Immunofluorescence Microscopy Findings Research Articles

The relationship between immun staining and progression markers in IgA nephropathy

Objective: To determine the relationship between immunofluorescence microscopy findings and progression markers at the time of diagnosis in immunoglobulin A (IgA) nephropathy. Methods: Fifty-two patients with pathological diagnosis of primary IgA nephropathy by showing mesangial and mesangiocapillary IgA-dominant immune deposits in immunofluorescence microscopy were included in the study. At the time of biopsy, biochemical and hematological data, Oxford MEST score and immunofluorescent staining findings were recorded. The serum IgA/C3 ratio was calculated. The immunofluorescence results of the total group were compared with the markers of progression at the time of diagnosis, estimated glomerular filtration rate (eGFR), hematuria, proteinuria, creatinine, and serum IgA/C3 ratio. Results: The mean age of the study group was 39.9±12.3 years and 55.8% were male. eGFR, albumin, hemoglobin, IgM were significantly lower, and uric acid and hematuria were significantly higher in those with proteinuria above 1 g compared to those with low proteinuria. A positive correlation was found between IgA, C3 and lambda staining and hematuria. There was a positive correlation between C3 staining and creatinine, and a positive correlation with hematuria. A correlation was found between Kappa staining and eGFR. Conclusion: Correlation was found between IgA, C3 and lambda staining and hematuria at the time of diagnosis in IgA nephropathy.

Epidemiological features of primary glomerular disease in Turkey: a multicenter study by the Turkish Society of Nephrology Glomerular Diseases Working Group

BackgroundThe largest data on the epidemiology of primary glomerular diseases (PGDs) are obtained from the databases of countries or centers. Here, we present the extended results of the Primary Glomerular Diseases Study of the Turkish Society of Nephrology Glomerular Diseases (TSN-GOLD) Working Group.MethodsData of patients who underwent renal biopsy and received the diagnosis of PGD were recorded in the database prepared for the study. A total of 4399 patients from 47 centers were evaluated between May 2009 and May 2019. The data obtained at the time of kidney biopsy were analyzed. After the exclusion of patients without light microscopy and immunofluorescence microscopy findings, a total of 3875 patients were included in the study.ResultsThe mean age was 41.5 ± 14.9 years. 1690 patients were female (43.6%) and 2185 (56.3%) were male. Nephrotic syndrome was the most common biopsy indication (51.7%). This was followed by asymptomatic urinary abnormalities (18.3%) and nephritic syndrome (17.8%). The most common PGD was IgA nephropathy (25.7%) followed by membranous nephropathy (25.6%) and focal segmental glomerulosclerosis (21.9%). The mean total number of glomeruli per biopsy was 17 ± 10. The mean baseline systolic blood pressure was 130 ± 20 mmHg and diastolic blood pressure was 81 ± 12 mmHg. The median proteinuria, serum creatinine, estimated GFR, and mean albumin values were 3300 (IQR: 1467–6307) mg/day, 1.0 (IQR: 0.7–1.6) mg/dL, 82.9 (IQR: 47.0–113.0) mL/min and 3.2 ± 0.9 g/dL, respectively.ConclusionsThe distribution of PGDs in Turkey has become similar to that in other European countries. IgA nephropathy diagnosed via renal biopsy has become more prevalent compared to membranous nephropathy.

Brunsting-Perry pemphigoid: A retrospective case series of a frequently unrecognized condition

Brunsting-Perry pemphigoid: A retrospective case series of a frequently unrecognized condition

Drug-induced Rowell syndrome, a rare and difficult to manage disease: A case report.

Rowell syndrome is defined as the association between lupus erythematosus, erythema multiforme-like lesions and characteristic immunological changes including positive tests for rheumatoid factor, speckled antinuclear antibody, positive anti-Ro or anti-La antibodies. The present report presents the case of a 45-year-old female patient who was previously diagnosed in January 2010 with terbinafine-induced subacute cutaneous lupus erythematosus and was admitted for a skin eruption consisting of erythematous-papular erythema multiforme-like lesions, primarily on the trunk and limbs. The associated symptoms consisted of fatigability, myalgia and gonalgia. In October 2015, the illness reoccurred ~1 week after the initiation of Helicobacter pylori eradication treatment. Anti-Ro antibodies, rheumatoid factor and antinuclear antibody tests were positive. Given the patient's medical history, clinical manifestations, and laboratory, histopathological and immunofluorescence microscopy findings, a diagnosis of Rowell syndrome was made. Systemic corticosteroids (methylprednisolone; 0.5 mg/kg/day) and immunomodulatory therapy (azathioprine; 50 mg/day) were administered with the associated medication (omeprazole, 20 mg/day; KCl, 1 g/day) and topical dermocorticoids (fluticasone propionate 0.05% cream; 1 application/day), with a favorable outcome. The major diagnostic criteria for Rowell syndrome are the presence of lupus erythematosus (acute, subacute or systemic), erythema multiforme-like lesions and positive testing for antinuclear antibodies. The minor diagnostic criteria for Rowell syndrome are chilblains, the presence of anti-Ro antibodies and positive testing for rheumatoid factor. A diagnosis of Rowell syndrome is made if the patient exhibits all major criteria and at least one minor criterion. The present case met all diagnostic criteria, excluding the presence of chilblains. Notably, in this case there was a co-occurrence of subacute lupus erythematosus and Rowell syndrome lesions, which was drug-induced.

10.5937/mckg48-3313 = Vasculitides of the small blood vessels of kidney: Etiopathogenesis, diagnosis and treatment

Vasculitis is an inflammation of blood vessel walls. Vasculitides of kidney's small blood vessels include microscopic polyangiitis, Wegener's granulomatosis and Churg-Strauss-syndrome. The main pathohistological characteristics of all three types of vasculitis are the inflammation of small blood vessels, glomeruli, with necrosis and glomerular crescents formation. Anti-neutrophil cytoplasmic antibodies (ANCA) play an important role in the development of vasculitis. Depending on the target antigen and immunofluorescence microscopy findings, there are two types of ANCA: cytoplasmic (cANCA) and perinuclear (pANCA). Immunopathogenesis of ANCA-associated vasculitis involves four main steps: regrouping of neutrophils and their approaching to the endothelial surface, interaction of ANCA with neutrophil antigens, adherence and penetration of neutrophils into the endothelium and, finally, activation of neutrophils and production of mediators that damage endothelial cells. Kidney damage is clinically manifested by appearance of microhematuria with nonnephrotic range proteinuria and development of acute nephritic syndrome. Diffuse alveolar hemorrhage (DAH) often develops, too. Kidney biopsy is the main diagnostic procedure that enables the right choice of treatment regime. The treatment comprises of two phases: the first is the induction of disease remission and the second is the maintenance of remission. The first phase consists of administering high doses of corticosteroids and cyclophosphamide while in the second phase azathioprine or methotrexate are administered instead of cyclophosphamide. Disease relapse is treated with rituximab.

Do C1q or IgM nephropathies predict disease severity in children with minimal change nephrotic syndrome?

It has been suggested that C1q and immunoglobulin M (IgM) nephropathy are variants of minimal change nephrotic syndrome (MCNS). Many researchers believe that these two conditions signify a worse prognosis for children with MCNS in comparison with immunofluorescence (IF)-negative MCNS. The aim of our study was to determine the prognostic significance of C1q nephropathy and IgM nephropathy in children with MCNS. Fifty-five children with MCNS who had been biopsied over the course of 24 years at our institution were retrospectively categorized into three groups on the basis of IF microscopy findings: IF-negative MCNS (29/55 patients), MCNS with IgM nephropathy (19/55 patients), and MCNS with C1q nephropathy (7/55 patients). Clinical characteristics at disease presentation, clinical course, and renal outcome were compared between groups during the median follow-up period of 16.9 years (minimum 1.0, maximum 31.1 years). No statistically significant differences in clinical characteristics at disease presentation, clinical course, and renal outcome were found. Children with IgM nephropathy, C1q nephropathy, and IF-negative MCNS were clinically indistinguishable. We concluded that C1q or IgM nephropathy variants do not seem to signify a worse prognosis in children with MCNS in comparison with IF-negative MCNS.

Targeted testing for DNAI1 hot spot-mutation utilizing immunofluorescence microscopy findings

Primary ciliary dyskinesia (PCD) is a rare (prevalence 1/20,000) genetic disease affecting motile cilia in the respiratory epithelium, spermatozoid flagella and primary cilia in the embryonic node. The most frequent (~60-70%) structural defect identified by TEM in the cilia of PCD patients are abnormal dynein arms. Several genes can cause PCD, but the majority of mutations were found in DNAH5 and DNAI1 genes (respectively ~28% and 4-10% of all cases), which encode heavy and intermediate chains of the outer dynein arms (ODAs), respectively. Mutations in both genes account collectively for almost 40% of PCD cases [Olbrich et al. 2002, Hornef et al. 2006, Zariwala et al. 2006, Zietkiewicz et al. 2010]. The hot-spot mutation in the DNAI1 gene appears in intron 1, with the frequency of the most popular mutation (IVS1+2_3insT, causes aberrant splicing) around 55% of all DNAI1 mutations. We have previously shown in a few PCD cases that proximal type-1 ODA complexes can be at least partially assembled in DNAI1– mutant cilia [Fliegauf et al. 2005]. We analysed the frequency of this hot spot mutation among 51 patients, in which immunofluorescence has identified abnormal ODA staining (proximal presence of DNAH5). The prevalence of the mutation in intron 1 of DNAI1 gene will be confirmed by PCR and restriction enzyme digestion. In addition, we analyzed respiratory cilia for the inner dynein arm component DNALI1 localization, which we expect not to be altered in DNAI1 mutant cilia, contrasting the findings present in KTU- mutant cilia.

Numerous Mucin Nodules in a Patient with Seropositive Wide Spread Discoid Lupus Erythematosus

Numerous Mucin Nodules in a Patient with Seropositive Wide Spread Discoid Lupus Erythematosus

Rab32 Modulates Apoptosis Onset and Mitochondria-associated Membrane (MAM) Properties

The mitochondria-associated membrane (MAM) has emerged as an endoplasmic reticulum (ER) signaling hub that accommodates ER chaperones, including the lectin calnexin. At the MAM, these chaperones control ER homeostasis but also play a role in the onset of ER stress-mediated apoptosis, likely through the modulation of ER calcium signaling. These opposing roles of MAM-localized chaperones suggest the existence of mechanisms that regulate the composition and the properties of ER membrane domains. Our results now show that the GTPase Rab32 localizes to the ER and mitochondria, and we identify this protein as a regulator of MAM properties. Consistent with such a role, Rab32 modulates ER calcium handling and disrupts the specific enrichment of calnexin on the MAM, while not affecting the ER distribution of protein-disulfide isomerase and mitofusin-2. Furthermore, Rab32 determines the targeting of PKA to mitochondrial and ER membranes and through its overexpression or inactivation increases the phosphorylation of Bad and of Drp1. Through a combination of its functions as a PKA-anchoring protein and a regulator of MAM properties, the activity and expression level of Rab32 determine the speed of apoptosis onset.

Crescentic glomerulonephritis associated with totally implantable central venous catheter-related Staphylococcus epidermidis infection

A 59-year-old woman was admitted to our hospital for treatment of acute renal insufficiency. She had been under home intravenous hyperalimentation therapy through a totally implantable central venous catheter for 2 years because of post-radiation enteritis. Clinical examination on admission revealed severe renal insufficiency complicated with hypocomplementemia, marked proteinuria and hematuria. Chest roentgenography demonstrated moderate pulmonary congestion. Hemodialysis was initiated and her pulmonary congestion improved. On the 14th and 21st hospital day, blood culture revealed Staphylococcus epidermidis colonization. Cefazolin was administered and C-reactive protein decreased, however, renal insufficiency and hypocomplementemia did not improve. To investigate the genesis of renal insufficiency, renal biopsy was performed. Light microscopic findings of the kidney revealed severe crescentic glomerulonephritis complicated with moderate tubulointerstitial damage. Immunofluorescence-microscopic findings of the kidney revealed positive IgG, IgM, C3 deposition along the capillary lumen. From these laboratory findings and the clinical course, we diagnosed her renal disease as crescentic glomerulonephritis induced by catheter-related bloodstream infection, and the central venous catheter was removed. After removal, urinary output and hypocomplementemia remarkably improved, however, unfortunately, her renal dysfunction did not improve and maintenance hemodialysis needed to be continued. Although her renal disease was not caused by ventriculo-atrial shunt but by central venous catheter-related bloodstream infection, we supposed that the pathogenesis was a closely similar entity to shunt nephritis.

Possible requirement of intercellular adhesion molecule-1 for invasion of gingival epithelial cells byTreponema medium

Oral treponemes are members of the spirochete family of bacteria associated with periodontal diseases. In the present study, we demonstrate that intercellular adhesion molecule-1 (ICAM-1) on human gingival epithelial cells (HGEC) contributed to the invasion of Treponema medium, a medium-sized oral Treponema, into those cells. The quantity of T. medium in HGEC was found to peak at 2 h after inoculation and then decreased gradually. Immunofluorescence microscopy findings showed that the bacteria were colocalized with ICAM-1 on HGEC. Furthermore, knockdown of ICAM-1 in HGEC resulted in inhibition of T. medium invasion by RNA interference, whereas that of Toll-like receptor 2 did not. These results suggest that ICAM-1 may be required for the invasion of T. medium into HGEC, and they indicate that the molecule plays a principal role in the primary stages of the development and progression of chronic periodontitis.

Membranous Glomerulopathy With Spherules: An Uncommon Variant With Obscure Pathogenesis

Occasional case reports of membranous glomerulopathy described unique subepithelial accumulations of an unusual type of immune deposit composed of spherular structures. The identity of such structures as nuclear pores has been suggested, but not established. We identified a cohort of patients (n = 14, including 1 patient with disease recurrence in an allograft) who presented with nephrotic syndrome and had renal biopsy specimens with light and immunofluorescence microscopic findings characteristic of membranous glomerulopathy. These patients were distinguished by ultrastructural studies that showed glomerular capillary wall accumulations of subepithelial immune deposits composed of uniform spherular structures, while lacking the typical granular electron-dense deposits seen in membranous glomerulopathy. The molecular identity of these spherular structures as nuclear pores was tested by using immunofluorescence microscopy and immunohistochemistry with mouse monoclonal antinuclear pore antibodies (Covance, Princeton, NJ) and anti-Nuclear Pore-O-Linked Glycoprotein (Affinity BioReagents Inc, Golden, CO) antibodies. Measurement of spherular structures by using high-magnification electron microscopy showed an average diameter of 84.5 nm, which correlated well with accepted diameters of nuclear pores (80 to 120 nm). Immunofluorescence microscopy and immunoperoxidase staining with both antibodies showed characteristic beaded staining of nuclear membranes of multiple cell types within normal control kidney, but no staining of immune-type deposits within glomerular basement membranes. These cases form a rare, but distinctive, morphological subclass of membranous glomerulopathy. The antigenic specificity of immune deposits in these cases remains elusive.

Protective Immunity againstStreptococcus pyogenesChallenge in Mice after Immunization with Fibronectin‐Binding Protein

Surface-associated fibronectin (Fn)-binding proteins of Streptococcus pyogenes play an important role in the bacterial invasion of epithelial cells. We examined the functional domain and protective antigenicity of the Fn-binding protein FbaA. To investigate the functional domain of FbaA and its localization on S. pyogenes, a series of recombinant glutathione S-transferase (GST)-truncated FbaA proteins was used for immunofluorescent microscopy, ligand blotting, and Biacore analyses. Mice were immunized with the truncated proteins for the determination of the immunogenic domains that contribute to protection against S. pyogenes infection. Ligand-blotting and Biacore analyses revealed that the FbaA fragments harboring a proline-rich repeat domain (RD), but not the N- and C-terminal regions, possessed Fn-binding activity. Immunofluorescent microscopy findings showed that the N terminus and RD were exposed to external regions, which suggests that the RD serves as a Fn-binding element on live organisms. Specific antibodies were efficiently induced in N terminus- and RD-immunized mice and demonstrated bactericidal activity against S. pyogenes in vitro. FbaA-immunized mice survived significantly longer than GST-immunized mice after infection with serotype M1 and M49 strains expressing FbaA. The Fn-binding RD and N terminus of FbaA are potential vaccine candidates for M1 strains of S. pyogenes infection.

Elastic fiber proteins in the glomerular mesangium in vivo and in cell culture

Glomerular capillaries of the mammalian kidney are exposed to high intraluminal hydrostatic pressures and require elastic constraint to maintain size, shape, and integrity. Previous morphological and functional studies indicated that the extracellular matrices of glomeruli, that is, basement membrane and mesangial matrix, contribute to glomerular resilience and mechanical stability. Immunofluorescence microscopy findings demonstrated elastic fiber components to be located in the renal vasculature, including glomeruli. The aim of this study was to clarify the exact glomerular localization, composition, and cellular production of these proteins. We examined the renal distribution of the elastic fiber proteins fibrillin-1, emilin, microfibril-associated glycoproteins (MAGPs) 1 and 2, latent transforming growth factor-binding protein-1 (LTBP-1), and elastin using immunohistology and immunoelectron microscopy of human, rat, and mouse kidneys. In mesangial cell cultures, we also studied the expression and extracellular deposition of such proteins by use of Northern blotting and immunocytochemistry. Fibrillin-1, emilin, MAGPs 1 and 2, and LTBP-1 were present in glomeruli of mouse, rat, and human kidney, where they were located predominantly in the mesangial extracellular matrix underlying glomerular endothelium and basement membrane. Several of these proteins, as well as elastin, were also expressed in the renal vasculature. While elastin localized to the glomerular vascular pole in afferent and efferent arterioles extending to Bowman's capsule, it was not found in the glomerular capillary tuft. Cultured mesangial cells of rat, mouse, and human kidneys expressed mRNAs of fibrillin-1, emilin, MAGP-2, and elastin, and the respective proteins localized within and outside of mesangial cells, as shown by immunocytochemistry. mRNA expression of fibrillin-1, emilin, and elastin was strong in quiescent mesangial cells; their gene expression was further up-regulated by transforming growth factor-beta1, while it was transiently reduced when cells were exposed to mitogenic 10% fetal calf serum and platelet-derived growth factor. These findings demonstrate that specific elastic fiber proteins are produced and secreted by mesangial cells. This process is regulated by growth factors. Their abundance in the extracellular matrix of the mesangium is in keeping with the concept that elastic fiber proteins contribute to the mechanical stability and elastic strength of the glomerular capillary tuft.

Renal immunofluorescence and the prediction of renal outcome in patients with proliferative lupus nephritis.

The risk for endstage renal failure in patients with proliferative lupus nephritis (PLN) depends largely on the severity and reversibility of the inflammatory process as determined by light microscopy (LM). As the intrarenal formation of immune complexes is thought to initiate this inflammation, we studied whether renal immunofluorescence microscopy (IFM) provides clinical or prognostic information in addition to LM findings. Clinical data at the time of renal biopsy and during a mean follow-up of 46 months were extracted from the records of 69 SLE patients with proliferative LN (WHO class III/IV). Biopsy specimens were analyzed by LM for AI and CI, while IFM was performed on cryostat sections with the use of antisera against IgG, IgM, IgA, C3, C1q and fibrin. IFM findings were recorded in terms of the localization (glomerular, tubular or vascular) and intensity of fluorescence (score from zero to three). IFM findings were then related to clinical and LM findings and its prognostic value studied by survival analysis. Glomerular immune deposits were present in 99% of patients, tubular deposits in 38% and vascular deposits in 17%. A 'full-house' pattern (all three Ig classes) was found in 67% of biopsies and C3 and C1q deposits in 93% and 74% respectively. Median scores for AI and CI were 6 (1-18) and 3 (0-10); aside from a negative correlation between IgA deposits and CI, we found no other correlation between the amount or type of immune deposits and AI or CI. IgM deposits were associated with high serum levels of anti-dsDNA, while IgG deposits correlated with high ESR and serum creatinin levels. IFM scores were not related to steroid dose at the time of biopsy and neither type of glomerular, tubular or overall renal immune deposits had prognostic value for renal survival. Renal immunofluorescence does not reflect light microscopy findings in patients with PLN and does not contribute prognostic information in patients with PLN. Lupus (2000) 9, 504-510.

Lesions in donor kidneys: nature, incidence, and influence on graft function.

The aim of this study was to assess the influence of kidney-donor transmitted pathology on graft function. Light and immunofluorescent microscopic findings from a surgical biopsy taken prior to transplantation from 114 cadaveric kidney donors were analyzed. Moderate to severe mesangial IgA deposits were considered consistent with IgA nephropathy. Pathological abnormalities were correlated together with donor age, number of mismatches, and type of immunosuppression by multivariate statistical analysis with the serum creatinine values from patients who experienced no acute rejection at 1 year. Serum creatinine values (n = 52) were not correlated with either non-specific light microscopic lesions or immunofluorescent deposits found in the majority of kidney donors or with changes consistent with IgA nephropathy observed in 9% of the cases. There was, however, a correlation with donor age, which was also correlated with the extent of chronic lesions (P < 0.001).

Lesions in donor kidneys: nature, incidence, and influence on graft function

Abstract The aim of this study was to assess the influence of kidney-donor transmitted pathology on graft function. Light and immunofluorescent microscopic findings from a surgical biopsy taken prior to transplantation from 114 cadaveric kidney donors were analyzed. Moderate to severe mesangial IgA deposits were considered consistent with IgA nephropathy. Pathological abnormalities were correlated together with donor age, number of mismatches, and type of immunosuppression by multivariate statistical analysis with the serum creatinine values from patients who experienced no acute rejection at 1 year. Serum creatinine values (n = 52) were not correlated with either nonspecific light microscopic lesions or immunofluorescent deposits found in the majority of kidney donors or with changes consistent with IgA nephropathy observed in 9 % of the cases. There was, however, a correlation with donor age, which was also correlated with the extent of chronic lesions (P < 0.001).

Localized cicatricial pemphigoid (Brunsting-Perry): Electron microscopic study

Electron microscopic findings are presented in a patient with localized cicatricial pemphigoid (Brunsting-Perry). These findings revealed subepidermal separation below the basal lamina. The basal lamina and anchoring fibrils were well preserved and attached to the intact epidermis to form the roof of the blister. These findings support the concept that localized cicatricial pemphigoid, cicatricial pemphigoid, and disseminated cicatricial pemphigoid are closely related diseases and may explain the occurrence of scar formation in localized cicatricial pemphigoid. A review of 42 cases reported in the world literature is included, with emphasis on clinical, histologic, and immunofluorescence microscopic findings.

Herpes Gestationis in Blacks

To our knowledge, only two cases of herpes gestationis (HG) have been previously reported in black patients. We describe herein two black women with confirmed HG. Both had typical historical and clinical features of the disease. Direct immunofluorescence microscopy demonstrated complement deposition along the dermoepidermal junction in both women. Where HLA typing was possible (in one patient), the combination of HLA-DR3 and HLA-DR4 was found. In addition, anti-HLA-DR2 antibodies were present in serum samples taken from this patient. The occurrence of the HLA-DR3-DR4 combination has been reported to be greatly increased in whites with HG (43%, as compared with a 3% incidence in control subjects). The HLA-DR4 antigen is uncommon in American blacks, which may explain the infrequent occurrence of HG in this population. The occurrence of the DR2 antigen is increased in the husbands of women with HG, and this increase is most pronounced in the husbands of patients with the DR3-DR4 combination. The occurrence of positive immunofluorescence microscopy findings, together with the presence of a rare histocompatibility antigen combination previously associated with HG and the presence of anti-DR2 antibodies in the serum of one of our patients all suggest that HG is pathogenically identical in both blacks and whites.

Herpes Gestationis Associated With Choriocarcinoma

Herpes gestationis (HG) is a rare, vesicobullous dermatosis of pregnancy or the puerperium. It has been associated with hydatid mole and the use of oral contraceptives. Reports of HG and choriocarcinoma are extremely rare: only two cases have been reported. Metastatic choriocarcinoma, followed by a pruritic, bullous eruption, developed in a 36-year-old woman. A biopsy specimen of the lesions showed subepidermal bullae, with an eosinophilic infiltrate. Direct immunofluorescent microscopic findings showed IgG and C3 deposition at the basement membrane zone, establishing the diagnosis of HG. Methotrexate therapy produced a cure of the carcinoma and a simultaneous resolution of the skin lesions. To our knowledge, this is the first report of the association of HG and choriocarcinoma with immunofluorescent microscopic findings.