For blocking of perforin/granzyme-mediated cytotoxicity, DN T cells were incubated O/N with CMA (115 nM; Sigma), washed twice, and added to the MLR. CFSE-labeled CD4+ T cells (2.5×105/well) were stimulated with allogeneic DC (1.25×105/well) in a 24-well tissue culture plate (Corning/Costar, NY, USA). DN T cells were Caspase activity assay added to the top chamber (2.5×105/well) together

with allogeneic DC (1.25×105/well). Top and bottom chambers were separated by a 0.4-μm membrane that allows soluble factors, but not T cells, to pass through. After 5 days, proliferation of CD4+ T cells in the bottom chamber was measured by flow cytometry. Data were compared using 2-tailed Student’s t-test. p-value less than 0.05 was considered significant. The authors thank Jana Berger and Dorothea Gebhardt for excellent technical assistance, Uwe Appelt for FACS sorting and

Thomas Hünig, Edward Kim, Jacobus Bosch, and Evelyn Ulrich for critical reading of the manuscript. This work was supported by the see more Deutsche Forschungsgemeinschaft (MA 1351/7-1, KFO 146). Conflict of interest: The authors declare no financial or commercial conflict of interest. Detailed facts of importance to specialist readers are published as ”Supporting Information”. Such documents are peer-reviewed, but not copy-edited or typeset. They are made available as submitted by the authors. “
“Citation Groer M, El-Badri N, Djeu J, Harrington M, Van Eepoel J. Suppression of natural killer cell cytotoxicity in postpartum women. Am J Reprod Immunol 2010; 63: 209–213 Problem  Natural Killer (NK) cell numbers and cytotoxicity are suppressed during pregnancy. Little is known about postpartum NK GPX6 number and function. Method of study  Postpartum women (n = 39) were studied at one week and then

monthly over the first six postpartum months. The standard natural killer cell cytotoxicity assay (NKCA) was performed. This is a Cr51 release assay from K562 cells cultured with peripheral blood mononuclear cells (PBMCs). Results  Data indicate suppression of NK cytotoxicity in postpartum women. Cytotoxicity at each effector:target (E:T) ratio showed a drop from 1 week postpartum, reaching a nadir at around 2 months, and a trend towards recovery of cytotoxicity from 3 to 6 months. Lytic units (LUs) from pre-incubated cells from postpartum women were lower than age-matched, non-pregnant, non-postpartum controls through the fifth postpartum month. Conclusion  These data suggest that the postpartum period, like pregnancy, is characterized by decreased NK cytotoxicity activity. This suppressed NK cytotoxic effect may result as a response to interaction with tolerized fetal microchimeric cells accumulated during pregnancy in maternal blood and tissues. “
“In cell culture, Rickettsia felis grows only at low temperatures (< 31 °C).