Abstract
The invention relates to a pharmaceutical and/or dietetic composition for increasing the impact of the immune defense of higher living beings, wherein bacteria of the species Lactobacillus fermentum from at least one of the strains K1-Lb1 or K1-Lb6 or K2-Lb4 or K6-Lb4 or K7-Lb1 or K8-Lb1 or K9-Lb6 are contained in order to control the adaptive and natural immune defense by means of T helper 1 and T helper 2 cells and/or bacteria of the species Lactobacillus fermentum from at least one of the strains K2-Lb6 or K11-Lb3 are contained in order to strengthen the native immune defense.
Claims
1. A method of strengthening the immune system in a human comprising: administering a composition comprising one or more isolated bacterial strains from Kimere to a human orally, wherein the isolated bacterial strains are strains of Lactobacillus fermentum and/or Lactobacillus plantarum, and wherein the administration of the composition provides an immunomodulating effect in the human, wherein the one or more strains are selected from: Lactobacillus fermentum K1-Lb1 (deposition number DSM 22837), or Lactobacillus plantarum K4-Lb6 (deposition number DSM 22830), wherein the administration of K1-Lb1 or K4-Lb6 changes the Th1/Th2 balance towards Th1; Lactobacillus fermentum K7-Lb1(deposition number DSM 22831), or Lactobacillus fermentum K8-Lb1 (deposition number DSM 22832), wherein the administration of K7-Lb1 or K8-Lb1 changes the Th1/Th2 balance towards Th2, or Lactobacillus fermentum K2-Lb6 (deposition number DSM 22829), or Lactobacillus fermentum K11-Lb3 (deposition number DSM 22838), wherein the administration of K2-Lb6 or K11-Lb3 increased the release of defensin from intestinal cell.
2. The method according to claim 1, wherein the isolated bacterial strains tolerate up to 0.3% (w/v) of bile acid.
3. The method according to claim 1, wherein the isolated bacterial strains are selected from Lactobacillus fermentum K7-Lb1, or Lactobacillus fermentum K8-Lb1, or Lactobacillus plantarum K4-Lb6 wherein the strain K4-Lb6 tolerates up to 3.0% (w/v) of bile acid and the strains K7-Lb1 and K8-Lb1 show survival rates of over 80% at a pH of 3.
4. A method of strengthening the immune system in a human comprising: administering a composition comprising one or more isolated bacterial strains from Kimere to a human orally, wherein the isolated bacterial strains are strains of Lactobacillus fermentum and/or Lactobacillus plantarum, and wherein the administration of the composition provides an immunomodulating effect in the human, wherein the one or more strains are selected from: Lactobacillus fermentum K11-Lb3 (deposition number DSM 22838), of Lactobacillus fermentum K7-Lb1(deposition number DSM 22831), or Lactobacillus fermentum K8-Lb1 (deposition number DSM 22832), wherein the immune system is strengthened by an inhibition of inflammations.
Description
(1) Further details and features of the invention are explained below in greater detail with reference to graphical descriptions. However, they are not intended to limit the invention but only explain it. In schematic, view:
(2) FIG. 1 shows an overview of the properties of the individual strains
(3) FIG. 2 phylogenetic tree of the Lactobacilli presented here in comparison with known bacteria
(4) FIG. 3 band pattern of all 10 strains according to the PFGE process
(5) FIGS. 4A-4C tolerance of the strains to bile salt
(6) FIG. 4A: shows the number of the surviving strains on exposure to different concentrations of cattle bile.
(7) FIG. 4B: shows the measurement of the absorption of different strains at 620 nm in 0.3% concentration of cattle bile solution.
(8) FIG. 4C: shows the measurement of the absorption of different strains at 620 nm in 3.0% concentration of cattle bile solution.
(9) FIG. 5 resistance of the strains to very acid milieu
(10) FIGS. 6A-6C Th1 and Th2 cytokine production of human blood cells (PBMCs) after coincubation with the strains
(11) FIG. 6A: shows the production of interferon-γ in different strains.
(12) FIG. 6B: shows the production of interleukin-4 in different strains.
(13) FIG. 6C: shows the resulting ratio of interferon-γ and interleukin-4.
(14) FIG. 1 shows, as a key property of the strains presented here, characteristic effects and their intensities. It is clear that almost all strains, with the exception of K11-Lb3 (deposition number DSM 22838), show three characteristic properties that distinguish them from one another. All of these strains have a more or less high bile salt tolerance and a greater or lesser pH tolerance. All the strains have an immunomodulating effect. This immunomodulating effect belongs either to the natural or innate immunity, such as the elevated defensin release due to the strains, in this case K2-Lb6 (deposition number DSM 22829) and K11-Lb3 (deposition number DSM 22838), or is part of the adaptive or acquired immunity, such as the influencing of the Th1/Th2 response.
(15) FIG. 1 shows that, of the strains presented here, K1-Lb1 (deposition number DSM 22837), K1-Lb6, K2-Lb4 and K4-Lb6 (deposition number DSM 22830), strengthen the Th helper reaction and reduce the Th2 influence. The strains K2-Lb6 (deposition number DSM 22829). K6-Lb4. K7-Lb1 (deposition number DSM 22831) and K8-Lb1 (deposition number DSM 22832), on the other hand, strengthen the Th2 response and reduce the Th1 influence.
(16) The intensity is shown in three stages. The highest of the observed values in each case is designated. Correspondingly, ++ indicates approximately [⅔] and + corresponds to about [⅓].
(17) In FIG. 2, some of the found strains are entered in a phylogenetic tree, in which some other, adequately well know bacterial strains have been entered. This phylogenetic tree is based on partial sequences of the 16S rDNA of selected examples of ARDRA-PCR grouping of isolates of the Lactobacilli from Kimere compared with the BLAST database. The evolutionary distances were derived using the UPMGA method, the bootstrap loader program being based on 500 repetitions. To calculate the evolutionary distances, the method of maximum combined probability was used. This phylogenetic analysis was performed in MEGA4 according to Tamura et. al. 2007.
(18) FIG. 3 shows the band patterns of all 10 strains presented here obtained by the PFGE process-pulse field gel electrophoresis. From FIG. 3, it immediately becomes clear that the individual band patterns differ from one another significantly. All the bacteria that are analysed by the PFGE method, and which show precisely this band pattern, are thereby clearly identified as this strain of the respective genus.
(19) The method of typifying microorganisms by PFGE has been known since 1984 and is part of the recognized prior art. With the extremely low effort of depositing a single image, it permits the unambiguous identification of the respective bacterial strain.
(20) FIGS. 4A to 4C show the tolerance of the strains to bile salt.
(21) FIG. 4A shows the number of the surviving strains on exposure to cattle bile with increasing concentration of 0.3%, via 0.5%. 1% and 2%, up to 3% cattle bile concentration (w/v).
(22) In FIG. 4B, by measurement of the absorption of the samples at 620 nm, the specific growth of the strains in 0.3% cattle bile solution is plotted against time. A somewhat similar behaviour is shown for all strains.
(23) FIG. 4C shows the results of an, in principle, identical measurement, but with 3.0% concentration of cattle bile. A particularly rapid growth of the strain K4-Lb6 (deposition number DSM 22830), which is very obviously highly robust with respect to bile salt, is shown here.
(24) In FIG. 5, the resistance of the strains to very acid milieu is shown, specifically for pH 2 and pH 3. The cells were incubated in 0.35% NaCl (containing 3 g I<−1> pepsin) and adjusted to pH values of 2, 3 and 6.5 for 3 h at 37[deg.] C. in each case. The surviving cells were counted on an MRS agar plate at 37[deg.] C. for 48 to 72 h. The values at pH 6.6 defined as reference value with 100% and the cells surviving at pH 2 and pH3 were compared with this reference value.
(25) FIGS. 6A to 6C show Th1 and Th2 cytokine production of human blood cells (PBMCs) after co-incubation in vitro with Kimere Lactobacilli strains, specifically with and without stimulation by superantigenic Staphylococcus enterotoxin A (SEA). As an effect of the bacterial strains on the imnmune system and on the shift of the Th1/Th2 ratio, FIG. 6A shows the production of interferon-[gamma] and FIG. 6B shows the production of interleukin-4 and FIG. 6C shows the resulting ratio of interferon-[gamma] and interleukin-4.
