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Biomedical subjects

L Forni

Publications and source records attributed to L Forni.

At least 37 records · Page 2Linked to original sources
PubMed1986

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+). At 2 weeks of infection most cells in these organs are enlarged and more than half are dividing. By 2 and 6 months after infection (chronic phase of resistant strains), large numbers of activated B lymphocytes and, to a lesser extent, of Lyt 2+ T cells are still detected. Similar results were obtained in C57BL/6 (resistant) and C3H/HeJ (susceptible) mouse strains. The implications of this massive polyclonal lymphocyte response to the parasite for the physiopathology of acute and chronic infection are discussed.

Animals↗
PubMed1985

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. No effector cell cytolytic activity could be detected among naturally activated T cells. The significance of these findings for the internal activity in the normal immune system is discussed.

Animals↗
PubMed1985

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. Inhibition of platelet cyclooxygenase activity by these agents is associated with variable effects on prostaglandin (PG) synthesis in the gastric mucosa and the kidney. Aspirin acetylates platelet cyclooxygenase and permanently inhibits thromboxane (TX) A2 production in a dose-dependent fashion when single doses of 0.1 to 2.0 mg/kg are given. Acetylation of the enzyme by low-dose aspirin is cumulative on repeated dosing. The fractional dose of aspirin necessary to achieve a given level of acetylation by virtue of cumulative effects approximately equals the fractional daily platelet turnover. Serum TXB2 measurements obtained during long-term dosing with 0.11, 0.22, and 0.44 mg/kg aspirin in four healthy subjects could be fitted by a theoretical model assuming identical acetylation of platelet (irreversible) and megakaryocyte (reversible) cyclooxygenase. For a given dose within this range, both the rate at which cumulative acetylation occurs and its maximal extent largely depend upon the rate of platelet turnover. Continuous administration of low-dose aspirin (20 to 40 mg/day) has no statistically significant effect on urinary excretion of either 6-keto-PGF1 alpha or 2,3-dinor-6-keto-PGF1 alpha, i.e., indexes of renal and extrarenal PGI2 biosynthesis in vivo. Whether a selective sparing of extraplatelet cyclooxygenase activity by low-dose aspirin will result in increased antithrombotic efficacy, fewer toxic reactions, or both remains to be established in prospective clinical trials.

Aspirin↗
PubMed1985

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. Of the Ig-secretors, 80% produced IgM molecules carrying either kappa or lambda chains in a ratio of 9:1. A surprisingly high frequency of clones (roughly 20%) were found to produce only light chains and in this group, the kappa/lambda ratio was altered to around 1. The indications from the primary screening were confirmed by isolating a large number of these hybrid cell lines and extensive testing of both culture supernatants and ascitic fluids in solid phase RIA and ELISA assays. In some cases, the absence or presence of heavy chain synthesis and the isotype of the light chain was also confirmed by biosynthetic labeling of the hybridoma clones, specific immuno-precipitation of cell lysates or supernatants, followed by analysis in SDS polyacrylamide gels. We conclude from these results that the absence of heavy chain expression is accompanied by frequencies of lambda chain expression, which are much higher than those observed in normal immunoglobulin producing cells, and equal to those of kappa chains. This conclusion can be interpreted as to suggest that the low frequency of lambda producing cells in mouse is in part determined by restrictions in the heavy-light chain interactions at the protein level.(ABSTRACT TRUNCATED AT 250 WORDS)

Alleles↗
PubMed1984

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. Cross-linking and removal of one of the molecules always resulted in the modulation of at least one other, often two membrane molecules. Concomitantly, an additional molecule was expressed in higher amount. The results are compatible with the existence of specific interactions at the level of the B cell membrane, involving the four molecules in the order I-A----IgM----I-E----IgD. Such interactions, probably reversible in the unperturbed state, are likely to change in affinity when one molecule interacts with a ligand. The possible relevance of membrane interactions between immunoglobulin and class II antigens in the regulation of the B cell is discussed.

Animals↗
PubMed1984

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. Interleukin 1-containing P388 supernatant, or recombinant DNA-derived murine interferon-gamma, did not reconstitute the PFC response in cell cultures suppressed by mu-specific antibodies, indicating that other factors are responsible for these effects. When spleen cell cultures, pre-activated with either lipopolysaccharide or monoclonal mouse mu-specific antibodies coupled to Sepharose, were exposed to EL4 supernatants in the presence of soluble mu-specific antibodies, maturation to secretion was inhibited while proliferation was not. The implications of these findings on assay systems for B cell growth and maturation factors are discussed.

Animals↗
PubMed1984

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. The characterization of this clone (BA.N 4:4.57) revealed its anti-trinitrophenyl specificity and demonstrated specific binding to five different monoclonal anti-Ia.7 antibodies but not to other anti-H-2 antibodies. The variable region specificity of these interactions was shown by the use of pepsin Fab fragments of the IgM antibody. Anti-Ia.7 antibodies were shown to specifically inhibit plaque formation by the hybridoma cells, and dinitrophenylglycine was shown to inhibit the reaction between the IgM antibody and anti-Ia.7 molecules. We interpret these results as to indicate that BA.N 4:4.57 expresses an idiotope or idiotopes which mimic Ia.7 determinants. This idiotypic family is naturally expressed in both newborn and adult BALB/c mice, as shown by the presence in normal serum of IgM molecules that specifically react with the F(ab')2 fragment of the 14-4-4S antibody. We speculate on the importance of idiotypic mimicry with major histocompatibility complex determinants, for both the selection of natural antibody repertoires and the evolution of antibody genes.

Animals↗
PubMed1983

A monoclonal antibody against a new differentiation antigen of thymocytes.

B14-2-14 is a monoclonal cytotoxic IgM antibody which reacts with thymocytes of all mouse strains tested. The fraction of positive cells (by visual immunofluorescence) varies between strains from about 25-45% in A.CA to 65-85% in C57BL/6, and high levels are dominant in F1 hybrids. In the periphery, the antigen is found on a few percent of lymph node and not on splenic T cells, and it is absent in nude mice. Among thymocytes, the distribution of the B14 determinant largely overlaps with that of the TL antigen and of molecules binding peanut agglutinin. The B14 antibody reacts only minimally with hydrocortisone-resistant thymus cells. Biochemical analysis shows that B14 antibody, anti-TL antibody and peanut agglutinin bind to separate molecules. The target of the B14 antibody may be either an immature, thymic form of Thy-1, or another molecule associated with it. Two polypeptides, of 40 and 35 kDa are precipitated by both B14 and anti-Thy-1 antibodies from biosynthetically labeled thymus cell lysates, and two others, of 27 and 17 kDa, from surface-iodinated thymus cell preparations. B14-2-14 offers an additional method for identification and selection of thymocytes at different stages of differentiation, and should also be useful for studies of the Thy-1 antigen.

Animals↗
PubMed1983

Studies on the differentiation of T lymphocytes in sheep. I. Recognition of a sheep T-lymphocyte differentiation antigen by a monoclonal antibody T-80.

The results presented in this paper demonstrate that a mouse IgM monoclonal antibody (T-80) recognizes an antigen on cells of the T-lymphocyte lineage of sheep. However, this antibody does not identify all T cells, as 10-20% of thymocytes and some peripheral-blood T cells are negative. T-80- thymocytes reside in the medulla. The majority of cortical thymocytes are T-80+ and classified as dull cells on the basis of antigen density per cell as measured by flow microfluorometry. In contrast, T-80+ cells in the periphery can be categorized into two populations, i.e., dull cells and bright cells. Suggestive evidence was obtained that bright T-80+ cells are fast recirculating T cells, whereas dull cells are sessile or less easily mobilizable T cells in the periphery. In foetal environment, over 90% of thymocytes and approximately 5% of spleen cells are T-80+ at 54 days of gestation (gestation period = 150 days), which may indicate that T-cell emigration from the thymus commences well before mid-gestation in sheep.

Animals↗
PubMed1982

The production of membrane or secretory forms immunoglobulins is regulated by C-gene-specific signals.

Selective patterns of antibody isotypes are produced in response to thymus-dependent and thymus-independent antigens and mitogens. Together with information on the organization of immunoglobulin CH genes in myelomas, various models on the control of C-gene expression and switch in normal B cells have been proposed but only secreted products or secretory cells have been considered. We report here a dissociation between expression of membrane-bound immunoglobulins and secretion of the same isotype, and in this case describe C-gene specific signals which regulate the production of membrane versus secretory forms of immunoglobulin. These results indicate that regulation of isotypic patterns operate at levels other than immunoglobulin gene structure, and suggest that the secretory phenotype alone is inadequate as the measure of C-gene expression.

Animals↗
PubMed1982

Antigen-independent, IgM-induced antibody responses: requirement for "recurrent" idiotypes.

Anti-idiotypic antibodies (a-Id) were produced in syngeneic mice against two monoclonal IgM antibodies of BALB/c origin, TNP. 11 and SP/603. In plaque inhibition tests, using IgM-secreting hybridoma cells and anti-idiotypic antibodies, these two IgM proteins, as well as the anti-TNP myeloma protein MOPC 460 (IgA) were found to carry non-cross-reactive idiotypes. Analysis of the anti-trinitrophenyl (TNP) plaque-forming cells (PFC) in BALB/c mice, either normal or immunized with TNP-horse red blood cells, revealed that in addition to the MOPC 460 Id, also the SP/603 Id is recurrent and expressed by a fraction of the anti-TNP antibody-secreting cells in all individuals tested. In contrast, the TNP. 11 Id could not be detected in any BALB/c mouse studied. TNP. 11 and SP/603 antibodies were then characterized by their ability to induce an antigen-independent anti-TNP response in normal BALB/c mice. While TNP. 11 was found to be inactive, the same titers of SP/603 IgM induced antigen-specific PFC all of which expressed the SP/603 Id, and increased titers of circulating IgM molecules carrying the same Id, suggesting that "recurrent" but not "nonrecurrent" Id are competent in this respect. A fraction of these SP/603-induced, SP/603-positive anti-TNP antibodies also carried MOPC 460 Id, suggesting expression on the same molecule of idiotypic determinants found on independent non-cross-reactive "recurrent" idiotypes.

Animals↗
PubMed1982

Intraclonal diversification in immunoglobulin isotype secretion: an analysis of switch probabilities.

The production of all immunoglobulin isotypes except IgD was studied in a large number of single lipopolysaccharide (LPS)-reactive B cell clones. The majority, but not all, of the IgM-producing clones were found to secrete one or more other isotypes. IgG3 and IgG2b were most frequently found while IgA secretion was extremely rare. Many clones produced all four IgG subclasses and the statistical analysis of the data indicates, with a high degree of significance, that single clonal precursors give rise to progenies producing multiple isotypes. By assuming that intraclonal diversification follows the C-gene order in chromosome 12, the absolute switch probabilities of normal, unprimed LPS-reactive B cells can be calculated. The multi-potentiality of C-gene expression was further analyzed at the single cell level: a sizeable fraction of all activated B cells express two different IgG isotypes in the membrane-bound form, indicating consecutive switch events. In contrast, the majority of IgE and IgA secreting cells appear to switch directly from IgM. These results might reflect the functional relevance of S-region homologies in the control of C-gene expression.

Animals↗
PubMed1982

Idiotypic characterization of antibody-induced antibody responses.

Anti-idiotypic antisera were produced in syngeneic (C57BL/6) mice against a monoclonal anti-Dextran B512 (Dex) antibody (38-13). In radioimmunoassays, anti-idiotypic antibodies were shown to react with the homologous idiotype, while failing to recognize another monoclonal anti-Dex antibody, independently derived from C57BL/6 mice (D-16). Plaque inhibition tests confirmed the specificity of the anti-idiotypic antibodies and revealed that the 38-13 idiotype is expressed by about half of all anti-Dex antibodies produced in C57BL/6, but not in CBA mice. Injection of normal (but not athymic) C57BL/6 mice with low doses of 38-13 monoclonal antibodies, contained culture supernatants or ascitic fluids, resulted in a 10-20 fold increase in the numbers of anti-Dex PFC detected in the spleen 5 days later, the majority of which carried the 38-13 idiotype.

Animals↗