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Biomedical subjects

L Adorini

Publications and source records attributed to L Adorini.

At least 145 records · Page 8Linked to original sources
PubMed1988

Enhancement of suppressor T cell activity by injection of anti-IFN-gamma monoclonal antibody.

In the present study, the contribution of IFN-gamma to the generation of helper activity in mice was investigated by use of anti-mouse IFN-gamma rat mAB (AN 18.17.24). This mAb was alum precipitated and injected i.p. before or after carrier priming. Results show that spleen cell helper activity is markedly inhibited by anti-IFN-gamma mAb injection. This inhibition is time and dose dependent, and counteracted by IFN-gamma administration. Thus, the anti-IFN-gamma mAb appears to inhibit helper cell activity by neutralization of the IFN-gamma required for the antibody response. Moreover, AN 18.17.24 mAb injection results in increased activation of Lyt-2+ T cells which markedly suppress Th activity. These findings altogether indicate that besides the activation of macrophages and Th, IFN-gamma seems to exert a negative interference in suppressor T lymphocyte circuits and, as a consequence, to inhibit immunosuppression.

Animals↗
PubMed1988

Interaction of an immunodominant epitope with Ia molecules in T-cell activation.

The amino acid sequence corresponding to residues 107-116 of hen egg-white lysozyme (HEL) has been identified as containing an immunodominant T-cell epitope recognized in association with the I-Ed molecule. The immunodominance of this epitope in HEL-primed H-2d mice was demonstrated by analysis of the T-cell proliferative response induced by synthetic peptides covering almost the entire HEL sequence. All the T-cell hybridomas from H-2d mice analyzed recognize the HEL sequence 107-116 in association with the I-Ed molecule. Correlating with the restriction of T-cell recognition, HEL-(105-120)-peptide binds to I-Ed but not to I-Ad molecules. Conservative or semiconservative substitutions at positions 113 (Asn----Lys), 114 (Arg----His), or 115 (Cys----Ala) abrogate the ability of HEL-(105-120) to activate T cells. Substitutions at residues 113 and 115 affect T-cell recognition but not the binding to I-Ed molecules, whereas, as shown by binding data and competition experiments, an Arg----His substitution at position 114 profoundly impairs the capacity of the peptide to interact with I-Ed molecules. In agreement with these results, [Lys113]HEL-(105-120)-peptide but not [His114]HEL-(105-120)-peptide was found to be immunogenic in H-2d mice. Thus, a single semiconservative substitution drastically reduces binding capacity and abolishes immunogenicity, suggesting that a strict correlation exists between binding of a peptide to Ia molecules and its immunogenicity.

Amino Acid Sequence↗
PubMed1988

Lack of influence of cyclosporine on antigen presentation to lysozyme-specific T cell hybridomas.

Cyclosporine (CsA) was tested for its ability to inhibit antigen presentation by spleen cells or the B lymphoma line A20 to T cell hybridomas specific for hen egg-white lysozyme (HEL). Antigen-presenting cells (APC) were treated with CsA or nonimmunosuppressive derivatives thereof during or prior to encounter with antigen. The APC were then washed extensively and incubated in CsA-free medium for 6 hr before the T hybridoma cells were added. Under these conditions, CsA had no effect on antigen presentation up to the cytostatic regimen (1 microgram/ml). Omission of the 6-hr interval between CsA treatment of APC and the addition of T hybridoma resulted in inhibition of interleukin 2 production, although the CsA concentrations required were 10-75-fold higher than the ones inhibiting T cells directly (IC50: 100-150 ng/ml vs. 2-10 ng/ml). The responses to both HEL and a synthetic peptide of HEL sequence 105-120 were inhibited, indicating that the step influenced by the drug was not antigen-processing. The nonimmunosuppressive derivatives remained ineffective under these conditions. The results illustrate that the carryover of CsA from APC to T cells can mimic a drug effect on antigen presentation. Therefore, the demonstration of a CsA effect on antigen presentation can only be considered as conclusive when the readout of APC function is not a T cell response.

Animals↗
PubMed1987

Age restriction in antigen-specific immunosuppression.

Age-related alterations of antigen-specific T cell-mediated suppression have been examined in the 4-hydroxy-3-nitrophenyl acetyl (NP) system. Inducer suppressor T cells (Tsi) were activated in mice at the age of 3 mo (young) or 18 mo (old) by i.v. injection of NP-conjugated syngeneic spleen cells (SC). Spleen cells from the NP-SC-injected mice were subcultured in vitro with spleen cells from normal young or old mice to generate transducer suppressor T cells (Tst). Four days later subcultured cells were added to responder cell cultures 1 day before the PFC assays to trigger effector suppressor T cells (Tse). Responder cell cultures, containing NP-conjugated horse red blood cells (HRBC) and spleen cells from HRBC-primed young or old mice, were assayed on day 4 for anti-NP and anti-HRBC PFC. Suppression was found to be antigen specific and age restricted. NP-specific suppressor cells are easily induced in subculture if the Tsi and Tst cell populations are both derived from young or old mice. Conversely, if Tsi cells from young or old mice are subcultured with Tst cells from mice of a different age, suppression of the anti-NP PFC response is hardly observed. Age restriction was also found to operate in the interactions between subcultured and responder cell populations, indicating that age-matching is required for effective triggering of Tse cells by Tst cells. These results altogether suggest that aging may affect the recognition repertoire expressed in suppressor T cell subsets. Moreover, the finding that suppression is less efficient when exerted on responder spleen cells from old than from young mice provides an explanation for the increased frequency of autoimmune disorders in aging.

Aging↗
PubMed1987

T cell receptor-homologous mRNA from a suppressor T cell clone directs the synthesis of antigen-specific suppressive products.

The LH8-105 T cell clone, obtained by radiation leukemia virus-induced transformation of hen egg-white lysozyme (HEL)-specific mouse suppressor T lymphocytes, constitutively releases in the culture supernatant products able to induce specific suppression of the immune response to HEL. LH8-105 cells rearrange and express the genes encoding the alpha and beta chains of the antigen-specific T cell receptor and display membrane T cell receptor structures. LH8-105 mRNA specific for the alpha and beta chains of the T cell receptor were positively selected on filter-bound cDNA encoding constant regions of alpha and beta chains, eluted and translated into Xenopus laevis oocytes. Translation products were then tested in vivo for specific suppression of the anti-HEL antibody response. LH8-105 culture supernatant and translation products of LH8-105 poly(A)+ RNA or a mixture of alpha and beta chain-homologous mRNA induce HEL-specific suppression whereas translation products of LH8-105 mRNA eluted from irrelevant DNA or unrecombined mRNA eluted from alpha and beta cDNA do not exert suppressive activity. These results indicate that alpha and beta chain-homologous mRNA expressed in LH8-105 cells direct the synthesis of molecules able to induce HEL-specific suppression.

Animals↗
PubMed1987

Enhanced frequency of mitogen-responsive T cell precursors in old mice injected with thymosin alpha 1.

Injection of old mice with thymosin alpha 1, a synthetic peptide consisting of 28 amino acid residues and exhibiting thymic hormone-like activity, increases the splenic frequency of T cell precursors. Young (3-month-old) or old (19-20-month-old) mice received a single i.p. injection of thymosin alpha 1 or of an equimolar amount of the N14 (N-terminal amino acid residues 1-14) or C14 (C-terminal amino acid residues 15-28) synthetic fragment of the thymosin alpha 1 molecule and their spleen cells were assayed 3 days later under limiting dilution conditions to assess the frequency of mitogen-responsive and interleukin 2-producing T cells. Injection of thymosin alpha 1 or of its N14 fragment increases the frequency of responsive T lymphocytes in old, but not in young mice whereas injection of the C14 fragment has no demonstrable effect in either young or old mice. These data are consistent with our previous observation that the biological activity of thymosin alpha 1 is restricted to the N-terminal half of the molecule and suggest that this peptide amplifies the pool of mitogen-responsive and interleukin 2-producing T cells in immunodeficient old mice.

Aging↗
PubMed1986

A microcomputer program for probit analysis of interleukin-2 (IL-2) titration data.

IL-2 activity is commonly estimated in cell culture supernatants by an IL-2-dependent cell proliferation assay. This method is both reproducible and sensitive. However, it often appears from the literature that statistical analysis of the titration data either is disregarded or, when performed, is based on statistically incorrect assumptions. The proposed method is based on the principle of biological assay by parallel lines as applied to probit analysis of quantitative responses. The procedure has been embodied in a simple and interactive computer program which automatically estimates the IL-2 concentration in the biological sample, in terms of U/ml, and provides its standard error and confidence limits. This program is also suitable for quantitative determination of other biologically active substances that show a sigmoid dose/response relationship.

Animals↗
PubMed1986

Analysis of lysozyme-specific immune responses by synthetic peptides. I. Characterization of antibody and T cell-mediated responses to the N-terminal peptide of hen egg-white lysozyme.

The immunological reactivity against the N-terminal region of hen egg-white lysozyme (HEL) has been investigated by a synthetic peptide (PHEL) comprising residue 1-18 of HEL and by an analogue peptide (PREL) in which phenylalanine at position 3 is substituted by tyrosine. Both peptides are immunogenic in (C57BL/10 X DBA/2)F1 mice genetically responder to HEL. In C57BL/6 mice, genetically nonresponder to HEL, PREL induces anti-peptide antibodies that also bind to PHEL whereas PHEL is not immunogenic. Thus, a single amino acid substitution in a synthetic peptide converts a nonresponder mouse strain into a responder one. Anti-PHEL antibodies demonstrate a higher binding to HEL than anti-PREL antibodies, indicating that phenylalanine at position 3 is important for induction of anti-peptide antibodies able to recognize native HEL. At the T cell level the two peptides show very high bidirectional cross-reactivity between themselves and with HEL for interleukin 2 production, antigen-specific proliferation and delayed-type hypersensitivity response, whereas conservation of phenylalanine at position 3 is required for induction of suppressor cells cross-reactive with HEL. This indicates that the N-terminal region of HEL contains epitope(s) able to induce the same level of helper T cell activity as the native HEL molecule. However, helper T cells do not discriminate between PHEL and PREL whereas phenylalanine at position 3 is critical for HEL-specific suppressor T cell induction.

Animals↗
PubMed1986

Immunoregulation of lysozyme-specific suppression. III. Epitope-specific amplification of immunosuppression induced by monoclonal suppressor-T-cell products.

The hen egg-white lysozyme (HEL)-specific suppression induced by soluble molecules produced by a monoclonal T-cell lymphoma line (LH8-105) obtained from HEL-specific suppressor T lymphocytes has been examined. Injection of I-J+ molecules from LH8-105 cell culture supernatant (TsFa) in HEL-primed mice during the afferent phase of the response induced Lyt-2+ second order suppressor T (Ts) cells which, upon transfer into HEL-CFA-primed syngeneic recipients, inhibit the delayed-type hypersensitivity (DTH) response to HEL. Transfer of spleen cells from TsFa-injected mice primed with HEL or human lysozyme suppresses the DTH response to HEL in recipient mice whereas this response is not affected by cell transfer from ring-necked pheasant egg-white lysozyme (REL)-primed and TsFa-injected mice, indicating that induction of second order Ts by TsFa is specific for a lysozyme epitope including phenylalanine at position 3. Fine antigenic specificity of second order Ts-cell induction is confirmed by similar results obtained upon injection of TsFa in mice primed with HEL N-terminal synthetic peptide or with an analog in which, as in REL, phenylalanine has been substituted by tyrosine at position 3. The same fine antigenic specificity observed in the induction of second order Ts cells is also present in the expression of TsFe suppressive activity. The similar antigenic specificity of Tsa and Tse suggests that Tse cells could result from amplification of the Tsa cell population or these two cell subsets could reflect different maturation stages of the same cell type rather than distinct T-cell populations activated in cascade.

Animals↗
PubMed1986

A microcomputer program for hydrophilicity and amphipathicity analysis of protein antigens.

A simple and ready-to-use program to analyze hydrophilicity and amphipathicity characteristics of protein sequences is described. Three of the most commonly used hydrophilicity scales can be selected and the block length can also be varied. The program is written in BASIC language, can be run on any microcomputer and does not require computer experience. The program output is exemplified by a sample of lysozyme hydrophilicity and amphipathicity analysis.

Computers↗
PubMed1986

Reconstitution of T cell functions in aging mice by thymosin alpha 1.

Helper T cell activity of spleen cells from BDF1 mice is impaired by aging but is restored to a large extent by injection of thymosin alpha 1, a synthetic peptide consisting of 28 amino acid residues. Injection of an equimolar amount of the N14 (N-terminal half of thymosin alpha 1) synthetic fragment is at least as effective as the entire alpha 1 molecule in increasing helper T cell activity of spleen cells from old (6-18 months) mice but not from young (3 months) mice. Conversely, injection of the C14 (C-terminal half of thymosin alpha 1) synthetic fragment is devoid of any effect in both young and old mice. Since helper T cell activity of spleen cells from old mice is also increased by injection of interleukin-2, the observed enhancement of interleukin-2 production by mitogen-activated spleen cells from old mice upon injection of thymosin alpha 1 or the N14 fragment suggests that these peptides amplify helper T cell activity by increasing the cell precursor frequency of interleukin-2-producing T cells. This conclusion is further supported by the finding that injection of thymosin alpha 1, or its N14, but not C14, fragment enhances the expression of interleukin-2 receptors on mitogen-activated spleen cells from old but not from young mice.

Aging↗
PubMed1985

Immunosuppression by cell-free translation products from monoclonal antigen-specific suppressor T cell mRNA.

Polypeptides synthesized in a rabbit reticulocyte lysate system directed by mRNA from the T cell line LH8-105, obtained by radiation leukemia virus-induced transformation of hen egg-white lysozyme (HEL)-specific suppressor T lymphocytes, are able, when injected into mice, to specifically suppress the antibody response and delayed-type hypersensitivity to HEL. The suppressive activity exerted by in vitro translated proteins appears to be independent from post-translational modifications. These in vitro translated polypeptides display fine antigenic specificity in immunosuppression and bind to HEL but not to the closely related ring-necked pheasant egg-white lysozyme immunosorbents. Suppressive molecules obtained by cell-free translation of LH8-105 mRNA or by culture supernatant of LH8-105 cells display, by gel filtration, a similar molecular mass of about 82-90 kDa.

Animals↗
PubMed1985

Disulfide-linked surface molecules of monoclonal antigen-specific suppressor T cells: evidence for T cell receptor structures.

By two-dimensional polyacrylamide gel electrophoresis analysis under nonreducing/reducing conditions, five proteins with interchain disulfide bridges are revealed on the surface of the suppressor T cell lymphoma line LH8-105 obtained by radiation leukemia virus-induced transformation of hen egg-white lysozyme-specific suppressor T lymphocytes. Two disulfide-linked surface proteins expressed by LH8-105 cells have been positively identified by immunoprecipitation with specific antisera. The major labeled membrane protein of LH8-105 cells is the murine leukemia virus env glycoprotein gp70. The second disulfide-linked molecule identified on LH8-105 cells has a molecular mass of 84 kDa under nonreducing conditions and 42 kDa after reduction, and is immunoprecipitated by an antiserum which recognizes the T cell receptor for antigen. A disulfide-linked molecule of a similar molecular mass is also immunoprecipitated from surface-labeled LH8-105 cells by a rabbit antiserum directed against a synthetic peptide predicted from the nucleotide sequence of a cDNA clone encoding the beta chain constant region of a helper T cell hybridoma. Therefore, a dimeric structure comparable to the T cell receptor expressed by cytotoxic and helper T cells is present on the cell surface of these monoclonal antigen-specific suppressor T cells.

Animals↗
PubMed1985

Enhancement of helper and suppressor T cell activities by thymosin alpha 1 injection in old mice.

A single injection of immunodeficient old (15-24 month old) BDF1 mice with 1-100 micrograms of synthetic thymosin alpha 1 (T alpha 1), a 28-amino acid residue peptide, shortly before horse erythrocyte (HRBC)-priming, enhances the helper activity of their spleen cells. Helper activity of spleen cells from uninjected or T alpha 1-injected HRBC-primed old mice was titrated by adding graded numbers of these primed cells to cultures containing a constant number of normal spleen cells from unprimed young (3-month-old) mice and the conjugate 2,4,6-trinitrophenyl (TNP)-HRBC. As evaluated from the in vitro anti-TNP antibody response, spleen cells exhibit higher helper activity when derived from T alpha 1-injected, as compared to uninjected, HRBC-primed old mice. No effect is observed when T alpha 1 is injected in young mice before HRBC-priming. Old mice were also injected with the N14 (N-terminal amino acid residues 1-14) or the C14 (C-terminal amino acid residues 15-28) synthetic fragments of the T alpha 1 molecule. Injection of T alpha 1 or the N14 fragment restores the helper activity of old mice to levels comparable to that displayed by helper cells from young mice. Conversely, injection of the C14 fragment in old mice induces only a negligible increase in helper activity of their spleen cells. T cells from HRBC-primed mice were separated by nylon wool filtration into Th1 (non-adherent) and Th2 (adherent) T cells and these cell subpopulations were assayed for helper activity. Helper activity of Th1 and Th2 cells is found to be impaired by aging when they are tested either separately or upon their recombination.(ABSTRACT TRUNCATED AT 250 WORDS)

Aging↗
PubMed1985

Rearrangement and expression of the alpha- and beta-chain genes of the T-cell antigen receptor in functional murine suppressor T-cell clones.

Two different antigen-specific radiation leukemia virus (RadLV)-transformed suppressor T-cell clones, LH8.105 and LA41, exhibiting anti-lysozyme and anti-acetylcholine-receptor suppressor activity, respectively, have been examined for rearrangement and expression of genes encoding the alpha and beta chains of the T-cell receptor for antigen. LH8.105 cells express the T-cell-receptor polypeptides, as shown by specific immunoprecipitation. In both cell lines, potentially functional transcripts of alpha- and beta-chain genes are detected by RNA blot analysis. These suppressor T-cell clones exhibit alpha-chain gene rearrangements, deletion of both alleles of the constant-region (C) gene segment C beta 1, and rearrangement of the two alleles of C beta 2 when analyzed by Southern blot hybridization. Restriction analysis suggests that the DNA rearrangement is beyond the second joining-region (J) minigene of the J beta 2 cluster. These results establish that at least some mouse suppressor T-cell clones, like helper and cytotoxic T lymphocytes, rearrange and transcribe the genes coding for the alpha and beta chains of the antigen-specific T-cell receptor.

Animals↗
PubMed1985

Synthetic peptides in the analysis of the induction and regulation of delayed-type hypersensitivity to lysozyme.

T cells mediating hen egg lysozyme (HEL)-specific delayed hypersensitivity can be activated by synthetic peptides of the 1-18 amino acid residues of hen egg lysozyme. The N-terminal 1-18 peptides of hen egg (PHEL) and ring-necked pheasant lysozyme (PREL) are highly cross-reactive in the induction of T cells mediating delayed hypersensitivity. The N-terminal 1-18 peptides of hen egg and ring-necked pheasant lysozyme (PHEL and PREL) are not cross-reactive in the induction of suppressor T cells, demonstrating that phenylalanine at amino acid residue 3 is critical for the formation of an epitope recognized by T suppressor cells.

Amino Acid Sequence↗