Monoclonal Antibodies in Clostridium Difficile Infection

Not Applicable

Completed

• Conditions: Clostridium Infection
• Interventions: Other: Collection of peripheral blood mononuclear cells

• Registration Number: NCT04874623
• Lead Sponsor: Assistance Publique - Hôpitaux de Paris

Brief Summary

Immunoglobulin A (IgA), the major mucosal antibody, plays a key role in maintaining diversity of the intestinal microbiota and eliminating intestinal pathogens. Dysbiosis is an important risk factor for Clostridium difficile infection, which is the leading cause of nosocomial diarrhea in industrialized countries. This study aims to develop IgA monoclonal antibodies targeting C. difficile surface proteins.

Detailed Description

Immunoglobulin A (IgA), the major mucosal antibody, plays a key role in maintaining the diversity of the intestinal microbiota and eliminating intestinal pathogens. They modulate microbiota composition and also commensal bacteria homeostasis, thus promoting a symbiotic relationship. These observations, derived from mouse studies, open promising therapeutic perspectives: IgA could be used to restore or maintain a healthy microbiota in individuals suffering from intestinal dysbiosis. Specific to a pathogen, IgA can also be considered as an alternative to antibiotics by mimicking a physiological elimination.

Abnormalities in microbial diversity (i.e. dysbiosis) have been associated in humans with various diseases such as Clostridium difficile infection, which is the leading cause of nosocomial diarrhea in industrialized countries. The incidence of CDI has dramatically raised since the early 2000s, with an increasing severity. Current treatments are limited to the administration of antibiotics that unbalance the intestinal microbiota, a risk factor for relapse. These frequent relapses contribute to the severity and chronicity of the infection.

This study aims to generate human IgA-type antibodies targeting C. difficile surface proteins with neutralizing and/or protective activity. These antibodies will be selected against surface proteins involved in the early stages of colonization. After injection or ingestion, these IgA antibodies should reproduce physiological mucosal immunity, treat severe forms and prevent the occurrence of C. difficile relapses while limiting deleterious effects on the intestinal microbiota.

C. difficile-specific B cells will be selected from infected patients. After selection of the most neutralizing IgA antibodies in vitro, these will be administered to C. difficile infected mice.

Study Timeline

• Study First Submitted: 2020-09-22
• Study First Submitted QC: 2021-04-30
• Study First Posted: 2021-05-06
• Study Start: 2021-11-29
• Primary Completion: 2021-11-29
• Status Verified: 2022-09
• Study Completion: 2022-09-28
• Last Update Submitted: 2022-10-03
• Last Update Posted: 2022-10-04

Recruitment & Eligibility

• Status: COMPLETED
• Sex: All
• Target Recruitment: 15

Inclusion Criteria

• adult patient, 18 years of age
• diarrhea and/or abdominal pain.
• Presence of C. difficile toxins in feces
• Patient having dated and signed informed consent.

Exclusion Criteria

• pregnant or nursing women
• adult under guardianship
• Ileus
• Peritonitis
• Pseudomembranous colitis
• Hemodynamic instability
• Fever ≥ 38.5°C
• Shivers
• Respiratory failure
• Leukocytosis > 15x109/L
• Serum creatinin >50% reference values)
• Serum lactate > 5mM
• Serum Albumine < 30g/l
• Other diarrhea cause
• Humoral immunodeficiency

Study & Design

• Study Type: INTERVENTIONAL
• Study Design: SINGLE_GROUP

Arm && Interventions

Group	Intervention	Description
CDI	Collection of peripheral blood mononuclear cells	-

Primary Outcome Measures

Name	Time	Method
Generate C. difficile specific antibodies from circulating memory B lymphocytes of C. difficile patients.	From the blood samples taken between Day 1 and Day 7 after the diagnostic of CDI	The presence of SlpA and Cwp84 specific memory B cells will be assessed from a single sample per patient. After separation of blood mononuclear cells, specific memory B cells will be selected using microfluidic drop technology. The variable VH and VL fragments of the immunoglobulins, which define the specificity of the antibodies, will be sequenced for each selected B cell. The corresponding antibodies will then be produced in a well-established cell line.

Secondary Outcome Measures

Name	Time	Method
Neutralizing capacity evaluation affinity	From the blood samples taken between Day1 and Day7 after the diagnostic of CDI	The neutralizing activity will be evaluated in vitro: the capacity of the antibodies to decrease the proliferation of the bacteria (in liquid culture), to decrease its mobility (solid culture), to inhibit biofilm formation and to inhibit its adhesion to the surface of intestinal epithelial cells (quantified using the Caco-213 line).
Antibody affinity assessment	From the blood samples taken between Day1 and Day7 after the diagnostic of CDI	The affinity of the antibodies will be evaluated by surface plasmon resonance (Biacore®).

Trial Locations

Locations (1)

Maladies infectieuses et tropicales, Hôpital Pitié-Salpêtrière; 🇫🇷 Paris, France