HUMAN B CELL DIFFERENTIATION: DEPENDENCE ON INTERACTIONS WITH MONOCYTES AND T LYMPHOCYTES VIA CD40, CD80 (B7.1), AND THE CD2-LIGANDS CD48 AND CD58 (LFA-3)

B cell differentiation depends on cellular interactions with T lymphocytes and monocytes via adhesion molecules (AM). In order to characterize AM which are required for B cell differentiation immunoglobulin production using unseparated peripheral blood mononuclear cells (PBMC) was studied. Unstimulated human PBMC were cultured for 9 days with mAb directed at CD2/CD48, /CD58, CD59, CD5/CD72, CD11a—CD18/CD54, CD28/CD80, CD86, CD40/CD40L, or rat CD2 (control). B cell differentiation was quantified measuring IgM and in some cases IgA, IgG, and IgE production. IgM levels were significantly reduced by mAb against CD40, CD48, CD58 and CD80. The reduction was not due to isotype switching to IgA, IgG or IgE. The role of CD40, CD48, CD58 and CD80 was further investigated after depletion of different cell types. Depletion of monocytes and NK cells resulted in no detectable IgM production irrespective of added mAbs. In contrast, IgM production was still present after depletion of T cells and NK cells. Only mAb against CD80 and CD48 significantly reduced IgM production, the reduction of IgM production by anti-CD40 mAb was less than in the presence of T cells. Importantly, anti-CD58 mAb had no effect on IgM production after T cell and NK cell depletion. Taken together, the AM CD40, CD48, CD58, and CD80 are involved in Ig production of unseparated PBMCs. In this model of B cell differentiation only the AM CD58 depend on the presence of T cells while CD48 and CD80 help was found to be T cell independent.     

Palaeoecological preferences and geographical distribution of Late Maastrichtian deep-sea ostracods in the South Atlantic

The distributional patterns of deep-sea ostracods in the terminal Cretaceous South Atlantic were examined from seven DSDP/ODP holes (356, 525A, 527, 528, 529, 698A and 689B). Many of the genera recorded in this study are previously known from Campanian-Maastrichtian deep-sea sediments. However, the recovery of Aversovalva, Hemiparacytheridea, Profundobythere, Mayburya and Legitimocythere reveals considerable similarities with the Tertiary deep-sea ostracod fauna of the Pacific and the North Atlantic. The underlying causes controlling the variation in faunal density among the various sites are likely to be related to variation in food supply and, to a lesser extent, to variation in oxygen levels, since all sites quantitatively assessed are regarded to represent oligotrophic, well-oxygenated conditions. A Q -mode principal component analysis with varimax rotation yielded a first component largely dominated by Bythocypris and significantly correlated with palaeotemperature; there is an increasing abundance of Bythocypris towards higher latitudes with a maximum abundance at Site 689 where the palaeotemperature was around 9°C. A species of the thermophilic genus Cytherelloidea occurs also at this site, probably as a result of a slow adjustment to colder conditions in the absence of a thermocline at that time. The second component is largely dominated by Krithe and significantly correlated with palaeotemperature and palaeodepth. Among the sites studied, Krithe is most common at Site 356, where the palaeotemperature was around 15°C; today, most representatives of this genus are psychrospheric and adapted to temperatures below 10°C.     

The relation between wave exposure and distribution of emergent vegetation in a eutrophic lake

SUMMARY 1. Maximum water depth penetration and changes in horizontal distribution during 39 years of the emergent vegetation in a eutrophic lake in southern Sweden were investigated.
2. The capacity of the emergent vegetation to penetrate into deeper water areas was found to be higher at wave exposed than al sheltered sites.
3. Differences in biomass and biomass allocation of the dominant speeies. Phragmites australis , between an exposed and a sheltered site suggest that horizontal expansion towards deeper water at sheltered sites is limited by unfavourable substrate conditions.     

A comparative study of Lower Cambrian Halkieria and Middle Cambrian Wiwaxia

Two Cambrian lepidote metazoans known from different respective types of preservation have been compared in order to elucidate their biology and affinities. The widely distributed Lower Cambrian Halkieria is represented by isolated hollow sclerites, probably of originally calcareous composition. The Middle Cambrian Wiwaxia is known from the Burgess Shale as isolated sclerites (scales and spines) and as more or less complete individuals. Although Halkieria sclerites were mineralized and those of Wiwaxia were probably not, there are fundamental structural and morphological similarities between the two. Both bad an imbricating scaly and spiny armour consisting of hollow sclerites with a longitudinally fibrous structure. The sclerites did not grow, but were probably moulted during the course of ontogenetic growth. Halkieria and Wiwaxia are regarded as closely related. Both are referred to the Order Sachitida He 1980. The sclerite armour of Halkieria is reconstructed on the template provided by Wiwaxia. The interpretation of sachitid sclerites as protective armour is an alternative to the interpretation by Jell (1981, Alcheringa 5 )that sachitid sclerites were respiratory organs in an animal of probable annelid affinities. Sachitids are interpreted as sluggish, benthic deposit feeders that do not belong to any recognized phylum.     

THE ATOMIC FORCE MICROSCOPE DETECTS ATP-SENSITIVE PROTEIN CLUSTERS IN THE PLASMA MEMBRANE OF TRANSFORMED MDCK CELLS

Plasma membrane proteins are supposed to form clusters that allow ‘functional cross-talk’ between individual molecules within nanometre distance. However, such hypothetical protein clusters have not yet been shown directly in native plasma membranes. Therefore, we developed a technique to get access to the inner face of the plasma membrane of cultured transformed kidney (MDCK) cells. The authors applied atomic force microscopy (AFM) to visualize clusters of native proteins protruding from the cytoplasmic membrane surface. We used the K⁺channel blocker iberiotoxin (IBTX), a positively charged toxin molecule, that binds with high affinity to plasma membrane potassium channels and to atomically flat mica. Thus, apical plasma membranes could be ‘glued’ with IBTX to the mica surface with the cytosolic side of the membrane accessible to the scanning AFM tip. The topography of these native inside-out membrane patches was imaged with AFM in electrolyte solution mimicking the cytosol. The plasma membrane could be clearly identified as a lipid bilayer with the characteristic height of 4.9±0.02nm. Multiple proteins protruded from the lipid bilayer into the cytosolic space with molecule heights between 1 and 20nm. Large protrusions were most likely protein clusters. Addition of the proteolytic enzyme pronase to the bath solution led to the disappearance of the proteins within minutes. The metabolic substrate ATP induced a shape-change of the protein clusters and smaller subunits became visible. ADP or the non-hydrolysable ATP analogue, ATP-γ-S, could not exert similar effects. It is concluded that plasma membrane proteins (and/or membrane associated proteins) form ‘functional clusters’ in their native environment. The ‘physiological’ arrangement of the protein molecules within a cluster requires ATP.     

Cyclic Stretch up-regulates Proliferation and Heat Shock Protein 90 Expression in Human Melanocytes

Human skin is repeatedly exposed to mechanical stretching in vivo, but in an ordinary culture of skin cells this prominent feature has been neglected. In order to study whether mechanical stretching plays a role for human melanocytes, we have established a culture technique to mimic this physical stretching: primary cultures of human melanocytes were plated on silicon supports, which undergo a stretching of about 10% of the initial length. After application of repeated stretching and relaxation for 4 days, cell count was significantly (about 40%) enhanced. In addition, we found ～ 2-fold increase in heat shock protein (HSP) 90, both at the protein and mRNA level. HSP 90 is known to bind to Raf-1 and, therefore, may contribute to the Raf-1-MEK (mitogen-activated protein-kinase kinase)-MAPK (mitogen-activated protein-kinase) signaling pathway. Disruption of the Raf-1-HSP 90 multimolecular complex by geldanamycin lead to a considerable decrease in melanocyte cell count. However, geldanamycin did not reverse the stretch-induced growth stimulation. Therefore, the stretch-mediated up-regulation of HSP 90 expression in melanocytes appears to be independent of stretch-mediated growth stimulation. These findings have strong implications for the in vitro cultivation of melanocytes for transplantation purposes.