Methods of isolating and purifying CD8 T cells
Methods are provided for isolating CD8+ T cells. Furthermore, methods for purifying a subset of IL-13 expressing CD8 T cells are provided, such methods comprising the steps of marking the CD8+ T cells by labeling CD8, or selectively removing non-CD8 cells, and then purifying a subset of IL-13 expressing CD8+ T cells by marking a human biomarker such as C10orf128. Related antibodies and antiserums are also described, such antibodies related to a cell surface domain peptide for biomarker C10orf128, and human homologs of related mouse “activated” CD8IL-13 cell surface biomarkers Tm4sf19 and 1830127L07Rik.
1. A method of isolating a subset of CD8 T cells that produce interleukin-13 upon activation, the method comprising the steps of:
obtaining a sample of lymphocytes from a mammal;
activating the sample of lymphocytes with immune-irradiated splenocytes to obtain a polyclonal T cell clone population, wherein the immune-irradiated splenocytes present an antigen recognized by the sample of lymphocytes;
expanding the polyclonal T cell clone population;
selectively depleting the CD4 T cell clones present in the polyclonal T cell clone population to isolate an activated CD8 T cell clone population;
measuring gene expression in the activated CD8 T cell clone population;
determining if any of the activated CD8 T cell clone population expresses an elevated level of a biomarker as compared to a healthy control, the biomarker selected from a group consisting of: 1810011H11Rik and C10orf128; and
isolating a subset of CD8 T cell clones that express the elevated level of the biomarker from the activated CD8 T cell population;
wherein the elevated level of the biomarker is indicative of interleukin-13 production by the subset of CD8 T cells.
2. The method of claim 1 , wherein:
the biomarker comprises C10orf128;
the mammal comprises a human; and
the steps of determining if any of the activated CD8 T cell clone population expresses an elevated level of the biomarker and isolating a subset of CD8 T cell clones that express the elevated level of the biomarker from the activated CD8 T cell clone population further comprise:
using an anti-C10orf128 antiserum to identify the subset of CD8 T cell clones that express the elevated level of C10orf128;
wherein the anti-C10orf128 antiserum is made against a peptide having an amino acid sequence comprising SEQ ID NO: 1, or a functional equivalent, variant, or fragment thereof, attached to a carrier protein.
3. The method of claim 2 , wherein the carrier protein attached to the peptide comprises a keyhole limpet hemocyanin.
4. The method of claim 1 , wherein the antigen comprises a UV-inactivated Chlamydia muridarum or elementary body-depleted soluble Chlamydia muridarum antigen preparation.
5. The method of claim 1 , wherein the step of determining if any of the activated CD8 T cell clone population expresses an elevated level of the biomarker as compared to a healthy control is applicable to the diagnosis of one or more disease states.
6. The method of claim 5 , wherein at least one of the disease states comprises scarring associated with a Chlamydia infection or systemic sclerosis.