Effect of Probiotics on Intestinal Bacterial Population and Immune Modulation

This study has been completed.
Sponsor:
Information provided by (Responsible Party):
Chien-Chang Chen, MD, Chang Gung Memorial Hospital
ClinicalTrials.gov Identifier:
NCT00763399
First received: September 23, 2008
Last updated: July 16, 2012
Last verified: July 2012
  Purpose

The balance between immunogenic and tolerogenic activities in human immune system strongly depends on microflora-induced pro-and anti-inflammatory activities. Probiotics are important components of microflora. The interactions of the different strains of probiotics and the cells of immune system are largely unknown.

There are many mechanisms by which probiotics enhance intestinal health, including stimulation of immunity, competition for limited nutrients, inhibition of epithelial and mucosal adherence, inhibition of epithelial invasion and production of antimicrobial substances.

Fecal immunoglobulin A(IgA), lactoferrin and calprotectin were determined by enzyme-linked immunosorbent assay(ELISA) and compared in different groups. Other clinical symptoms or signs, including fever, vomiting, diarrhea, abdominal pain, bloating abdomen, daily intake and body weight were also assessed.

The first aim of our study is to evaluate the role of probiotics and their preparation products on the restoration of intestinal bacterial population. The second aim of our study is determining the immunomodulating effects or anti-inflammatory effects of probiotics on the host (human being). We try to seek to gain an advanced understanding of probiotics versus intestinal microorganism and host interactions, as well as mucosal immune responses to probiotics in the intestine.


Condition Intervention Phase
Diarrhea
Constipation
Dietary Supplement: probiotics (antibiophilus(Lactobacillus casei), bio-three)
Phase 4

Study Type: Interventional
Study Design: Allocation: Randomized
Intervention Model: Crossover Assignment
Masking: Double Blind (Subject, Outcomes Assessor)
Primary Purpose: Treatment
Official Title: Phase 4 Study of Probiotics on Intestinal Bacterial Population and Immune Modulation

Resource links provided by NLM:


Further study details as provided by Chang Gung Memorial Hospital:

Primary Outcome Measures:
  • Duration of diarrhea, number of stool passage, bacterial culture for intestinal or cytokine [ Time Frame: within 4 weeks ] [ Designated as safety issue: Yes ]

Secondary Outcome Measures:
  • body weight change, appetite and daily intake, Fecal IgA, lactoferrin and calprotectin levels [ Time Frame: Within first two weeks ] [ Designated as safety issue: Yes ]
    secondary outcome including body weight change, appetite and daily intake, bloating or abdominal distension, abdominal pain or colic, constipation, fever, and vomiting were also assessed. IgA levels and bacterial culture were performed on homogenized fecal samples. Fecal lactoferrin and calprotectin levels were measured with samples of feces.


Estimated Enrollment: 300
Study Start Date: October 2008
Study Completion Date: May 2010
Primary Completion Date: May 2010 (Final data collection date for primary outcome measure)
Arms Assigned Interventions
Experimental: 97-0549B Dietary Supplement: probiotics (antibiophilus(Lactobacillus casei), bio-three)
probiotics (antibiophilus(Lactobacillus casei), bio-three) 4x 10^8CFU/day
Other Name: probiotics (antibiophilus(Lactobacillus casei), bio-three)

Detailed Description:

Some clinical parameters were evaluated according to the following: primary outcome (severity of diarrhea), and secondary outcome including body weight change, appetite and daily intake, bloating or abdominal distension, abdominal pain or colic, constipation, fever, and vomiting were also assessed.

Peripheral blood isolated by Lymphoprep, washed twice in normal saline and once in medium, and suspended in medium [RPMI 1640] to a density of 1 x 106/mL. PBMCs will be isolated from blood donor buffy coats by density gradient centrifugation. The concentration of PBMCs will be adjusted to 106 cells per ml in complete medium, and the cells will be transferred to 24-well plates.Cell surface phenotype expression and intracellular staining. Cells will be stained using a panel of monoclonal antibody (MAb) directed against surface antigens expressed by lymphocytes, monocytes and the appropriate species-specific immunoglobulin G isotype controls. Cells will be acquired using an FACScan (Becton Dickinson) and analyzed with Cell Quest software.

To assess the colonization of intestinal bacteria, fecal samples were collected from each patient on day 0 (the day when patients were enrolled), day 3 and day 7 after probiotics or placebo treatment. The fecal specimens were weighed, homogenized, and serially diluted and plated on selective agar for analysis of bacteria. Fecal bacteria count was expressed as log10 CFU/g feces.

Fecal samples were collected during the treatment period. IgA levels were performed on homogenized fecal samples. Total IgA was determined using goat anti-human IgA-HRP conjugate. The reaction was developed with tetramethyl benzidine (TMB; Zymed Labs.) and read at 450 nm. OD values were converted to ng/g feces of total IgA by comparison with a standard curve developed with anti-human IgA.

The stool samples were prepared and analyzed for lactoferrin. A polyclonal antibody specific for lactoferrin has been pre-coated onto a microplate. Lactoferrin in standards and samples is sandwiched by the immobilized antibody and a biotinylated polyclonal antibody specific for lactoferrin, which is recognized by a streptavidin-peroxidase conjugate. Absorbance is read at OD 450 nm. Lactoferrin was expressed as μg/g feces.

The stool samples were prepared and analyzed for calprotectin. The supernatant was collected and frozen at -20°C. The supernatants were thawed and calprotectin was analyzed with the quantitative calprotectin ELISA and read at OD 450 nm. Calprotectin was expressed as μg/g feces.

  Eligibility

Ages Eligible for Study:   3 Months to 10 Years
Genders Eligible for Study:   Both
Accepts Healthy Volunteers:   Yes
Criteria

Inclusion Criteria:

  • Diarrhea
  • constipation

Exclusion Criteria:

  • Shock
  • Sepsis
  • Past history with GI tract surgery
  • Immunodeficiency
  Contacts and Locations
Choosing to participate in a study is an important personal decision. Talk with your doctor and family members or friends about deciding to join a study. To learn more about this study, you or your doctor may contact the study research staff using the Contacts provided below. For general information, see Learn About Clinical Studies.

Please refer to this study by its ClinicalTrials.gov identifier: NCT00763399

Locations
Taiwan
Chang Gung Memorial Hospital
Taoyuan, Taiwan, 333
Sponsors and Collaborators
Chang Gung Memorial Hospital
Investigators
Principal Investigator: Chien-Chang Chen, MD Pediatric Department, Chang Gung Memorial Hospital, Taiwan
  More Information

No publications provided

Responsible Party: Chien-Chang Chen, MD, Department of pediatrics, Chang Gung Memorial Hospital
ClinicalTrials.gov Identifier: NCT00763399     History of Changes
Other Study ID Numbers: 97-0549B
Study First Received: September 23, 2008
Last Updated: July 16, 2012
Health Authority: Taiwan: Institutional Review Board

Keywords provided by Chang Gung Memorial Hospital:
probiotics

Additional relevant MeSH terms:
Constipation
Diarrhea
Signs and Symptoms, Digestive
Signs and Symptoms

ClinicalTrials.gov processed this record on July 20, 2014