilia-associated diseases. The patients with hypereosinophilia-associated diseases had a significantly higher count, and a higher incidence of counts of more than 2.052 x 10(9)/l, than others, including patients with malignancies and symptoms conventionally referred as "atopic diseases". CONCLUSION: Hypereosinophilia-associated diseases are associated with a very high eosinophile count of more than 2.052 x 10(9)/l, which was observed rarely.

Peroxisome proliferator-activated receptor-gamma haploinsufficiency enhances B cell proliferative responses and exacerbates experimentally induced arthritis.

Peroxisome proliferator-activated receptor-gamma (PPAR gamma) controls adipogenesis and glucose metabolism. It was reported recently that PPAR gamma activation by its agonistic ligands modifies lymphocyte function. Since synthetic ligands are known to exert their effect via PPAR gamma-dependent and -independent pathways, we examined the physiological role of PPAR gamma in lymphocytes by using heterozygote mutant mice in which one allele of PPAR gamma is deleted (PPAR gamma(+/-)). In contrast to T cells, which did not exhibit a significant difference, B cells from PPAR gamma(+/-) showed an enhanced proliferative response to stimulation by either lipopolysaccharide or cross-linking of antigen receptors. Dysregulation of the NF-kappa B pathway in B cells from PPAR gamma(+/-) was indicated by spontaneous NF-kappa B activation, as well as increased I kappa B alpha phosphorylation and gel-shift activity following LPS stimulation. Mice primed with either ovalbumin or methylated BSA also showed enhanced antigen-specific immune response of both T and B cells, an immunological abnormality that exacerbated antigen-induced arthritis. These findings indicate that PPAR gamma plays a critical role in the control of B cell response and imply a role in diseases in which B cell hyperreactivity is involved, such as arthritis and autoimmunity.