Effect of Probiotics on Intestinal Bacterial Population and Immune Modulation
Condition: Diarrhea; Constipation
MeSH terms: Constipation; Diarrhea
Study type: Interventional
Phase: Phase 4

Summary: 
      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.
    

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.
    

Intervention name(s): probiotics (antibiophilus(Lactobacillus casei), bio-three)
Intervention type(s): Dietary Supplement
Intervention description(s): probiotics (antibiophilus(Lactobacillus casei), bio-three) 4x 10^8CFU/day
Allocation: Randomized
Intervention model: Crossover Assignment
Masking: Double (Participant, Outcomes Assessor)
Primary purpose: Treatment
Keywords: probiotics