Proteome analysis of mitochondrial outer membrane from Neurospora crassa.

The mitochondrial outer membrane mediates numerous interactions between the metabolic and genetic systems of mitochondria and the rest of the eukaryotic cell. We performed a proteomic study to discover novel functions of components of the mitochondrial outer membrane. Proteins of highly pure outer membrane vesicles (OMV) from Neurospora crassa were identified by a combination of LC-MS/MS of tryptic peptide digests and gel electrophoresis of solubilized OMV proteins, followed by their identification using MALDI-MS PMF.

Differential expression of cytokines in human blood monocyte subpopulations.

Cytokine expression was analyzed in CD14++ regular monocytes and in the novel subset of CD14+/CD16+ small monocytes. Biologic activity for tumor necrosis factor (TNF), interleukin-1 (IL-1), and IL-6 in the supernatant of elutriator-enriched, cell sorter-purified small monocytes was about 10-fold lower compared with regular monocytes when stimulated with lipopolysaccharide (LPS) for 12 hours. In CD14++ regular monocytes levels were 1,157 U x 10(-3)/mL, 158 U/mL, and 1,337 U/mL for TNF, IL-1, and IL-6, respectively.

Splenectomy does not influence outcome of pneumococcal septicemia in a porcine model.

The existence of the overwhelming postsplenectomy infection syndrome in adults after traumatic splenectomy is controversial. Due to the similarity of the porcine immune system to man we chose the pig to study subsets of peripheral mononuclear cells after splenectomy and resistance to experimental Pneumococcal infection after splenic surgery and specific immunization. Female miniature pigs were assigned to four operative groups: sham operation, splenectomy, splenic resection, and heterotopic splenic autotransplantation.

Small (CD14+/CD16+) monocytes and regular monocytes in human blood.

Compared to the obvious phenotypic and functional heterogeneity of tissue macrophages, little information is available on subsets of blood monocytes. We have employed two-color immunofluorescence and flow cytometry for the definition of regular and small monocytes, the latter characterized by the low-density expression of CD14 and the strong expression of the CD16 (Fcy-RIII) antigen. These cells comprise 15% of the blood monocytes and they appear to be similar in phenotype to the alveolar macrophage.

Inverse relationship of CA2+ mobilization and cell proliferation in CD8+ memory and virgin T cells.

Memory T cells can now be defined with various monoclonal antibodies but little is known about the functional properties of these cells as compared to virgin T cells. We have studied Ca2+ mobilization and proliferation of memory and virgin cells in both the CD4 and CD8 subsets in response to phytohemagglutin-in. Using two-color fluorescence (yellow, red) for cell surface staining combined with two-color fluorescence for the Ca2+ chelator indo-1 (violet, blue) we can demonstrate that CD8+UCHL1+ memory cells exhibit a 1,5-fold higher influx than the CD8+UCHL1- virgin cells.