Macrophage-T cell interactions. III. Production from T cells, under the influence of macrophages, of a factor which complexes Ig and antigen.

Soluble complexes of Ig and antigen have been detected in the serum of mice within 6 hr after immunization. Such complexes are taken up by a subpopulation of T cells. We present evidence which suggests that the complexes are formed through the mediation of a factor released from T cells, tentatively called Ig-antigen complexing factor or IACF. IACF is produced as a result of a macrophage/T-cell interaction, when macrophages are present in an optimal proportion in relation to T cells (4%). Particulate or aggregated substances stimulate macrophages to release a mediator which subsequently acts on Fc receptor-negative T cells to produce IACF. Free-SH groups are important for the activity of the macrophage mediator. Mercaptoethanol and l-cysteine can also release IACF from T cells in the absence of macrophages. Protein synthesis is necessary for the production of this factor, the activity of which is abolished by trypsin digestion. It is postulated that the complexes of Ig and antigen formed under the influence of IACF represent a mechanism of presentation of antigen to T cells.     

INTRACELLULAR PHYTOCHROME DISTRIBUTION AS A FUNCTION OF ITS MOLECULAR FORM AND OF ITS DESTRUCTION

The immunocytochemically observed intracellular redistribution of phytochrome as a function of its molecular form is described by utilizing color photomicrography. The reversible change from a diffuse to a discretely localized distribution following photoconversion of the red-absorbing Pr form to the far-red-absorbing Pfr form observed with etiolated oat (Avena sativa L., cv. Garry) coleoptile parenchyma cells is not seen with etiolated wheat (Triticum sativum L., cv. unknown), barley (Hordeum vulgare L., cv. Harrison), or rye (Secale cereale L., cv. Balbo). Whether redistribution in these latter cases does not occur or is below the limit of detection is not known. Upon continuous actinic irradiation, phytochrome, which is discretely localized as Pfr, rapidly disappears by both immunocytochemical and spectral assay. However, after about 90 min irradiation, a new association of phytochrome with nuclei is evident which is more pronounced after 4 or 8 h of irradiation. With longer irradiation times there is a total loss of antigenically detectable phytochrome at the resolution employed in these experiments.     

delta5 desaturation of fatty acids in L-M cells

L-M cells grown in a lipid-free medium containing ¹⁴C-labeled 9,12-linoleic acid incorporated most of this acid into glycerolipids as linoleic acid. Only a small amount (3%) was elongated to eicosadienoic acid. No Δ6 desaturation occurred. When the cells were incubated with ¹⁴C-labeled 8, 11, 14-eicosatrienoic acid, 22% of the activity was found in 5,8,11,14-eicosatetraenoic acid. Treatment of the cells for 24 hr with N-isopropylethanolamine, a choline analog, depressed this desaturation reaction to about 60% of control values. The identity of the tetraene product was established by two different chromatographic analyses of the fatty acid methyl esters. Location of the double bond at position C-5 was determined by ozonolysis and subsequent reduction of the ozonides to aldesters followed by gas-liquid chromatography. These results prove that L-M cells have a Δ5 desaturase and an elongation enzyme converting 18:2 to 20:2, but lack a Δ6 desaturase.     

Purification and characterization of two isozymes of 3-phosphoglycerate kinase from the mouse.

Two isozymes of 3-phosphoglycerate kinase (ATP:3-phospho-D-glycerate 1-phosphotransferase, EC 2.7.2.3), designated PGK-A and PGK-B, were purified from separate extracts of muscle and testicular tissue of DBA/2J mice, respectively. A similar procedure was used to purify the corresponding isozymes from C57BL/6J mice in order to make inter-strain comparisons. The purification involved the use of affinity chromatography with an 8-(6-aminohexyl)amino-ATP-Sepharose column and DEAE-Sephadex chromatography. Lactate dehydrogenase isozyme LDH-X was also co-purified from extract of mouse testes by this two-step procedure. The same isozyme isolated from either mouse strain was found to be identical in physical and biochemical properties. Both isozymes are monomeric as determined by gel filtration chromatography and by sodium dodecyl sulfate polyacrylamide gel electrophoresis. Furthermore, the isozymes have similar molecular weights, of 47 000 +/- 2000 and exhibit similar Km values for both coenzymes and substrate, as well as temperature dependence of enzyme activity. However, it was observed that the B isozyme is more labile than the A isozyme by denaturation at high temperature, urea and acidic pH.     

Localization of 5S RNA and rRNA genes in the Norway rat

The genes coding for the two classes of ribosomal RNA molecules, 5S RNA and 18+28S RNA, have been localized in the Norway rat (Rattus norvegicus). The 18+28S RNA cistrons are found on three chromosomes, at secondary constrictions on the short arms of chromosomes 3 and 12 and at the telomere of the short arm of chromosome 11. These sites were confirmed using the silver staining technique for nucleolar organizer regions. Two sites were found for the 5S RNA genes; one is closely linked to the 18+28S gene site on chromosome 12. The second site is at or near the telomere of the long arm of chromosome 19.     

Transport of amino acids in intact 3T3 and SV3T3 cells. Binding activity for leucine in membrane preparations of ehrlich ascites tumor cells

Transport of amino acids into 3T3 and SV3T3 (SV40 virus-transformed 3T3) cells was measured on glass cover slips. The 3T3 and SV3T3 cells contain both A (alanine preferring) and L (leucine preferring) systems for neutral amino acid transport. Initial rates of uptake of amino acids are about twofold higher in SV3T3 than in 3T3 cells. Other parameters measured, however, do not indicate marked differences in the transport of amino acids by the two cell types. L-system amino acids, such as leucine, are subject to trans-stimulation in both cell lines, whereas A-system amino acids, such as alanine and glycine, are not. Leucine was transported to higher levels in confluent cells than in nonconfluent cells. Glycine, however, shows distinctly less transport activity as the cells become confluent. Ehrlich ascites cell plasma membranes were prepared and assayed for amino acid-binding activity. Leucine-binding activity was detected by equilibrium dialysis in Triton X-100-treated membrane preparations.     

Application of step-wise discriminant analysis and Bayesian classification procedure in determining prognosis of acute myocardial infarction.

A retrospective study was carried out to assess the feasibility of computer-assisted prognostication by discriminant analysis and the Bayesian classification procedure based on clinical information collected on patients with acute myocardial infarction. The overall accuracy was 94.2% in predicting hospital death but the prediction of late death after discharge was less accurate. It was found that not all of the 44 variables used for analysis were necessary to reach the same level of predictive accuracy--16 to 20 variables would result in almost the identical prediction. The Bayesian classification procedure was applied to estimate probabilities of individual patients belonging to the different prognostic categories.