Listeria-induced immunorecruitment and activation, and methods of use thereof
Provided are reagents and methods for administering an attenuated bacterium for use in treating a cancerous or infectious condition. Reagents and methods for administering an attenuated bacterium for use in inducing an immune response against a tumor, cancer cell, or infective agent are further provided. Also provided are methods of diagnosis and kits.
1 . A method for treating a mammal having a cancerous or non-listerial infectious condition, wherein the cancerous or infection condition is in the liver of the mammal, comprising administering to the mammal an effective amount of a metabolically active, attenuated Listeria , wherein the Listeria does not comprise a nucleic acid encoding a non-listerial antigen capable of stimulating a specific immune response against the condition, and wherein the attenuated Listeria is administered to the mammal in multiple doses.
2 . The method of claim 1 , wherein the cancerous or infectious condition is inhibited or reduced in the mammal by the administration of the effective amount of the attenuated Listeria.
3 . The method of claim 1 , wherein survival of the mammal is enhanced by the administration of the effective amount of the attenuated Listeria.
4 . The method of claim 1 , wherein the attenuated Listeria is attenuated in one or more of:
a. growth;
b. cell to cell spread;
c. binding to or entry into a host cell;
d. replication; or
e. DNA repair.
5 . The method of claim 4 , wherein the attenuated Listeria is attenuated in:
a. cell to cell spread; or
b. both cell-to-cell spread and entry into nonphagocytic cells.
6 . The method of claim 1 , wherein the Listeria is attenuated by one or more of:
a. an actA mutation;
b. an inlB mutation;
c. a uvrA mutation;
d. a uvrB mutation;
e. a uvrC mutation;
f. a nucleic acid targeting compound; or
g. a uvrAB mutation and a nucleic acid targeting compound.
7 . The method of claim 6 , wherein the Listeria is attenuated by:
a. an actA mutation; or
b. both an actA mutation and an inlB mutation.
8 . The method of claim 7 , wherein the nucleic acid targeting compound is a psoralen.
9 . The method of claim 1 , wherein the Listeria cannot do one or more of:
a. form colonies;
b. replicate; or
c. divide.
10 . The method of claim 1 , wherein the Listeria is killed, but metabolically active (KBMA).
11 . The method of claim 1 , wherein the attenuated Listeria is administered intravenously.
12 . The method of claim 1 , wherein the attenuated Listeria is administered in three or more doses.
13 . The method of claim 1 , wherein the attenuated Listeria is one or both of:
a. not administered orally to the mammal, or
b. administered as a composition that is at least 99% free of other types of bacteria.
14 . The method of claim 1 , wherein the attenuated Listeria is administered to the mammal in a pharmaceutical composition.
15 . The method of claim 1 , wherein the mammal has not previously been administered a vaccine against the cancerous or infectious condition.
16 . The method of claim 1 , wherein the method does not further comprise administering a vaccine against the cancerous or infectious condition to the mammal.
17 . The method of claim 1 , wherein the mammal comprises the cancerous condition.
18 . The method of claim 17 , wherein the condition comprises a tumor or cancer.
19 . The method of claim 18 , wherein the condition comprises a cancer that has metastasized to the liver.
20 . The method of claim 19 , wherein the cancer is colorectal cancer.
21 . The method of claim 1 , wherein the mammal comprises the non-listerial infection.
22 . The method of claim 1 , wherein the infectious condition comprises one or more of:
a. hepatitis B;
b. hepatitis C;
c. human immunodeficiency virus (HIV);
d. cytomegalovirus (CMV);
e. Epstein Barr virus (EBV); or
f. leishmaniasis.
23 . The method of claim 1 , wherein the administering stimulates an innate immune response against the condition.
24 . The method of claim 1 , wherein the administering stimulates an acquired immune response against the condition.
25 . The method of claim 1 , wherein the administering stimulates one, or any combination, of a:
a. NK cell;
b. NKT cell;
c. dendritic cell (DC);
d. monocyte or macrophage;
e. neutrophil; or
f. toll like receptor (TLR) or nucleotide binding oligomerization domain (NOD) protein,
as compared with immune response in the absence of the administering of the effective amount of the attenuated Listeria.
26 . The method of claim 1 , wherein the administering stimulates increased expression of any one, or any combination, of:
a. CD69;
b. interferon-gamma (IFNgamma);
c. interferon alpha (IFNalpha) or interferon beta (IFNbeta);
d. interleukin 12 (IL 12);
e. monocyte chemoattractant protein (MCP 1); or
f. interleukin 6 (IL 6),
as compared with expression in the absence of the administering of the effective amount of the attenuated Listeria.
27 . The method of claim 1 , wherein the mammal is human.
28 . The method of claim 1 , wherein the Listeria is Listeria monocytogenes.
29 . The method of claim 1 , further comprising administering one, or any combination of:
a. an agonist or antagonist of a cytokine;
b. an inhibitor of a T regulatory cell (Treg); or
c. a tumor cell attenuated in growth or replication.
30 . The method of claim 29 , wherein the inhibitor of a Treg is cyclophosphamide (CTX).
31 . The method of claim 1 , wherein the effective amount comprises at least about 1×10 3 CFU/kg or at least about 1×10 3 Listeria cells/kg.
32 . A method for inducing an immune response against a cancer cell, tumor, or non-listerial infective agent in a mammal, wherein the mammal comprises the cancer cell, tumor, or non-listerial infective agent in its liver, comprising administering to the mammal an effective amount of a metabolically active, attenuated Listeria , wherein the Listeria does not comprise a nucleic acid encoding a non-listerial antigen capable of stimulating a specific immune response against the condition, wherein the attenuated Listeria is administered to the mammal in multiple doses, and wherein the attenuated Listeria is one or both of:
a. not administered orally to the mammal, or
b. administered as a composition that is at least 99% free of other types of bacteria.
33 . The method of claim 32 , wherein the attenuated Listeria is attenuated in one or more of:
a. growth;
b. cell to cell spread;
c. binding to or entry into a host cell;
d. replication; or
e. DNA repair.
34 . The method of claim 33 , wherein the attenuated Listeria is attenuated in:
a. cell to cell spread; or
b. both cell-to-cell spread and entry into nonphagocytic cells.
35 . The method of claim 32 , wherein the Listeria is attenuated by one or more of:
a. an actA mutation;
b. an inlB mutation;
c. a uvrA mutation;
d. a uvrB mutation;
e. a uvrC mutation;
f. a nucleic acid targeting compound; or
g. a uvrAB mutation and a nucleic acid targeting compound.
36 . The method of claim 35 , wherein the Listeria is attenuated by:
a. an actA mutation; or
b. both an actA mutation and an inlB mutation.
37 . The method of claim 35 , wherein the nucleic acid targeting compound is a psoralen.
38 . The method of claim 32 , wherein the Listeria cannot do one or more of:
a. form colonies;
b. replicate; or
c. divide.
39 . The method of claim 32 , wherein the Listeria is killed, but metabolically active (KBMA).
40 . The method of claim 32 , wherein the attenuated Listeria is administered intravenously.
41 . The method of claim 32 , wherein the attenuated Listeria is administered in three or more doses.
42 . The method of claim 32 , wherein the mammal is not administered a vaccine capable of stimulating a specific immune response against the cancer cell, tumor, or non-listerial infective agent.
43 . The method of claim 32 , wherein the mammal comprises the cancer cell or tumor.
44 . The method of claim 32 , wherein the mammal comprises the non-listerial infective agent in its liver.
45 . The method of claim 32 , wherein the immune response inhibits or reduces one, or any combination, of the:
a. number or tumors or cancer cells;
b. tumor mass; or
c. titer of an infectious agent,
in the mammal.
46 . The method of claim 32 , wherein the administering stimulates an innate immune response against the cancer cell, tumor, or non-listerial infective agent.
47 . The method of claim 32 , wherein the administering stimulates an acquired immune response against the cancer cell, tumor, or non-listerial infective agent.
48 . The method of claim 32 , wherein the administering stimulates one, or any combination, of a:
a. NK cell;
b. NKT cell;
c. dendritic cell (DC);
d. monocyte or macrophage;
e. neutrophil; or
f. toll like receptor (TLR) or nucleotide binding oligomerization domain (NOD) protein,
as compared with immune response in the absence of the administering of the effective amount of the attenuated Listeria.
49 . The method of claim 32 , wherein the administering stimulates increased expression of any one, or any combination, of:
a. CD69;
b. interferon-gamma (IFNgamma);
c. interferon alpha (IFNalpha) or interferon beta (IFNbeta);
d. interleukin 12 (IL 12);
e. monocyte chemoattractant protein (MCP 1); or
f. interleukin 6 (IL 6),
as compared with expression in the absence of the administering of the effective amount of the attenuated Listeria.
50 . The method of claim 32 , wherein the mammal is human.
51 . The method of claim 32 , wherein the Listeria is Listeria monocytogenes.
52 . The method of claim 32 , wherein the immune response comprises stimulating one or both of:
a. an increase in the percent of hepatic leukocytes that is NK cells, compared to the percent without the administering of the attenuated Listeria ; or
b. an increase in expression of an activation marker by a hepatic NK cell, compared to the expression without the administering of the attenuated Listeria.
53 . The method of claim 32 , wherein the effective amount of attenuated Listeria comprises at least about 1×10 3 CFU/kg or at least about 1×10 3 Listeria cells/kg.
54 . A method for inducing an immune response against a cancer cell, tumor, or non-listerial infective agent in a mammal, wherein the mammal comprises the cancer cell, tumor, or non-listerial infective agent in its liver, comprising administering to the mammal an effective amount of a metabolically active, attenuated Listeria , wherein the Listeria does not comprise a nucleic acid encoding a non-listerial antigen capable of stimulating a specific immune response against the condition, wherein the attenuated Listeria is administered to the mammal in multiple doses, and wherein the attenuated Listeria is one or both of:
a. administered in a pharmaceutical composition; or
b. a non-naturally occurring strain.
55 . A method for treating a mammal having a cancerous or non-listerial infectious condition, wherein the cancerous or infectious condition is in the liver of the mammal, comprising administering to the mammal an effective amount of a metabolically active, attenuated Listeria , wherein the Listeria does not comprise a nucleic acid encoding a non-listerial antigen capable of stimulating a specific immune response against the condition, and wherein the Listeria is administered to the mammal in the absence of a separately generated, vaccine-induced immune response to the cancerous or infectious condition in the mammal.