Cytokine 56 (2011) 1820

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Cytokine
journal homepage: www.elsevier.com/locate/issn/10434666

Poster Session 1: Cytokines in Lymphocyte Biology

PS1-008
Differential requirement of N-ras for IFN-g expression in CD4+ and CD8+ effector
T cells
Edgar Fernndez-Malav a, Paloma Martn Acosta b, Jess Rein a, Sabela Daz
Castroverde a, Ana V. Marn-Marn a, Eugenio Santos c, Salvador Iborra d,
a
Inmunologa, Facultad de Medicina, Universidad Complutense, Madrid, Spain,
b
Hospital Puerta de Hierro, Madrid, c Centro de Investigacin del Cncer (CSIC-USAL),
Salamanca, Spain, d Centro Nacional de Microbiologa, Instituto de Salud Carlos III, Madrid
Enhanced and sustained IFN-g production is a trait common to CD4+ (Th1) and
CD8+ (CTL) effector lymphocytes. However, signals emanating from the TCR that control IFN-g expression in these functional T cell lineages are poorly understood. Using
mice decient for the small GTPase N-ras, we demonstrate that N-ras downstream of
the TCR is critical in CD4+ but dispensable in CD8+ effectors for effective IFN-g
expression. Accordingly, in vitro and in vivo TCR-activated CD4+ T cells from N-rasdecient mice exhibited markedly decreased production of IFN-g, concomitantly with
impaired induction of transcription factor T-bet. By contrast, N-ras-decient CD8+ T
cells produced normal amounts of IFN-g, and displayed unaffected T-bet expression
upon exposure to viral antigen in vitro or following infection. Of interest, effector
CD8+ T cells lacking N-ras were impaired in induction of Eomesodermin (Eomes),
another T-box containing transcription factor, suggesting that, unlike T-bet, Eomes
is not involved in the early regulation of IFN-g expression in effector CD8+ T cells.
Of note, N-ras-decient CD4+ T cells showed increased IL-17 production upon TCR
stimulation, although this was not seemingly related with defective IFN-g expression.
Thus, our study identies N-Ras as a key regulator of signalling pathways controlling
the early production of IFN-g in CD4+ but not CD8+ effector T cells. Further, it suggests that isoform-specic Ras inhibitors could be of value for selectively modulating
CD4+ and CD8+ effector functions in the clinical setting.
doi:10.1016/j.cyto.2011.07.067

PS1-009
TLR responsiveness of B cells is modulated in patients with multiple sclerosis
following IFN-b therapy
Severa M. Giacomini Elena a, F. Rizzo a, M.E. Remoli a, V. Gafa a, S. Romano b, R.
Mechelli b, M. Salvetti b, E.M. Coccia b, a Istituto Superiore di Sanit, Roma, b Universit di
Roma La Sapienza
Although multiple sclerosis (MS) is traditionally considered as a T-cell mediated
disease, B cells are increasingly recognized as important players in MS immunopathology. IFN-b has been used over the past 15 years as primary therapy for relapsing
remitting (RR) MS. However, the immunomodulatory mechanisms that provide a
therapeutic effect against this inammatory disease are not yet completely elucidated. In the attempt to characterize the response of B cells to IFN-b, we rstly analyzed the expression of TLR7 and TLR9 genes, which represent two key molecules
involved in the activation, proliferation and differentiation of B cells. Blood samples
from RRMS patients were collected at baseline and 1 month after the beginning of
IFN-b. TLR7 gene was clearly induced in in vivo IFN-b-conditioned B cells isolated
from MS patients, indicating an effective response to this immunomodulatory treatment. This effect was specic, since no modulation was observed in patients undergoing glatiramer acetate (GA) therapy. Conversely, TLR9 expression was not
modied upon both GA or IFN-b therapy.
Based on these results, we sought to investigate the response to TLR triggering of
in vivo IFN-b-conditioned B cells. We found that along IFN-b therapy, the expression
of CD38 and CD86, respectively markers of cell activation and maturation, was significantly enhanced on B cells following in vitro stimulation with a specic TLR7 agonist
of peripheral blood mononuclear cells (PBMC), while this phenomenon did not occurr
when TLR9 was engaged or when patients followed GA therapy. Moreover, IFN-b signicantly enhanced the capacity of TLR7 to promote IgM and IgG secretion, while

doi:10.1016/S1043-4666(11)00267-5

poorly inuenced TLR9 mediated B cell differentiation after 7 days of culture. To

investigate the mechanisms responsible for the effects induced by IFN-b, we studied
soluble factors that could contribute to B cell differentiation/activation, namely IL-10
and IL-6. Interestingly, following TLR7 agonist stimulation of PBMC we found a significant enhancement of the release of both these cytokines after 1 month from the
beginning of the therapy with IFN-b when compared to the baseline.
A possible explanation of TLR7-mediated effects observed after IFN-b therapy
could rely on a reduced expression of TLR7 gene in PBMC from MS patients compared
to that of healthy donors, which is partly restored after 1 month of IFN-b treatment.
In conclusion, these results suggest that IFN-b treatment can exert its therapeutic
effects also by acting on the differentiation and/or activation status of B lymphocytes
and in particular by modulating the responsiveness to TLR7 and, in turn, B cells
functions.
This work was supported by FISM grant (#2009/R/7).
doi:10.1016/j.cyto.2011.07.068

PS1-010
CD8 T cell priming in the presence of IFNa improves memory differentiation and
the efcacy of adoptive immunotherapy
Sandra Hervas-Stubbs a, Uxua Mancheo b, Ana Larraga a, Carlos Alfaro a, Maria C.
Ochoa a, Aitziber Echeverria a, Iranzu Gonzalez b, Jose-Ignacio Riezu-Boj a, Esther
Larrea a, Ignacio Melero a,c, a Division of Gene Therapy and Hepatology, Centre for
Applied Medical Research (CIMA), University of Navarra, Pamplona, 31008, Spain, b Digna
Biotech, Madrid, 28003, Spain, c Clinica Universidad de Navarra. University of Navarra
Previous studies on mouse and human CD8 T cells have demonstrated that Interferons (IFN)- a/b enhance the acquisition of effector functions, while there are data in mice
supporting a role for IFNa/b in memory CD8 T-cell differentiation. Here we show that
IFNa directly provides a critical signal to human nave CD8 T cells supporting their differentiation into T central Memory-like cytotoxic T cells (CTLs). Such CTLs are endowed
with an enhanced ability to respond to homeostatic cytokines and to eventual secondary antigen stimulation. The ability to mount a robust recall response was also found in
an in vivo model in which ex vivo primed murine CTLs were adoptively transferred into
mice and re-challenged with antigen 90 days later. These experiments show that IFNasignaling during priming of nave CD8 T cells imprints decisive information that is lastingly remembered. Importantly, exposure to IFNa during in vitro priming of nave HLAA2+ CD8 T cells with autologous DC loaded with Melanoma Antigen Recognized by T
cells (MART1)26-35 peptide enhanced the capacity of the resulting CTLs to specically
lyse MART1 expressing-melanoma cells. Moreover, in a mouse model of melanoma,
adoptive transfer of tumor-specic CTLs primed in vitro with IFNa exhibited an
improved ability to contain tumor progression. Our results indicate that IFNa plays
an important role in evoking steadfast memory CTLs and that the effects of this cytokine
might be exploited to enhance the efcacy of adoptive T-cell therapy.
doi:10.1016/j.cyto.2011.07.069

PS1-011
The Role Of Type I Interferon Signaling Via T Cells In The Early Phase Of Experimental Autoimmune Encephalomyelitis (EAE)
Nadia Kavrochorianou a,d, Maria Evangelidou b, Michael Tovey c, George
Thyphronitis d, Sylva Haralambous a, a Laboratory of Transgenic Technology, Hellenic
Pasteur Institute, Athens, Greece, b Laboratory of Molecular Genetics, Hellenic Pasteur
Institute, Athens, Greece, c Laboratory of Biotechnology and Applied Pharmacology, CNRS
UMR8113, Ecole Normale Suprieure de Cachan, Cachan, France, d Department of
Biological Applications and Technologies, University of Ioannina, Ioannina, Greece