Benedita Rocha

Response of naïve and memory CD8+ T cells to antigen stimulation in vivo.

We studied the influence of memory T cell properties on the efficiency of secondary immune responses by comparing the in vivo immune response of the same numbers of T cell receptor (TCR) transgenic (Tg) naïve and memory T cells. Compared to naïve Tg cells, memory cells divided after a shorter lag time; had an increased division rate; a lower loss rate; and showed more rapid and efficient differentiation to effector functions. We found that initial naïve T cell priming resulted in cells expressing mutually exclusive effector functions, whereas memory T cells were multifunctional after reactivation, with each individual cell expressing two to three different effector functions simultaneously. These special properties of memory T cells ensure the immediate control of reinfection.

A thymic pathway of mouse natural killer cell development characterized by expression of GATA-3 and CD127

Natural killer (NK) cell development is thought to occur in the bone marrow. Here we identify the transcription factor GATA-3 and CD127 (IL-7Rα) as molecular markers of a pathway of mouse NK cell development that originates in the thymus. Thymus-derived CD127+ NK cells repopulated peripheral lymphoid organs, and their homeostasis was strictly dependent on GATA-3 and interleukin 7. The CD127+ NK cells had a distinct phenotype (CD11bloCD16−CD69hiLy49lo) and unusual functional attributes, including reduced cytotoxicity but considerable cytokine production. Those characteristics are reminiscent of human CD56hiCD16− NK cells, which we found expressed CD127 and had more GATA-3 expression than human CD56+CD16+ NK cells. We propose that bone marrow and thymic NK cell pathways generate distinct mouse NK cells with properties similar to those of the two human CD56 NK cell subsets.

Alternatively spliced NKp30 isoforms affect the prognosis of gastrointestinal stromal tumors

The natural killer (NK) cell receptor NKp30 is involved in the recognition of tumor and dendritic cells (DCs). Here we describe the influence of three NKp30 splice variants on the prognosis of gastrointestinal sarcoma (GIST), a malignancy that expresses NKp30 ligands and that is treated with NK-stimulatory KIT tyrosine kinase inhibitors. Healthy individuals and those with GIST show distinct patterns of transcription of functionally different NKp30 isoforms. In a retrospective analysis of 80 individuals with GIST, predominant expression of the immunosuppressive NKp30c isoform (over the immunostimulatory NKp30a and NKp30b isoforms) was associated with reduced survival of subjects, decreased NKp30-dependent tumor necrosis factor-α (TNF-α) and CD107a release, and defective interferon-γ (IFN-γ) and interleukin-12 (IL-12) secretion in the NK-DC cross-talk that could be restored by blocking of IL-10. Preferential NKp30c expression resulted partly from a single-nucleotide polymorphism at position 3790 in the 3′ untranslated region of the gene encoding NKp30. The genetically determined NKp30 status predicts the clinical outcomes of individuals with GIST independently from KIT mutation.

The extrathymic T-cell development pathway

In normal mice, not all T-lineage cells are generated and selected in the thymus; an alternative, extrathymic, development pathway exists. Extrathymic T cells are rare in the spleen and lymph nodes, but are abundant in some tissues, such as the gut. Here, Benedita Rocha, Pierre Vassalli and Delphine Guy-Grand discuss the rules of selection of extrathymic T cells, assess the possible role of these cells in the defence of epithelial integrity and their potential role in autoimmune disease.

Lymphocyte Population Kinetics in the Mouse

In a normal dynamic equilibrium, at least half of the peripheral T-cell pool is constituted by lymphocytes which have divided 24-48 h previously, and are therefore rapidly renewed. The renewal of peripheral T cells occurs partly by influx of cells from the thymus and, more importantly, by cell division at the periphery. The cyclic pattern of decay observed for T cells after HU treatment suggests the presence of progenitor-descendent relationships within the peripheral T-cell pool. Peripheral progenitors must contain both cycling and non-cycling cells to account for cell recovery after HU administration in ATx mice. T-cell production at the periphery involves both organized (spleen or lymph nodes) as well as non-organized lymphoid tissue (GALT). The latter may in fact provide the major contribution. Expansion of mature T lymphocytes contributes to clonal persistence at the periphery and to the choice of T-cell repertoires. The importance of post-thymic selection of T-cell repertoires is suggested by the considerable expansion potential revealed by peripheral T cells.

Modifications of CD8+ T Cell Function during In Vivo Memory or Tolerance Induction

Naive monoclonal T cells specific for the male antigen can be stimulated in vivo to eliminate male cells and become memory cells or to permit survival of male cells and become tolerant. Memory cells responded to TCR ligation by cyclic oscillations of calcium levels and immediate secretion of very high levels of IL-2 and interferon-gamma. Tolerant cells did not proliferate in response to ionomycin and phorbol myristate acetate, failing to mobilize calcium to produce IL-2 or express IL-2R, but survived for long time periods in vivo and secreted IL-10. These results emphasize that tolerance is not an absence of all functional activity and may be associated with modifications of behavior conferring important regulatory functions on tolerant T cells.

Extrathymic T cell differentiation

In the mouse, the gut mucosa is a major site of extrathymic differentiation of T cells. Recent data in this past year show that this process differs from the main thymic differentiation pathway not only in its location, but also in its use of costimulatory molecules, signal transduction modules, and mechanisms of repertoire selection. The thymus exerts an influence on the expansion of the extrathymically differentiated gut intraepithelial lymphocytes that appears to be varied in nature, including acting as a source of TCR- progenitors. All gut intraepithelial lymphocytes, whatever their extrathymic or thymic site of differentiation, have common features of activated and specialized cytotoxic cells. Other T cells may differentiate extrathymically, in particular in the liver; these later cells appear to have a very restricted, probably autoreactive repertoire, and also display natural killer cell features.

Peripheral T cell survival

T cell survival in the periphery is an active process, depending on continuous TCR engagement by peptide-MHC complexes and/or response to environmental cytokines. Naive T cells require interactions with the MHC restricting element. The survival requirements of memory T cells are as yet insufficiently characterized, but MHC-restricted interactions are not necessary.

The organization of mature T-cell pools.

To deal with exogenous pathogens the peripheral T-cell compartment requires diverse repertoires (as those of naive cells) and efficient responses, the latter dependent on the persistence of memory cells. In the present work we show that (i) naive and memory cells differ in the type of interactions required for survival and division; (ii) they are segregated into independent ecological niches; (iii) that the size of each niche is controlled by independent homeostatic mechanisms; and (iv) that naive T cells do not have intrinsic life spans, surviving in the absence of thymus output but being continuously substituted by thymus export. The independent homeostatic regulation of the naive and memory T-cell pools guarantees the maintenance of versatile and efficient repertoires throughout life as well as the persistence of the naive T-cell pool after the thymus atrophies at puberty.

High expression of active CDK6 in the cytoplasm of CD8 memory cells favors rapid division

Antigen-driven CD8 memory T cell proliferation is more rapid than that of naive T cells, ensuring efficient control of the pathogen after reinfection. We show here that naive and memory cells are in different states of G0/G1 arrest. Naive cells are in a classical state of G0/G1 arrest, with high expression of p27Kip1 and low CDK6 and CDK2 kinase activity. In contrast, memory cells have low expression of p27Kip1 and high CDK6 kinase activity. This preactivated kinase is associated with cyclin D3 in the cytoplasm, and neutralization of these complexes with antibody to cyclin D3 blocks the rapid division of memory cells. Therefore G0/G1 memory cells are at a unique preactivated state that favors rapid division after antigen stimulation.