A bispecific antibody targeting HLA-DQ2.5-gluten peptides potently blocks gluten-specific T cells induced by gluten ingestion in patients with celiac disease.

The gluten-free diet for celiac disease (CeD) is restrictive and often fails to induce complete symptom and/or mucosal disease remission. Central to CeD pathogenesis is the gluten-specific CD4+ T cell that is restricted by HLA-DQ2.5 in over 85% of CeD patients, making HLA-DQ2.5 an attractive target for suppressing gluten-dependent immunity. Recently, a novel anti-HLA-DQ2.5 antibody that specifically recognizes the complexes of HLA-DQ2.5 and multiple gluten epitopes was developed (DONQ52). OBJECTIVE: To assess the ability of DONQ52 to inhibit CeD patient-derived T-cell responses to the most immunogenic gluten peptides that encompass immunodominant T cell epitopes. METHODS: We employed an in vivo gluten challenge model in patients with CeD that affords a quantitative readout of disease-relevant gluten-specific T-cell responses. HLA-DQ2.5+ CeD patients consumed food containing wheat, barley, or rye for 3 days with collection of blood before (D1) and 6 days after (D6) commencing the challenge. Peripheral blood mononuclear cells were isolated and assessed in an interferon (IFN)-γ enzyme-linked immunosorbent spot assay (ELISpot) testing responses to gluten peptides encompassing a series of immunodominant T cell epitopes. The inhibitory effect of DONQ52 (4 or 40 µg/mL) was assessed and compared to pan-HLA-DQ blockade (SPVL3 antibody). RESULTS: In HLA-DQ2.5+ CeD patients, DONQ52 reduced T cell responses to all wheat gluten peptides to an equivalent or more effective degree than pan-HLA-DQ antibody blockade. It reduced T cell responses to a cocktail of the most immunodominant wheat epitopes by a median of 87% (IQR 72-92). Notably, DONQ52 also substantially reduced T-cell responses to dominant barley hordein and rye secalin derived peptides. DONQ52 had no effect on T-cell responses to non-gluten antigens. CONCLUSION: DONQ52 can significantly block HLA-DQ2.5-restricted T cell responses to the most highly immunogenic gluten peptides in CeD. Our findings support in vitro data that DONQ52 displays selectivity and broad cross-reactivity against multiple gluten peptide:HLA-DQ2.5 complexes. This work provides proof-of-concept multi-specific antibody blockade has the potential to meaningfully inhibit pathogenic gluten-specific T-cell responses in CeD and supports ongoing therapeutic development.

Actions of porcine pancreatic and Bacillus subtilis alpha-amylases and Aspergillus niger glucoamylase on phosphorylated (1--4)-alpha-D-glucan.

Porcine pancreatic alpha-amylase (1,4-alpha-D-glucan glucanohydrolase, EC 3.2.1.1) produced O-6-phosphoryl-alpha-D-glucopyranosyl-(1--4)-O-alpha-D-glucopyranosyl -(1--4)-D-glucopyranose (6(3)-phosphoryl maltotriose) and O-alpha-D-glucopyranosyl-(1--4)-O-alpha-D-glucopyranosyl- O-3-phosphoryl-alpha-D-glucopyranosyl-(1--4)-D-glucopyranose (3(2)-phosphoryl maltotetraose) from potato starch upon exhaustive hydrolysis, while Bacillus subtilis alpha-amylase (liquefying type) yielded O-alpha-D-glucopyranosyl-(1--4)-O-6-phosphoryl-alpha-D- -glucopyranosyl-(1--4)-D-glucopyranose (6(2)-phosphoryl maltotriose) and 3(2)-phosphoryl maltotetraose. Thus, the two alpha-amylases cleave different sites in the vicinity of the phosphate group at C-6 of the glucosyl residue but the same site in the vicinity of that at C-3. Aspergillus niger glucoamylase (1,4-alpha-D-glucan glucohydrolase, EC 3.2.1.3) hydrolyzed the (1--4)-alpha-linkage of the 6-phosphoryl residue at the non-reducing site (Abe, J., Takeda, Y. and Hizukuri, S. (1982) Biochim. Biophys. Acta 703, 26-33) but not that of the 3-phosphoryl residue, leaving a glucosyl residue attached to the 3-phosphoryl residue. These results indicate that the phosphate group at C-3 is more obstructive than that at C-6 for the actions of these three amylases.

Involvement of collagen-binding heat shock protein 47 and procollagen type I synthesis in idiopathic pulmonary fibrosis: contribution of type II pneumocytes to fibrosis.

The most common pathologic form of idiopathic pulmonary fibrosis is usual interstitial pneumonia, which is characterized by patchy fibrotic areas, marked increase in the number of fibroblasts and type II pneumocytes, and excessive deposition of extracellular matrix proteins, especially collagen. Heat shock protein 47 is a collagen-binding stress protein and has a specific role in intracellular processing of procollagen molecules as a collagen-specific molecular chaperone. However, its role in the causation of fibrosis in usual interstitial pneumonia is unknown. In this study, we examined the expression of heat shock protein 47 and type I procollagen in 12 patients with usual interstitial pneumonia by immunohistochemistry on sequential sections. Heat shock protein 47 was localized predominantly in alpha-smooth muscle actin-positive myofibroblasts and surfactant protein-A-positive type II pneumocytes in active fibrotic areas of usual interstitial pneumonia. Type I procollagen was also expressed in those cells. In contrast, heat shock protein 47 and type I procollagen were weakly or not at all expressed in myofibroblasts and type II pneumocytes in bronchiolitis obliterans organizing pneumonia and normal lung tissue samples obtained from excised lung cancer tissues. The numbers of heat shock protein 47- and type I procollagen-positive cells to type II pneumocytes or myofibroblasts were significantly higher in usual interstitial pneumonia than in bronchiolitis obliterans organizing pneumonia and normal lung tissue specimens. Our results suggest that myofibroblasts and type II pneumocytes play an important role in the progression of fibrosis through the induction of heat shock protein 47, which regulates the synthesis/assembly of type I procollagen in usual interstitial pneumonia. HUM PATHOL 31:1498-1505.

Glomerulocystic kidney disease (GCKD) associated with Henoch-Schoenlein purpura: a case report and a review of adult cases of GCKD.

We report on a 71-year-old male with Henoch-Schoenlein purpura (HSP) who developed glomerulocystic kidney disease (GCKD) without congenital abnormality. He had mild renal dysfunction. Renal biopsy findings showed mild proliferation of mesangial cells and matrixes, and tubular atrophy, interstitial fibrosis, cystic dilation of Bowman's capsule and atrophy of the glomerular tuft. The deposition of IgA and C3 in the mesangial area was observed with the fluorescent antibody technique. Therefore he was diagnosed with GCKD-associated HSP. This was the oldest patient among the previous case reports and the patient was the first case to be reported for concurrent GCKD and HSP. In this study, we also reviewed the patient to previously reported adult patients with GCKD.

Successful treatment with cyclosporine in a case of hemophagocytic syndrome manifesting as severe liver dysfunction.

Forms of hemophagocytic syndrome, which affects mainly children, vary from mild to very severe and often fatal. We describe an adult patient with hemophagocytic syndrome in whom severe liver dysfunction developed. The condition continued to deteriorate despite treatment with plasma exchange, high-dose gamma globulin, and corticosteroid therapy. Treatment with cyclosporine (2.3 mg/kg/day) dramatically improved the condition and normalized liver function. Cyclosporine reduced the serum levels of ferritin, interferon-tau, interleukin-6, and soluble interleukin-2 receptor. These findings suggest that hemophagocytic syndrome accompanied with severe liver dysfunction results from hypercytokinemia, and cyclosporine is useful in preventing a fatal outcome during the acute phase.

Acute tubular necrosis with loin pain and persistent multiple wedge-shaped contrast enhancement on CT.

A 38-year-old man had severe loin pain and computed tomography performed 48 hours later, after drip infusion pyelography (DIP), revealed wedge-shaped contrast enhancement. He showed mild impairment of renal function with no evidence of rhabdomyolysis. The loin pain lasted for 5 days and the wedge-shaped contrast enhancement on CT persisted for 14 days and improved. The case was compatible with the new syndrome, loin pain and persistent wedge-shaped contrast enhancement on CT, proposed by Ishikawa et al (Nephron 27: 31, 1981).

Type II diabetes mellitus and primary Sjögren's syndrome complicated by pleural effusion.

A 73-year-old man was admitted to our hospital because of pleural effusion and nephrotic syndrome. Sjogren's syndrome (Sjs) was diagnosed based on a positive test for antibodies to Ro and La, and the result of labial salivary gland biopsy. The pleural effusion showed a high number of lymphocytes and high titers of antibodies to Ro and La. By immunohistochemistry, it was determined that infiltrating CD3+ cells predominated over infiltrating CD20+ cells in the pleura. Nephrotic syndrome was also present, which, as confirmed by renal biopsy was due to advanced diabetic nephropathy. Here, we report a case of Type II diabetes mellitus and primary Sjs complicated by pleural effusion, discuss the available treatment for pleural effusion.

The "nutcracker" phenomenon in combination with IgA nephropathy.

The "nutcracker" phenomenon results from compression of the left renal vein between the superior mesenteric artery and the aorta. The main features of this phenomenon are non-glomerular haematuria on urinalysis and stenosis of the left renal vein with dilatation of the vein distal to the stenosis. The characteristics of two patients with the "nutcracker" phenomenon complicated by immunoglobulin A nephropathy were compared with those of 10 patients showing only the "nutcracker" phenomenon. Patients with the "nutcracker" phenomenon complicated by immunoglobulin A nephropathy showed aggravation of haematuria after upper respiratory infections, urinary red cell morphology indicating haematuria of glomerular origin, elevation of serum IgA, persistence of proteinuria, and granular casts in the urine. The coexistence of immunoglobulin A nephropathy was suspected when these characteristics were observed in patients with the "nutcracker" phenomenon. In such cases, a renal biopsy is needed for a final diagnosis.

Evaluation of anti-HTLV-1 antibody in primary glomerulonephritis.

Human T-cell leukaemia virus type-1 (HTLV-1) is known to cause adult T-cell leukaemia. The prevalence of anti-HTLV-1 antibody in haemodialysis patients has been reported to be higher than that in the general population. The anti-HTLV-1 antibody-positive rate in patients with primary glomerulonephritis in the Nagasaki district, an endemic area of HTLV-1, was evaluated. The antibody-positive rates in patients with primary glomerulonephritis (9.9%) and in haemodialysis patients (18.4%) were significantly higher than the rate in general blood donors (6.6%). Of 142 patients with primary glomerulonephritis, 14 (9.9%) were positive for the antibody; histological evaluation of these patients showed minor glomerular abnormality in one, mesangial proliferative glomerulonephritis in eight (IgA nephropathy in six and non-IgA nephropathy in two), membranous nephropathy in three, and crescentic glomerulonephritis in two. Evaluation of 10 antibody-positive patients by immunofluorescent microscopy showed immunocomplex-type nephritis in nine, suggesting the involvement of HTLV-1-associated antigen in the development and progression of glomerulonephritis.

Prostaglandin E1 for renal papillary necrosis in a patient with diabetes mellitus.

We report a case of renal papillary necrosis with diabetes mellitus which was treated with prostaglandin E1. An intravenous infusion of 40 mg/day prostaglandin E1 was given for 14 days in an attempt to improve renal circulation. Treatment resulted in an improved creatinine clearance, renal plasma flow and renogram, and proteinuria was decreased. The administration of prostaglandin E1 produced an improvement in renal haemodynamics and can be considered as a possible therapy for renal papillary necrosis in diabetic patients.

[Cytomegalovirus retinitis in a patient with systemic lupus erythematosus].

We report a rare case of cytomegalovirus (CMV) retinitis in association with systemic lupus erythematosus (SLE). A 36-year-old female was admitted to our hospital because of nephrotic syndrome and renal insufficiency due to lupus nephritis. After steroid and immunosuppressant treatment for lupus nephritis, she began to complain of floaters and visual disturbance. She was diagnosed as having CMV retinitis based upon the depressed immune status and the clinical appearance of fundus. As soon as CMV retinitis was diagnosed, antiviral treatment with intravenous ganciclovir was instituted and maintained for 7 months. She was treated successfully and almost no progression or relapse of CMV retinitis was documented without ganciclovir maintenance treatment during a 2 year follow-up period. It is suggested that CMV retinitis can occur in patients with SLE treated with steroid and immunosuppressant, indicating the importance of early diagnosis and treatment for CMV retinitis.

[Tuberculosis in patients undergoing hemodialysis].

The incidence of tuberculosis is very high in patients treated by hemodialysis particularly in the early stage of hemodialysis. The diagnosis of tuberculosis in dialysis patients was obscured as symptoms were nonspecific and extrapulmonary involvement was seen frequently. We investigated cell mediated immunity in dialysis patients in relation to the period of dialysis. The data indicate that dialysis patients show the following immunological impairments; 1) lymphopenia, 2) decreased B cell, 3) alteration of T cell subset, 4) decreased reaction of PPD skin test, 5) decreased T cell activity, 6) decreased IL-2 production, 7) decreased PHA induced lymphocyte blastogenesis, 8) decreased NK cell. Decreased immunologic host defence mentioned above may contribute to the high incidence of tuberculosis in the early stage as well as in the maintenance phase of dialysis.

Effect of tonsillectomy on peripheral blood T cell surface markers and cytokine production in patients with IgA nephropathy accompanied by chronic tonsillitis.

The present study investigated changes in peripheral blood T cell surface markers and cytokines (interleukin-2, IL-2; tumor necrosis factor-alpha, TNF-alpha; and interferon-gamma, IFN-gamma) following tonsillectomy in patients with IgA nephropathy accompanied by chronic tonsillitis. Peripheral blood CD8+ cells, CD45RA+CD4+ cells and CD8+CD11b- cells increased significantly after tonsillectomy, compared with their preoperative values. In some cases, the preoperative serum TNF-alpha and IFN-gamma levels were higher than normal before surgery, but decreased after surgery. These results suggest that tonsillectomy suppresses a decrease in suppressor T cells in patients with IgA nephropathy and corrects abnormal cell-mediated immune responses in these patients.

Nephrotic syndrome in the elderly--clinicopathological study.

Clinical and pathological findings and the effects of therapy were investigated in 90 cases of nephrotic syndrome (NS) in elderly patients aged over 60 years. Membranous nephropathy was the most frequent type of primary NS. Amyloidosis and malignancy were common causes of secondary NS. Damage to the interstitium in the kidney, such as focal mononuclear cell infiltration, fibrosis and thickening of the small arterial wall in membranous cases, was often observed. Stage I and II based on electron-microscopy, were mainly observed in the patients, with membranous nephropathy. Prednisolone and immunosuppressive agent were most effective in these patients with membranous nephropathy. Prednisolone alone was the most effective on minimal change NS in the elderly. In the course of therapy, side effects such as pneumonia, sepsis due to fungus infections, such as aspergillus and candida, and infection, such as cytomegalovirus and herpes zoster, were more frequently observed, especially in the cases of MPGN, DPGN with moderate to severe mesangial proliferation, with a decline in renal function (Ccr < 50 L/day) and secondary NS. In secondary NS, the prognosis of amyloidosis was very poor and the findings pointed to a relationship between malignancy and nephrotic syndrome.

A case of acute renal failure due to ethylene glycol intoxication.

We describe an 18-year-old man with acute renal failure due to inadvertent ingestion of antifreeze that contained ethylene glycol (EG). A relatively small amount of EG was ingested, but nausea and vomiting were observed soon after ingestion. During admission to a local hospital, consciousness became impaired and generalized convulsion was noted. He was transferred to our hospital because of rapid deterioration of renal function. Emergency hemodialysis was begun. The patient underwent one treatment session of hemodialysis each day, for a total of 8 hemodialytic sessions before his renal function recovered. Examination of the renal biopsy specimen revealed degeneration of the renal tubular epithelium and presence of intratubular calcium oxalate crystals. The clinical features of the patient were mild except for acute renal failure. These findings suggest that even a small amount of EG will have toxic effects on the kidney.

[A case of systemic lupus erythematosus associated with minimal change nephrotic syndrome].

A case of systemic lupus erythematosus (SLE) associated with minimal change nephrotic syndrome (MCNS) in a 25-year-old female is described. The patient suddenly manifested butterfly rash and proteinuria was first pointed out on March, 1994. On admission, her skin biopsy indicated SLE. Subsequently, she developed nephrotic syndrome. Urinalysis showed heavy proteinuria (4.1 g/day), with no other abnormalities in the urinary sediment. Immunological examination revealed positive antinuclear antibody at a titer of 1:80 with a speckled pattern. Anti-ssDNA and anti-SS-A antibodies were positive, but other antibodies were negative. Serum complement (CH50) was within the normal range (30.5 U/ml). The renal biopsy showed no apparent cellular proliferation or increase of extracellular matrices in glomeruli by light microscopy. Slight deposition of IgG, IgM, C3 and C1q was focally seen in the mesangium and capillary wall by immunofluorescence. Electron microscopic examination revealed small and scattered dense deposits in the mesangium, subepithelium and subendothelium, associated with diffuse fusion of the foot processes of epithelial cells along the glomerular basement membrane. According to the WHO classification, the histological features were compatible with those of lupus nephritis (LN), class Ib. The patient was treated with PREDNISOLONE, Mizorbine and Dilazep, resulting in the disappearance of proteinuria and a normal serum level of total protein. The association of LN and MCNS is very rare. We also investigated the relationship between the intensity of proteinuria and histological types of 53 cases with LN examined in our laboratory. The cases with heavy proteinuria were mostly classified as WHO-Class IV and Class V. We report here a case of LN associated with MCNS and also review the literatures.