CHLAMYDIA-SPECIFIC CD8+ T CELLS AND METHODS OF ISOLATING
Isolated, Chlamydia -specific, IL-13 expressing CD8+ T cell clones are provided for understanding the biology of Chlamydia infection and screening of therapeutics. Furthermore, methods are provided for isolating CD8+ T cells comprising the steps of infecting or identifying a naturally infected mammal with at least one species of Chlamydia causing bacteria, allowing the bacteria to clear or for a T cell immune response to the natural infection, collecting immune splenocytes from the mammal, providing at least one antigen from a Chlamydia -specific bacteria, and expanding and depleting the CD4+ T cell population or purifying the CD8 T cell population to isolate IL-13 expressing CD8+ T cell clones.
1 . A method for isolating Chlamydia -specific CD8+ T cells comprising the steps of:
recovering immune splenocytes from a mammal previously infected with at least one species of a Chlamydia -causing bacteria;
providing the immune splenocytes with at least one antigen to the Chlamydia -causing bacteria;
culturing a population of the immune splenocytes under conditions which bring about an increase in the number of CD8+ T cells in the a population and utilizes immune antigen presenting cells;
isolating CD8+ T cells from the population; and
deriving individual CD8+ T cell clones from the isolated CD8+ T cells, wherein the clones comprise interleukin-13 expressing CD8+ T cell clones.
2 . The method of claim 1 , wherein the Chlamydia -causing bacteria comprises a Chlamydia serovar.
3 . The method of claim 1 , wherein the immune splenocytes comprise immune lymphocytes.
4 . The method of claim 1 , further comprising the steps of:
infecting a mammal with at least one Chlamydia -causing bacteria or identifying a mammal with a naturally acquired infection; and
allowing a period of time to pass sufficient for clearance of the bacteria from the infected mammal or to allow for a T cell immune response in the mammal with the naturally acquired infection.
5 . The method of claim 1 , wherein the step of isolating CD8+ T cells from the population comprises depleting the CD4+ T cells present in the population.
6 . The method of claim 1 , wherein the step of isolating CD8+ T cell from the population comprises purifying the CD8+ T cells from the population.
7 . The method of claim 1 , wherein the mammal comprises a mouse or a human and the interleukin-13 comprises murine or human interleukin-13, respectively.
8 . The method of claim 1 , wherein the CD8+ T cell clones do not express Plac8.
9 . The method of claim 1 , wherein the immune antigen presenting cells comprise irradiated immune antigen presenting cells.
10 . The method of claim 4 , wherein the period of time sufficient to allow for a T cell immune response in the mammal with the naturally acquired infection comprises at least about two weeks.
11 . An isolated, Chlamydia -specific, interleukin-13 expressing CD8+ T cell clone cultured from a population of immune splenocytes utilizing immune antigen presenting cells.
12 . The isolated cell clone of claim 11 , wherein the cell clone does not express Plac8.
13 . The isolated cell clone of claim 12 , isolated using a method comprising the steps of:
recovering immune splenocytes from a mammal previously infected with at least one species of a Chlamydia -causing bacteria;
providing the immune splenocytes with at least one antigen to the Chlamydia -causing bacteria;
culturing the population of the immune splenocytes under conditions which bring about an increase in the number of CD8+ T cells in the a population and utilizes the immune antigen presenting cells;
isolating CD8+ T cells from the population; and
deriving the CD8+ T cell clone from the isolated CD8+ T cells.
14 . The isolated cell clone of claim 13 , wherein the immune antigen presenting cells comprise irradiated immune antigen presenting cells.
15 . A method for determining a T cell subset comprising the step of identifying a Chlamydia -specific, interleukin-13 expressing CD8+ T cell subset applicable to the diagnosis and/or treatment of one or more disease states.
16 . The method of claim 15 , wherein at least one of the disease states comprises scarring associated with a Chlamydia infection.
17 . The method of claim 15 , wherein at least one of the disease states comprises systemic sclerosis.
18 . The method of claim 15 , wherein the step of identifying a Chlamydia -specific, interleukin-13 expressing CD8+ T cell subset applicable to the diagnosis and/or treatment of one or more disease states comprises employing a gene expression microarray.
19 . The method of claim 15 , wherein the step of identifying a Chlamydia -specific, interleukin-13 expressing CD8+ T cell subset applicable to the diagnosis and/or treatment of one or more disease states comprises employing a proteomics-based methodology.
20 . The method of claim 19 , wherein the proteomics-based methodology comprises MALDI-TOF mass spectroscopy.