Fine-needle aspiration biopsy sampling in renal transplantation: interstitial cellular infiltrates and major histocompatibility complex class-II antigen expression in renal tubular cells

The dynamics of major histocompatibility complex (MHC) class-II (DR) antigen products in renal tubular cells and cellular infiltrations in the interstitium of the kidney following renal transplantation without immunosuppressive therapy are presented. DR antigen in 27 normal kidneys and in 18 of 19 kidneys transplanted as autograft showed no DR-antigen expression on their tubular cells. These DR-antigen-negative tubular cells could be induced to express the antigen during courses of untreated rejection both in primary (n = 6) and repeat (n = 4) allografts. Parallel to the increasing DR-antigen expression in the allograft, the autologous kidney expressed this antigen with the same time dependency and with the same intensity. Graft infiltrating cells were evaluated by daily fine-needle aspiration biopsy and core biopsy. Induction of DR antigen was associated with a significant infiltration of blastogenic cells and monocytes/macrophages in the allograft. During rejection of the allograft no cellular infiltrate was noted in the autograft, but during an initial time period after allograft removal cellular infiltrates were seen. Following both courses of rejection DR antigen returns to negative by 6-8 days as did blastogenic cells and monocytes/macrophages in the autograft interstitium. Inducible antigen expression in normally DR-antigen-negative allograft parenchymal cells during rejection is shown to depend on inflammatory cells in the graft. The antigen expression is not restricted to the tissue transplanted, but also affects autologous tissue of the host organism.     

Production of interleukin 1 in glomerular cell cultures from patients with rapidly progressive crescentic glomerulonephritis

Interleukin 1 (IL-1) activity was measured in glomerular culture supernatants from 3 patients with rapidly progressive crescentic glomerulonephritis (RPGN). Macrophages were present in both capillary tufts and cellular crescents as identified by OKM1-positive cells on immunoperoxidase labelling. Glomeruli from 4 rejecting renal cadaver allografts were used as a disease control, in addition to glomeruli from a normal kidney. IL-1 activity as measured by the thymocyte proliferation assay was greater in the supernatants from cultured glomerular outgrowths of patients with crescentic GN than in those from rejected renal allografts and glomeruli isolated from the normal tissue. IL-1 production from cultured glomerular cells from patients with RPGN was detectable in the serum-free conditioned media harvested after 3 days of culture and increased in a stepwise fashion over 28 days of culture. The prominent feature of the glomerular outgrowth of the glomeruli in the RPGN patients was the presence of large numbers of macrophages, which were not present in cultured control glomeruli. These findings indicate that the immunoregulatory aberration in patients with RPGN may in part be due to IL-1 production by activated macrophages.     

Leukocyte analysis using monoclonal antibodies in human glomerulonephritis

The leukocyte subpopulations were analyzed within both the glomeruli and the interstitium in renal biopsies from 145 patients with various forms of glomerulonephritis. Cells were identified by monoclonal antibodies to leukocyte cell-surface antigens and immunoperoxidase labelling. Leukocytes, as defined by a monoclonal antibody to the leukocyte common antigen (PHM1), were present in normal, human renal tissue in both glomeruli (2.8 +/- 0.6 cells/glom. cross section) and interstitium (102 +/- 18 cells/mm2). Monocytes constituted the predominant infiltrating cell type in normal glomeruli (1.3 +/- 0.2) and T cells were rarely found (0.3: range 0 to 0.8), whereas both monocytes (34 +/- 10/mm2) and T lymphocytes (33 +/- 14/mm2) were found in the normal interstitium. In the non-proliferative forms of glomerulonephritis there was no significant increase in the number of glomerular inflammatory cells when compared with normal glomeruli. However, significantly increased numbers of T lymphocytes were seen in the interstitium of biopsies with minor non-specific changes (67 +/- 15/mm2), membranous nephropathy (134 +/- 30/mm2), focal glomerulosclerosis (207 +/- 53/mm2), and diabetic nephropathy (198 +/- 81/mm2). In the proliferative forms of glomerulonephritis only crescentic GN and post-infectious GN demonstrated significantly-increased glomerular monocytes and granulocytes. There was no significant increase in the number of glomerular T cells when compared with normal glomeruli. However, there was a significant increase in the number of interstitial T lymphocytes in all forms of proliferative glomerulonephritis when compared with the normal interstitial cell population.(ABSTRACT TRUNCATED AT 250 WORDS)     

The role of monocyte chemotactic and activating factor (MCAF)/monocyte chemoattractant protein (MCP)-1 in subgroups of rapidly progressive glomerulonephritis

To elucidate the role of monocyte chemotactic and activating factor (MCAF)/monocyte chemoattractant protein(MCP)-1 in the pathogenesis of rapidly progressive glomerulonephritis (RPGN), we determined the urinary levels of MCAF/MCP-1 in 20 healthy subjects, 30 patients showing RPGN with crescents, and 39 patients with various types of renal diseases without crescents. We divided RPGN into two subgroups, the acute type and the insidious type, with regard to the declination rate of reciprocals of serum creatinine with time as previously reported. In addition, we divided the patients with RPGN into anti-neutrophil cytoplasmic antibody(ANCA)-related diseases and immune complex(IC)-mediated diseases with regard to etiology. Urinary levels of MCAF/MCP-1 were significantly higher in patients with RPGN as compared with those of other renal diseases and healthy volunteers(21.8 +/- 4.5 vs. 11.6 +/- 3.5, 1.0 +/- 0.1 pg/ml creatinine, respectively, p < 0.01, mean +/- SEM). There was no difference in the urinary levels of MCAF/MCP-1 between the acute and insidious types of RPGN patients. In addition, there was no difference in the urinary levels of MCAF/MCP-1 between the patients with ANCA-related and IC-mediated diseases. Urinary levels of MCAF/MCP-1 in patients with RPGN were correlated well with the percentage of both total crescents and fibrocellular/fibrous crescents and the number of CD68-positive infiltrating cells in the interstitium. Immunohistochemical examinations revealed that MCAF/MCP-1 positive cells were detected in tubular epithelial and endothelial cells and mononuclear infiltrated cells in the interstitium. Moreover, elevated urinary MCAF/MCP-1 levels in patients with RPGN, regardless of subgroups, were dramatically decreased during methylprednisolone pulse therapy induced convalescence. These results suggest that MCAF/MCP-1 may be involved in the pathogenesis of RPGN via macrophage recruitment and activation.     

Intraglomerular T cells and monocytes in nephritis: study with monoclonal antibodies

Intraglomerular T cells, monocytes, total leucocytes and other mononuclear subsets were sought in renal biopsies from patients with glomerulonephritis, using monoclonal antibodies and immunoperoxidase techniques. Twenty-four biopsies with no significant glomerular proliferation on optical microscopy, thirty-two with only endocapillary hypercellularity, and twenty-one with extra capillary crescentic glomerular disease were studied. Few intra-glomerular leucocytes were seen in the non-proliferative group. In contrast, when compared with this group, biopsies with glomerular hypercellularity and particularly those with crescents showed increased numbers of intra-glomerular total leucocytes and monocytes/macrophages, as well as an excess of T lymphocytes and T cytotoxic/suppressor cells; T helper/inducer lymphocytes were significantly increased only in the crescentic group. Only small numbers of B lymphocytes and NK cells were found in all groups. The numbers of total glomerular T-cells and monocytes per glomerular cross section were highly correlated in the crescentic group. Only idiopathic IgA nephropathy failed to show a significant increase in the numbers of intra-glomerular leucocytes, in comparison with the non-proliferative group, Henoch-Sch?nlein purpura biopsies in contrast had an excess of both monocytes and T cell subsets. The finding of T lymphocytes as well as monocytes in glomeruli of proliferative nephritis suggests that cellular immune mechanisms may play a role in their pathogenesis, especially when crescents are present.     

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.     

Detection of urinary macrophages expressing the CD16 (Fc gamma RIII) molecule: a novel marker of acute inflammatory glomerular injury

BACKGROUND: The CD16 antigen is the Fc gamma receptor III. CD14+CD16+ cells are proinflammatory monocytes/macrophages (Mo/M phi) that constitute a minor population in the peripheral blood of healthy individuals. Little is known about the expression of CD16 antigen on Mo/M phi in glomerulonephritis. METHODS: Flow cytometric analyses were performed on urine and blood samples obtained from 209 patients with various renal diseases. Patients variously suffered from rapidly progressive crescentic glomerulonephritis (RPGN), membranoproliferative glomerulonephritis (MPGN), postinfectious acute glomerulonephritis (AGN), Henoch-Sch?nlein purpura nephritis (HSPN), IgA nephropathy (IgAN), membranous nephropathy (MN), minimal change nephrotic syndrome (MCNS), lupus nephritis (LN), acute interstitial nephritis, hereditary nephropathy, idiopathic renal hematuria (IRH), and renal stone. RESULTS: The CD16+ M phi population of cells was present in the urine of hematuria-positive patients with proliferative glomerulonephritis, including AGN, IgAN, RPGN, MPGN, and LN with acute inflammatory lesions, such as endocapillary proliferation, tuft necrosis, and cellular crescents. In contrast, the urinary CD16+ M phi population was negligible in hematuria-positive patients with nonproliferative renal disease, including hereditary nephropathy, IRH, and renal stone and also in patients with proliferative glomerulonephritis lacking acute inflammatory lesions. Total urinary M phi of these patients were much less than those of patients having proliferative glomerulonephritis with acute inflammatory lesions. Transient expansion of the CD16+ M phi population in urine was observed during the acute exacerbation of urinary abnormalities, whereas the disappearance of CD16+ M phi closely preceded the amelioration of urinary abnormalities in patients with proliferative glomerulonephritis. In 38 of the 98 patients positive for CD16+ M phi population in urine, the CD16+ Mo population was negligible in peripheral blood. Immunohistochemically, CD16+ M phi were present in the glomeruli of active proliferative glomerulonephritis, whereas such cells were absent in inactive proliferative glomerulonephritis or nonproliferative glomerular diseases. CONCLUSION: CD16+ M phi may be effector cells involved in the acute inflammation common to all types of proliferative glomerulonephritis. Furthermore, the detection of CD16+ M phi in urine, as well as urinary M phi counts, may serve as a useful indicator of the active stage of proliferative glomerulonephritis.     

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.     

中性粒细胞细胞外网络与B淋巴细胞在狼疮肾炎患者肾组织的表达及其意义

目的 探讨中性粒细胞细胞外网络(NET)在狼疮肾炎(LN)患者肾组织中的表达与狼疮活动的关系.方法 采用免疫组织化学技术,分别检测NET(以瓜氨酸化的组蛋白H3为标志)在20例LN、8例微小病变(MCD)患者和3例健康对照的肾组织的表达,同时检测相应部位B淋巴细胞(CD19标志)的浸润.分析LN患者肾组织活动指数和慢性指数评分,以及它们与NET的关系.结果 LN患者肾小球中NET表达显著高于健康对照组和MCD组(1.056±0.985比0±0、0±0,均P＜0.01).在中、重度增生的系膜细胞、细胞新月体、有炎性细胞浸润的肾小管间质,NET表达显著上调;系膜细胞中、重度增生肾小球的NET表达显著高于其他类型肾小球,差异均有统计学意义(均P＜0.01).中、重度系膜细胞增生肾小球的平均NET阳性细胞数与LN病理活性指数呈正相关(r=0.620,P=0.004);与SLE-DAI评分呈正相关(r=0.492,P=0.027);与肾小管中NET阳性细胞数呈正相关(r=0.558,P=0.011).肾间质NET阳性细胞数与肾间质阳性B淋巴细胞数呈正相关(r=0.573,P=0.008);与LN病理慢性指数呈正相关(r=0.645,P=0.002).结论 NET在LN患者肾组织广泛表达,肾小球中浸润的NET可能参与了LN患者肾组织的活动性损伤.     

Cellular and non-cellular compositions of crescents in human glomerulonephritis

The composition of glomerular crescents was examined on the frozen kidney sections obtained from 10 patients (5 patients with IgA nephropathy, two with Henoch-Sch?nlein purpura nephritis and three with glomerulonephritis due to undetermined etiology) using well-defined monoclonal and polyclonal antibodies to coagulation proteins, extracellular matrices, intermediate filament proteins and immune cells. Fibrinogen/fibrin related antigens (FRA), which were stained with anti-fibrinogen serum, were positive in the crescents of all the patients, but monoclonal antibody to crosslinked fibrin or von Willebrand factor (factor VIII related) antigen did not bind to the crescents. This suggests that the FRA deposited in the crescents is fibrinogen or its degradation products rather than fibrin. Staining for intrinsic components of renal basement membrane, including type IV and V collagens, laminin and fibronectin, were consistently positive in all stages of the crescents. Cytokeratin, showing cytoplasmic staining of the glomerular parietal epithelium and tubular epithelium in the normal kidney, was demonstrated in three patients with cellular crescents. Vimentin, which is normally distributed in parietal and visceral epithelial cells in the glomeruli and interstitial cells, was found at all stages of the crescents. These findings suggest that in the early stage of crescent formation, glomerular epithelial cells play an important role, and that the accumulation of intrinsic basement membrane constituents is associated with the formation and progression of the crescents. None of the crescent cells reacted with either of two monoclonal antibodies (Mo2 and FMC 32) to monocytes/macrophages or with nonspecific esterase staining. It seems that, at least in our patients, monocytes are a minor factor contributing to the formation of glomerular crescents.     

Mononuclear leukocytes, expression of HLA class II antigens and intercellular adhesion molecule 1 in focal segmental glomerulosclerosis.

Immunophenotyping of infiltrating glomerular and interstitial mononuclear leukocytes performed in renal tissues of 15 patients with focal segmental glomerulosclerosis (FSGS) was compared to 15 normal kidneys in order to investigate a possible role for cell-mediated immunity (CMI) in FSGS. In addition, distribution of HLA class II (-DQ, -DR, -DP and -DY) antigens and of the intercellular adhesion molecule 1 (ICAM-1) as well as the expression of well-defined renal antigens along the human nephron was analyzed. In comparison to normal kidneys, a reduction in HLA class II antigens of ICAM-1 and of renal antigens defined by the monoclonal antibodies TN8-TN10 was observed in sclerotic glomeruli. Furthermore, an increased number of T lymphocytes was found in glomeruli of FSGS with slight predominance of the CD8+ subset. Interstitial inflammation was present in all FSGS cases except 1 with T lymphocytes and monocytes/macrophages constituting the predominant infiltrating cell types. In contrast to the glomerular T cells, the number of interstitial CD4+ cells was greater than the number of CD8+ cells in almost all cases. As a sign of activation, most interstitial inflammatory cells carried HLA class II antigens and some of them also expressed ICAM-1. Proximal tubular epithelial cells often presented an abnormal expression of HLA-DQ and HLA-DP antigens associated with aberrant expression of ICAM-1 and TN8. The number of interstitial mononuclear leukocytes was correlated to serum creatinine levels at the time of renal biopsy. The present results provide further support for the involvement of CMI in the pathogenesis of FSGS.     

Contribution of mononuclear leucocytes to the progression of experimental focal glomerular sclerosis

Uninephrectomized Sprague-Dawley rats repeatedly administered with aminonucleoside of puromycin and protamine sulfate developed progressive focal glomerular sclerosis (FGS). The contribution to disease progression of both glomerular and interstitial infiltrating leucocytes was studied throughout the disease evolution. Leucocyte subsets were quantitated with an immunoperoxidase technique using monoclonal antibodies for rat leucocyte surface antigens: OXI (total leucocytes) OX6 (Ia positive cells), OX8 (suppressor/cytotoxic T cells), OX19 (total T cells), OX22 (B cells and subsets of T cells), and ED1 (macrophages/monocytes). In the glomeruli, macrophages and Ia positive cells were significantly increased when sclerotic lesions appeared, but T lymphocytes and subsets of T lymphocytes were not found. However, in the interstitium, all leucocytes were identified and increased in number throughout the disease evolution. Early in the disease, monocytes and lymphocytes were both present in large numbers, but at the end stage of the process, the predominant infiltrating leucocytes were CD4+ve T cells. In FGS rats treated throughout the disease with oral prednisolone (begun after disease induction), renal function was significantly better than in the untreated group, whereas the sclerosis and leucocyte accumulation in the glomeruli were unchanged. However, prednisolone treatment resulted in significantly fewer interstitial leucocytes and especially reduced the numbers of CD4+vc cells. These results suggest that the glomerular sclerotic lesions are related to the participation of macrophages independent of T cells, and that immune mechanisms mediated by T cells in the interstitium have an important role in the progression of this disease to end-stage renal failure.(ABSTRACT TRUNCATED AT 250 WORDS)     

Proteinase-3 mRNA expressed by glomerular epithelial cells correlates with crescent formation in Wegener's granulomatosis

BACKGROUND: Wegener's granulomatosis (WG) is characterized by systemic vasculitis with crescentic glomerulonephritis (CGN) and circulating autoantibodies directed against neutrophil cytoplasmic antigens (ANCA). Proteinase 3 (PR-3), a neutral serine proteinase in neutrophils implicated in the growth control of myeloid cells, has been identified as the target antigen for ANCA in WG. Since the kidneys are frequently involved in WG, we studied the in situ expression of PR-3 by renal parenchymal cells. METHODS: We assessed the expression of PR-3 in kidney biopsies of 15 patients with WG by immunohistochemistry (IHC) and in situ hybridization (ISH). Normal kidney tissue served as the control. RESULTS: We detected PR-3 mRNA and PR-3 protein in distal tubular epithelial cells (TECs) and glomerular epithelial cells (GECs) in normal kidney tissue and in CGN. Furthermore, a strong glomerular PR-3mRNA expression restricted to the site of cellular crescents was detected in patients with WG. The analysis of 144 glomeruli with cellular or sclerotic crescents revealed a positive correlation of glomerular PR-3mRNA expression with the percentage of cellular crescents per glomerulus. The capability of human TECs and GECs to synthesize PR-3 was confirmed by Northern blot and ISH on cultured cells. CONCLUSION: These data provide evidence that nonhematopoetic renal parenchymal cells express PR-3 and that glomerular expression of PR-3 is associated with crescent formation in WG. Our findings suggest that renal parenchymal cells may directly be involved in the pathogenesis of CGN in WG.     

Definition, genetics, and possible significance of a newly defined endothelial antigen in the rat

The purpose of the present experiments was to define a non-major-histocompatibility-complex (MHC) endothelial antigen system in the rat and to study the genetics of these antigens as well as their significance in renal transplantation. Several MHC-identical rat strain combinations underwent reciprocal immunization using spleen and lymph node cells and complete Freund's adjuvant. In one combination (MAXX anti-BN) alloantibodies were found against antigens on peritubular and venous endothelium of the kidney from the immunizing strain as well as from two other strains. Preliminary results suggested that the endothelial antigen is present on monocytes but not on nonstimulated T and B lymphocytes. With kidneys from 7 MHC-congenic lines it was shown that the endothelial antigens are encoded outside the MHC-region. The antigen seems to be expressed as a dominant trait. In an F2 population of 32 animals, segregation of the endothelial antigen or antigens appeared to be independent of the MHC, AgF, and tubular basement membrane antigens--as well as the locus for albinism. Transplantation of MHC-identical but endothelial-antigen-incompatible kidneys into nonimmunized recipients did not provoke acute rejection. Pretransplant immunity against donor endothelial antigens was, however, associated with accelerated acute rejection. The rejection was donor-specific because third-party MHC-incompatible but endothelial-antigen-compatible kidneys were rejected like first-set grafts. This model shows that graft rejection in presensitized recipients of an MHC-identical kidney can be mediated through immunity against non-MHC antigens.     

Expression of CD44 and major histocompatibility complex class II antigens correlate with renal scarring in primary and systemic renal diseases

OBJECTIVE: Phenotypical changes in the tubular epithelial cells (TEC) seem to be important in the progression of renal diseases. The present study was designed to identify the relation between the expression of major histocompatibility complex (MHC) class II antigens and CD44 by TEC, with parameters of renal scarring in primary and systemic renal diseases. MATERIAL AND METHODS: Expression of MHC class II and CD44 antigens was determined immunohistochemically in 71 renal biopsies and eight nephrectomy specimens with chronic pyelonephritis (CP). RESULTS: CD44 expression was increased in renal diseases compared with autopsy cases and was strongly correlated with parameters of renal scarring and MHC class II antigen expression in primary and systemic renal diseases. CD44 expression was demonstrated in chronic pyelonephritis, postinfectious glomerulonephritis, diabetic nephropathy and hypertensive nephropathy, as well as other diseases described previously. Similar results were obtained for MHC class II antigen expression by TEC and these results were correlated with serum creatinine values. CONCLUSIONS: CD44 expression by TEC is a common pathway in renal scarring, like MHC class II antigen expression, and both of these may be important in renal scarring in CP cases as well as other primary and systemic renal diseases.     

A case of atypical congenital nephrotic syndrome

We present a female newborn with the nephrotic syndrome of intrauterine onset and a unique set of extrarenal abnormalities, as well as atypical renal lesions. The extrarenal anomalies comprised a soft tissue hemangioma in the frontotemporal region, unilateral microphthalmia (with persistent hyperplastic corpus vitreous and detachment of the retina), and glaucoma in the other eye. Immature glomeruli and/or glomeruli with large cellular crescents were found in renal biopsy specimens in the 3rd week of life. On autopsy, 7 weeks later, diffuse mesangial sclerosis (DMS) was the predominant type of glomerular lesion. In addition, dilations of tubules, forming microcysts, as well as clusters of infiltrating cells in the interstitium, were found both in renal biopsy and autopsy specimens. Although the symptoms observed in our patient did not match any reported in association with the known forms of the congenital nephrotic syndrome (CNS), the most probable diagnosis seemed to be CNS due to DMS of intrauterine onset, with superimposed drug-related tubulointerstitial nephritis.     

The relationship of infiltrating renal leucocytes to disease activity in lupus and cryoglobulinaemic glomerulonephritis

In order to evaluate the contribution of cellular immune mechanisms in the pathogenesis of immune complex-mediated glomerulonephritis, renal biopsies from 18 patients with lupus glomerulonephritis and 26 with cryoglobulinaemic glomerulonephritis were studied. Leucocyte profiles including T cell subsets and 'activated' macrophages within both glomeruli and interstitium were determined, using a panel of monoclonal antibodies as markers, and a sensitive 4-layer peroxidase technique to localize these within tissues. The infiltrating leucocytes were correlated with clinical, histological and immunological parameters of disease activity. Normal glomeruli contained few leucocytes though normal interstitium did (145 +/- 30 mm2), made up predominantly of T lymphocytes and macrophages. There was a significant increase in intraglomerular leucocytes in both systemic lupus erythematosus 4-fold, and essential mixed cryoglobulinaemia 7-fold, as compared to normal. These leucocytes consisted mainly of macrophages, and particularly in cryoglobulinaemia of 'activated' macrophages as demonstrated by their surface expression of the procoagulant tissue factor recognized by the A13 monoclonal antibody. In cryoglobulinaemic glomerulonephritis (GN) there was also a significant increase in T lymphocytes due to a predominance of suppressor-cytotoxic cells (OKT8+). There was a significant increase in interstitial leucocytes in both diseases, lymphocytes (mainly OKT8+ve), and macrophages (mainly 'activated' A13+ve). There were significant positive correlations between disease activity and interstitial leucocyte infiltration including, in lupus nephritis, degree of proteinuria and total leucocytes, hypocomplementaemia and T lymphocytes, increased numbers of monocytes and lymphocytes with a higher histological index of activity, and in cryoglobulinaemic GN of T lymphocytes and proliferative lesions, and T lymphocytes and C1q deposition. This study has demonstrated the importance of the interstitium in the pathogenesis of both diseases, delineated the presence of both T lymphocytes and activated monocytes which make cell-mediated immune mechanisms feasible, and linked the presence of immune mediators to disease activity.     

Effect of cyclosporine on MHC class II antigen expression on arterial and venous endothelium in vitro

MHC class II antigens play a crucial role in immunological responses. The expression of MHC class II antigens on monocytes and endothelial cells is reported to be variable and able to be induced by gamma-interferon. In this study we report on MHC class II antigen expression in vitro by arterial and venous canine endothelial cells, as detected with FACS analysis and indirect immunofluorescence with a monoclonal antibody against canine MHC class II antigens. It appears that cultured endothelial cells do not express MHC class II antigens. Their expression could be induced during a three-day incubation period in lymphokine-containing supernatant produced in mixed leukocyte culture (MLC). Cyclosporine (CsA) added to allogeneically stimulated or unstimulated canine lymphocytes in MLC inhibited the induction of expression by the MLC supernatant. The addition of CsA to MLC supernatant did not have an inhibitory effect. It is concluded that CsA inhibits the production of an MHC class-II-antigen-inducing lymphokine produced by lymphocytes in mixed cultures; allogeneic stimulation is not necessary for production of the lymphokine. It is postulated that a possible mode of action of CsA in prolongation of allograft survival is based on prevention of the induction of MHC class II antigen expression by endothelial cells.     

Polymorphonuclear neutrophils in Wegener's granulomatosis acquire characteristics of antigen presenting cells.

BACKGROUND: Constitutive expression of major histocompatibility complex (MHC) class II antigens and of the co-stimulatory receptors CD80 and CD86 is restricted to professional antigen presenting cells. Polymorphonuclear neutrophils (PMN) of healthy donors are negative for those antigens. Our recent study, however, found that PMN of patients with active Wegener's granulomatosis acquired MHC class II antigens. METHODS: To continue and extend the previous study results, PMN and monocytes of 60 patients with Wegener's granulomatosis, 24 patients with microscopic polyangiitis (MPA), 20 patients with acute bacterial infection, and 53 healthy donors were analyzed for the expression of MHC class II antigens as well as of CD80 and CD86. Moreover, induction on PMN of MHC class II expression was studied, as was antigen presentation as a possible functional consequence. RESULTS: PMN of patients with acute, active Wegener's granulomatosis expressed MHC class II antigens, CD80 and CD86; on monocytes up-regulation of MHC class II was seen. In contrast, PMN of patients with inactive disease, or with relapse, patients with microscopic polyangiitis or with bacterial infections expressed neither MHC class II, nor CD80 or CD86. PMN of healthy donors acquired these antigens when cultured in the presence of T cells or T cell-derived cytokines. The PMN were then able to present to T cell antigens in a MHC-class II restricted manner. CONCLUSION: During active disease, the PMN of patients with Wegener's granulomatosis acquire characteristics of antigen presenting cells, whereas the PMN of patients with MPA or bacterial infection do not. The finding reflects differences in the pattern of the respective inflammatory response and suggests new effector functions of PMN. Moreover, MHC class II expression on PMN could serve as a novel marker for active Wegener's granulomatosis.     

Association of glomerular and interstitial mononuclear leukocytes with different forms of glomerulonephritis

Immunophenotyping of mononuclear leukocytes was performed in renal tissue obtained from 69 patients with different forms of glomerulonephritis (GN) and from ten donors' kidneys for transplantation used as controls. A panel of monoclonal antibodies was used in the immunoperoxidase technique on frozen sections to define B- and T-lymphocyte subpopulations, NK cells and monocytes/macrophages, as well as the expression of HLA class II antigens-DQ, -DR and -DP. Quantification of labelled leukocytes revealed a significant increase of CD4+ and CD8+ T-cells in glomeruli of rapidly progressive glomerulonephritis, membranoproliferative glomerulonephritis and even of focal gomerulosclerosis. The number of glomerular monocytes/macrophages was significantly increased only in rapidly progressive glomerulonephritis, whereas in membranoproliferative glomerulonephritis it was decreased. No differences to normal tissue were detected in glomeruli for all other types of inflammatory cells. Interstitial cells were mostly T-lymphocytes in all forms of glomerulonephritis. In all groups the CD4+/CD8+ ratio was somewhat greater than 1 and even about 2 in rapidly progressive glomerulonephritis. Only in particular case was this ratio inversed. High expression of HLA class II antigens was observed on interstitial mononuclear leukocytes, as a sign of their activation. The excess of HLA-DQ-positive cells over the sum of CD14+ and CD20+ cells provides evidence not only for presence of activated T-lymphocytes but perhaps also for accumulation of renal dendritic cells in the interstitium in glomerulonephritis associated with interstitial infiltration.