Isolation of third, fourth, and fifth instar larval midgut epithelia of the moth, Trichoplusia ni

A new tissue isolation technique was used to create intact midgut epithelial wholemounts from three Trichoplusia ni (Lepidoptera: Noctuidae) larval instars. The protease, dispase, removed the basal lamina and associated connective tissue and allowed for high resolution light microscopy of entire epithelia. Columnar, goblet, differentiating, and stem cells were characterized by double fluorescent labelling of f-actin and nuclei. A comparison of cell populations by digital image analysis revealed significant regional and temporal changes in the density and number of differentiating and stem cells. Growth of the midgut epithelium from third to fourth instar, and from fourth to fifth instar, was accomplished by both cell differentiation and cell division. Cell division however, was greatly reduced from fourth to fifth instar with a concomitant sharp decrease in the stem cell population.     

Pigment and lipid compositions of algal and bacterial communities in Ace Lake,Vestfold Hills,Antarctica

The compositions of carotenoids, chlorophylls and lipids at four depths in Ace Lake have been determined as a means of studying the vertical zonation of species in the lake and for comparison with the lipids found in the bottom sediments. The four major species of phytoplankton found in the lake were identified by electron microscopy. The most abundant phytoplankter was Pyramimonas gelidicola McFadden (Chlorophyta, Prasinophyceae) which occurred in greatest numbers at 10 m, the base of the oxylimnion. The pigments and lipids at this depth were mainly derived from this alga. At 11 m (the top of the anoxylimnion) only traces of lipids and pigments attributable to P. gelidicola were found, indicating only limited settling of algal cells through to the anoxylimnion, at least in summer. The pigments at 11 m were dominated by bacteriochlorophylls c derived from green photosynthetic bacteria Chlorobium spp. These pigments were also abundant at 23 m suggesting the presence of intact bacterial cells which had settled out from higher in the water column. Major non-polar lipid classes in the sediments included sterols, alcohols, hydrocarbons and an unusual suite of very long-chain unsaturated ketones and esters which have not previously been reported from antarctic environments. Several novel compounds, not found previously in either sediments or organisms, are reported. These include tri- and tetra-unsaturated straight-chain C₃₉ methyl ketones and C₄₀ ethyl ketones and the methyl ester of a tetra-unsaturated straight-chain C₃₆ fatty acid. The distributions of lipids in the sediment were markedly different from those in the water column indicating extensive bacterial degradation and recycling of labile lipids.     

Activated human T cells can present alloantigens but cannot present soluble antigens

Human T cells, when activated by antigen or mitogen, express Ia antigens. We have examined the capacity of activated T cells to stimulate autologous and allogeneic T cells and their ability to present soluble antigen. Interleukin 2-dependent T-cell lines (TCL), free of accessory cells, were used for antigen-presenting cells. These activated T cells were potent stimulators in an autologous mixed lymphocyte reaction (AMLR), more so than autologous irradiated non-T mononuclear cells. Activated T cells were also able to stimulate proliferation of allogeneic T cells in the absence of any other accessory cells, and this stimulation was blocked by anti-Ia antibodies. Resting unstimulated T cells were unable to stimulate autologous or allogeneic responses. Thus, activated T cells were able to present self antigens and alloantigens. However, activated T cells could not present soluble antigens to autologous T cells or to antigen-specific TCL even if exogenous interleukin 1 was added to cultures. The ability of activated T cells to stimulate an AMLR in vitro may reflect an important immunologic amplification mechanism in vivo. The ability of activated T cells to present alloantigens but not soluble antigens suggests an inability to process antigen, and this may provide further insights into the complexities of antigen presentation.     

Characterization of a T4+/Leu-8+ T cell clone that directly helps B cell Ig production by secreting B cell differentiation factor

A human T4+/Leu-8+ T cell clone (YA2) was established by phytohemagglutinin activation and interleukin 2 (IL 2) propagation. Functional characterization of this clone demonstrated that it provided potent help towards Ig production by pokeweed mitogen-stimulated B cells in the presence of small numbers of autologous T cells or by Staphylococcus aureus Cowan I (SAC)-activated B cells in the presence of B cell growth factor (BCGF). YA2 provided no help to resting B cells and minimal help to either unactivated B cells cultured with BCGF or SAC-activated B cells. Supernatant generated from clone YA2 by IL 2 stimulation had significant B cell differentiation activity but no BCGF or IL 2 activity. Thus, YA2 is a T4+/Leu-8+ potent direct helper only to B cells that are activated and proliferating due to its selective secretion of a differentiation factor, and not an activation and growth factor. The availability of phenotypically defined cloned populations of T cells with restricted functional helper activity related to the secretion of selected B cell tropic factors should prove useful in the dissection of the role of individual T cell subsets in the regulation of the human B cell cycle.     

Insulin-like growth factor-I supports proliferation of autocrine thymic lymphoma cells with a pre-T cell phenotype

We have studied the phenotypic characteristics and growth properties of murine T lymphoma cell lines derived from primary x-ray-induced thymic lymphomas at the earliest stage at which they can be detected, and well before spreading to other organs has occurred. These cell lines serve as model systems for the earliest events in T cell lymphoma induction, before tumor cell progression and spreading to other organs. We find that primary x-ray-induced T cell lymphoma lines have phenotypic characteristics of thymic pre-T cells and show no proliferative response to any of the IL tested nor to other hematopoietic growth factors. However, they do proliferate in response to insulin-like growth factor I (IGF-I) and to a small autocrine peptide distinct from IGF-I, which we term lymphoma growth factor. One of the earliest lesions in T cell lymphoma induction may therefore be an inhibition of differentiation at one of several specific points. In its early stages, T lymphoma cell growth may be restricted to an environment where local concentrations of specific growth factors such as IGF-I or lymphoma growth factor are sufficiently high.     

OCCURRENCE OF AN ISOPRENOID C25 DIUNASATURATED ALKENE AND HIGH NEUTRAL LIPID CONTENT IN ANTRACTIC SEA-ICE DIATOM COMMUNITIES1

The lipid and hydrocarbon composition of natural populations of diatom communities collected during the austral spring bloom of 1985 in the sea-ice at McMurdo Sound, Antartica was analyzed by TLC-FID, GC and GC-MS. Sea-ice diatom communities were dominated by Amphiprora sp., Nitzschia stellata Manguin and Berkeleya sp. at Cape Armitage; N. stellata, Amphiprora, Pleurosigma, N. kerguelensis (O'Meara) Hasle and some small centric diatoms adjacent to the Erebus Ice Tongue; and Porosira pseudodenticulata (Hustedt) Jouse at Wohlschlag Bay. Lipid distributions of the sea ice diatom communities from the Cape Armitage and Ereus sites were characterized by high concentrations of tracylaglecycerol (triacylglycerolplar lipid = 1.0 to 1.5). The hydrocarbon n-C_21:6, common in temperate diatoms, and an isoprenoid C₂₅ diunsaturated alkene were the dominant hydrocarons detected at these two sites. Hydrogenation of the C₂₅ diene produced the known alkane 2, 6, 10, 14-tetramethyl-7- (3-methylpentyl)-pentadecane. The C₂₅ diene is one of several structurally related hydrocarbons reported in many estuarine, coastal and ocean ic sediments. We propose that certain species of diatoms are a likely source of these alkenes in sediments. The first reported biological occurrence of the C₂₅ diene in the green seaweed Enteromorpha prolifera may have been due to the presence of epiphytic microalgae in the field sample analysed.     

A comparison of lipid components of the fresh and dead leaves and pneumatophores of the mangrove Avicennia marina

The component hydrocarbons, sterols, alcohols, monobasic, α,ω-dibasic and ω-hydroxy acids of the fresh hand decayed leaves and the pneumatophores of the mangrove Avicennia marina are reported in detail. From the quantitative comparisons which can be drawn, relative changes in the lipid classes occurring during leaf decay can be highlighted. These base-line data are important to our understanding of inputs to marine intertidal sediments. During leaf decay the only significant changes were a reduction in the total absolute concentrations of monobasic acids due largely to a decrease in concentration of the C₁₈ polyunsaturated fatty acids, and an enhancement of the concentrations of the long-chain monobasic acids, ω-hydroxy acids and α,ω-dibasic acids. This resistance to degradation shown by the cutin derived acids (α,ω-dibasic, ω-hydroxy and long-chain monobasic acids) relative to the cellular and wax derived lipids may allow these cutin components to be used as quantitative markers of A. marina in mangrove associated sediments.     

Activation of human B lymphocytes. XVII. Synergy between nonspecific and specific signals in the antigen-specific responses of human B cells

The incubation of human peripheral blood lymphocytes (PBL) with the natural killer (NK)-sensitive MOLT-4 cell line results in PBL-target cell conjugate formation by certain lymphocyte subpopulations. Following velocity sedimentation, the PBL depleted of these conjugate-forming subpopulations are markedly diminished in the ability to mediate either antibody-dependent cellular cytotoxicity (ADCC) or NK activity. The immediate testing of highly pure PBL subpopulations isolated from the NK target conjugates does not reveal the expected recovery of augmented ADCC or NK levels. Following in vitro incubation, however, the PBL NK target-binding subpopulations do manifest augmented levels of both NK and ADCC, whereas the depleted PBL continue to display diminished NK and ADCC levels. In addition, the degree of augmented NK and ADCC levels recovered by the NK target-binding PBL subpopulations appears dependent on both the time and the temperature of in vitro incubation. Moreover, the ADCC recovery patterns are identical to those observed for NK activity regardless of the time and temperature of in vitro incubation. These results directly demonstrate that the PBL subpopulations isolated from certain NK target cells are functionally enriched in the ability to mediate from ADCC and NK activity.