Critical role of extracellular vesicles in modulating the cellular effects of cytokines

Under physiological and pathological conditions, extracellular vesicles (EVs) are present in the extracellular compartment simultaneously with soluble mediators. We hypothesized that cytokine effects may be modulated by EVs, the recently recognized conveyors of intercellular messages. In order to test this hypothesis, human monocyte cells were incubated with CCRF acute lymphoblastic leukemia cell line-derived EVs with or without the addition of recombinant human TNF, and global gene expression changes were analyzed. EVs alone regulated the expression of numerous genes related to inflammation and signaling. In combination, the effects of EVs and TNF were additive, antagonistic, or independent. The differential effects of EVs and TNF or their simultaneous presence were also validated by Taqman assays and ELISA, and by testing different populations of purified EVs. In the case of the paramount chemokine IL-8, we were able to demonstrate a synergistic upregulation by purified EVs and TNF. Our data suggest that neglecting the modulating role of EVs on the effects of soluble mediators may skew experimental results. On the other hand, considering the combined effects of cytokines and EVs may prove therapeutically useful by targeting both compartments at the same time.     

The mammalian sodium channel BNC1 is required for normal touch sensation

Of the vertebrate senses, touch is the least understood at the molecular level The ion channels that form the core of the mechanosensory complex and confer touch sensitivity remain unknown. However, the similarity of the brain sodium channel 1 (BNC1) to nematode proteins involved in mechanotransduction indicated that it might be a part of such a mechanosensor. Here we show that disrupting the mouse BNC1 gene markedly reduces the sensitivity of a specific component of mechanosensation: low-threshold rapidly adapting mechanoreceptors. In rodent hairy skin these mechanoreceptors are excited by hair movement. Consistent with this function, we found BNC1 in the lanceolate nerve endings that lie adjacent to and surround the hair follicle. Although BNC1 has been proposed to have a role in pH sensing, the acid-evoked current in cultured sensory neurons and the response of acid-stimulated nociceptors were normal in BNC1 null mice. These data identify the BNC1 channel as essential for the normal detection of light touch and indicate that BNC1 may be a central component of a mechanosensory complex.     

Immunological sensitization following inapparent infection with dengue virus type 3 in rhesus monkeys.

Rhesus monkeys previously given dengue virus type 3, without apparent viremia or antibody response, exhibited a secondary-type response upon reinoculation. These data suggest that monkeys can be immunologically sensitized to dengue virus without detectable antibody production.     

Atomic structure of a fragment of human CD4 containing two immunoglobulin-like domains.

The structure of an N-terminal fragment of CD4 has been determined to 2.4 A resolution. It has two tightly abutting domains connected by a continuous beta strand. Both have the immunoglobulin fold, but domain 2 has a truncated beta barrel and a non-standard disulphide bond. The binding sites for monoclonal antibodies, class II major histocompatibility complex molecules, and human immunodeficiency virus gp120 can be mapped on the molecular surface.     

Preparation of isolated single cardiac cells from adult frog atrial tissue.

Isolated cardiac cells from bullfrog atrial tissue can be readily prepared by digestion of intact fragments of atrial tissue with trypsin and collagenase. These isolated cells have dimensions of about 5 mum in width and range in length from 300 mum to over 500 mum. Such isolated cells may prove useful for the investigation of contractile activity of cardiac muscle at the single cell level and at the sarcomere level within the single cell.     

Immunological sensitization following inapparent infection with dengue virus type 3 in rhesus monkeys

Summary Rhesus monkeys previously given dengue virus type 3, without apparent viremia or antibody response, exhibited a secondary-type response upon reinoculation. These data suggest that monkeys can be immunologically sensitized to dengue virus without detectable antibody production.The authors wish to express their deepest gratitude to Alma Till man, Carlie White, Annie McCoy, Francis Brisset, Albert Jackson and Becky Kagarise for their devoted technical assistance. This research was supported in part by the U.S. Army Medical Research and Development Command under Contract No. DADA-16-69-C-9048 and DAMD-17-74-C-4005 and by the National Institute of Allergy and Infectious Diseases, National Institutes of Health, U.S. Public Health Service, under Research Grant No. 5-R22-AI02686-13,-14 and-15, and Training Grant No. 5-T01-AI-00110-14.     

Preparation of isolated single cardiac cells from adult frog atrial tissue

Summary Isolated cardiac cells from bullforg atrial tissue can be readily prepared by digestion of intact fragments of atrial tissue with trypsin and collagenase. These isolated cells have dimensions of about 5 m in width and range in length from 300 m to over 500 m. Such isolated cells may prove useful for the investigation of contractile activity of cardiac muscle at the single cell level and at the sarcomere level within the single cell.This investigation was supported by U.S. Public Health Service, National Institutes of Health Grant No. HL 12426 and a Kansas Heart Association Grant-in-Aid.We wish to thank Dr.J. Sommers of Duke University Medical Center for doing electron microscopy of the isolated cells to confirm the single cell nature of the preparations.     

Chemically mediated cell-to-cell contractile activation in isolated frog atrial cardiac cells

Summary A dying single frog atrial cardiac cell liberates an unknown substance which diffuses away from the dying cell and activates contractile activity in other isolated intact single cardiac cells within the vicinity of the dying cell.This investigation was supported by US Public Health Service, National Institutes of Health Grant HL 18943 and a Kansas Heart Association Grant-In-Aid.We wish to thank Dr Alan Chapman of the Department of Anatomy for doing the scanning electron microscopy of the cardiac cells.