Proteinase 3. A distinct human polymorphonuclear leukocyte proteinase that produces emphysema in hamsters.

Kao, R C; Wehner, Nancy; Skubitz, Keith M.; Gray, Beulah H.; Hoidal, John R.

doi:10.1172/jci113816

Cited by 392 publications

(204 citation statements)

References 49 publications

Supporting

Mentioning

195

Contrasting

Unclassified

Order By: Relevance

“…Granules of human PMN were isolated [20] and PR3 was purified using an aprotinin-Sepharose column [21] followed by Matrex gel orange A chromatography [5]. The eluate was then immunoadsorbed using a PR3 monoclonal antibody (CLB, Clone 12.8, Amsterdam) [221.…”

Section: Purification Of Pr3 From Pmn Granulesmentioning

confidence: 99%

“…Proteinase 3 (PR3) belongs to the broad family of neutrophil-derived cationic glycoproteins involved in the defense against pathogens via the non-oxygen-dependent pathway [I], Described first by Baggiolini ia 1978 [2], PR3 is an elastase-like serine pretense packed in azurophil granules from neutrophili¢ polymorphonuclear leukocytes tPMN) [3] which degrad~ a variety of extracellular matrix proteins [4] and can cause emphysema in hamsters [5].…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti‐neutrophil cytoplasmic autoantibodies

et al. 1996

View full text Add to dashboard Cite

Using the bnculovlrus/Insect cells system, we have expmmed n recombinant protelnase 3 (PR3) --the neutrophil.derived sedne protease autonntlgen In Wegener's gntnulomatosis --as a illyemyiated Intrncellular and membrane-associated protein, OIIgosnochnrldes accounted for the difference in molecular weights between recombinant (34 kDa) and neutrophiI-PR3 (29 kDn), Whereas rabbit-anti-PR3 igG recognized both recombinant and nentrophll-derlved PR3, autoantibodies from Wegener patient sent reeognl~l only neutrophil-derived PR3, Although ollgosaeeharides were not involved in PR3 epitope recognition, autoantibodles did not recognize the amino acid primary structure of recombinant PR3, Improper disulfide bond formation and/or lack of post-translational events in insect cells, may affect the conformation of PR3, precluding its reactivity with sera from WG patients.

show abstract

Section: Purification Of Pr3 From Pmn Granulesmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti‐neutrophil cytoplasmic autoantibodies

et al. 1996

View full text Add to dashboard Cite

show abstract

“…emphysema [4,9] and Wegener's granulomatosis (WG) [10]. WG is a specific form of vasculitis associated with circulating anti-neutrophil cytoplasmic autoantibodies directed against PR3 (c-ANCA) [3].…”

Section: Introductionmentioning

confidence: 99%

Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC‐1 cells is enzymatically active and recognized by c‐ANCA

Specks¹,

Fass²,

Fautsch³

et al. 1996

FEBS Letters

View full text Add to dashboard Cite

We developed a stable expression system for conformationally intact recombinant human PR3 (rPR3) using the human mast cell line HMC-I. Like in U937 cells, the rPR3 is processed from a 34 kDa precursor to the 29 kDa mature form, primarily as the result of oligosaccharide trimming. The rPR3 binds [3H]DFP and hydrolyzes the snbstrate N-methoxysuccinylAla-Ala-Pro-Val-pNA. The enzymatic activity is inhibited by greater than 95% by al-PI. The rPR3 and the enzymatically inactive mutant rPR3-S176A are both packaged in granules. Thus, proteolytic autoprocessing is not required for PRYs targeting to granules. This rPR3 is the first to be recognized by most c-ANCA from WG patients and all anti-PR3 ANCA that were detected by standard anti-PR3 specific ELISA. This expression system for rPR3 represents a versatile tool for the analysis of its intraceHular processing, structure-function relationships and interaction with autoantibodies.

show abstract

“…Purification of PR3 PR3 was purified as described by Kao et al [16] and affinitypurified as described by Lüdemann et al [7] using an extract of granulocytes. For inhibition experiments, serum antibodies were mixed with purified PR3 v/v diluted to 0 .…”

Section: Preparation Of Cell Extractsmentioning

confidence: 99%

“…RNA was diluted in double distilled (dd) water (1 g/ 14 l) and heated at 658C for 5-10 min. Reverse transcription (RT) mix (16 l/ g sample RNA) consisting of 3 l oligo (dT) 16 , 0 . 5 mg/ml (Sigma, St Louis, MO), 150 U M-MLV reverse transcriptase (BRL, Gaithersburg, MD) and 40 U Rnasin (Promega, Madison, WI) in 50 mM Tris-HCl pH 8 .…”

Section: Rna Extraction and Semiquantitative Polymerase Chain Reactionmentioning

confidence: 99%

Antibodies to proteinase 3 mediate expression of vascular cell adhesion molecule-1 (VCAM-1)

Mayet¹,

Schwarting²,

Orth³

et al. 1996

Clinical and Experimental Immunology

View full text Add to dashboard Cite

VCAM‐1 was first identified as an adhesion molecule induced on human endothelial cells (HEC) by inflammatory cytokines such as IL‐1, tumour necrosis factor (TNF), and lipopolysaccharide (LPS). The molecule binds to a variety of leucocytes, including B cells, T cells, basophils, eosinophils and monocytes. Vascular expression of VCAM‐1 has been associated with a number of disease states, including rheumatoid arthritis and vasculitis. The detection of antineutrophil cytoplasmic antibodies (ANCA), especially to proteinase 3 (PR3), has become important in the diagnosis of Wegener’s granulomatosis (WG) and related vasculitides. Recently we were able to demonstrate a direct effect of anti‐PR3 antibodies on neutrophil–endothelial interactions (Blood 1993; 82:1221). Binding of anti‐PR3 antibodies to their antigen translocated into the membrane of HEC leads to an enhanced adhesion of neutrophils via induction of E‐selectin (Clin Exp Immunol 1993; 94:440). The aim of this study was to investigate the effect of anti‐PR3 antibodies on the expression of VCAM‐1. HEC was isolated from umbilical vein and cultured on microtitre plates. After preincubatiion with purified anti‐PR3 antibody, purified control antibodies (SS‐A, SS‐B, RNP) (IgG and F(ab′)2 fragments) or different cytokines (controls), VCAM‐1 was detected on the surface of unfixed HEC by cyto‐ELISA and polymerase chain reaction analysis. Incubation of HEC with anti‐PR3 antibodies led to a marked increase of endothelial VCAM‐1 expression with a peak after 8 h. Incubation with TNF‐α also led to maximal VCAM‐1 expression after 4–6 h (control). Increased adhesion of T lymphocytes to HEC after binding of anti‐PR3 antibodies to their antigen could be confirmed by performing adherence assays. This effect could be inhibited by antibodies to VLA‐4. In conclusion, we have been able to show that cytokine‐like effects of anti‐PR3 antibodies on HEC are not limited to induction of neutrophil adhesion. Anti‐PR3 antibodies may thus contribute to the regulation of T lymphocyte migration from the blood by HEC in ANCA‐related vasculitides.

show abstract

Proteinase 3. A distinct human polymorphonuclear leukocyte proteinase that produces emphysema in hamsters.

Cited by 392 publications

References 49 publications

Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti‐neutrophil cytoplasmic autoantibodies

Characterization of a recombinant proteinase 3, the autoantigen in Wegener's granulomatosis and its reactivity with anti‐neutrophil cytoplasmic autoantibodies

Recombinant human proteinase 3, the Wegener's autoantigen, expressed in HMC‐1 cells is enzymatically active and recognized by c‐ANCA

Antibodies to proteinase 3 mediate expression of vascular cell adhesion molecule-1 (VCAM-1)

Contact Info

Product

Resources

About