C4d/CD34 double-immunofluorescence staining of renal allograft biopsies for assessing peritubular capillary C4d positivity

Jen, Kuang-Yu; Nguyen, Thu Quynh; Vincenti, Flavio; Lászik, Zoltán

doi:10.1038/modpathol.2011.168

Cited by 8 publications

(7 citation statements)

References 9 publications

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“…In this circumstance, double immunofluorescence with C4d and CD34 can facilitate more accurate assessment of the true percent positive. 20 By less sensitive immunohistochemistry, anything ≥ 0% peritubular capillary staining is considered positive. These thresholds for positivity with either immunoflourescence or immunohistochemistry represent a reduction compared with prior Banff publications.…”

Section: C4dmentioning

confidence: 99%

Transplant glomerulopathy

Filippone¹,

McCue

Farber

2018

Modern Pathology

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In the renal allograft, transplant glomerulopathy represents a morphologic lesion and not a specific diagnosis. The hallmark pathologic feature is glomerular basement membrane reduplication by light microscopy or electron microscopy in the absence of immune complex deposits. Transplant glomerulopathy results from chronic, recurring endothelial cell injury that can be mediated by HLA alloantibodies (donor-specific antibodies), various autoantibodies, cell-mediated immune injury, thrombotic microangiopathy, or chronic hepatitis C. Clinically, transplant glomerulopathy may be silent, detectable on protocol biopsy, or present with overt manifestations, including up to nephrotic range proteinuria, hypertension, and declining glomerular filtration rate. In either case, transplant glomerulopathy is associated with reduced graft survival. This review details the morphologic features of transplant glomerulopathy found on light microscopy, immunofluorescence microscopy, and electron microscopy. The pathophysiology of the causes and risk factors are discussed. Clinical manifestations are emphasized and potential therapeutic modalities are examined.

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Section: C4dmentioning

confidence: 99%

Transplant glomerulopathy

Filippone¹,

McCue

Farber

2018

Modern Pathology

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“…These data, along with the rarity of observed insulitis in humans, suggest the possibility of an alternative explanation of β-cell destruction; namely, that islet autoantibodies, present before disease onset, may fix C on the surface of β-cells and promote their lysis. Therefore, we quantified C4d, a marker of antibody-mediated C activation, in pancreata from individuals with T1D (including long-standing disease) and type 2 diabetes (T2D), as well as autoantibody-positive and autoantibody-negative nondiabetic subjects (7–9). …”

mentioning

confidence: 99%

Increased Complement Activation in Human Type 1 Diabetes Pancreata

Rowe¹,

Wasserfall

Croker

et al. 2013

Diabetes Care

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OBJECTIVEEvidence supporting an association between complement (C) and type 1 diabetes (T1D) includes the identification of C-fixing islet cell autoantibodies in T1D sera and genetic associations with the major histocompatibility complex III C4 region on chromosome 6. Therefore, we investigated whether C activation was present in pancreata from those with or at increased risk (positive for T1D associated autoantibodies) for T1D.RESEARCH DESIGN AND METHODSImmunohistochemical techniques were used to measure the C degradation product C4d in organ donor pancreata from patients with T1D and type 2 diabetes and autoantibody-positive and autoantibody-negative subjects.RESULTSMedian C4d antigen density differed across the groups (P < 0.0001) and was highest in patients with T1D. C4d immunostaining localized to the blood vessel endothelium and extracellular matrix surrounding blood vessels and exocrine ducts. Receiver operating characteristic analysis resulted in 81.8% sensitivity and 94.4% specificity for C4d staining.CONCLUSIONSThese data suggest that C activation is occurring within pancreata from patients with T1D and C4d may be a biomarker for T1D.

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“…However, in many published date researcher mostly used indirect IF only to confirm the presence of protein without further analysis and quantification. 22 , 28 , 32 , 39 A few studies over improvements of articular cartilage functions or differentiation of bone marrow cells towards chondrocytes have used evaluation of IF images to count percentage of positively stained cells or measure the mean intensity by manual drawing area of the signal. 3 , 59 In presented study by using threshold of RGB converted images into 16-bit picture, we can describe the exact specific-signal area and count mean gray value.…”

Section: Discussionmentioning

confidence: 99%

Bioimaging: An Useful Tool to Monitor Differentiation of Human Embryonic Stem Cells into Chondrocytes

et al. 2015

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To improve the recovery of damaged cartilage tissue, pluripotent stem cell-based therapies are being intensively explored. A number of techniques exist that enable monitoring of stem cell differentiation, including immunofluorescence staining. This simple and fast method enables changes to be observed during the differentiation process. Here, two protocols for the differentiation of human embryonic stem cells into chondrocytes were used (monolayer cell culture and embryoid body formation). Cells were labeled for markers expressed during the differentiation process at different time points (pluripotent: NANOG, SOX2, OCT3/4, E-cadherin; prochondrogenic: SOX6, SOX9, Collagen type II; extracellular matrix components: chondroitin sulfate, heparan sulfate; beta-catenin, CXCR4, and Brachyury). Comparison of the signal intensity of differentiated cells to control cell populations (articular cartilage chondrocytes and human embryonic stem cells) showed decreased signal intensities of pluripotent markers, E-cadherin and beta-catenin. Increased signal intensities of prochondrogenic markers and extracellular matrix components were observed. The changes during chondrogenic differentiation monitored by evaluation of pluripotent and chondrogenic markers signal intensity were described. The changes were similar to several studies over chondrogenesis. These results were confirmed by semi-quantitative analysis of IF signals. In this research we indicate a bioimaging as a useful tool to monitor and semi-quantify the IF pictures during the differentiation of hES into chondrocyte-like.Electronic supplementary materialThe online version of this article (doi:10.1007/s10439-015-1443-z) contains supplementary material, which is available to authorized users.

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C4d/CD34 double-immunofluorescence staining of renal allograft biopsies for assessing peritubular capillary C4d positivity

Cited by 8 publications

References 9 publications

Transplant glomerulopathy

Transplant glomerulopathy

Increased Complement Activation in Human Type 1 Diabetes Pancreata

Bioimaging: An Useful Tool to Monitor Differentiation of Human Embryonic Stem Cells into Chondrocytes

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