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NCT Number: NCT03842787

Anti-ficolin-3 Antibodies in Lupus Nephritis

Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by the production of multiple autoantibodies and accumulation of immune complexes resulting in systemic inflammatory response and tissue damage. Although the underlying mechanisms are complex, defects in dying cells elimination are likely to contribute to autoantigen overload and development of autoimmunity. Molecules important in damaged cell clearance, such as early complement components, may thus have a protective role. According to this hypothesis, deficiencies in C1q and MBL, the recognition proteins of the classical and lectin pathways of complement; are associated with increased susceptibility to SLE. In the proposed project, the investigators will investigate the involvement of another related recognition protein, ficolin-3, which activates the complement lectin pathway and recognizes necrotic cells. The investigators have shown in a recent study a significant association between the presence of anti-ficolin-3 antibodies and active nephritis in patients with SLE. However, the possible involvement of anti-ficolin-3 antibodies in the pathogenesis of SLE and particularly in lupus nephritis (LN) remains to be elucidated. This project plans to investigate the role of ficolin-3 and ficolin-3 autoantibodies in LN. The study associates two aspects, aiming at deciphering the role of anti-ficolin-3 antibodies in dying cells recognition and investigating the role of ficolin-3 in renal tissue damage. This pilot study will be performed for 14 patients with active LN on serum and renal biopsy, realized for routine patient care. The investigators will explore the effect of anti-ficolin-3 antibodies purified from the patient serum on ficolin-3-dependent necrotic cells recognition, in relation with possible altered clearance of dead cells, which is an important hypothesis of the pathogenesis of SLE. The investigators will also investigate ficolin-3 deposition in renal biopsy, which may contribute to the local formation of immune complexes, leading to complement activation and subsequent inflammation and tissue injury.

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Key information

Conditions

Systemic Lupus Erythematosus Nephritis Autoimmune Diseases Connective Tissue Diseases Female Urogenital Diseases Female Urogenital Diseases and Pregnancy Complications Glomerulonephritis Immune System Diseases Kidney Diseases Lupus Erythematosus, Systemic Lupus Nephritis Male Urogenital Diseases Nephritis Skin and Connective Tissue Diseases Urogenital Diseases Urologic Diseases

Age range

18 year and older

Sex eligibility

All sexes

Study type

Observational

Primary location

JOUVE

La Tronche, France

About this study

PRIMARY OUTCOME MEASURE Exploration of the inhibition of anti-ficolin-3 antibodies purified from the serum of 14 patients with active lupus nephritis in ficolin-3-dependent necrotic cells recognition.

The criterion used is the shift of MFI (Mean Fluorescence Intensity) measured after addition of these antibodies to necrotic Jurkat cells incubated with ficolin-3.

The study has a single visit approach with serum collection, so every outcome is measured at T0, which is the only visit for the patient.

SECONDARY OUTCOME MEASURES

  • Investigation of ficolin-3 deposition in renal biopsy of the same 14 patients with active LN.

Analysis: deposition of ficolin-3 will be evaluated by immunostaining on renal biopsy.

  • Quantification of anti-ficolin-3 antibodies. Analysis: Anti-ficolin-3 antibodies are quantified by ELISA.
  • Quantification of serum levels of ficolin-3. Analysis: Ficolin-3 is quantified by ELISA.
  • Correlation between anti-ficolin-3 antibodies and serum levels of ficolin-3. Analysis: Anti-ficolin-3 antibodies and ficolin-3 are quantified by ELISA.
  • Correlation between serum levels of anti-ficolin-3 antibodies and ficolin-3 deposition in the kidney.
  • Correlation between serum levels of ficolin-3 and ficolin-3 deposition in the kidney.
  • Exploration of the inhibition of anti-ficolin-2 antibodies purified from the serum of 14 patients with active lupus nephritis in ficolin-2-dependent necrotic cells recognition.

The criterion used is the shift of MFI (Mean Fluorescence Intensity) measured after addition of these antibodies to necrotic Jurkat cells incubated with ficolin-2.

  • Investigation of ficolin-2 deposition in renal biopsy of the same 14 patients with active LN.

Analysis: deposition of ficolin-2 will be evaluated by immunostaining on renal biopsy.

  • Quantification of anti-ficolin-2 antibodies. Analysis: Anti-ficolin-2 antibodies are quantified by ELISA.
  • Quantification of serum levels of ficolin-2. Analysis: Ficolin-2 is quantified by ELISA.
  • Correlation between anti-ficolin-2 antibodies and serum levels of ficolin-2. Analysis: Anti-ficolin-2 antibodies and ficolin-2 are quantified by ELISA.
  • Correlation between serum levels of anti-ficolin-2 antibodies and ficolin-2 deposition in the kidney.
  • Correlation between serum levels of ficolin-2 and ficolin-2 deposition in the kidney.

Who can participate

Healthy volunteers accepted: No

Only the study team can determine whether someone qualifies for participation.

Inclusion criteria

  • Age ≥ 18 years old
  • Patients who have valid health insurance
  • Non-opposition to participation obtained
  • Diagnostic de lupus according to SLICC 2012, performed more than 3 months ago.
  • Active lupus nephritis defined by :

elevated SLEDAI indexes (≥ 4), the presence of a significant proteinuria (≥ 0.5 g/day) and/or the presence of hematuria, aseptic leukocyturia or urinary casts, and documented by renal biopsy and classified according to the ISN/RPS classification.

Non-inclusion Criteria:

  • Patient with a known progressing cancer
  • Patient who had started lupus nephritis flare treatment
  • Participant involved in another interventional clinical study
  • Person deprived of liberty by judicial order
  • Person under guardianship or curatorship
  • Hemoglobin level < 7 g/dL

Treatment and study plan

Biological analysis

Other

Biological and research analysis:

Quantification of ficolin-3, anti-ficolin-3 antibodies, ficolin-2, anti-ficolin-2 antibodies

Purification of patients' antibodies (anti-ficolin-3 and -2)

Evaluation of effects of anti-ficolin-3 and anti-ficolin-2 purified antibodies Investigation of ficolin-3 and ficolin-2 deposition in renal biopsy

Primary outcomes

  1. Exploration of the inhibition of anti-ficolin-3 antibodies purified from the serum of 14 patients with active lupus nephritis in ficolin-3-dependent necrotic cells recognition.

    Time frame: Measure at day of inclusion = TO

    In order to investigate the possible interference of the anti-ficolin-3 antibodies purified from patients'sera with ficolin-3 dependent necrotic cells recognition, recombinant ficolin-3 will be preincubated with the purified specific autoantibodies before addition to Jurkat necrotic cells. Ficolin-3 binding will be measured using the flow cytometry and immunofluorescence assays described above and quantified using the mean fluorescence intensity (MFI). The criterion used is the shift of MFI (Mean Fluorescence Intensity) measured after addition of these antibodies to necrotic Jurkat cells incubated with ficolin-3.

Secondary outcomes

  1. Investigation of ficolin-3 deposition in renal biopsy of the same 14 patients with active LN.

    Time frame: Measure at day of inclusion = TO

    The investigators will investigate the presence of ficolin-3 in the glomeruli by direct immunofluorescence analysis. They search deposits in the interstitial vessels, interstitium and glomeruli (mesangium, extra-membranous, glomerular basement membrane) and quantify them semi-quantitatively (+, ++ or +++).

  2. Quantification of anti-ficolin-3 antibodies.

    Time frame: Measure at day of inclusion = TO

    Anti-ficolin-3 antibodies are quantified by ELISA. Results are given in Arbitrary Units (AU)

  3. Quantification of serum levels of ficolin-3.

    Time frame: Measure at day of inclusion = TO

    Ficolin-3 is quantified by ELISA. Results are given in µg/mL.

  4. Correlation of anti-ficolin-3 antibodies and serum levels of ficolin-3.

    Time frame: Measure at day of inclusion = TO

    Anti-ficolin-3 antibodies and ficolin-3 are quantified by ELISA.

  5. Correlation between serum levels of anti-ficolin-3 antibodies and ficolin-3 deposition in the kidney.

    Time frame: Measure at day of inclusion = TO

    Serum levels of anti-ficolin-3 antibodies and ficolin-3 deposition in the kidney are quantified by ELISA.

  6. Correlation between serum levels of ficolin-3 and ficolin-3 deposition in the kidney.

    Time frame: Measure at day of inclusion = TO

    Serum levels of ficolin-3 and ficolin-3 deposition in the kidney are quantified by ELISA.

  7. Exploration of the inhibition of anti-ficolin-2 antibodies purified from the serum of 14 patients with active lupus nephritis in ficolin-2-dependent necrotic cells recognition.

    Time frame: Measure at day of inclusion = TO

    In order to investigate the possible interference of the anti-ficolin-2 antibodies purified from patients'sera with ficolin-2 dependent necrotic cells recognition, recombinant ficolin-2 will be preincubated with the purified specific autoantibodies before addition to Jurkat necrotic cells. Ficolin-2 binding will be measured using the flow cytometry and immunofluorescence assays described above and quantified using the mean fluorescence intensity (MFI). The criterion used is the shift of MFI (Mean Fluorescence Intensity) measured after addition of these antibodies to necrotic Jurkat cells incubated with ficolin-2.

  8. Investigation of ficolin-2 deposition in renal biopsy of the same 14 patients with active LN.

    Time frame: Measure at day of inclusion = TO

    The investigators will investigate the presence of ficolin-2 in the glomeruli by direct immunofluorescence analysis. They search deposits in the interstitial vessels, interstitium and glomeruli (mesangium, extra-membranous, glomerular basement membrane) and quantify them semi-quantitatively (+, ++ or +++).

  9. Quantification of anti-ficolin-2 antibodies.

    Time frame: Measure at day of inclusion = TO

    Anti-ficolin-2 antibodies are quantified by ELISA. Results are given in arbitrary units (AU).

  10. Quantification of serum levels of ficolin-2.

    Time frame: Measure at day of inclusion = TO

    Ficolin-2 is quantified by ELISA. Results are given in µg/mL.

  11. Correlation between anti-ficolin-2 antibodies and serum levels of ficolin-2.

    Time frame: Measure at day of inclusion = TO

    Anti-ficolin-2 antibodies and ficolin-2 are quantified by ELISA.

  12. Correlation between serum levels of anti-ficolin-2 antibodies and ficolin-2 deposition in the kidney.

    Time frame: Measure at day of inclusion = TO

    Serum levels of anti-ficolin-2 antibodies and ficolin-2 deposition in the kidney are quantified by ELISA.

  13. Correlation between serum levels of ficolin-2 and ficolin-2 deposition in the kidney.

    Time frame: Measure at day of inclusion = TO

    Serum levels of ficolin-2 and ficolin-2 deposition in the kidney are quantified by ELISA.

Sponsors and collaborators

Lead sponsor

University Hospital, Grenoble

Other

Collaborators

  • Institut de Biologie Structurale Grenoble

Registry information

Official study title

Role of Anti-ficolin-3 Antibodies in the Pathogenesis of Lupus Nephritis

Acronym: IgFicoLupus

Important dates

Study start
2019
Primary completion
2020
Study completion
2020
First posted
Feb 15, 2019
Registry last updated
Sep 28, 2021

OpenTrials presents study information sourced from ClinicalTrials.gov. The official registry record should be consulted for the latest information.

View the official ClinicalTrials.gov record (opens in a new tab)

This listing is for discovery and informational purposes only. It is not medical advice, does not guarantee that a study is recruiting, and does not determine eligibility. Contact the study team and a qualified healthcare professional when considering participation.

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