Adverse effect of exercise on immune complex-mediated glomerulonephritis

To assess the effect of strenuous daily exercise on immune complex-mediated glomerulonephritis (GN), rabbits were randomly assigned to one of three experimental groups: Group I (n = 12): treadmill exercise for 28 days plus twice weekly intravenous injections of saline. Group II (n = 10): treadmill exercise for 28 days plus twice weekly intravenous bovine serum albumin (BSA) injection. Group III (n = 9): intravenous doses of BSA, as in group II, but no exercise. Blood and urine samples were collected from each animal periodically during the 28-day experimental period. On the 29th day of the study all animals were sacrificed and tissue taken for renal histopathologic studies. We found that in group II (exercise + GN) abnormal albuminuria was more frequent (p less than 0.001), blood urea nitrogen (BUN) levels rose significantly with time (p less than 0.02) and hematuria (blood in renal tubules) was more common (p less than 0.05), compared to group III (GN only). The differences between groups II and III could not be explained by the effect of exercise alone since group I (exercise only) developed no abnormal albuminuria, BUN levels or hematuria during the course of the study. These findings suggest that strenuous exercise superimposed on active immune complex-mediated GN results in worsening of the abnormal glomerular function.     

The influence of MHC and non-MHC genes on the nature of murine cardiac allograft rejection. I. Kinetic analysis of mononuclear cell infiltrate and MHC-class I/class II expression in donor tissue

While normal cardiac tissue expresses low levels of MHC-class I, undetectable levels of MHC-class II antigens, and no mononuclear cell infiltrates, posttransplantation allogeneic donor cardiac tissue demonstrates dramatic increases of MHC-class I/class II expression coincident with the infiltration of the tissue with mononuclear cells. Results of this study demonstrate that the kinetics of MHC-class I/II antigen expression and the phenotype of mononuclear cell infiltrate are influenced, to a great degree, by the genetic H-2, intra-H-2 and non-H-2 incompatibility between donor and recipient strains of mice. Increases of MHC-class I precede class II expression in cells from donor cardiac tissue from completely allogeneic BALB/c, H-2-disparate B10.D2, B10.BR, and K, I-A and I-E-disparate B10.T (6R) strains of mice implanted in B10 recipients. In contrast, increase in the level of MHC-class II precedes MHC-class I increases in donor cardiac tissue from H-2-identical but non-H-2-incompatible A.By and the I-E + H-2D end-different B10.A(5R) donor tissue. The completely allogeneic, H-2-disparate or K, I-A, I-E-disparate donor cardiac tissue induced the infiltration of predominantly CD8+ T cells, whereas the non H-2 and I-E + H-2D end-different donor cardiac tissue induced the infiltration of predominantly CD4+ T cells. Finally, whereas bm1 donor cardiac tissue is rejected by B6 recipients by day 32, the (bm1 x bm12)F1 allografts are rejected by day 20, and both express MHC-class I antigens followed by MHC-class II antigens, and contain predominantly CD8+ T cells. In contrast, bm12 allografts are not rejected by B6 recipients, express chronic low levels of both MHC-class I and II antigens, and contain predominantly CD4+ T cells. Of interest is our preliminary finding that bm12 allografts placed in one ear of B6 recipients appear to modify the kinetics of MHC antigen expression and the predominant phenotype of mononuclear cell infiltrates in bm1 allografts placed in the opposite ear. Cumulatively, these data suggest that the type of genetic disparity between cardiac donor and recipient greatly influences the quantitative and qualitative host responses.     

Expression of HLA-DQ, -DR, and -DP antigens in normal kidney and glomerulonephritis

Expression of the defined subtypes of HLA-class II antigens DQ, DR, DP, as well as of a putatively new HLA-class II determinant DY was evaluated with specific monoclonal antibodies on frozen sections of 15 normal kidneys, as well as of renal tissue of 65 patients with different forms of glomerulonephritis (GN). In normal kidney HLA-DR and/or -DY versus DQ or DP antigens were shown to be differentially expressed on subpopulations of glomerular and interstitial cells, as well as vascular endothelia. Normal proximal tubular epithelia lacked HLA-DQ and -DP antigens, but carried -DY and variably -DR products constitutively. In comparison, aberrant presence of HLA-DQ and/or -DP antigens was found on proximal tubular cells in the majority of patients with rapidly progressive (RPGN), membranoproliferative GN (MPGN), or focal glomerular sclerosis (FGS), but more rarely observed in other forms of proliferative or non-proliferative GN. In addition all cases with RPGN revealed reduction of HLA-DQ, -DR, -DP or -DY+ glomerular cells. Decline of HLA-DP and/or -DR+ glomerular cells was variably seen in mesangioproliferative glomerulonephritis (MesPGN) and MPGN, whereas in FGS HLA-DQ antigens appeared to be increased in glomeruli. HLA-DQ, -DR, -DY+ interstitial cellular infiltrates were present in RPGN, FGS and MPGN and only occasionally occurred in other forms of GN. Altered renal expression of HLA-class II antigens may indicate specific sites of immunologically-mediated kidney injuries in GN.     

A comparison of the clinical and laboratory features of thin basement membrane disease (TBMD) and IgA glomerulonephritis (IgA GN).

AIM: The aim of this study was to determine the clinical and laboratory characteristics that distinguished thin basement membrane disease (TBMD) from IgA glomerulonephritis (IgA GN) at presentation and at follow-up. PATIENTS AND METHODS: Seventy-one patients with TBMD and 31 with IgA GN were studied. Males accounted for 11/71 (15%) patients with TBMD, and 20/31 (65%) of those with IgA GN (p < 0.001). RESULTS: At presentation, patients with TBMD had hematuria (42%) or proteinuria (42%), and sometimes both (24%), while those with IgA GN usually had both hematuria and proteinuria (71%, p < 0.0001). Furthermore, patients with IgA GN were more likely to have higher urinary RBC counts (p < 0.001), and more proteinuria (p < 0.001) than those with TBMD. An elevated serum creatinine or blood pressure did not distinguish between TBMD and IgA GN at presentation. At review, fewer individuals with IgA GN had elevated levels of urinary RBC and protein, and the proportions were not different from those in patients with TBMD. This was presumably because the acute episode had resolved. CONCLUSION: The outcome was worse in patients with IgA GN, with 8/31 (26%) having an elevated serum creatinine and 3/31 (10%) with end-stage renal failure, compared with an elevated serum creatinine in 3/61 (5%) patients with TBMD (p < 0.02) and no patients with renal failure (p < 0.05).     

Prognostic significance of T cell associated surface antigen density changes during OKT3 therapy of renal allograft rejection

We describe 12 acute rejection episodes in 11 cadaver donor renal allograft recipients who required OKT3* rescue treatment for steroid-resistant acute rejection (9) or for severe vascular (antibody-mediated) rejection (3). There were 3 treatment failures with subsequent graft loss. Using 2-color flow cytometry the total T (CD3), B (DR+), activated T (CD3DR), T helper/inducer (CD4), T cytotoxic/suppressor (CD8) and activated T cytotoxic cell (CD8DR) subsets were analyzed before, in mid course (5 to 7 days) and at the end of 12 to 14 days of therapy with 5 mg. OKT3 intravenously daily. In parallel changes in the density of such T cell associated antigens were analyzed. Significant decreases in the mean levels of the CD3 (p less than 0.001), CD3DR (p less than 0.05), CD4 (p less than 0.05), CD8 (p less than 0.05) and CD8DR (p less than 0.05) subsets were observed at mid course. A significant decrease in the density of CD3 was observed (p less than 0.0001). The surface antigen density of CD3DR, CD4 and CD8 had decreased by 160% (p less than 0.002), 383% (p less than 0.001) and 260% (p less than 0.001), respectively. At the end of treatment CD3 and CD4 subset levels increased by 425% and 240% (p less than 0.001 and p less than 0.005), respectively. In contrast, the CD3DR and CD8DR subset levels continued to decrease (p less than 0.05). A higher pre-treatment level of CD3DR and a less sharp decrease in CD3, CD4 and CD8 subsets were associated with a higher risk of treatment failure (p less than 0.05, p less than 0.01, p less than 0.05 and p less than 0.05, respectively). The mean decrease in the density of the CD3 marker in the lost grafts was significantly smaller compared to successful outcomes (p less than 0.001). The results of this preliminary study suggest that OKT3 affects T cell associated antigens other than CD3. Such may provide a sensitive prognostic index for the effectiveness of OKT3 therapy, and permit the identification of those patients who might require higher doses and/or duration of OKT3 therapy to enhance renal allograft salvage rates.     

Association between activated renin angiotensin system and bone formation in hemodialysis patients: is the bone mass genetically determined by ACE gene polymorphism?

BACKGROUND: Angiotensin II (ang II) receptor subtype I binding sites has been recently demonstrated on bone cell precursors. Ang II stimulates DNA and collagen synthesis in human adult bone cells. The aim of this study is to evaluate the role of renin angiotensin system in the bone metabolism and to address the genetic influence of angiotensin converting enzyme (ACE) gene polymorphism on bone mass in hemodialysis patients. METHODS: Forty-eight end-stage renal disease patients (28 male, 20 female mean age 42+/-13 years,) on maintenance hemodialysis were included in the study. Bone mineral density (BMD) was estimated at lumbar spine and T score worse than -1.5 were considered as osteopenia. Serum parathyroid hormone (iPTH) and osteocalcin (OC), bone alkaline phosphatase (bAP) and carboxy terminal propeptide type 1 collagen (PICP) levels were measured as markers of bone metabolism. Plasma renin activity (PRA), serum ACE activity and ACE gene polymorphism (II, ID, DD) were determined. RESULTS: Bone mineral density and T score of the hemodialysis patients were 0.92+/-0.17 g/cm2 and -1.36+/-1.50, respectively. Twenty-one patients (43,7%) were osteopenic (T score worse than -1.5) and mean T score of osteopenic patients was -2.72+/-0.72. T score of nonosteopenic group was -0.29+/-0.99. Serum calcium, serum, phosphorus, serum OC, serum bAP, serum PCIP, serum PTH levels were similar in osteopenics and nonosteopenics. No difference was observed in predialysis PRA and in both pre- and postdialysis serum ACE activity of patients in both groups. PRA after hemodialysis in nonosteopenic group was higher than osteopenics (p<0.05). Percent increment in PRA in hemodialysis patients was correlated with T score (R=0.48 p <0.05). Serum ACE activity was positively correlated with serum iPTH (R=0.29, p=0.02), serum OC (R=0.35, p=0.01), serum bAP (R=0.34, p=0.01), serum PCIP (R=0.36, p=0.01). T score (-0.7+/-1.5, vs -1.7+/-1.3 p <0.05) was higher in DD group (n=19) compared to II+ID group (n=29). CONCLUSIONS: Association of biochemical and radiological signs of increased bone formation with activated RAS in hemodialysis patients might be an evidence for the involvement of this system in the regulation of bone metabolism.     

Elevated dietary protein intake impairs the renal hemodynamic response to hyperaminoacidemia in patients with primary glomerular diseases

We have evaluated the renal hemodynamic response to a mixed amino acid infusion in 7 control subjects and in 8 patients with primary glomerulonephritis (GN). In order to evaluate the role of dietary protein intake in this response, GN patients were maintained for 3 weeks on two separate dietary regimens providing 130 +/- 5 g of protein/day (study 1) and 60 +/- 3 g of protein/day (study 2), respectively. Normal subjects were studied while consuming a free diet. In GN patients, following the reduction in dietary protein intake basal RPF and GFR decreased from 589 +/- 109 to 422 +/- 81 ml/1.73 m2/min (p less than 0.01, vs. study 1) and from 75 +/- 7 to 70 +/- 8 ml/1.73 m2/min (p = NS). Filtration fraction rose from 0.14 +/- 0.02 to 0.19 +/- 0.03 (p less than 0.05). In study 1, during amino acid infusion GFR and RPF did not change significantly from baseline (75 +/- 7 vs. 66 +/- 8 ml/1.73 m2/min at 180 min and 589 +/- 109 vs. 567 +/- 102 ml/1.73 m2/min, respectively). These results are at variance with data obtained in normal controls in whom both GFR and RPF rose significantly following hyperaminoacidemia. In contrast, when dietary protein intake was reduced, a normal renal hemodynamic response to amino acid infusion was restored (GFR went from 70 +/- 8 to 90 +/- 18 ml/1.73 m2/min and RPF from 422 +/- 81 to 517 +/- 90 ml/1.73 m2/min, both p less than 0.05 vs. basal), both absolute and percentage increases were similar to what was observed in controls.(ABSTRACT TRUNCATED AT 250 WORDS)     

Expression of complement 3 receptors (CR1 and CR3) on neutrophils and erythrocytes in patients with IgA nephropathy

Expression of C3 receptors (CR1 and CR3) on neutrophils (PMN) was measured in 26 patients with IgA nephropathy (IgA N), 17 normal persons, and 8 patients with non-IgA glomerulonephritis (non-IgA GN) by fluorescence activated cell sorting after labeling with monoclonal antibodies. Mean channel fluorescence for both CR1 and CR3 on PMN was significantly higher in IgA N patients than in either control group (p less than 0.01). (CR1 mean +/- SD 42.5 +/- 10.4 in IgA N, 31.7 +/- 11.5 in normal controls, 27.8 +/- 9.0 in non-IgA GN; CR3 94.0 +/- 16.5, 75.0 +/- 16.6 and 76.7 +/- 15.6, respectively.) No differences were found between the two control groups or between IgA N patients with normal and impaired renal function. These results imply that PMN are activated in IgA N patients. The expression of CR1 and CR3 of PMN may be upregulated by immune complexes (ICs), enhancing both phagocytosis of C3b- or iC3b-coated particles, and absorptive pinocytosis of soluble ICs containing C3b or iC3b. Erythrocyte CR1 and CR3 expression was measured by ELISA and found to be slightly but significantly lower in IgA N patients than in the other 2 groups (CR1 85.3 +/- 28.4 in IgA N, 113.1 +/- 28.8 in normal controls, 109.4 +/- 28.2 in non-IgA GN, p less than 0.02; CR3 80.9 +/- 20.0, 100.4 +/- 19.9, 101.2 +/- 24.9, respectively, p less than 0.05).     

Apoptosis and myofibroblast expression in human glomerular disease: a possible link with transforming growth factor-beta-1

BACKGROUND/AIMS: The pathophysiological pathways involved in the pathogenesis and evolution of renal fibrosis, have not been fully elucidated. Transforming growth factor-beta(1) (TGF-beta(1)) is involved in the development of renal scarring. Apoptosis is responsible for intrinsic cell deletion observed in end-stage kidney disease. Myofibroblasts are involved in the development of renal fibrosis. This study investigates whether there is a potential relationship between apoptosis, myofibroblast infiltration and TGF-beta(1) expression in the kidney of patients with glomerulonephritis (GN). METHODS: Forty patients with various types of GN were included in the study. Myofibroblasts and TGF-beta(1) positive cells were detected in kidney biopsies by immunohistochemistry, while apoptotic cells were detected by the in situ end labelling of fragmented DNA. RESULTS: Myofibroblasts were identified in the glomeruli of some patients with severe mesangioproliferative GN and glomerulosclerosis but a more intensive myofibroblast expression was found in the renal interstitium. TGF-beta(1) was expressed in the cytoplasm of tubular epithelial cells, in the renal interstitium and in the glomeruli of patients with GN. Apoptotic cells were mainly detected in the tubules and interstitium and were present in areas with intense myofibroblast infiltration. Positive correlations were observed between the intensity of myofibroblast expression in the interstitium and apoptosis in the tubulointerstitial area (r = 0.521, p < 0.01) as well as TGF-beta(1) expression (r = 0.462, p < 0.05) and degree of renal impairment (r = 0.430, p < 0.05). CONCLUSIONS: These observations suggest that myofibroblast infiltration and apoptosis along with TGF-beta(1) expression are associated with the development of interstitial fibrosis in patients with glomerular disease.     

Expression of HLA-DR antigens on peripheral blood T lymphocytes and renal graft tubular epithelial cells in association with rejection

Since the expression of HLA-DR antigens on peripheral blood T lymphocytes and renal graft tubular epithelial cells may be associated with immunological stimulation, we investigated the expression of these antigens on blood T lymphocytes and graft tubular epithelium in 84 renal transplant patients. Peripheral blood T lymphocytes were monitored by flow cytometry during the first 3 months after transplantation. Since the DR+ lymphocytes were selected by a double-labeling technique used for the Leu2a phenotype, the majority of the DR+ lymphocytes were also Leu2a+ with a small percentage of unidentified Leu2a- lymphocytes. Forty-one patients were treated with cyclosporine (CsA) and 43 with azathioprine (Aza), while both groups received low-dose steroids. Frozen sections of 57 renal biopsies from 43 patients (21 on CsA and 22 on Aza) were stained for HLA-DR antigens. In the Aza group, clinical rejection episodes correlated with an increased percentage of DR+ peripheral lymphocytes (P = 0.0005), and the expression of DR antigens on graft epithelial cells (P less than 0.001). In the CsA group, no relation between the expression of DR antigens on blood lymphocytes and clinical rejection episodes was evident, and the correlation between tubular DR staining and clinical rejection episodes was weaker than in the Aza group (P = 0.03). In both the Aza and CsA group, an increase in DR+ peripheral lymphocytes correlated with positive staining of the renal tubular cells for HLA-DR antigens (P less than 0.001).     

Renal functional reserve in patients with a reduced number of functioning glomeruli

Renal functional reserve (RFR) has been reported to be either reduced or absent in patients with renal insufficiency. Our study consisted in measuring RFR by acute protein load (PL) in 3 groups of patients: the first one was composed of 20 patients (pts) with biopsy-proven glomerular disease (GN) and a varying percentage of sclerotic glomeruli (15-70%); the second one consisted of 10 patients with acquired single kidney (SK) and the third group contained 5 patients with surgical ablation of more than 50% renal tissue (LRRM). Twenty-four healthy volunteers were studied as control subjects. The GFR percentage increase (delta GFR%) after PL in CS did not differ from that of the three groups of patients, despite a significant difference in resting GFR (CS = 113 +/- 11 ml/min/1.73 m2: GN 72 +/- 28 ml/min/1.7, p less than 0.01 vs CS; SK 81 +/- 20 ml/min/1.73 m2, p less than 0.01 vs CS; LRRM 45 +/- 10 ml/min/1.7, p less than 0.01 vs CS; Moreover, an inverse correlation was not found either between GFR and the percentage of sclerotic glomeruli in GN (r = 0.01, p = NS) or between GFR and the extent of excised renal tissue in the other two groups (r = 0.38, p = NS). In conclusion, our data do not confirm that RFR is necessarily reduced or absent in patients with a reduced number of functioning glomeruli, nor do they uphold the hypothesis of constant hyperfiltration in the remaining glomeruli.     

Single-dose rATG induction at renal transplantation: superior renal function and glucoregulation with less hypomagnesemia

Stevens RB, Lane JT, Boerner BP, Miles CD, Rigley TH, Sandoz JP, Nielsen KJ, Skorupa JY, Skorupa AJ, Kaplan B, Wrenshall LE. Single‐dose rATG induction at renal transplantation: superior renal function and glucoregulation with less hypomagnesemia.
Clin Transplant 2012: 26: 123–132.
© 2011 John Wiley & Sons A/S. Abstract: Background: Rabbit anti‐thymocyte globulin (rATG) induction reduces reperfusion injury and improves renal function in kidney recipients by means of properties unrelated to T‐cell lysis. Here, we analyze intensive rATG induction (single dose, rATG_S, vs. divided dose, rATG_D) for improved renal function and protection against hyperglycemia. Methods: Patients without diabetes (n = 98 of 180) in a prospective randomized trial of intensive rATG induction were followed for six months for the major secondary composite end point of impaired glucose regulation (hyperglycemia and new‐onset diabetes after transplantation, NODAT). Prospectively collected data included fasting blood glucose and HbA_1c. Serum Mg⁺⁺ was routinely collected and retrospectively analyzed. Results: Induction with rATG_S produced less impaired glucose regulation (p = 0.05), delayed NODAT development (p = 0.02), less hyperglycemia (p = 0.02), better renal function (p = 0.04), and less hypomagnesemia (p = 0.02), a factor associated with a lower incidence of NODAT. Generalized linear modeling confirmed that rATG_S protects against a synergistic interaction between tacrolimus and sirolimus that otherwise increased hypomagnesemia (p = 0.008) and hyperglycemia (p = 0.03). Conclusions: rATG_S initiated before renal reperfusion improved early renal function and reduced impaired glucose regulation, an injury by diabetogenic maintenance agents (tacrolimus and sirolimus).     

Altered intragraft immune responses and improved renal function in MHC class II-deficient mouse kidney allografts

BACKGROUND: During renal allograft rejection, expression of MHC class II antigens is up-regulated on the parenchymal cells of the kidney. This up-regulation of MHC class II proteins may stimulate the intragraft alloimmune response by promoting their recognition by recipient CD4+ T cells. In previous studies, absence of donor MHC class II antigens did not affect skin graft survival, but resulted in prolonged survival of cardiac allografts. METHODS: To further explore the role of MHC class II antigens in kidney graft rejection, we performed vascularized kidney transplants using donor kidneys from A(beta)b-deficient mice that lack MHC class II expression. RESULTS: At 4 weeks after transplant, GFR was substantially depressed in control allografts (2.18+/-0.46 ml/min/kg) compared to nonrejecting isografts (7.98+/-1.62 ml/min/kg; P<0.01), but significantly higher in class II- allografts (4.38+/-0.60 ml/min/kg; P<0.05). Despite the improvement in renal function, class II- allograft demonstrated histologic features of acute rejection, not unlike control allografts. However, morphometric analysis at 1 week after transplantation demonstrated significantly fewer CD4+ T cells infiltrating class II- allografts (12.8+/-1.2 cells/mm2) compared to controls (25.5+/-2.6 cells/mm2; P=0.0007). Finally, the intragraft profile of cytokines was altered in class II- allografts, with significantly reduced expression of Th2 cytokine mRNA compared to controls. CONCLUSIONS: These results support a role of MHC class II antigens in the kidney regulating immune cells within the graft. Further, effector pathways triggered by class II antigens promote renal injury during rejection.     

The effect of steroids on the regulation of major histocompatibility complex-class II expression on nonlymphoid tissue

The induction of MHC-class II antigens on human nonlymphoid tissues plays an essential role during the inhibition and augmentation of the immune response. Steroids have long been shown to possess strong immunosuppressive properties and successful steroid treatment has been associated with the absence of MHC-class II antigens on grated tissues. In this study we more specifically investigated the effect of steroids on the regulation of MHC-class II expression on nonlymphoid tissue. First, the influence of prednisolone on the induction process of the MHC-class II antigens on nonlymphoid tissue was determined. For this purpose vascular endothelial cells, kidney epithelial cells, fibroblasts, and a human colon tumor cell line were incubated with rIFN-gamma or primary MLC supernatant in the absence or presence of different concentrations of prednisolone. It was demonstrated that the induction process of MHC-class II antigens on these cell types was not affected by the drug at the different concentrations tested (1, 10, and 100 micrograms/ml). Next, the effect of prednisolone on the production of MHC-class II inducing factors was investigated. The drug was added at initiation of culture to primary MLC and the MHC-class II-inducing capacity of the supernatants was determined on day 7. Prednisolone at concentrations of 0.5-100 micrograms/ml clearly inhibited the overall production of factors responsible for MHC-class II induction on nonimmunological cells. The drug inhibited the production of IFN-gamma as well as non-IFN-gamma MHC-class II-inducing mediators. At a concentration of 1 microgram/ml the production of IFN-gamma and non-IFN-gamma MHC-class II-inducing mediators was reduced by, respectively, 85 per cent and 75 per cent. At a concentration of 10 micrograms/ml the production of both IFN-gamma and non-IFN-gamma mediators was almost completely inhibited. It is concluded that steroids downregulate the expression of MHC-class II antigens on nonlymphoid tissue by the inhibition of the production of MHC-class II-inducing mediators. However, once these mediators are present, the induction process itself is not affected by the drug.     

The clinico-pathological studies of hepatitis B virus nephropathy in adults

The purpose of this study is to clarify the clinical and pathological features of glomerulonephritis associated with hepatitis B virus infection (HBV-GN) in adults. Of the 47 adult cases with HBV-GN, 7 cases were diagnosed as minor glomerular abnormalities, 19 as mesangial proliferative GN (mild 13, moderate 4, severe 2), 11 as membranous GN, 10 as membranoproliferative GN (type I 2, type III 8). Indirect immunoperoxidase method using monoclonal antibody raised against HBV related antigens (HBsAg and HBeAg) revealed glomerular deposition of only HBsAg in 10 cases, only HBeAg in 2, and both antigens in 10. Deposition of HBeAg was observed dominantly along the capillary walls in comparison with that of HBsAg (p less than 0.01). Additionally, in 4 cases diagnosed as the IgA-GN because of the IgA dominant mesangial deposition and normal liver function, HBV related antigens were detected in the mesangial areas. Aggravation of renal function with respect to serum creatinine level, only 0.6-0.8 mg/dl increased, was demonstrated in 4 of 27 cases followed up for more than a year. These results suggest that the high incidence of membranous GN and membrano-proliferative GN was observed in HBV-GN in adults. HBsAg as well as HBeAg may contribute to the pathogenesis of this glomerulonephritis, and then HBsAg may play in capillary walls and mesangial areas, whereas HBeAg in capillary walls mainly. And the possibility exists that HBV related antigens are the responsible antigenic agents in some cases of IgA-GN.     

Mast cells in rapidly progressive glomerulonephritis.

The role of mast cells (MC) in tubulointerstitial damage in glomerulonephritis (GN) is not fully understood. The distribution of MC was compared in renal biopsies from 50 patients with different stages of rapidly progressive GN (RPGN) and in 20 control samples. The immunoreactivity of renal MC with anti-tryptase and anti-chymase antibodies was studied. Interstitial myofibroblasts were stained with anti-alpha-smooth muscle actin (alpha-SMA) antibody, and inflammatory cells were identified by anti-CD3, -CD20, and -CD68 monoclonal antibodies. Positively stained cells were counted, and the relative interstitial and fractional areas of anti-alpha-SMA-stained cells were measured. MC were rarely found in control samples. In contrast, samples showing crescentic GN contained numerous tryptase-positive MC (MC(T)) (43.7+/-4.65 versus 7.14+/-1.3/mm2) and fewer tryptase- and chymase-positive MC (MC(TC)) (13.8+/-1.86 versus 1.89+/-0.86/mm2) in the renal interstitium but never in the glomerulus. Double immunostaining demonstrated the presence of both phenotypes of MC. Accumulation of MC was significantly correlated with the numbers of T lymphocytes (MC(T), r = 0.67) and interstitial macrophages (MC(T), r = 0.455). There was also a significant correlation between the number of MC(T) and the relative interstitial area. The number of MC(TC) was well correlated with the fractional area of alpha-SMA-positive interstitium (r = 0.749) and the percentage of the interstitial fibrotic area (r = 0.598). There was also a significant negative correlation between interstitial MC(TC) accumulation and creatinine clearance (r = 0.661). The density of MC(TC) was higher (1.4-fold) in advanced forms of GN associated with fibrocellular crescents and interstitial fibrosis. These results show the potential involvement of MC in the fibroproliferative process in the renal interstitium of patients with RPGN. The results indicate that these cells constitute part of the overall inflammatory cell accumulation in RPGN.     

Circulating immune complexes in glomerulonephritis: a longitudinal study

A longitudinal study of circulating immune complexes (CIC) was performed in 121 patients with biopsy verified glomerulonephritis (GN). 1286 blood samples were obtained during a mean observation period of 21 months. Two methods for detection of CIC were used, the Clq-binding activity and a PEG precipitation test. CIC were detected by both tests in 21% of all blood samples and detected in at least one blood sample from 57 patients. The presence of CIC was found to be either transient (34 patients), intermittent (11 patients) or permanent (12 patients). CIC were found transiently at the time of renal biopsy and disappeared within months in patients with idiopathic extracapillary GN (7 of 9 patients), endocapillary GN (2/2) and GN associated with polyarteritis nodosa (5/6), Wegener's granulomatosis (3/3) and Henoch-Schoenlein syndrome (3/6). CIC were detected either transiently, intermittently or permanently for years after renal biopsy in patients with SLE (12/14) and membranoproliferative GN type I (7/12). CIC were only occasionally detected in patients with minor change nephropathy (1/9), membranoproliferative GN type II (0/2), IgA nephropathy (6/17), focal segmental sclerosis (1/8) and membranous GN (2/11). In these patients CIC were often transiently present without apparent relationship to time since renal biopsy. Overall, a relationship was found between the presence of CIC and decreasing serum creatinine, but there was no correlation with changes in proteinuria or with increasing blood pressure. Serial measurements of CIC showed correlations with clinical events only in individual patients, but not in the population as a whole.     

长链非编码RNA氧化应激反应丝氨酸丰富1反义RNA 1对肾癌细胞增殖、迁移及侵袭的影响

目的探讨长链非编码RNA氧化应激反应丝氨酸丰富1反义RNA 1(lncRNA OSER1-AS1)对肾癌ACHN细胞的增殖、迁移、侵袭的影响及其对微小RNA(microRNA,miR)-612的调控作用。方法2017年1月至2020年3月,采用实时定量反转录聚合酶链反应(RT-qPCR)法检测肾癌组织、癌旁组织中OSER1-AS1、miR-612的表达量;体外培养肾癌细胞ACHN,分别将OSER1-AS1小分子干扰RNA(si-OSER1-AS1)及其阴性对照(si-NC)、miR-612寡核苷酸模拟物(miR-612 mimics)及阴性对照mimic NC序列(miR-NC)、si-OSER1-AS1与miR-612特异性寡核苷酸抑制剂的阴性对照(anti-miR-NC)、si-OSER1-AS1与miR-612特异性寡核苷酸抑制剂(anti-miR-612)转染至ACHN细胞;采用RT-qPCR法检测细胞中OSER1-AS1、miR-612的表达量;采用噻唑蓝(MTT)、Transwell小室实验分别检测细胞增殖、迁移及侵袭能力;双荧光素酶报告实验检测OSER1-AS1、miR-612的靶向关系;蛋白质印迹法(Western blot)检测细胞周期蛋白1(Cyclin D1)、基质金属蛋白酶(MMP)-2、MMP-9、p21蛋白表达量。两组间比较采用独立样本t检验,多组间比较采用单因素方差分析。结果肾癌组织OSER1-AS1的表达水平(1.00±0.08比3.37±0.28)高于癌旁组织(t=52.113,P<0.05),miR-612的表达水平(1.00±0.06比0.47±0.04)低于癌旁组织(t=47.062,P<0.05);si-OSER1-AS1组细胞活力(0.66±0.05比0.31±0.03)与Cyclin D1(0.63±0.05比0.25±0.02)、MMP-2(0.82±0.07比0.35±0.03)、MMP-9(0.76±0.06比0.28±0.03)蛋白水平低于si-NC组(t=18.007、21.169、18.514、21.466,P<0.05),si-OSER1-AS1组迁移细胞数[(115.56±9.52)个比(55.99±5.59)个]、侵袭细胞数[(93.41±6.09)个比(46.05±4.35)个]低于si-NC组(t=16.188、18.984,P<0.05),si-OSER1-AS1组p21蛋白水平(0.15±0.02比0.57±0.05)高于si-NC组(t=23.398,P<0.05);miR-612组细胞活力(0.68±0.05比0.39±0.03)与Cyclin D1(0.66±0.05比0.28±0.03)、MMP-2(0.85±0.06比0.46±0.03)、MMP-9(0.78±0.05比0.34±0.02)蛋白水平低于miR-NC组(t=14.920、19.551、17.441、24.512,P<0.05),miR-612组迁移细胞数[(118.76±9.87)个比(64.39±4.65)个]、侵袭细胞数[(99.65±9.12)个比(53.57±3.67)个]低于miR-NC组(t=14.950、14.062,P<0.05),miR-612组p21蛋白水平(0.14±0.02比0.51±0.04)高于miR-NC组(t=24.820,P<0.05);双荧光素酶报告实验证实OSER1-AS1可靶向结合miR-612;抑制miR-612表达可明显逆转干扰OSER1-AS1对细胞增殖、迁移及侵袭的作用。结论干扰OSER1-AS1可通过上调miR-612的表达从而抑制肾癌ACHN细胞增殖、迁移及侵袭。     

Expression of the intercellular adhesion molecule-1 (ICAM-1) in human renal allografts and cultured human tubular cells.

The in vivo distribution of the intercellular adhesion molecule ICAM-1 was investigated in renal tissue specimens obtained from 17 renal allotransplanted patients, nine normal kidneys (controls), seven native kidneys, nine patients with mesangioproliferative glomerulonephritis, and nine patients with active extracapillary glomerulonephritis. Biopsies from patients with signs of acute rejection showed a significant increase in ICAM-1 expression in the tubular epithelium (P less than 0.05). In normal kidneys (controls) ICAM-1 expression was found in endothelial cells; additional expression in the tubular epithelial cells was induced in patients with extracapillary glomerulonephritis. The in vitro expression of ICAM-1 was examined in cultured human tubular cells after stimulation with gamma-interferon and interleukin-1, treatment with cyclosporin and/or verapamil and coculture with allogenic mononuclear cells. An increased ICAM-1 expression was demonstrated by coculture with allogenic mononuclear cells and after stimulation with gamma-interferon and/or interleukin-1. Cyclosporin or verapamil induced no changes. Our results give support to the hypothesis that ICAM-1 upregulation is important in immune interactions such as allograft rejection. Furthermore, the in vitro model indicates that ICAM-1 expression is regulated by gamma-interferon and interleukin-1 produced by activated T lymphocytes and macrophages.