Lactobacillus johnsonii La1 NCC533 (CNCM I-1225) and immune disorders

Abstract

The present invention generally relates to the field of preventing and/or treating inflammatory and infectious disorders, in particular by boosting the endogenous antimicrobial defences. One embodiment of the present invention is the use of non-replicating L. johnsonii La1 NCC533 (deposit number CNCM I-1225) for use in the treatment or prevention of disorders related to the immune system including infections.

Claims

1. A method for treatment of a disorder selected from the group consisting of Crohn's disease, infectious diarrhea, antibiotic-associated diarrhea, gingivitis, and combinations thereof, the method comprising administering a composition comprising Lactobacillus johnsonii La1 (NCC533, deposit number CNCM I-1225) to an individual having the disorder, wherein the L. johnsonii La1 is rendered non-replicating by a heat treatment at a temperature of 110 C. to 140 C. for 1-30 seconds.

2. The method in accordance with claim 1, wherein the heat treatment is carried out for at least 10 seconds.

3. The method in accordance with claim 1, wherein the composition is administered to the individual in an amount that provides 10.sup.4 to 10.sup.12 cfu of the L. johnsonii La1 (NCC533, deposit number CNCM 1-1225) per day.

4. The method in accordance with claim 1, wherein the composition is administered to the individual in an amount that provides about 0.005 mg 1000 mg of the L. johnsonii La1 (NCC533, deposit number CNCM 1-1225) per day.

5. The method in accordance with claim 1, wherein the composition is selected from the group consisting of food compositions, food products, drinks, formulas for complete nutrition, nutritional supplements, nutraceuticals, food additives, pharmaceutical compositions, cosmetical compositions, topical compositions, and medicaments.

6. The method in accordance with claim 1, wherein the composition increases endogenous hBD1 expression.

7. The method of claim 1, wherein the heat treatment is at 120 C. for 15 seconds.

8. The method of claim 1, wherein the composition increases endogenous antimicrobial defences.

9. A method to increase the effectiveness of L. johnsonii La1 (NCC533, deposit number CNCM I-1225) in treatment of a disorder selected from the group consisting of Crohn's disease, infectious diarrhea, antibiotic-associated diarrhea, gingivitis, and combinations thereof, the method comprising: rendering the L. johnsonii La1 non-replicating by a heat treatment at a temperature of 110 C. to 140 C. for 1-30 seconds; and administering the non-replicating L. johnsonii La1 to a patient having the disorder.

10. The method in accordance with claim 9, wherein the L. johnsonii La1 (NCC533, deposit number CNCM 1-1225) are rendered non-replicating by the heat treatment for at least 10 seconds.

11. The method of claim 9, wherein the heat treatment is at 120 C. for 15 seconds.

12. The method of claim 9, wherein the heat treatment is performed for 10 to 20 seconds.

Description

(1) FIG. 1 shows that heat treated La1 (NCC533, deposit number CNCM I-1225) at 120 C.-15 sec strongly induces hBD1 mRNA in intestinal epithelial cells in vitro compared with other heat-treated strains. T84 cells were incubated for 4 h with the heat-treated strains. Gene expression of hBD1 was analyzed by real-time PCR. The bars represent the meanssem normalized to basal expression of non stimulated cells.

(2) FIG. 2 shows that a high temperature and short time treatment of La1 (NCC533, deposit number CNCM I-1225) tends to be the best to induce hBD1 mRNA expression. T84 cells were stimulated for 4 h with the live and heat-treated La1 (NCC533, deposit number CNCM I-1225) at 120 C.15 sec or 85 C.20 min. Gene expression of hBD1 was analyzed by real-time PCR. The bars represent the meanssem normalized to basal expression of non stimulated cells.

EXAMPLES

(3) Experimental Protocol:

(4) T84 cells were used from passage 30-40 and cultured in Dulbecco's modified essential medium/F-12 (Sigma D 6421) containing 5% of foetal calf serum (FCS) (Amined BioConcept) and 2 mM glutamine. Cells were seeded at a concentration of 210.sup.6 cell/well in 6-well culture plates and grown as monolayers at 37 C. in a 5% CO.sub.295% air atmosphere. Cells grown to 1 week after confluence were incubated with serum and antibiotic-free medium for at least 12H. This step was necessary to eliminate serum-induced defensin expression and prevent any influence of antibiotics on the probiotics and on the cell immune response. Cells were further incubated with probiotics or heat-treated strains for 4H. At the end of the incubation time, cells were washed with PBS and harvested with TriPure isolation reagent according to the supplier's protocol. Human hBD1 and hBD2 gene expression in the so-treated cells was assessed by quantitative PCR.

(5) Bacterial strains used in this experiment are B. longum (NCC 2705, deposit number CNCM I-2618), B. lactis (NCC 2818, deposit number CNCM I-3446), L. johnsonii (La1, NCC 533, deposit number CNCM I-1225), L. paracasei (ST11, NCC 2461, deposit number CNCM I-2116). These strains were tested live or heat-treated at either 120 C.15 sec or 85 C.20 min.

(6) Results:

(7) Heat-treated La1 (NCC533, deposit number CNCM I-1225) at 120 C., 15 sec induced strongly hBD1 mRNA expression after 4 h of incubation (FIG. 1) in contrast to the other tested heat-treated strains. These data are unique, as HBD1 expression, which is constituvely expressed, is currently thought by the scientific community as virtually non modulable by microbes, microbial products or inflammation.

(8) Both live and heat-treated La1 (NCC533, deposit number CNCM I-1225) strongly induced hBD1 mRNA expression, but the highest induction of hBD1 was elicited by heat-treated La1 (high temperature and short time treatment) (FIG. 2).