Antigen-specific murine T-cell lymphomas. II. H-2 restriction of primary and secondary responses

A stable clone of C57BL/6 (H-2b) radiation leukemia virus transformed ovalbumin (OVA)-specific murine T-cell lymphoma cells was able to mediate carrier-specific helper activity. The ability of these lymphoma cells to express helper activity for both primary and secondary hapten-specific B-cell responses was analyzed in nonirradiated normal or hapten-primed recipients. The lymphoma cells augmented anti-hapten responses in a carrier-specific manner; no bystander effects were noted. Helper activity was primarily noted in the IgG responses. The genetic restrictions affecting the expression of lymphoma-mediated helper activity were also analyzed. The pattern of restriction indicated that genes in the H-2 complex controlled the expression of helper activity; disparities at the Igh complex failed to influence helper activity. The cellular site of the H-2 restriction was between the antigen-presenting cells and the T-cell lymphoma not between the T and B cells. Precise intra-H-2 mapping of the gene(s) which control expression of lymphoma-mediated helper activity was attempted. Although most of the data were consistent with localization of the gene(s) to the I-A region, anomolous responses were noted in one strain.     

Immortalization of antigen specific, helper T cell lines by transformation with the radiation leukemia virus (RadLV)

Antigen-specific immune T lymphocytes of male C57BL/6 mice were enriched in vitro on monolayers of antigen-pulsed syngeneic macrophages. The cells were treated in vitro with RadLV and inoculated intrathymically into irradiated female C56BL/6 animals. Thymomas arising in the inoculated recipients were characterized as donor- (male) type according to their karyotype. In vivo and in vitro cell lines were established from the primary lymphomas, two of which (designated ROT/6.1 and ROT/6.2) were capable of providing antigen- (carrier) specific help in normal or preimmunized mice. None of the lymphomas could induce antigen-specific DTH reaction. Five months after their establishment, ROT/6.2 alone retained its carrier specificity. ROT/6.2 consisted mainly of Lyt-1+ cells, whereas the ROT/6.1 population was more heterogeneous and contained Lyt-1+, Lyt-2+, and Lyt-3+ cells. The carrier specificity of the latter may have been lost due to selection against the specific helper cells during prolonged passages.     

Specific binding of T lymphocytes to macrophages. III. Spontaneous dissociation of T cells from antigen-pulsed macrophages.

Peritoneal exudate lymphocytes (PEL) from immune guinea pigs that adhere to antigen-pulsed macrophages (MO) were cultured for 1 week to yield a population enriched in antigen-specific (selected) T cells. These cells bind specifically within hours to fresh autologous antigen-pulsed MO. Thd dissociation of these selected PEL from antigen-pulsed MO was studied. No evidence was obtained that factors in the culture medium play a role in dissociation. Lymphocytes that have dissociated from antigen-pulsed MO are usually fully capable of rebinding to MO freshly pulsed with antigen, suggesting that there is no deficiency in the lymphocytes ability to bind. In contrast, readding antigen to cultures during incubation prevents the predicted dissociation. Moreover, repulsing MO cultured without selected PEL restores their capacity to bind fresh selected PEL. These findings indicate that decay of antigen associated with with MO is the major mechanism underlying the observed dissociation.     

Structure-activity relationships of heparin-mimicking compounds in induction of bFGF release from extracellular matrix and inhibition of smooth muscle cell proliferation and heparanase activity

A series of nine synthetic polyaromatic compounds were synthesized by polymerization of aromatic ring monomers with formaldehyde, which yield substantially ordered backbones with different functional anionic groups (hydroxyl and carboxyl) on the phenol ring. These compounds were tested for their heparin-mimicking activity: (1) inhibition of heparanase activity; (2) inhibition of SMC proliferation; and (3) release of bFGF from the ECM. We demonstrate that compounds that have two hydroxyl groups para and ortho to the carboxylic group and a carboxylic group at a distance of two carbons from the phenol ring inhibit heparanase activity and SMC proliferation, as well as induced an almost complete release of bFGF from ECM. Addition of a methyl group next to the carboxylic group led to a preferential inhibition of heparanase activity. Similar results were obtained with a compound that contains one hydroxyl group para to the carboxylic group and an ether group near the carboxylic group on the phenol ring. Preferential inhibition of SMC proliferation was best achieved when the position of the hydroxyl group is para and ortho to the carboxylic group and the carboxylic group is at a distance of one carbon from the phenol ring. On the other hand, for maximal release of bFGF from ECM, the position of the carboxylic group should be three carbons away from the phenol ring. These new heparin-mimicking compounds may have a potential use in inhibition of tumor metastasis, arteriosclerosis, and inflammation.     

Electrical Sizing of Particles in Suspensions: III. Rigid Spheroids and Red Blood Cells

The processes involved during the passage of a suspended particle through a small cylindrical orifice across which exists an electric field are investigated experimentally for an approximate prolate spheroid in the form of two tangent, rigid spheres (ragweed pollen particles) and for fresh, human red blood cells. Oscillograms of current pulses produced by both types of particles are presented and discussed in terms of particle shape and orientation and the effects of the hydrodynamic field. It is concluded that all the particles enter the orifice with their major axes aligned parallel to the orifice axis (electric field), but that during their passage some are rotated by the hydrodynamic field. Cells with their equatorial plane perpendicular to a radius of the orifice change their orientation with respect to the electric field as they are rotated, the others do not; only in the former case is there any deformation. It is shown that the bimodal or skewed size distributions can be explained on this basis, and that size (shape factor × volume) is actually a normally distributed variable (P > 95%). The average size of samples from 10 healthy adults was found to be 102.7 μ³ with a coefficient of variation of 1.8%. For a volume of 87 μ³, this corresponds to a shape factor of 1.18, an axial ratio (assuming a perfect oblate spheroid) of 0.26, and an equivalent major axis of 8.6 μ. The effect of high electric fields on red cell size distributions is mentioned.     

A synthetic heparin-mimicking polyanionic compound binds to the LDL receptor-related protein and inhibits vascular smooth muscle cell proliferation

A synthetic heparin-mimicking polyaromatic anionic compound RG-13577 (polymer of 4-hydroxyphenoxy acetic acid and formaldehyde ammonium salt, Mr approximately 5800) exhibits specific binding to vascular smooth muscle cells (SMCs) and inhibits their proliferative response to growth promoting factors. Receptor binding of (14)C-RG-13577 was efficiently competed by apolipoprotein E3 (apoE), lactoferrin, and the LRP (LDL receptor-related protein) receptor associated 39 kDa protein (RAP). Unlike cell surface binding of apoE, binding of RG-13577 to SMCs was not affected by heparin, heparan sulfate degrading enzymes, or low density lipoprotein (LDL). Moreover, wild-type and heparan sulfate-deficient Chinese hamster ovary (CHO) cells, as well as normal- and LDL receptor negative- human skin fibroblasts bind RG-13577, but not apoE, to a similar extent. On the other hand, homozygous mouse embryonic fibroblasts deficient in the LDL receptor-related protein (LRP) expressed a markedly reduced binding of RG-13577 as compared to normal mouse embryonic fibroblasts. These results indicate that RG-13577 and related compounds bind to the LRP receptor on the surface of vascular SMCs. Addition of lactoferrin to cultured SMCs protected the cells against the antiproliferative effect of compound RG-13577, suggesting that this inhibition is mediated by RG-13577 binding to LRP receptors on the SMC surface. Altogether, we have identified a series of synthetic polyaromatic anionic molecules that exhibit specific binding to LRP and thereby exert an antiproliferative effect on vascular SMCs. These compounds are applied to suppress SMC proliferation associated with restenosis and accelerated atherosclerosis.     

IL-6 increases primed cell expansion and survival

Cytochrome c-specific CD4 T cells from transgenic donors transferred to syngeneic B10.A mice expand more vigorously upon immunization if exogenous IL-6 is provided during the initial phase of immunization. The resultant increase in the frequency and number of Ag-specific cells is observed in the blood, lymph nodes, spleen, liver, and lung and persists for at least 3 mo. Treatment of immunized recipients with anti-IL-6 or use of IL-6 knockout recipients reduced the frequency of Ag-specific CD4 T cells during a comparable period, indicating that IL-6 is physiologically involved in the expansion of memory and/or effector cells and thus in the persistence of memory. IL-6 did not alter the duration of Ag-presenting activity. Both CFSE dilution studies and labeling with BrdU indicated that IL-6 does not effect proliferative rates of responding CD4 T cells. By contrast, annexin V staining was diminished in responding cells from the IL-6-treated animals, particularly among those cells that had undergone five or more divisions. These results indicate that IL-6 reduces the level of apoptosis among Ag-stimulated cells; thus, it plays a central role in determining numbers of memory and/or effector CD4 T cells in response to immunization over extended periods.     

Fractional modeling dynamics of HIV and CD4<Superscript>+</Superscript> T-cells during primary infection

In this paper, we introduce fractional-order into a model of HIV-1 infection of CD4⁺ T cells. We study the effect of the changing the average number of viral particles N with different sets of initial conditions on the dynamics of the presented model. Generalized Euler method (GEM) will be used to find a numerical solution of the HIV-1 infection fractional order model.