The Effect of Probiotics on Type 1 Diabetes Mellitus in Children

Study Overview
=================
Brief Summary
-----------------
In this study, investigators try to administer probiotics (Lactobacillus salivarius + Lactobacillus johnsonii + Bifidobacterium lactis from glac biotech Co., Ltd.) to children T1DM patients for 6 months to observe if the inhibition effect of T1DM animal model could be discerned in a short-term period from both change of serum cytokines and beta cells insulin secretion ability.

Detailed Description
-----------------
Type 1 (insulin-dependent) Diabetes Mellitus (T1DM) is among the most well studied organ-specific autoimmune diseases which approximately 75% of newly diagnosed DM patients acquire this type before the age of 18. T1DM is well known for as the consequence of selective destruction of pancreatic insulin-producing beta cells within the islets of Langerhans. Basically, autoimmune reactions against beta cells may come from activation of the immune system in genetically susceptible individuals triggered by environmental factors that bear epitopes similar to those expressed by the beta cells. Several mechanisms such as molecular mimicry, metabolic stress on beta cells, cryptic epitope exposure and costimulatory molecule upregulation have been proposed but none of them could be solely responsible for the pathogenesis of T1DM. Recently, T1DM has been considered a consequence of dysregulated or over-activation of immune responses in genetically predisposed individuals, similar to other autoimmune diseases. The rapid increase in the incidence of T1DM in developed countries including Taiwan during recent decades refers to the role of environmental factors in this disease. Candidate environmental factors influencing T1DM include various microbial and food components encountered at mucosal surfaces as well as gut mucosal parameters such as gut permeability. However, difficulty exists in characterizing the environmental factors and mechanisms in T1DM because of their complexity of interaction, the long lag period between the induction of disease trigger factors and the clinical onset of the disease. Environmental factors in T1DM seem to prevent full penetration of the disease rather than trigger it. It had been reported that high diabetes incidence in germ-free mice and an involvement of innate immune mechanisms in the disease. In this study, investigators try to administer probiotics (Lactobacillus salivarius + Lactobacillus johnsonii + Bifidobacterium lactis from glac biotech Co., Ltd.) to children T1DM patients for 6 months to see if the inhibition effect of T1DM animal model could be discerned in a short-term period from both change of serum cytokines and beta cells insulin secretion ability. Subjects will collect blood before the test and every 3 months after the test for total 4 times. Each time the collected blood volume is about 5 8cc. A part of the blood sample will be given to the Department of laboratory medicine for the detection of hemoglobin A1c (HbA1c) and fasting blood glucose, and the other part will be centrifuged to separate serum. The serum macrophage inflammatory proteins-1beta (MIP-1β), regulated on activation, normal T cell expressed and secreted (RANTES), interleukin-8 (IL-8), interleukin-17 (IL-17), tumor necrosis factor alpha (TNF-α) and transforming growth factor beta1 (TGF-β1) concentrations will be measured by ELISA.

Official Title
-----------------
The Effect of Probiotics on Type 1 Diabetes Mellitus in Children

Conditions
-----------------
Type 1 Diabetes Mellitus

Intervention / Treatment
-----------------
* Other: L. johnsonii MH-68, B. animalis subsp. lactis CP-9 and L. salivarius AP-32
* Other: Placebo


Participation Criteria
=================
Eligibility Criteria
-----------------
Inclusion Criteria: Age between 6 to 18 years old. T1DM patients confirmed by glucagon tests and/or presence of autoantibody(ies). Exclusion Criteria: Significant cardiac, renal and hepatic disease. The physician diagnosed the immunodeficiency or the immune function was low. Currently using probiotics supplements or had ever taken probiotics for more than one month. Currently using antibiotics or gastrointestinal medicine. Ever allergic reaction(s) to probiotics or prebiotics regimen.

Ages Eligible for Study
-----------------
Minimum Age: 6 Years
Maximum Age: 18 Years

Sexes Eligible for Study
-----------------
All

Accepts Healthy Volunteers
-----------------
No

Study Plan
=================
How is the study designed?
-----------------

Design Details

Primary Purpose: Treatment
Allocation: Randomized
Intervention Model: Parallel Assignment
Interventional Model Description: Probiotics or placebo group
Masking: Double


Arms and Interventions

| Participant Group/Arm | Intervention/Treatment |
| --- | --- |
| Experimental: Probiotics capsule<br>Taking 1 L. johnsonii MH-68, B. animalis subsp. lactis CP-9 and L. salivarius AP-32 mix probiotics capsule twice a day before meals for six months. | Other: L. johnsonii MH-68, B. animalis subsp. lactis CP-9 and L. salivarius AP-32<br>* Taking 1 L. johnsonii MH-68, B. animalis subsp. lactis CP-9 and L. salivarius AP-32 mix probiotics capsule twice a day before meals for six months.<br>|
| Placebo Comparator: Placebo capsule<br>Taking 1 placebo capsule twice a day before meals for six months. | Other: Placebo<br>* Taking 1 placebo capsule twice a day before meals for six months.<br>|

What is the study measuring?
-----------------
Primary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in percentage of HbA1c | Subjects will draw blood once before the test. During the test, every 3 months will draw blood to 6th month, each time the blood volume is about 5   8cc to detect HbA1c and other blood biochemical values. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of blood glucose (AC) | The study will require subject to record their own daily fasting blood glucose. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 

Secondary Outcome Measures

| Outcome Measure | Measure Description | Time Frame |
| --- | --- | --- |
| Change in concentration of MIP-1β | Serum was isolated by extra blood draw, serum MIP-1β (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of RANTES | Serum was isolated by extra blood draw, serum RANTES (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of IL-8 | Serum was isolated by extra blood draw, serum IL-8 (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of IL-17 | Serum was isolated by extra blood draw, serum IL-17 (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of TNF-α | Serum was isolated by extra blood draw, serum TNF-α (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 
| Change in concentration of TGF-β1 | Serum was isolated by extra blood draw, serum TGF-β1 (pg/ml) concentrations were measured from first blood draw to third blood draw by ELISA. | From date of first blood draw after entering the trial until the date of third blood draw, assessed up to 6 months. | 

 Terms related to the study
=================
Keywords Provided by Centre Hospitalier Valida
-----------------
T1DM, Autoimmune diseases, Microbiota, Probiotics