thymosin and Uremia

Although influenza vaccination is widely recommended for immunosuppressed people, the same immune dysfunction that can increase the risk of contracting influenza might also compromise vaccine effectiveness, especially during pandemics. Clinical data have highlighted the role of adjuvants in improving vaccine efficacy. As uremic patients are especially vulnerable to infections, it is recommended that they should be vaccinated yearly against influenza. This paper presents the results of a pilot clinical trial, conducted in hemodialyzed patients with an adjuvated pandemic H1N1v influenza vaccine alone and combined with Thymosin-alpha 1.. Subjects were subdivided into 3 treatment groups receiving: the adjuvated pandemic influenza vaccine (Focetria) only (first treatment group), and the Vaccine+Thymosin alpha 1 (Zadaxin) at a dose of 3.2 and 6.4 mg (second and third treatment groups respectively). The immunoresponse was assessed on days 0, 21, 42, 84 and 168 after vaccine administration by means of Hemagglutination Inhibition (HI), Microneutralization (MN) and Single Radial Hemolysis (SRH) assays. The CHMP regards HI as the gold standard test to evaluate the immune response to influenza vaccines before influenza vaccines are licensed. The CHMP criteria are slightly different in adults (18-60-year-old subjects) and the elderly (>60 years old). Indeed, 40% of seroconversion, 70% of subjects seroprotected 21 days after vaccination, and a 2.5-fold increase in GMR (Geometric Mean Ratio) are required in adults, while in the elderly, the corresponding threshold values are: 30%, 60% and a 2-fold increase. All these criteria must be met for the licensing of a pandemic influenza vaccine. Safety evaluation was performed by means of Adverse Event (AE) recording, laboratory assays (hematology and chemistry), electrocardiogram, and assessment of vital signs.. Three populations were considered: Intention-To-Treat (ITT) (94 patients), Per Protocol (PP) (82 patients), and Safety population (99 patients). With regard to the Geometric Mean Titer (GMT) and the Geometric Mean Ratio (GMR) of HI on Day 21 in the ITT population, both "Vaccine+Thymosin alpha 1" groups presented better results than the "Vaccine only" group. A large proportion of ITT patients in the two Vaccine+Thymosin alpha 1 groups achieved seroconversion by Day 21. On Day 42, the decrease in the GMT of HI was greater in the Vaccine+Thymosin alpha 1 groups than in the vaccine only group. Similar results were obtained in the PP population. The CHMP criteria were fully met in the groups treated with Vaccine+Thymosin alpha 1. No AE was found to be related to Thymosin alpha 1 nor to the Focetria vaccine.. Although further studies in larger hemodialyzed populations are necessary, it can be concluded that Thymosin alpha 1 enhanced the immunogenicity of the pandemic influenza vaccine used. Moreover, it proved safe and well tolerated, and did not affect hematology or blood-chemistry values.

Topics: Adjuvants, Immunologic; Adolescent; Adult; Aged; Aged, 80 and over; Antibodies, Neutralizing; Antibodies, Viral; Female; Humans; Immunoassay; Influenza A Virus, H1N1 Subtype; Influenza Vaccines; Influenza, Human; Male; Middle Aged; Pilot Projects; Renal Dialysis; Thymalfasin; Thymosin; Uremia; Young Adult

Deacetyl-thymosin beta 12 was synthesized in a conventional manner by assembling six peptide fragments followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole (molar ratio, 1:1) in trifluoroacetic acid in the presence of m-cresol and dimethyl-selenium. Incubation of peripheral lymphocytes isolated from uremic patients with the synthetic deacetyl-thymosin beta 12 showed an enhancing effect on the reduced beta lymphocytes but had no restoring effect on the impaired blastogenic response of T lymphocytes.

Topics: Amino Acid Sequence; B-Lymphocytes; Chromatography, High Pressure Liquid; Humans; Lymphocyte Activation; Molecular Sequence Data; Sequence Homology, Amino Acid; T-Lymphocytes; Thymosin; Uremia

The complete murine prothymosin alpha molecule (110 residues) except for the N-terminal methionine deduced from the cloned cDNA has been synthesized by a solid-phase method. Peptide synthesis was performed manually by the stepwise solid-phase method using the base-labile Fmoc group for protecting the alpha-amino group. The peptide was assembled on a p-alkoxybenzyl alcohol resin. After the last coupling step, the Fmoc group was removed with 50% piperidine in DMF. The peptide resin was treated with thioanisole-o-cresol in TFA, and then purified by gel filtration, ion-exchange column chromatography and high-performance liquid chromatography. A 2.9-mg sample of a highly purified peptide was finally obtained. The overall yield of the synthesis was less than 1%, based on the amino acid content of the starting Fmoc-Asp (OtBu)-resin. The synthetic peptide was found to have a restoring activity on low-E-rosette-forming lymphocytes after incubation of peripheral blood from uremic patients with the synthetic peptide. This peptide exhibited far stronger restoring effect than that of our synthetic thymosin alpha 1.

Topics: Amino Acid Sequence; Animals; Chromatography, Gel; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Cloning, Molecular; DNA, Complementary; Humans; Mice; Molecular Sequence Data; Protein Precursors; Rosette Formation; T-Lymphocytes; Thymosin; Uremia

Two deacetyl-thymosin alpha 1 analogues containing Phe or Phe(4F) at position 21 were synthesized by the manual solid-phase method and their immunological effects on the low E-rosette-forming lymphocytes of uremic patients were studied. Fluorination of the p-position of Phe21 resulted in a marked restorative effect on the low E-rosette-forming lymphocytes of uremic patients compared with that of [Phe21]deacetyl-thymosin alpha 1. The synthetic [Phe21]deacetyl-thymosin alpha 1 was approximately equal in potency to our synthetic deacetyl-thymosin alpha 1 in uremic patients.

Topics: Amino Acid Sequence; Humans; In Vitro Techniques; Lymphocytes; Molecular Sequence Data; Rosette Formation; Structure-Activity Relationship; Thymalfasin; Thymosin; Uremia

Thymus lymphocyte subsets in uraemic rats were studied using monoclonal antibodies. Severe and moderate uraemia was induced in rats, and sham-operated and normal rats were used as the controls. As a result, the weight of the thymus decreased in uraemic rats. As for lymphocyte subsets, the frequency of W3/25+OX8+ decreased and those of W3/25-OX8-, W3/25+OX8- and W3/25- OX8+ relatively increased in uraemic rats. All these changes were more significant in severely uraemic than in moderately uraemic rats. When thymosin fraction 5 (TF5) was administered to severely uraemic rats, the weight of the thymus increased and the lymphocytes subsets normalized. These results suggest that uraemia may cause a maturational impairment of thymus lymphocytes by the depression of thymic hormone secretion.

Topics: Animals; Antibodies, Monoclonal; Antigens, Differentiation; Body Weight; Lymphocyte Subsets; Male; Organ Size; Rats; Rats, Inbred Strains; Thymosin; Thymus Gland; Uremia

A fragment corresponding to N-terminal octaeicosapeptide of rat parathymosin alpha was synthesized by assembling 5 peptide fragments, followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole (molar ratio 1:1) in trifluoroacetic acid in the presence of dimethylselenium. Incubation of impaired T-lymphocytes isolated from uremic patients with the synthetic parathymosin alpha fragment 1-28 showed no immunological restoring effect, but when it was administered together with thymosin alpha 1, it appeared to suppress the restoring effect of the thymosin alpha 1 on the impaired T-lymphocytes of uremic patients.

Topics: Amino Acid Sequence; Animals; Chromatography, High Pressure Liquid; Humans; Magnetic Resonance Spectroscopy; Molecular Sequence Data; Phytohemagglutinins; Protein Precursors; Rats; Sequence Homology, Nucleic Acid; Spectrometry, Mass, Fast Atom Bombardment; T-Lymphocytes; Thymalfasin; Thymosin; Uremia

Rat prothymosin alpha fragment 29-111, an 83-residue polypeptide corresponding to desthymosin alpha 1-prothymosin alpha, has been synthesized by a solid-phase method. Hydrogen fluoride was used to deprotect and cleave the peptide from the resin. The crude product was purified by gel-filtration, ion-exchange chromatography and high-performance liquid chromatography. A 3.2-mg sample of a ca. 96% pure peptide was finally obtained. The overall yield of the synthesis was less than 1%. An increase of E-rosette-forming lymphocytes was obtained after incubation of peripheral blood from uremic patients with the synthetic prothymosin alpha fragment 29-111. The restoring effect of the synthetic prothymosin alpha fragment 29-111 was greater than that of our synthetic thymosin alpha 1.

Topics: Amino Acid Sequence; Animals; Chromatography, High Pressure Liquid; Chromatography, Ion Exchange; Humans; Molecular Sequence Data; Peptide Fragments; Protein Precursors; Rats; Rosette Formation; Thymosin; Uremia

Cellular immunity of thymus lymphocytes in uraemic rats was studied. Severe and moderate uraemia was induced in rats, and sham-operated and normal rats were used as the controls. As a result, the response of thymus lymphocytes to concanavalin A (Con A) significantly decreased in severely uraemic rats, but did not change in moderately uraemic rats. However, when the thymus lymphocytes were pretreated with thymosin fraction 5, the response to Con A was ameliorated in severely uraemic rats. There was a significant correlation between the effect of thymosin fraction 5 on Con A response and Con A response of thymus lymphocytes. In addition, serum from severe uraemic rats suppressed the response of normal thymus lymphocytes to Con A. These results indicate that severe uraemia may cause an impairment in maturation of thymus lymphocytes, which can be improved by thymosin fraction 5 in vitro.

Topics: Animals; Cell Division; Cells, Cultured; Concanavalin A; Immune Tolerance; Immunity, Cellular; In Vitro Techniques; Male; Rats; Rats, Inbred F344; T-Lymphocytes; Thymosin; Thymus Gland; Uremia

An analog of thymosin beta Xen4 isolated from oocytes of Xenopus laevis, deacetyl-thymosin beta Xen4, was synthesized by assembling 6 peptide fragments, followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole (molar ratio, 1:1) in trifluoroacetic acid in the presence of dimethylselenium. Finally, the deprotected peptide was incubated with dithiothreitol to reduce sulfoxide on the methionine side chain. The synthetic tritetracontapeptide was found to have a restoring effect on the impaired blastogenic response of T-lymphocytes isolated from uremic patients.

Topics: Amino Acid Sequence; Humans; In Vitro Techniques; Lymphocyte Activation; Molecular Sequence Data; Phytohemagglutinins; Thymosin; Uremia

Topics: Adjuvants, Immunologic; Humans; Immunologic Deficiency Syndromes; Lymphocyte Activation; T-Lymphocytes; Thymosin; Uremia

The untetracontapeptide corresponding to the entire amino acid sequence of deacetylthymosin beta 11 was synthesized by assembling six peptide fragments via the azide followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole in trifluoroacetic acid in the presence of dimethylselenide. The synthetic peptide was tested for its effect on the impaired blastogenic response of phytohemagglutinin-stimulated T-lymphocytes of a uremic patient with common variable immunodeficiency. The synthetic peptide had some restoring activity on the impaired blastogenic response of T-lymphocytes in the one patient tested.

Topics: Amino Acid Sequence; Amino Acids; Fluorometry; Humans; Immunologic Deficiency Syndromes; Male; Molecular Sequence Data; T-Lymphocytes; Thymosin; Uremia

A tritetracontapeptide corresponding to the entire amino acid sequence of endo-Arg38a-deacetylthymosin beta 10 was synthesized by a conventional solution method. Seven peptide fragments were assembled, followed by deprotection with 1 M trifluoromethanesulfonic acid-thioanisole-Me2Se in trifluoroacetic acid. In preliminary experiments the synthetic tritetracontapeptide increased the entire peripheral T-cell population and a helper T-cell subset when incubated in vitro with blood which was obtained from a uremic patient with pneumonia, but a suppressor/cytotoxic T-cell subset was unaffected under these conditions. The synthetic endo-Arg38a-deacetylthymosin beta 10 was as active as synthetic deacetylthymosin beta 10 in this in vitro assay.

Topics: Amino Acid Sequence; Antibodies, Monoclonal; Chromatography, Thin Layer; Humans; Pneumonia; T-Lymphocytes; Thymosin; Uremia

Topics: Amino Acid Sequence; Humans; In Vitro Techniques; Lymphocyte Activation; Molecular Sequence Data; Peptide Fragments; Pneumonia; Spectrometry, Fluorescence; T-Lymphocytes; Thymosin; Uremia

Topics: Chemical Phenomena; Chemistry; Humans; In Vitro Techniques; T-Lymphocytes; Thymosin; Tuberculosis; Uremia

Changes in the circulating lymphocyte populations and thymus glands were studied in rats with experimental chronic renal insufficiency (CRI). Compared to normal or sham-operated animals, rats with CRI had significant reduction in the percentages of circulating T and B lymphocytes. CRI was also associated with marked thymic atrophy and reduction in the numbers of small cortical and medullary thymic lymphocytes. Quantitative microfluorometry revealed a significant reduction in the intrathymic concentration of a potent immunomodulator, thymosin alpha 1 in all uremic animals. There was a significant positive correlation between the percentages of circulating T lymphocytes and the intrathymic concentrations of thymosin alpha 1.

Topics: Animals; Atrophy; B-Lymphocytes; Fluorescent Antibody Technique; Fluorometry; Leukocyte Count; Male; Rats; Rats, Inbred Lew; T-Lymphocytes; Thymalfasin; Thymosin; Thymus Gland; Uremia

Topics: Humans; Lymphocyte Activation; T-Lymphocytes; Thymalfasin; Thymosin; Thymus Hormones; Uremia

Topics: Binding Sites, Antibody; Chronic Disease; Humans; Immune Adherence Reaction; Immunity, Cellular; T-Lymphocytes; Thymosin; Thymus Extracts; Uremia