Generation of C5-dependent bioactivity by tissue-bound anti-BMZ autoantibodies

W. R. Gammon, K. B. Yancey, Karen L. Mangum, John D. Hendrix, C. H. Hammer

Research output: Contribution to journalArticle

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Abstract

We previously reported that complement-binding antibasement membrane zone (BMZ) autoantibodies can mediate complement-dependent directed migration and adherence of leukocytes to the BMZ in cryostat skin sections and that there is heterogeneity in the ability of anti-BMZ autoantibodies to mediate that response. Those observations suggested that directed migration and adherence of leukocytes to the BMZ might be dependent on the amount of complement-activating autoantibody deposited at the BMZ and the extent to which those antibodies could activate complement and generate C5-derived peptides (C5a, C5a des arg). In this study, we have examined the role of autoantibody concentration and C5 in mediating the adherence response. When cryostat skin sections were pretreated with anti-BMZ autoantibodies and subsequently incubated with neutrophils suspended in fresh serum, neutrophils adhered to the BMZ. Adherence was anti-BMZ autoantibody specific and proportional to anti-BMZ autoantibody concentration. To determine the role of C5 in mediating adherence, neutrophils were suspended in increasing concentrations of: 1) fresh serum, 2)heat-inactivated serum, 3)serum pretreated with antihuman C5, 4) serum pretreated with antihuman IgG, 5) C5-depleted serum, 6) purified C5, and 7) C5-depleted serum reconstituted with increasing concentrations of purified C5. The suspensions were then incubated with autoantibody-treated skin sections. The results showed a dose-dependent requirement for fresh serum and for C5-depleted serum reconstituted with increasing doses of C5. Adherence could be detected with C5 concentrations less than 200 ng/ml, which correspond to a C5a/C5a des arg concentration of 10-8-10-9 molar. These results suggest that complement-dependent neutrophil adherence is a highly sensitive method for detecting and quantitating the ability of tissue-deposited anti-BMZ autoantibodies to activate complement and generate C5-derived bioactive peptides, for estimating the amount of C-activating anti-BMZ autoantibody deposited at the BMZ in vivo, and for evaluating the potential role of C-activating anti-BMZ autoantibodies in the pathogenesis of lesions.

Original languageEnglish (US)
Pages (from-to)195-200
Number of pages6
JournalJournal of Investigative Dermatology
Volume93
Issue number2
StatePublished - Aug 1989

Fingerprint

Bioactivity
Autoantibodies
Tissue
Membranes
Serum
des-Arginine Complement C5a
Neutrophils
Complement C5
Skin
Cryostats
Leukocytes
Peptides
Suspensions
Immunoglobulin G
Hot Temperature

ASJC Scopus subject areas

  • Dermatology

Cite this

Gammon, W. R., Yancey, K. B., Mangum, K. L., Hendrix, J. D., & Hammer, C. H. (1989). Generation of C5-dependent bioactivity by tissue-bound anti-BMZ autoantibodies. Journal of Investigative Dermatology, 93(2), 195-200.

Generation of C5-dependent bioactivity by tissue-bound anti-BMZ autoantibodies. / Gammon, W. R.; Yancey, K. B.; Mangum, Karen L.; Hendrix, John D.; Hammer, C. H.

In: Journal of Investigative Dermatology, Vol. 93, No. 2, 08.1989, p. 195-200.

Research output: Contribution to journalArticle

Gammon, WR, Yancey, KB, Mangum, KL, Hendrix, JD & Hammer, CH 1989, 'Generation of C5-dependent bioactivity by tissue-bound anti-BMZ autoantibodies', Journal of Investigative Dermatology, vol. 93, no. 2, pp. 195-200.
Gammon, W. R. ; Yancey, K. B. ; Mangum, Karen L. ; Hendrix, John D. ; Hammer, C. H. / Generation of C5-dependent bioactivity by tissue-bound anti-BMZ autoantibodies. In: Journal of Investigative Dermatology. 1989 ; Vol. 93, No. 2. pp. 195-200.
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