Potential role of multiple members of the kallikrein-related peptidase family of serine proteases in activating latent TGFβ1 in semen

2010 ◽  
Vol 391 (1) ◽  
Author(s):  
Nashmil Emami ◽  
Eleftherios P. Diamandis
Keyword(s):  
Conformational Changes ◽  
Seminal Plasma ◽  
Serine Proteases ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Transforming Growth Factor Β1 ◽  
Peptide Fragments ◽  
Biochemical Mechanisms

Abstract Transforming growth factor β1 (TGFβ1) has been implicated as a key contributor of immunosuppression in seminal plasma. The biochemical mechanisms that lead to production of active seminal TGFβ1 are not fully understood. It is plausible that TGFβ1 activation is partly induced simultaneously with the release of motile spermatozoa following liquefaction of the semen coagulum. Several members of the kallikrein-related peptidase (KLK) family are involved in the regulation of semen liquefaction. This study examines the involvement of these KLKs in TGFβ1 activation in vitro and ex vivo, in seminal plasma. Latent TGFβ1 was rapidly activated by KLK14. The latency-associated propeptide (LAP) was shown to be cleaved by KLK14 into small peptide fragments, providing a possible mechanism for TGFβ1 activation. KLK14 also cleaved the latent TGFβ binding protein 1 (LTBP1). KLK1, 2, and 5 might also contribute to TGFβ1 activation by nicking the LAP motif and inducing conformational changes that aid in subsequent processing of LAP or through LTBP1 cleavage. Our study provides strong evidence for the involvement of multiple members of the seminal KLK cascade in activation of latent TGFβ1 in seminal plasma. These findings might have clinical implications in infertility treatment of cases with concurrent delayed liquefaction and TGFβ1-related semen antigenicity.

Disease-dependent mechanisms of albuminuria

2008 ◽  
Vol 295 (6) ◽  
pp. F1589-F1600 ◽  
Author(s):  
Wayne D. Comper ◽  
Lucinda M. Hilliard ◽  
David J. Nikolic-Paterson ◽  
Leileata M. Russo
Keyword(s):  
Transforming Growth Factor ◽  
Degradation Pathway ◽  
Transforming Growth Factor Β1 ◽  
Renal Physiology ◽  
Peptide Fragments ◽  
Metabolic Factors ◽  
Charge Selectivity ◽  
Proximal Tubular ◽  
Glomerular Morphology

The mechanism of albuminuria is perhaps one of the most complex yet important questions in renal physiology today. Recent studies have directly demonstrated that the normal glomerulus filters substantial amounts of albumin and that charge selectivity plays little or no role in preventing this process. This filtered albumin is then processed by proximal tubular cells by two distinct pathways; dysfunction in either one of these pathways gives rise to discrete forms of albuminuria. Most of the filtered albumin is returned to the peritubular blood supply by a retrieval pathway. Albuminuria in the nephrotic range would arise from retrieval pathway dysfunction. The small quantities of filtered albumin that are not retrieved undergo obligatory lysosomal degradation before urinary excretion as small peptide fragments. This degradation pathway is sensitive to metabolic factors responsible for hypertrophy and fibrosis, particularly molecules such as angiotensin II and transforming growth factor-β1, whose production is stimulated by hyperglycemic and hypertensive environments. Dysfunction in this degradation pathway leads to albuminuria below the nephrotic range. These new insights into albumin filtration and processing argue for a reassessment of the role of podocytes and the slit diaphragm as major direct determinants governing albuminuria, provide information on how glomerular morphology and “tubular” albuminuria may be interrelated, and offer a new rationale for drug development.

scholarly journals IL-9 as a mediator of Th17-driven inflammatory disease

2009 ◽  
Vol 206 (8) ◽  
pp. 1653-1660 ◽  
Author(s):  
Elizabeth C. Nowak ◽  
Casey T. Weaver ◽  
Henrietta Turner ◽  
Sakhina Begum-Haque ◽  
Burkhard Becher ◽  
...  
Keyword(s):  
Inflammatory Disease ◽  
Th17 Cells ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Autoimmune Encephalomyelitis ◽  
Regional Lymph Nodes ◽  
Functional Consequences ◽  
The Central Nervous System

We report that like other T cells cultured in the presence of transforming growth factor (TGF) β, Th17 cells also produce interleukin (IL) 9. Th17 cells generated in vitro with IL-6 and TGF-β as well as purified ex vivo Th17 cells both produced IL-9. To determine if IL-9 has functional consequences in Th17-mediated inflammatory disease, we evaluated the role of IL-9 in the development and progression of experimental autoimmune encephalomyelitis, a mouse model of multiple sclerosis. The data show that IL-9 neutralization and IL-9 receptor deficiency attenuates disease, and this correlates with decreases in Th17 cells and IL-6–producing macrophages in the central nervous system, as well as mast cell numbers in the regional lymph nodes. Collectively, these data implicate IL-9 as a Th17-derived cytokine that can contribute to inflammatory disease.

scholarly journals Deceased donor kidney degradomics indicate cytoskeletal proteolytic alterations impact post-transplant function.

2021 ◽  
Author(s):  
Rebecca H Vaughan ◽  
Jean Kresse ◽  
Louise Farmer ◽  
Marie Thezenas ◽  
Benedikt M Kessler ◽  
...  
Keyword(s):  
Brain Death ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Human Kidney ◽  
Cytoskeletal Proteins ◽  
Cerebral Injury ◽  
Peptide Fragments ◽  
Post Transplant ◽  
Transplant Function

Background. In brain death, cerebral injury contributes to systemic biological dysregulation, causing significant cellular stress in donor kidneys that adversely impacts the quality of grafts. Here, we hypothesized that DBD kidneys may undergo proteolytic processes that renders grafts susceptible to post-transplant dysfunction. Material & Methods. Using mass spectrometry and immunoblotting analyses, we profiled degradation patterns of cytoskeletal proteins in deceased (n=55) and living (n=10) donor kidneys. Results. We found that in DBD kidneys, key podocyte cytoskeletal proteins had been proteolytically cleaved. Generated degradation profiles were independent to donor related demographic and clinical factors but were associated to suboptimal post-transplant function. Strikingly, α-actinin -4 and Talin-1 degradation profiles were not observed in circulatory-death or living-donor kidneys. As Talin-1 is a specific proteolytic target of Calpain-1, we investigated a potential trigger of Calpain activation and Talin-1 degradation using ex-vivo precision-cut human kidney slices and in-vitro immortalised human podocytes. Notably, we found that Transforming-Growth Factor-β (TGF-β) activated Calpain-1 and proteolytically cleaved Talin-1 to generate distinct peptide fragments. These peptide fragments were of similar size and matched the degradation patterns observed in DBD kidneys. Talin-1 degradation was prevented in-vitro by Calpain-1 inhibition. Conclusions. Here, we provide initial evidence that DBD kidneys are susceptible to cytoskeletal protein degradation that impacts posttransplant kidney function. Subsequent studies should aim to further investigate the link between brain death and activation of proteolytic pathways exploring new therapeutic opportunities.

ATL

2012 ◽  
Vol 22 (7) ◽  
pp. 1130-1137 ◽  
Author(s):  
Sreenivas Adurthi ◽  
Geetashree Mukherjee ◽  
H. Krishnamurthy ◽  
Krishna Sudhir ◽  
Uttamchand D. Bafna ◽  
...  
Keyword(s):  
T Cells ◽  
Growth Factor ◽  
Transforming Growth Factor ◽  
Ex Vivo ◽  
Interferon Γ ◽  
Transforming Growth Factor Β1 ◽  
Effector T Cells ◽  
Interleukin 4 ◽  
Autologous Tumor

ObjectiveAnalysis of tumor-infiltrating lymphocytes (TILs) is one of the cornerstones for the understanding of immune responses prevailing in the tumor microenvironment. We studied TILs from squamous cell carcinoma of the cervix ex vivo without proliferating them in vitro before analysis.MethodsWhereas TILs were magnetic activated cell separation enriched and flow sorted into CD4+CD25hi(regulatory T cells [Tregs]), CD4+CD25int(effector T cells [Teffs]) were directly purified by flow cytometry, and both these subsets were characterized phenotypically and functionally. Tissue sections were probed for interleukin 4 (IL-4) and interferon γ.ResultsEffector T cells constitutively expressed both interferon γ and IL-4 prototypical cytokines of TH1 and TH2, respectively, and were able to proliferate and secrete higher quantities of both cytokines in response to anti-CD3/anti-CD28 and autologous tumor lysates. Only 53% of cervical cancer Tregs were FOXP3+, elaborated transforming growth factor β1, and IL-10 and were able to inhibit both T helper subsets.ConclusionsIntratumoral Teffs represented functionally active subsets of both TH1 and TH2 that were not anergic but were suppressed by multiple Treg subsets, which comprised FOXP3 + Tregs and Tregs secreting transforming growth factor β1 and IL-10. These results imply that the microenvironment of cervical carcinomas harbored both TH1 and TH2 subsets of CD4+Teffs that were functionally active but were perhaps unable to perform because of the overpowering effect of Tregs.

Are all granzymes cytotoxic in vivo?

2014 ◽  
Vol 395 (2) ◽  
pp. 181-202 ◽  
Author(s):  
Lars T. Joeckel ◽  
Phillip I. Bird
Keyword(s):  
Serine Proteases ◽  
Ex Vivo ◽  
Cytotoxic Lymphocytes ◽  
Cytotoxic Effects ◽  
Knock Out ◽  
Granzyme A ◽  
Knock Out Mice

Abstract Granzymes are serine proteases mainly found in cytotoxic lymphocytes. The most-studied member of this group is granzyme B, which is a potent cytotoxin that has set the paradigm that all granzymes are cyototoxic. In the last 5 years, this paradigm has become controversial. On one hand, there is a plethora of sometimes contradictory publications showing mainly caspase-independent cytotoxic effects of granzyme A and the so-called orphan granzymes in vitro. On the other hand, there are increasing numbers of reports of granzymes failing to induce cell death in vitro unless very high (potentially supra-physiological) concentrations are used. Furthermore, experiments with granzyme A or granzyme M knock-out mice reveal little or no deficit in their cytotoxic lymphocytes’ killing ability ex vivo, but indicate impairment in the inflammatory response. These findings of non-cytotoxic effects of granzymes challenge dogma, and thus require alternative or additional explanations to be developed of the role of granzymes in defeating pathogens. Here we review evidence for granzyme cytotoxicity, give an overview of their non-cytotoxic functions, and suggest technical improvements for future investigations.

Effect of Curcumin Against the Transforming Growth Factor β1-Induced Myocardial Fibrosis and Mechanism

2019 ◽  
Vol 9 (10) ◽  
pp. 1435-1440
Author(s):  
Li-Ping Chen ◽  
Zai-Tao Wu
Keyword(s):  
Myocardial Fibrosis ◽  
Potential Role ◽  
Transforming Growth Factor ◽  
Cardiac Fibroblasts ◽  
Mrna Level ◽  
Transforming Growth Factor Β1 ◽  
Good Growth ◽  
Growth Status

This study investigates the mechanism of curcumin in myocardial fibrosis-mediated inhibition of myocardial fibrosis by observing the roles of curcumin in TGF-β1/smads pathway in mouse cardiac fibroblasts. The logarithmic stage cells with good growth status were selected from in vitro cultured mouse myocardial fibroblasts (CFs). Cells, at were categorized: control, vehicle control (1% DMSO), TGF-β1 (10 ng/ml) group, and low curcumin group (TGF-β1 + 1 ug/ml curcumin), medium curcumin (TGF-β1 + 3 ug/ml curcumin), and high curcumin group (TGF-β1 + 6 ug/ml curcumin). qRT-PCR was applied to calculated the mRNA level. The protein level was determined by western blot. The regulatory role of TGF-β1 in the expression of collagen I/III, Smad1, Smad2/3, Smad4, TGF-β1, and Smad7 was reversed by curcumin. Curcumin alleviates the cardiomyocytes fibrosis of mouse. The regulatory roles of curcumin in mouse cardiomyocytes and modulating TGF-β1/Smads pathways predicted its potential role in inhibiting pulmonary fibrosis and anti-fibrosis. This may provide a therapeutic strategy for myocardial fibrosis.

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