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OwnerNicole Neeltje Speelman-Van Der Wel, Michael Barry Brenner +1inventors
InventorsNicole Neeltje Speelman-Van Der Wel, Michael Barry Brenner, Jacobus Peter Johannes Peters
Estimating the term…
Mycobacteria such as M. tuberculosis and M. leprae are considered to be prototypical intracellular bacilli that have evolved strategies to enable growth in the intracellular phagosomes of the host cell. By contrast, we show that lysosomes rapidly fuse with the virulent M. tuberculosis and M. leprae— containing phagosomes of human monocyte-derived dendritic cells and macrophages. After 2 days, M. tuberculosis progressively translocate from phagolysosomes into the cytosol where they replicate. Cytosolic entry is also observed for M. leprae but not for the vaccine strain, M. bovis BCG, or killed mycobacteria, and is dependent upon secretion of the mycobacterial gene products CFP-IO and ESAT-6 of the RDI region. The present invention further provides means and methods for using these findings in therapeutic and immunogenic compositions.
A method for determining whether a product of a gene of a mycobacterium is involved in translocation of said mycobacterium from the phagosome to the cytosol of a host cell, said method comprising altering said gene product and/or expression of said gene product in said mycobacterium and determining whether said translocation of said mycobacterium in said host cell is affected.
A method for reducing the phago-cytosolic translocation of a mycobacterium comprising at least reducing the expression of CFPIO, ESAT6 or EspA in said mycobacterium.
A method for enhancing phago-cytosolic translocation of a CFPIO, ESAT6 and/or EspA deficient mycobacterium, said method comprising providing said mycobacterium with CFPIO, ESAT6 and/or EspA.
A method for generating a recombinant BCG strain comprising providing BCG or a derivative thereof with CFPIO, ESAT6 and/or EspA.
A method for producing a mycobacterium that is substantially deficient in phago-cytosolic translocation comprising functionally reducing the expression of CFPIO, ESAT6 and/or EspA in said mycobacterium.
10-11. (canceled)
An attenuated mycobacterium comprising a nucleic acid encoding CFPIO, ESAT6 and/or EspA further comprising a heterologous nucleic acid for inhibiting cytosolic replication and/or cytosolic translocation of said mycobacterium.
14-18. (canceled)
(canceled)
23-25. (canceled)
(canceled)
A method for providing a mycobacterium with the capacity to translocate from a phagosome to the cytosol of a host cell, or to enhance said capacity, comprising infecting the mycobacterium with a nucleic acid encoding CFPIO, ESAT6 and/or EspA.
29-32. (canceled)
A method for selecting a mycobacterium for the preparation of a vaccine comprising infecting cells permissive for said mycobacterium in vitro with said collection and selecting from said collection a mycobacterium which translocates to the cytosol of infected cells.
35-43. (canceled)
Dependencies are inferred from the claim text.