Multiple sclerosis is an inflammatory, demyelinating disease of the central nervous system (CNS) characterized by a wide range of clinical signs 1 . The location of lesions in the CNS is variable and is a crucial determinant of clinical outcome. Multiple sclerosis is believed to be mediated by myelinspecific T cells, but the mechanisms that determine where T cells initiate inflammation are unknown. Differences in lesion distribution have been linked to the HLA complex, suggesting that T cell specificity influences sites of inflammation2. We demonstrate that T cells that are specific for different myelin epitopes generate populations characterized by different T helper type 17 (T H 17) to T helper type 1 (T H 1) ratios depending on the functional avidity of interactions between TCR and peptide-MHC complexes. Notably, the T H 17:T H 1 ratio of infiltrating T cells determines where inflammation occurs in the CNS. Myelin-specific T cells infiltrate the meninges throughout the CNS, regardless of the T H 17:T H 1 ratio. However, T cell infiltration and inflammation in the brain parenchyma occurs only when T H 17 cells outnumber T H 1 cells and trigger a disproportionate increase in interleukin-17 expression in the brain. In contrast, T cells showing a wide range of T H 17:T H 1 ratios induce spinal cord parenchymal inflammation. These findings reveal critical differences in the regulation of inflammation in the brain and spinal cord.Experimental autoimmune encephalomyelitis (EAE) is an animal model that shows many similarities to multiple sclerosis3. However, rodent EAE differs from multiple sclerosis by manifesting as ascending flaccid paralysis, reflecting unexplained preferential targeting of inflammation to the spinal cord (described as classic EAE). In a small number of antigenspecific models, brain inflammation occurs (described as atypical EAE)4 -8. Interferon-γ (IFN-γ) deficiency also causes certain myelin-specific T cells to preferentially induce brain

CONCLUSION BIBLIOGRAPHY during a heated political time (and featuring a country with a sordid history with the U.S.) is certainly something impressive, something worth taking a look at. This exploration of Hijuelos's work will allow readers a new perspective about U.S. definitions of "immigrant" and "Latino." A writer such as Hijuelos, with his mainstream appeal yet culturally specific style, content, and narrative devices, makes U.S. audiences rethink the boundaries of American literature. The U.S. has had a tumultuous relationship with the island of Cuba for decades. Cuba has both social and political significance for the United States for many reasons, but the major issues keeping Cuba on the minds of U.S. politicians and citizens are geographical proximity and Fidel Castro. Cuba is located just 112 miles from the coast of Florida (and about 90 miles from Key West) and yet for the most part remains untouchable by many U.S. Americans initially because of the dictator Batista's fall to Castro in 1958 3 and later because of the travel embargo firmly in place since 1962 under John F. Kennedy's presidency. 4 Previous to Castro's takeover, the U.S. was a large presence in Cuba physically, with many American families living there to work for U.S. companies, and in commerce. Jacqueline Kaye writes, "From 1898 onwards the US freely intervened in Cuban affairs, sometimes at the invitation of Cuban politicians" and, it is safe to say, sometimes without invitation (133). 5 Since JFK's travel embargo in 1962, tension between the two countries has not lessened much, if at all. Probably because Cuba is a kind of forbidden experience, an untouchable 3 Batista, in power partly through a coup staged by the U.S. government, had historically favored U.S. companies in Cuba. When Castro took power, he forced all U.S. companies out. 4 The Bay of Pigs invasion likely did not breed amicable feelings between the U.S. and Cuba, either.

Murine models of Autologous Cellular Immunotherapy (ACI) have been useful tools for understanding the mechanisms by which human immunotherapy may work. Sipuleucel-T is an FDA approved therapeutic for the treatment of asymptomatic or minimally symptomatic, metastatic castration-resistant metastatic prostate cancer; Phase 3 studies demonstrated a statistically significant prolongation in overall survival when compared to a control group. Sipuleucel-T is manufactured by activating peripheral blood mononuclear cells (PBMC), including antigen presenting cells, with a recombinant protein fusion of prostatic acid phosphatase and granulocyte macrophage colony-stimulating factor (GM-CSF). We have previously reported a method for the use of cultured PBMC in a pre-clinical setting to elicit a T cell response against a tumor associated antigen, and to protect mice from tumors expressing that antigen. The model antigen we used was human Carbonic Anhydrase IX (hCA9). Mouse PBMCs were cultured with the antigen fused to murine GM-CSF (hCA9-GM) or with GM-CSF alone. Mice were immunized 3 times with these cells at 2 week intervals. Satellite mice were then harvested for evaluation of in vitro T cell response (antigen recall assay) and the remaining mice were inoculated with hCA9-expressing tumor cells, and tumor growth followed over time. Immunized mice mounted a robust T cell response in vitro to hCA9 in a dose dependent manner, a result that correlated with protection from tumor challenge in mice immunized with PBMC cultured in hCA9-GM. Herein, we investigate the cellular requirements for protection against tumor challenge and ability of a selective Toll-like receptor (TLR) 9 agonist to enhance the potency of this cellular immunotherapy. To investigate the importance of T cells for anti-tumor responses, we demonstrate that selective depletion of either CD4 or CD8 T cells results in a diminution or loss of protection toward tumor challenge; indicating T cells are necessary in vivo to prevent tumor growth. Next, to stimulate antigen specific T cell activity in the ACI therapy, relative to hCA9 antigen alone, cells were cultured with hCA9-GM antigen in combination with the TLR9 agonist CpG1826. Addition of CpG1826 resulted in significant accumulation of pro-inflammatory cytokines (IL-1b, IL-6, IL-10, TNFa, KC and IFNg) in the culture medium, and to higher surface expression of activation and co-stimulatory markers (CD25, CD69, CD80, CD86, and MHC Class II) on cultured cells. Moreover, inclusion of CpG1826 to cultures enhanced protection against tumor cell challenge. Interestingly, this enhancement in potency was not associated with increased antigen specific T cell responses, as measured in the in vitro antigen recall assay. In conclusion, we show that, in mice, unfractionated white blood cells collected from peripheral blood can be harnessed to mount effective anti-tumor immune responses and that TLR9 agonists complement this approach. Furthermore, this active immunotherapy triggers a balanced immune response relying on the activity of both CD4 and CD8 T cells. Future studies will be aimed at evaluating the role of various PBMC subsets (T cell, B cell and myeloid cell) in protecting mice from tumor cell challenge. Citation Format: Kenneth A. Brasel, Craig Meagher, Marykay Ligocki, Lauren Cerretti, Felecia Wagener, Sam Li, James Trager. Protective immunizations using cultured peripheral blood cells. [abstract]. In: Proceedings of the AACR Special Conference on Tumor Immunology: Multidisciplinary Science Driving Basic and Clinical Advances; Dec 2-5, 2012; Miami, FL. Philadelphia (PA): AACR; Cancer Res 2013;73(1 Suppl):Abstract nr B71.