Strain differences in the postnatal development of the mouse splenic lymphoid system.

Four strains of mice, CBA/J, BALB/c, C57BL/6J and B10.BR, were studied for cellular composition of the developing spleen in postnatal life. Considerable strain differences were found in the absolute numbers of splenic lymphoid cells at various ages, the frequencies of B and T cells, the L3T4/Lyt-2 ratio and the time of appearance of IgD and class II antigen on B cells.

Natural lymphocyte activation in postnatal development of germ-free and conventional mice.

The degree of activation of B and T cells in the developing spleen during postnatal life was studied in germ-free (GF) and specific-pathogen-free (SPF) BALB/c mice of the same breeding stock. We found that the progeny of GF mothers up to 3 weeks of age contain higher numbers of activated splenic cells than baby SPF mice, thus suggesting qualitative differences in maternally-derived antibodies. This "advantage" of GF mice is also indicated by an anticipated maturation of the splenic lymphoid compartment and is reflected in higher frequencies of B and T lymphocytes in adult life.

Membrane expression of IgG but not maturation to secretion requires DNA replication.

In this work we have addressed two questions. Does switch recombination occur before membrane expression or only concomitantly with induction to high rate synthesis and secretion of IgG? Does interleukin-4 induce switch to IgG1 or maturation of already switched cells? To answer these questions we used the DNA synthesis inhibitor hydroxyurea (HU) to analyze the requirements for DNA replication in the induction of membrane expression or high rate secretion of all IgG subclasses by cultures of mouse spleen cells stimulated by lipopolysaccharide (LPS) and supplemented with IL-4, the absolute numbers of cells bearing membrane bound IgG was always decreased by HU, indicating that immunoglobulin class switching requires DNA replication. IL-4 did not augment the numbers of cells expressing any IgG isotype.

Treatment of severe psoriasis with somatostatin: four years of experience.

Over a period of 4 years, 20 patients suffering from severe forms of psoriasis (erythrodermic, sub-erythrodermic, resistant generalized forms and/or forms associated with acute arthropathy) were treated with 96 h of continuous i.v. infusion of somatostatin (Stilamin, Serono) diluted in D5W at 250 micrograms/h.

On the reaction of molecular oxygen with thiyl radicals: a re-examination.

Absolute rate constants for the addition of oxygen to thiyl radicals, i.e. RS.

"In vivo" activated splenic T cells are refractory to interleukin 2 growth "in vitro".

Naturally activated T lymphocytes present in normal mouse spleen were studied for direct reactivity to interleukin 2 (IL 2) and for binding of anti-IL 2 receptor (IL 2R) antibodies or radiolabeled IL 2. The majority of large-sized splenic T lymphocytes are IL 2R-; thus, at the most one third of large L3T4+ T cells and of large Lyt-2+ T lymphocytes bind (weakly) anti-IL 2R antibodies; furthermore, most IL 2R+ cells in the normal spleen are actually Lyt2-, L3T4-. Total large splenic lymphocytes do not express more than an average of 150 high-affinity IL 2R/cell.

T-cell-dependent modulation of the polyclonal B-lymphocyte responses in normal spleen cell cultures stimulated by lipopolysaccharide.

The in vitro polyclonal B-cell proliferative and plaque-forming cell (PFC) responses to the T-independent (TI) mitogen lipopolysaccharide (LPS) are increased by the addition of normal syngeneic splenic T cells. Normal irradiated Lyt-2- T cells also alter the IgG subclass distribution from the typical predominance of IgG3 and IgG2b PFC to the appearance of IgG1, IgG2a and IgA PFC in T-cell-depleted spleen cell (SC) cultures. Furthermore, secondary LPS blast cultures yield increased PFC responses when co-cultured which syngeneic fresh normal T cells which, even in the absence of mitogen, induce PFC responses in such activated B cells.

Electron spin resonance and pulse radiolysis studies on the spin trapping of sulphur-centered radicals.

Thiyl radicals are shown to be readily trapped with the spin traps 5,5-dimethyl-1-pyrroline-N-oxide (DMPO) and 3,3,5,5-tetramethyl-1-pyrroline-N-oxide (TMPO) giving characteristic spin adducts with hyperfine coupling constants aN 1.52-1.58, aH 1.

Thiyl and phenoxyl free radicals and NADH. Direct observation of one-electron oxidation.

Absolute rate constants for the reaction of NADH with thiyl free radicals derived from various sulphur-containing compounds of biological significance were measured by using the technique of pulse radiolysis. These and related reactions with phenoxyl free radicals are believed to occur through one-electron-transfer processes. Further evidence comes from studies with deuterated NADH.

Thiyl free radicals and the oxidation of ferrocytochrome c. Direct observation of coupled hydrogen-atom- and electron-transfer reactions.

Absolute rate constants for the reaction of ferrocytochrome c with the thiyl radicals derived from cysteine, GSH, penicillamine and N-(2-mercaptopropionyl)glycine were measured by using the technique of pulse radiolysis. The reaction is believed to occur through a one-electron-transfer process, in agreement with the hypothesis that thiols may act as catalysts linking hydrogen-atom- and electron-transfer reactions.

Polyclonal lymphocyte responses to murine Trypanosoma cruzi infection. I. Quantitation of both T- and B-cell responses.

Lymphoid activity was studied in spleen and lymph node cells from Trypanosoma cruzi-infected mice. Blast transformation in each lymphocyte class was assessed by dual parameter analysis for size and surface markers by both FACS and conventional immunofluorescence, while proliferative activity was measured by tritiated thymidine uptake, autoradiography, and analysis of DNA content in single cells. Acute infection results in rapid blast transformation and proliferative activity of all three lymphocyte classes (Ig+, L3T4+, and Lyt 2+).

Autonomous activation of B and T cells in antigen-free mice.

The spleen of adult antigen-free mice contains a sizable proportion (5-15%) of activated cells in all lymphocyte sets, as marked by the membrane expression of immunoglobulins, L3T4 and Lyt-2 antigens. The frequency of activated cells is very high in early post-natal life, and reaches adult levels by 6 weeks of age when it is comparable to that observed in healthy unmanipulated mice raised in conventional conditions. The effector B cell compartment is quantitatively similar in antigen-free mice and specific pathogen-free mice, but the former is deficient in isotype diversification, since IgG- and IgA-secreting cells are drastically reduced.

Interferon-gamma, mitomycin C, and cycloheximide as regulatory agents of MHC class II-associated invariant chain expression.

The murine and human major histocompatibility complex class II-associated invariant chain genes are expressed in mature B cells and in antigen-presenting cells. Several pre-B cell lines and fibroblasts do not naturally contain invariant chain mRNA. Expression is inducible, however, by interferons and other agents interfering with proliferation.

Organic free radicals and proteins in biochemical injury: electron- or hydrogen-transfer reactions?

The reactions of organic free radicals, acting as either reductants or oxidants, have been studied by pulse radiolysis in neutral aqueous solution at room temperature. Manyhydroxyl-substituted aliphatic carbon-centred radicals and one-electron adducts have been shown to be good one-electron reductants, while several oxygen-, sulphur- and nitrogen- (but not carbon-) centred free radicals have been shown to be good one-electron oxidants. Several carbon-centred radicals can be reduced rapidly by hydrogen transfer, from undissociated thiol compounds which can thus act as catalysts facilitating the overall reduction of a carbon-centred radical by an electron-donating molecule.

Clinical pharmacology of platelet cyclooxygenase inhibition.

Nonsteroidal anti-inflammatory drugs and sulfinpyrazone compete dose-dependently with arachidonate for binding to platelet cyclooxygenase. Such a process closely follows systemic plasma drug concentrations and is reversible as a function of drug elimination. Peak inhibition and extent of its reversibility at 24 hr varies consistently with individual pharmacokinetic profile.

Natural effector T lymphocytes in normal mice.

The "natural" T-cell activity in normal unimmunized mice was studied. By double-parameter fluorescence-activated cell sorter analysis, it was found that 5-10% of all splenic Lyt-2+ and L3T4+ lymphocytes are large, of which more than half are in mitotic cycle. In contrast with small resting cells of the same phenotype, activated (large) T cells isolated from normal mice are functional effector cells: L3T4+ large cells induce normal B lymphocytes into proliferation and antibody secretion, while large Lyt-2+ cells efficiently suppress B-lymphocyte responses.

Absence of immunoglobulin heavy chain expression results in altered kappa/lambda light chain ratios.

A few hundred hybridoma cell lines derived from spleen cells of normal, nonimmunized, 6-day-old BALB/c or BALB.B10 mice were screened for H- and L-chain production. Roughly half of these hybridomas produced no Ig chain into the culture supernatants.

Oxygen-centred free radicals can efficiently degrade the polypeptide of proteoglycans in whole cartilage.

Bovine nasal cartilage slices, biosynthetically labelled in their proteoglycan with 35SO4, were used as substrate for the attack of free radicals generated on exposure to a Co60 source (which allows study of single radical species), and by chemical and enzymatic means. Systems generating hydroxyl (OH.) and superoxide (O2.

Effects of mu-specific antibodies on B cell growth and maturation.

Over a wide range of concentrations affinity-purified rabbit anti-mouse mu chain antibodies, or their F(ab')2 fragments, inhibit the appearance of immunoglobulin-secreting cells (plaque-forming cells; PFC) in lipopolysaccharide-stimulated murine spleen cell cultures without affecting proliferation. Both IgM and IgG PFC are inhibited although the number of blasts bearing surface IgG remains unaltered. The IgM and IgG PFC response could be reconstituted to normal levels in cell cultures suppressed by mu-specific antibodies by the addition of supernatants from in vitro propagated helper T cell clones, or from EL4 lymphoma cells induced with phorbol ester.

Interactions between immunoglobulins and I region antigens on the B lymphocyte membrane.

Modulation and co-modulation of membrane-bound IgM and IgD and of I region-encoded antigens (I-A and I-E) were studied by immunofluorescence with the use of monoclonal antibodies. The two sets of molecules displayed different ability to be redistributed, by divalent ligands: immunoglobulins could be easily redistributed, while this was not the case for I-A and I-E molecules. After short periods of activation, however, this property was found to change drastically: I-region antigens were redistributed into polar caps, and immunoglobulins were modulated without clear polar redistribution.

The membrane receptor complex regulating induction and clonal expansion of normal murine B lymphocytes.

Induction of resting B lymphocytes results from the interaction of competent ligands or helper cells with "triggering receptors." Subsequent clonal expansion and performance are thought to be regulated by the interaction of selective growth or maturation factors with specific receptors on induced B cells. A set of membrane molecules of B lymphocytes, including IgM, IgD, IA, IE, lipopolysaccharide receptors, receptors for Fc and C3b, and other non-immunoglobulin structures recognized by some antiidiotypic antibodies, display ligand-induced relationships.

Idiotypic determinants of natural IgM antibodies that resemble self Ia antigens.

A collection of immunoglobulin-secreting B-cell hybridomas was derived from normal neonatal BALB/c spleen and searched for reactivity against a panel of monoclonal anti-H-2 antibodies. We describe here one IgM antibody which was found to react with the monoclonal anti-Ia.7 antibody 14-4-4S.