Nafamostat to Stabilise Plasma Samples Taken for Complement Measurements

Watkins, J.; Wild, Graeme; Smith, S.E.

doi:10.1016/s0140-6736(89)92883-3

Cited by 18 publications

(11 citation statements)

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“…The recent development, however, of a synthetic protease inhibitor FUT-175, nafamstat mesilate (Aoyama et ai, 1984) ('Futhan'), which when combined with disodium ethylene diamine tetra acetate (EDTA) is capable of minimizing the spurious in vitro activation of complement that occurs when EDTA alone is used, now enables reliable investigation of m vivo Correspondence; Dr P. Hughes, Department of Medicine, Clinical Sciences Centre, Northern General Hospital, Sheffield S5 7AU, UK. complement activation to be made (Watkins, Wild & Smith, 1989). In the present study, such an approach has been used to examine the usefulness of C3a and C4a anaphylatoxin levels as a measure of disease 'activity' by simultaneously estimating total C3 and C4 complement levels, circulating Clq binding immune complexes and DNA antibodies in patients with SLE whose disease was treated either purely by prednisolone or was sufficiently severe to require both azathioprine and prednisolone.…”

Section: Introductionmentioning

confidence: 99%

C4a anaphylatoxin levels as an indicator of disease activity in systemic lupus erythematosus

Wild

Watkins

Ward

et al. 1990

Clinical and Experimental Immunology

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SUMMARYThe use of a synthetic protease inhibitor, nafamstat mesilate, has enabled reliable estimations of in vivo complement activation to be made in systemic lupus erythematosus (SLE). Elevation of C3a anaphylatoxins was found in two out of 24 patients and elevation of C4a anaphylatoxins was found in 20 out of 24 patients, confirming that complement activation, predominantly by the classical pathway, is a common occurrence in the disease. Significantly higher levels of C4a anaphylatoxin were found in 16 patients, with more aggressive disease requiring supplementary treatment with azathioprine, while the remaining eight patients, with less severe disease, required purely steroid therapy. Very strong associations between elevated C4a anaphylatoxins and raised DNA antibody titres, Clq binding activity and low complement C4 levels were also observed, suggesting that anaphylatoxin measurement may be a sensitive additional method for monitoring disease activity in SLE.

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Section: Introductionmentioning

confidence: 99%

C4a anaphylatoxin levels as an indicator of disease activity in systemic lupus erythematosus

Wild

Watkins

Ward

et al. 1990

Clinical and Experimental Immunology

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“…which, when combined with EDTA. is capable of minimizing the spurious in vitro activation of comple ment that occurs when EDTA alone is used, now ena bles reliable investigation of in vivo complement activa tion to be made [15,18,19], In the extraction of horny tissues, we used a buffer that contained a cocktail of protease inhibitors as well as EDTA to prevent an in vitro complement activation by proteases which might have been released from kcratinocytes and infiltrated leukocytes.…”

Section: Discussionmentioning

confidence: 99%

Generation of Terminal Complement Complexes in Psoriatic Lesional Skin

Takematsu

Tagami

1992

Dermatology

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The complement system is thought to play an important role in the recruitment of neutrophils within the epidermis. In the present study we examined whether or not complement activation in psoriatic lesional skin results in the deposition of terminal complement complexes within the epidermis by measuring levels of SC5b-9 in the plasma and horny tissues of psoriatic patients. The levels of SC5b-9 in psoriatic plasma were significantly higher than those of controls or those of patients with atopic dermatitis. However, when the levels of SC5b-9 in the psoriatic plasma were compared before and after successful treatment of psoriasis, a significant reduction was observed after treatment. Studies of total protein extracts from lesional skin showed that, while no SC5b-9 was detected in the noninflammatory horny tissues, there were high levels of SC5b-9 in lesional horny tissues of psoriasis. By immunofluroescence using a monoclonal antibody to the C5b-9 neoantigen, deposition of C5b-9 was observed only in the stratum corneum of psoriatic skin. Thus the results of the present study suggests that in psoriatic lesional skin, the complement system is activated and that this complement activation proceeds all the way to the terminal step, generating membrane attack complex.

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“…Venous blood samples were drawn into tubes containing EDTA and Futhan [8]; following separation, plasma was stored at -20°C until assayed for C3, C4, C3a and C4a within 1 week o f collection.…”

Section: Methodsmentioning

confidence: 99%

“…Previously, accurate measurement of these components was made difficult by the ease with which complement is activated in vitro when blood samples are collected in EDTA tubes. However, addition of the recently developed synthetic protease inhibitor FUT-175, nafamstat mesilate (Futhan; Amersham International pic, Amersham, UK) to EDTA in the sample collection tube has proved reliable in minimizing spurious in vitro complement activation, thus enabling a more sensitive estimation of in vivo complement activation to be made [8].…”

Section: Introductionmentioning

confidence: 99%

Plasma Levels of the Anaphylatoxins C3a and C4a in Patients with IgA Nephropathy/Henoch-Schönlein Nephritis

Abou-Ragheb

Williams

Brown

et al. 1992

Nephron

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Measurements of plasma anaphylatoxins C3a and C4a were carried out as an indicator of in vivo complement activation in 46 patients suffering from IgA nephropathy/Henoch-Schönlein nephritis. There was a significant correlation between plasma levels of C4a and plasma creatinine and urea. We also found a significant correlation of plasma levels of C3a with plasma creatinine. We propose that the measurement of anaphylatoxin levels provides a sensitive indicator of in vivo complement activation and may serve as an additional method for monitoring the progress of disease in these patients.

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Nafamostat to Stabilise Plasma Samples Taken for Complement Measurements

Cited by 18 publications

References 0 publications

C4a anaphylatoxin levels as an indicator of disease activity in systemic lupus erythematosus

C4a anaphylatoxin levels as an indicator of disease activity in systemic lupus erythematosus

Generation of Terminal Complement Complexes in Psoriatic Lesional Skin

Plasma Levels of the Anaphylatoxins C3a and C4a in Patients with IgA Nephropathy/Henoch-Schönlein Nephritis

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