Bifidobacterium longum able to beneficially modulate immune response to respiratory virus infection
A technology of bifidobacteria longum and respiratory tract, applied in the field of bifidobacteria, can solve problems such as damage
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Embodiment 1
[0369] Example 1: Cell Wall Precipitate Formation
[0370] Bacteria Harvesting / Washing
[0371] method:
[0372] 1. by centrifugation (14000rpm, 4 ℃, 20min; Rotor JA-20 (Avanti J-26x P BeckmanCoulter) harvests the original bacterial biomass (total cell count=1.5X10) that is equivalent to 250ml 11 ). Bacterial pellets were washed with sterile PBS, the supernatant was discarded and washed again (repeated twice).
[0373] 2. Live freeze-dried bacteria and non-live freeze-dried bacteria (0.5g of 3.0X 10 11 Powder or total cell count = 1.5X10 11 ) was resuspended in 50ml and harvested by centrifugation (14000rpm, 4°C, 20min; rotor JA-20 (Avanti J-26x PBeckman Coulter). Wash the bacterial pellet with sterile PBS, discard the supernatant and wash again ( Repeat twice).
[0374] 3. Finally the pellet was resuspended in 50ml sterile PBS and the bacterial solution was divided into two 25ml aliquots.
[0375] cell disruption
[0376] The purpose of this process is to produce cell...
Embodiment 2
[0421] Example 2 In specific cell types (monocytes and dendritic cells), from The cell wall component of the strain favorably blocks type I interferons and the resulting induction of IP-10 (pro-inflammatory chemokine)
[0422] An exaggerated immune response of monocytes and dendritic cells to viral infection elicits a pro-inflammatory response in the lung. To determine whether 30 mg / ml of cell wall fraction from 35624 produced as in Example 1 had beneficial antiviral effects, isolated cells of human CD14+ monocytes from peripheral blood were exposed to rhinovirus (RV) and monitored IP-10 Response to RV. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors using density gradient centrifugation. Peripheral blood mononuclear cells (PBMCs) were isolated from healthy donors using density gradient centrifugation. Human peripheral blood mononuclear cells were isolated using CD14 positive isolation using the MACS system (Miltenyi Biotec, 130-050-201). Cell...
Embodiment 3
[0427] Example 3: Not all cell wall components from bifidobacterial species have the same effect
[0428] Cell wall components from another bifidobacterium, Bifidobacteria breve (Bif UCC2003), were also tested using the method described in Example 2 and did not show similarly significant effects. The Bif UCC2003 fraction reduced IP-10 production after virus challenge, but not to the same extent as the Bifidobacterium longum 35624 cell wall fraction in the 24-hour assay ( Figure 5 ).
[0429] Furthermore, in inflamed mucosa, not only the virus itself induces IP-10 secretion; other cytokines also induce IP-10 production. Cytokines such as IFN-γ, IFN-β and IFN-λ are all produced as part of the primary antiviral host response. In particular IFN-γ and IFN-β are potent inducers of IP-10. We examined the effect of cell wall components on the secretion of IP-10 in response to IFN-γ, IFN-β and IFN-λ ( Image 6 ). Cell wall fraction (30mg / ml) was pretreated for 1 hour and then sti...
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