Lactic Acid Bacteria for Coeliac Disease

Abstract

The present invention discloses strains of Lactobacillus and Streptococcus which have a capacity to degrade gliadin peptides involved in coeliac disease and which peptide degrading activity is stable under low pH and in the presence of mammalian digestive enzymes. These strains are suitable in a product for use in prevention and/or treatment of celiac disease.

1 . A method of treating or reducing the likelihood of acquiring coeliac disease in a subject in need thereof, the method comprising administering to the subject a strain of lactic acid bacterium or a medium fermented by a strain of lactic acid bacterium, wherein said lactic acid bacterium or medium fermented by a lactic acid bacterium is capable of degrading the 33-mer peptide LQLQPFPQPQLPYPQPQLPYPQPQLPYPQPQPF (SEQ ID NO: 1), the 20-mer peptide QQLPQPQQPQQSPFQQQRPF (SEQ ID NO: 2), the 13-mer peptide LGQQQPFPPQQPY (SEQ ID NO: 3) or the 18-mer peptide PQLPYPQPQLPYPQPQPF (SEQ ID NO: 4), and wherein the lactic acid bacterium is selected from the group consisting of the genus  Lactobacillus  and  Streptococcus.    
     
     
         2 . The method of strain  claim 1 , wherein said lactic acid bacteria or wherein said medium fermented by said lactic acid bacteria are capable of degrading the 33-mer peptide of SEQ ID NO: 1 and the 18-mer peptide of SEQ ID NO: 4. 
     
     
         3 . The method of  claim 1 , wherein said lactic acid bacterium produces protein elongation factors or wherein the medium fermented by said lactic acid bacteria comprises protein elongation factors, and wherein said protein elongation factors have a peptidase activity capable of degrading the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 and/or the 18-mer peptide of SEQ ID NO: 4. 
     
     
         4 . The method of  claim 1 , wherein the degradation of said peptides occurs in the small intestine of a subject. 
     
     
         5 . The method of  claim 3 , wherein at least 45% of the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 or the 18-mer peptide of SEQ ID NO: 4 is hydrolyzed in the presence of a culture supernatant of said strains. 
     
     
         6 . The method of  claim 1 , wherein said lactic acid bacteria is selected from the group consisting of  Lactobacillus casei  CNCM I-1518, Lactobacillus helveticus CNCM I-4279,  L. delbrueckii  subsp  lactis  CNCM I-4280,  Lactobacillus casei  CNCM I-4270 and  Streptococcus thermophilus  CNCM I-4269. 
     
     
         7 . A method of treating or reducing the likelihood of acquiring coeliac disease in a subject in need thereof, the method comprising administering to the subject a mixture of two strains of lactic acid bacteria or mixture of a medium fermented by two strains of lactic acid bacteria, wherein said lactic acid bacteria or medium fermented by lactic acid bacteria are capable of degrading the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 or the 18-mer peptide of SEQ ID NO: 4, and wherein the lactic acid bacteria are selected from the group consisting of the genus  Lactobacillus  and  Streptococcus.    
     
     
         8 . The method of  claim 7 , wherein the lactic acid bacteria are  S. thermophilus  CNCM I-2776 and  L. bulgaricus  CNCM I-2787. 
     
     
         9 . (canceled) 
     
     
         10 . A composition comprising at least one strain of lactic acid bacterium selected from the group consisting of  Lactobacillus helveticus  CNCM I-4279,  L. delbrueckii  subsp  lactis  CNCM I-4280,  Lactobacillus casei  CNCM I-4270,  Streptococcus thermophilus  CNCM I-4269 and a mixture of  S. thermophilus  CNCM I-2776 and  L. bulgaricus  CNCM I-2787. 
     
     
         11 . The composition according to  claim 10  where in the composition is administered to a subject to prevent or reduce the likelihood of acquiring coeliac disease. 
     
     
         12 . The composition of  claim 10 , wherein the composition is a liquid product. 
     
     
         13 . The composition of  claim 10 , wherein the composition is a dairy product. 
     
     
         14 . The composition of  claim 10 , wherein the composition is a fermented product. 
     
     
         15 . The composition of  claim 10 , wherein the composition comprises 10 5  to 10 13  cfu per g dry weight. 
     
     
         16 . The composition according to  claim 10 , wherein the lactic acid bacteria are administered to the subject at a dose of at least 10 10  cfu/day. 
     
     
         17 . A method of producing a protein elongation factor for degradation of the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 and/or the 18-mer peptide of SEQ ID NO: 4, the method comprising fermenting at least one strain of lactic acid bacteria selected from the group consisting of the genus  Lactobacillus  and  Streptococcus.    
     
     
         18 . The method of  claim 17  wherein the lactic acid bacteria are selected from the group consisting of  Lactobacillus casei  CNCM I-1518,  Lactobacillus casei  CNCM I-4270,  Lactobacillus helveticus  CNCM I-4279,  Lactobacillus delbrueckii  subsp  lactis  CNCM I-4280, and a mixture of  Streptococcus thermophilus  CNCM I-2776 and  Lactobacillus bulgaricus  CNCM I-2787. 
     
     
         19 . A lactic acid bacteria or a mixture of two strains of lactic acid bacteria selected from the group consisting of  Lactobacillus casei  CNCM I-4270,  Lactobacillus helveticus  CNCM I-4279,  Lactobacillus delbrueckii  subsp  lactis  CNCM I-4280,  Streptococcus thermophilus  CNCM I-2776,  Lactobacillus bulgaricus  CNCM I-2787, and the mixture of  Streptococcus, thermophilus  CNCM I-2776 and  Lactobacillus bulgaricus  CNCM I-2787. 
     
     
         20 . A method of selecting strains of lactic acid bacteria for use in treatment of coeliac disease comprising the steps of selecting the strains with the capacity of degrading the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 or the 18 mer peptide of SEQ ID NO: 4, when at least 45% of said peptide is hydrolyzed in the presence of a culture supernatant of said strains. 
     
     
         21 . The method according to  claim 20  further comprising the step of selecting the strains wherein the capability to degrade the 33-mer peptide of SEQ ID NO: 1, the 20-mer peptide of SEQ ID NO: 2, the 13-mer peptide of SEQ ID NO: 3 or the 18-mer peptide of SEQ ID NO: 4 is stable at pH between 4 and 6 or stable in the presence of digestive enzymes selected from the group consisting of lysozyme, pepsin, chymotrypsin and trypsin.