CD8 T Cell Tumor Infiltration Following Tc1 or Tc2 Therapy Public Deposited

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  • June 7, 2019
  • Burgents, Meredith Lynn
    • Affiliation: School of Dentistry, Oral and Craniofacial Biomedicine PhD Program
  • Type I (Tc1) CD8+ T cells have been shown to be more effective than type II (Tc2) CD8+ T cells for adoptive cell transfer therapy in several tumor models. Migration differences between Tc1 and Tc2 cells were previously proposed to contribute to this difference in therapeutic efficacy. In order to evaluate Tc1 and Tc2 migration in vivo, we developed a model using transfected EL-4 thymoma tumor cells expressing the p33 peptide antigen from lymphocytic choriomeningitis virus (p33.EL-4). We used P14 mice, which are transgenic for the T cell receptor specific for p33 peptide that is expressed by these p33.EL-4 tumors. We crossed UBI.GFP mice that ubiquitously express GFP with the P14 mice, in order to generate mice that express both GFP and the p33-specific TCR in the CD8+ T cell population. Splenocytes from these mice were cultured to generate Tc1 and Tc2 cells, which were injected intraveinously into tumor-bearing mice. Donor cells were phenotyped before transfer and on days 3 and 7 after transfer. The CD8+ T cells were examined for GFP and adhesion molecule expression by flow cytometry. We examined gene expression of T-bet and enzymes important for selectin ligand glycosylation in Tc1 and Tc2 cultures, as well as the gene expression of cytokines, chemokines, and chemokine receptors in Tc1 and Tc2 treated mice. We found significantly more Tc1 than Tc2 cells in TDLNs and tumors on days 3 and 7 after transfer. Both Tc1 and Tc2 donor cells were found in TDLN and tumor sites of p33 positive-positive tumors compared to sites of p33 antigen-negative tumors. More importantly, all CD8+ T cells isolated from these tumors on days 3 and 7 after therapy, regardless of host or donor origin and Tc1 or Tc2 phenotype, expressed high levels of adhesion molecules important for T cell migration. This suggests that antigen does not alter the adhesion molecule expression of tumor infiltrating CD8+ T cells. These cells expressed high levels of CD44, leukocyte function-associated antigen-1 (LFA-1), and P-selectin glycoprotein ligand 1 (PSGL-1, CD162), suggesting a required “tumor infiltrating phenotype”. Before transfer and 3 days after transfer, Tc1 cells expressed higher levels of this tumor infiltrating phenotype compared to Tc2 cells. Thus, increased Tc1 cell migration to TDLN and infiltration of tumors may be due to higher expression of a tumor infiltrating phenotype compared to Tc2 cells. We found that T-bet expression is higher in cells from Tc1 vs. Tc2 cultures, which may promote the type I phenotype, including higher adhesion molecule expression. However, we did not find a significant difference between the gene expression of selectin ligand glycosylating enzymes or PSGL-1 gene expression in Tc1 vs. Tc2 cells. In addition, when we examined the gene expression of chemokines in the tumors of Tc1 vs. Tc2 treated mice, we found that type I interferon (IFN)−γ inducible protein (IP)-10 is more highly expressed compared to macrophage derived chemokine (MDC) or macrophage inducing protein (MIP)-1α in either treatment group. Also, donor and host CD8+ cells in Tc1 and Tc2 treated mice express chemokine receptor CXCR3 and cytokine IFN−γ, but no interleukin (IL)-4. This data suggest that in addition to surface expression of adhesion molecules CD44, LFA-1, PSGL-1, expression of type I cytokine IFN-γ and chemokine receptor CXCR3 is also a characteristic of a “tumor infiltrating phentoype.”
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  • Flood, Patrick M.
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