Simian immunodeficiency virus (SIV)-specific CD8+ T-cell responses in vervet African green monkeys chronically infected with SIVagm

African green monkeys (AGM) do not develop overt signs of disease following simian immunodeficiency virus (SIV) infection. While it is still unknown how natural hosts like AGM can cope with this lentivirus infection, a large number of investigations have shown that CD8(+) T-cell responses are critical for the containment of AIDS viruses in humans and Asian nonhuman primates. Here we have compared the phenotypes of T-cell subsets and magnitudes of SIV-specific CD8(+) T-cell responses in vervet AGM chronically infected with SIVagm and rhesus monkeys (RM) infected with SIVmac. In comparison to RM, vervet AGM exhibited weaker signs of immune activation and associated proliferation of CD8(+) T cells as detected by granzyme B, Ki-67, and programmed death 1 staining. By gamma interferon enzyme-linked immunospot assay and intracellular cytokine staining, SIV Gag- and Env-specific immune responses were detectable at variable but lower levels in vervet AGM than in RM. These observations demonstrate that natural hosts like SIV-infected vervet AGM develop SIV-specific T-cell responses, but the disease-free course of infection does not depend on the generation of robust CD8(+) T-cell responses.     

A MICROSCOPIC TECHNIQUE TO MEASURE MESOPHYLL SUCCULENCE

Mesophyll succulence (S_m), the ratio of water content to chlorophyll content of a cell or tissue, has been proposed as an index of CAM potential. In plant tissues where all cells contain chloroplasts, S_m may be measured easily. If water tissue (water-storing parenchyma lacking chloroplasts) is present, however, severe technical difficulties arise. We have developed a comparable index, morphological mesophyll succulence (S_mm), which allows rapid determination of vacuole to chloroplast volumes in cells. There is a high correlation between the results obtained by these two techniques. Advantages of the microscopic technique include: more rapid determinations, fewer equipment needs, ability to be applied in the field, and no need to physically separate water tissues from photosynthetic tissues.     

Histoincompatibility reactions in the hydrocoral Millepora dichotoma

The xenogeneic- and allogeneic immunological specificity of the hydrocoral Millepora dichotoma has been investigated. Xenogeneic histoicompatibility reactions have been observed between this hydrocoral and a series of species belonging to the Demospongiae and to the Anthozoa (both Hexacorallia and Octocorallia). The xenogeneic histoincompatibility reactions proceed in the following sequence: (a) Species-unspecific sensitization; (b) necrosis formation, which is very likely due to an autolytic process; (c) callus formation, due to an hyperplastic growth of stolons; and (d) formation of a contact barrier in form of a barrier layer or a restored stolonial layer. Allogeneic histoincompatibility reactions are restricted to those regions of the coral which are interspersed with polyps; allogenic fusion is observed between branches, with a high density of polyps and the “foot”-region, which is characterized by a low polyp density.     

The biological activity of bacteriophage DNA, prepared by the cationic detergent dilution technique

The preparation of phage lambda DNA infecting E. coli K 12 with cationic detergent is described. This DNA infects E. coli spheroblasts with the same efficiency as DNA prepared by phenol methods.     

Human serum deoxyribonuclease assay in (3H)DNA-polyacrylamide gels without staining artifacts

A method is described for detecting and evaluating serum deoxyribonuclease (DNase) activities after their separation by disc electrophoresis in polyacrylamide gels containing radioactively labeled DNA.After the electrophoretic run the gels are sliced, incubated in an appropriate buffer, and the amount of diffusible radioactive DNA fragments formed by the action of DNases in the incubation buffer is determined.The method has a high sensitivity as well as a quantitative reproducibility within ±5% even at low enzyme activities down to 10 pg Worthington DNase I equivalents.This method has been found superior to procedures that use staining of the gels for unhydrolyzed DNA, where irrelevant stained bands may invalidate the results. Thus, we get meaningful results even with human serum.     

Lectin, a possible basis for symbiosis between bacteria and sponges.

From the marine sponge Halichondria panicea a lectin was isolated and characterized. The homogeneous lectin (composed of protein to 80.7% and of neutral carbohydrates to 14.1%) had a molecular weight of 78,000 (determined by gel filtration) and consisted of four subunits with a molecular weight of 21,000 each (determined by gel electrophoresis in the presence of sodium dodecyl sulfate). The hemagglutinating activity was only slightly dependent upon ionic strength and incubation temperature and did not require divalent cations, but it was inhibited by reagents for thiol groups. The Halichondria lectin was completely inhibited in hemagglutination competition experiments in the presence of fetuin, D-galacturonic acid, D-glucuronic acid, polygalacturonic acid, or L-fucose. The purified Halichondria lectin did not cause reaggregation of dissociated H. panicea cells. From the same sponge species bacteria were isolated and identified as Pseudomonas insolita. These bacteria were cultivated in marine broth 2216. Under these culture conditions the bacteria grew only in the presence of the homologous lectin; the lectin-caused effect was not abolished by D-glucuronic acid or D-galacturonic acid. However, after addition of a polysaccharide-containing fraction isolated from P. insolita, the lectin-caused, growth-promoting effect was abolished. Other lectins were found to exhibit no growth-promoting effect. On the basis of colony counts, P. insolita was the predominant bacterial species in the sponge extract; 1.9 X 10(6) Pseudomonas colonies were measured in extracts isolated from 1 g of sponge. The assumption of an interrelationship between the sponge and the bacterium is supported by the results indicating that the Halichondria lectin has no effect on the growth of such bacteria isolated from six other marine sponge species. Evidence is presented which indicates that the Halichondria lectin is not utilized during growth of the Pseudomonas species. Lectin activity was detected on the surface of mucoid cells from H. panicea. From the data obtained the possibility is discussed that the Halichondria lectin is a basis for a symbiotic relationship between the sponge and the bacterium.