We describe the isolation of a protein, SWAP-70-like adapter of T cells (SLAT), which is expressed at high levels in thymocytes and differentiated Th2 cells. SLAT expression was upregulated in differentiating Th2 cells and downregulated in Th1 cells. Ectopic SLAT expression exerted positive or negative effects on IL-4 versus IFNgamma induction, respectively. TCR signaling induced translocation of SLAT to the immunological synapse and its association with ZAP-70 kinase. SLAT reduced the association of ZAP-70 with TCR-zeta and interfered with ZAP-70 but not Lck signaling. Consistent with these results, pharmacological inhibition of ZAP-70 also induced Th2 skewing. Thus, SLAT is a protein which plays a role in Th2 development and/or activation, perhaps by interfering with ZAP-70 signaling.

The molecular basis of T cell anergy is not completely understood. We show that in antigen-primed anergic murine CD4(+) T cells the linker for activation of T cells (LAT) is hypophosphorylated upon CD3/CD28 restimulation. Signaling events downstream of LAT (PLCgamma1 phosphorylation and p85 [PI3-K] association) were impaired, whereas upstream events (CD3zeta and ZAP-70 phosphorylation) remained intact. LAT recruitment to the immunological synapse and its localization in detergent-resistant membrane (DRM) fractions were defective in anergic T cells. These defects resulted from impaired palmitoylation of LAT and were selective since the DRM localization and palmitoylation of Fyn were intact. This LAT defect was independent of Cbl-b and did not reflect enhanced LAT degradation. These results identify LAT as the most upstream target of anergy induction; moreover, they suggest that regulation of the amount of LAT in the immunological synapse and DRM by posttranslational palmitoylation contributes to the induction of T cell anergy.

Protein kinase C-h (PKCh) plays an important role in T-cell activation via stimulation of AP-1 and NF-jB. Here we report the isolation of SPAK, a Ste20-related upstream mitogen-activated protein kinase (MAPK), as a PKCh-interacting kinase. SPAK interacted with PKCh (but not with PKCa) via its 99 COOH-terminal residues. TCR/CD28 costimulation enhanced this association and stimulated the catalytic activity of SPAK. Recombinant SPAK was phosphorylated on Ser-311 in its kinase domain by PKCh, but not by PKCa. The magnitude and duration of TCR/CD28-induced endogenous SPAK activation were markedly impaired in PKCh-deficient T cells. Transfected SPAK synergized with constitutively active PKCh to activate AP-1, but not NF-jB. This synergistic activity, as well as the receptor-induced SPAK activation, required the PKChinteracting region of SPAK, and Ser-311 mutation greatly reduced these activities of SPAK. Conversely, a SPAKspecific RNAi or a dominant-negative SPAK mutant inhibited PKCh-and TCR/CD28-induced AP-1, but not NF-jB, activation. These results define SPAK as a substrate and target of PKCh in a TCR/CD28-induced signaling pathway leading selectively to AP-1 (but not NF-jB) activation.

PurposeTo compare the effects of endurance exercise performed in the morning and evening on inflammatory cytokine responses in young men.MethodsFourteen healthy male participants aged 24.3 ± 0.8 years (mean ± standard error) performed endurance exercise in the morning (0900–1000 h) on one day and then in the evening (1700–1800 h) on another day with an interval of at least 1 week between each trial. In both the morning and evening trials, the participants walked for 60 minutes at approximately 60% of the maximal oxygen uptake () on a treadmill. Blood samples were collected to determine hormones and inflammatory cytokines at pre-exercise, immediately post exercise, and 2 h post exercise.ResultsPlasma interleukin (IL)-6 and adrenaline concentrations were significantly higher immediately after exercise in the evening trial than in the morning trial (P < 0.01, both). Serum free fatty acids concentrations were significantly higher in the evening trial than in the morning trial at 2 h after exercise (P < 0.05). Furthermore, a significant correlation was observed between the levels of IL-6 immediately post-exercise and free fatty acids 2 h post-exercise in the evening (r = 0.68, P < 0.01).ConclusionsThese findings suggest that the effect of acute endurance exercise in the evening enhances the plasma IL-6 and adrenaline concentrations compared to that in the morning. In addition, IL-6 was involved in increasing free fatty acids, suggesting that the evening is more effective for exercise-induced lipolysis compared with the morning.

Objectives The objectives of the present study were to translate the English version of the Pregnancy Physical Activity Questionnaire into Chinese (PPAQ-C) and to determine its reliability and validity for use by pregnant Chinese women. Methods The study included 224 pregnant women during their first, second, or third trimesters of pregnancy who completed the PPAQ-C on their first visit and wore a uniaxial accelerometer (Lifecorder; Suzuken Co. Ltd) for 7 days. One week after the first visit, we collected the data from the uniaxial accelerometer records, and the women were asked to complete the PPAQ-C again. Results We used intraclass correlation coefficients to determine the reliability of the PPAQ-C. The intraclass correlation coefficients were 0.77 for total activity (light and above), 0.76 for sedentary activity, 0.75 for light activity, 0.59 for moderate activity, and 0.28 for vigorous activity. The intraclass correlation coefficients were 0.74 for "household and caregiving", 0.75 for "occupational" activities, and 0.34 for "sports/exercise". Validity between the PPAQ-C and accelerometer data was determined by Spearman correlation coefficients. Although there were no significant correlations for moderate activity (r = 0.19, P > 0.05) or vigorous activity (r = 0.15, P > 0.05), there were significant correlations for total activity [light and above; r = 0.35, P < 0.01)] and for light activity (r = 0.33, P < 0.01). Conclusions for Practice The PPAQ-C is reliable and moderately accurate for measuring physical activity in pregnant Chinese women.