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Treatment of combined immunodeficiency with thymic extract (Thymostimulin).

A 14-day-old Chinese male baby was admitted with extensive skin lesions. A wound culture grew Staphylococcus aureus, Acinetobacter anitratus, Enterobacter cloacae, and Candida albicans and a blood culture grew group A beta-Streptococcus hemolyticus. The patient's lymphocyte counts were low and his lymphocytes were unable to produce IgG and IgA in vitro. The immunoglobulin-bearing cell studies also failed to demonstrate IgG and IgA bearing cells. Active Tac+ T cells, total T cells, and T cell subsets were at very low levels. Lymphoproliferative response to mitogens was also poor. Migration inhibitory factor production to Candida antigen was also decreased. The initial lymph node biopsy demonstrated no follicular formation and extensive depletion of lymphocytes in both thymic-dependent and thymic-independent areas. After Thymostimulin (a specific bovine thymic extract, TP-1) treatment, the second lymph node biopsy demonstrated germinal centers containing IgA-bearing cells and IgM-bearing cells and, subsequently, cortical and medullary differentiation. Serum IgG, IgA, and IgM became detectable at low levels and IgG-, IgA-, and IgM-bearing lymphocytes appeared in the peripheral blood. This also correlated with in vitro immunoglobulin synthesis. Active Tac+ T cells, total T cells, T cell subsets and lymphoproliferative response to mitogens increased gradually after thymostimulin therapy. This investigation demonstrated the therapeutic effectiveness of Thymostimulin in combined immunodeficiency both histologically and immunologically and the successful reconstitution of B cell function that did not require continued therapy.

Biopsy↗

[In vitro reactivity of lymphocytes after the addition of immunostimulatory drugs].

The in vitro immunomodulating, effects of two thymic extracts--Thymex L and Thymomodulin--on lymphocytes of lung cancer patients, were studied. The number of total and active T cell and PHA-induced lymphoproliferative response were evaluated before and after the addition of two different concentrations (5 mg/ml and 0.5 mg/ml) of these agents. The in vitro incubation with Thymex L and Thymomodulin had no effect on the number of total T lymphocytes, but the active T cells and proliferative responses were significantly higher. This increases was observed in the majority of patients with diminished baseline values. These results indicated that several immunological parameters should be used in vitro testing of an immunomodulatory drug. They also indicated the need for in vitro testing selection of candidates for immunotherapy among immunodepressed cancer patients.

Adjuvants, Immunologic↗

In vitro effect of a bovine thymic extract (thymostimulin) on T-cell differentiation in cord blood lymphocytes.

Cord blood lymphocytes were isolated from 40 normal newborns. The initial 20 samples were used to determine the dose-response curve of bovine thymic extract (Thymostimulin) by the measurement of active T cells. Results showed that the active T cells increased significantly when the Thymostimulin concentration was increased to 50 ng/ml and 100 ng/ml. The remaining 20 samples were divided into three portions and preincubated either with 50 ng and 100 ng of Thymostimulin or without Thymostimulin. The total T cells, active T cells, B cells, T-cell subsets, and lymphoproliferative responses to phytohemagglutinin, concanavalin A, and pokeweed mitogen were determined. The results showed that the active T cells, total T cells, B cells, OKT4 cells, and OKT8 cells were significantly increased after Thymostimulin treatment. The lymphoproliferative responses were also significantly increased. These data strongly support our conclusion that Thymostimulin has a marked stimulating effect on human lymphocyte proliferation and differentiation.

Animals↗

Treatment of cell-mediated immunodeficiency with calf thymic hormone (T.P.I.).

Eight children, four with combined immunodeficiencies and four with isolated T cell defects were treated with TPI, a crude calf thymic extract. Assessment of response was made on clinical grounds and by monitoring of immune function (T and B cell numbers, immunoglobulin levels, PHA response and delayed cutaneous reaction to Candida antigen). One child with severe combined immunodeficiency showed marked clinical improvement together with reconstitution of T cell numbers and, surprisingly, B cell numbers and immunoglobulin production. Two further children, one with a form of combined immunodeficiency and the other with a T cell defect and extensive cutaneous herpes simplex infection, showed benefit from therapy though in the latter we could not demonstrate that this was important in clearing the virus infection. None of the other children showed any consistent improvement either clinically or on testing. We found no evidence of toxicity, allergy or other adverse effects of therapy.

Animals↗

Attempt to treat acute type B hepatitis with an orally administered thymic extract (thymomodulin): preliminary results.

A double-blind trial with thymomodulin (TM) was performed in a consecutive series of 50 inpatients affected with acute type B hepatitis. Twenty-six randomly selected patients received TM (Leucotrofina, Ellem), 1 ampoule b.i.d. per os for 30 days, and 24 patients received the same amount of placebo for the same period. TM-treated patients showed accelerated AST and ALT decrease and an earlier HBsAg clearance. However, only the difference in ALT decrease was statistically significant in comparison with the controls (p less than 0.02). Before the treatment was started, lymphocyte subsets, as determined by monoclonal antibodies, showed a different pattern in the two groups despite strict randomization. Nevertheless, by the end of the trial, mean T4+/T8+ ratios were increased in the treated group, but remained unchanged in the control group. The trends in the two groups were significantly different (p less than 0.005). Further information is expected from a long-term follow-up.

Acute Disease↗

T-dependent immunity in aged humans. II. Clinical and immunological evaluation after three months of administering a thymic extract.

We investigated the effect of Thymostimulin (TP-1) (a bovine thymic extract) on some clinical and immunological parameters in hospitalized aged humans. Forty aged patients were randomized into two groups, one received no therapy and the other was treated with TP-1 for 3 mth. The Ig and complement levels, sheep rosette-forming cells, absolute lymphocyte numbers and sedimentation rate were measured on days 0, 90 and 180. Moreover, the incidence of infections was determined. The baseline immunological data of the patients studied were within the normal range. These data remained unchanged after TP-1 treatment. In the TP-1 treated group we observed a significant reduction of infections (P=0.023) in comparison to the control group. A significant reduction of the sedimentation rate was also observed in the treated group.

Aged↗

Treatment with thymic extract TFX for chronic active hepatitis B.

Eighteen patients with biopsy-proven chronic active hepatitis B (CAHB) and a significantly lowered CD4+/CD8+ cell number ratio were treated with thymic factor X (TFX): group I (n = 12) - for 12 months, group II (n = 6) for 6 months. As early as 14 days after starting the treatment a lowering of CD8+ cell numbers with a rise in CD4+/CD8+ cell number ratio were found. These changes continued to progress during the next months and were accompanied by decreased in NK cell numbers with enhancement of NK cell activity. Normalization of the biochemical and immunological parameters occurred after 5-6 months of the treatment. In both groups of patients seroconversion in the HBe system was observed after 9-12 months of the treatment. After two years complete clinical remission persists with normal biochemical and immunological findings and without reversion in the HBe system. The results seem to indicate that TFX has an immunostimulatory action and exerts beneficial effects on the course of CAHB.

Adolescent↗

Enhancement of interleukin-2 and gamma-interferon production in vitro on cord blood lymphocytes and in vivo on primary cellular immunodeficiency patients with thymic extract (thymostimulin).

Cord blood mononuclear cells (MNC) were isolated from 20 normal full-term newborns. These MNC were preincubated with either 50, 100, or 200 micrograms/ml Thymostimulin or without Thymostimulin. The interleukin-2 (IL-2) and gamma-interferon (gamma-IFN) production, cytotoxicity, and lymphoproliferation and IL-2 receptor (Tac) expression were all significantly increased after Thymostimulin treatment. For evaluation of the in vivo effect, two combined-immunodeficiency patients defective on the thymic level, one with progressive BCG infection, and one with DiGeorge syndrome were used. Before Thymostimulin treatment, the patient's MNC did not produce sufficient amounts of IL-2 and gamma-IFN. The cytotoxicity and lymphoproliferation were also low. After Thymostimulin treatment, the IL-2 and gamma-IFN production, cytotoxicity, and lymphoproliferative response were enhanced. These results suggest that Thymostimulin may be beneficial in the clinical treatment of primary cellular immunodeficiency. The improved immune reactivity including cytotoxicity and enhanced IL-2 and gamma-IFN production in the Thymostimulin treatment also indicates that there may be a beneficial effect on the combination of chemotherapy and Thymostimulin.

Adjuvants, Immunologic↗

Purification and biochemical characterization of nuclear ribonucleoprotein antigen using purified antibody from serum of a patient with mixed connective tissue disease.

We have achieved a high degree of purification of nuclear ribonucleoprotein antigen from calf thymus nuclear extract through antibody affinity chromatography. Antibody to nuclear ribonucleoprotein was purified from the serum of a patient with mixed connective tissue disease and Sepharose 4B was covalently coupled with the purified human antibody. The sodium thiocyanate eluate from the affinity column contained active ribonucleoprotein antigen and the specific activity of the antigen in this eluate was 488 times higher than the original nuclear extract. The protein component of the eluate consisted of six polypeptides determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Two of them were shown to be antigenic by a hemagglutination inhibition test. The molecular weights of these two peptides were approximately 13,000 and those of the other four were 13,000, 13,000, 30,000, and 65,000. The ribonucleic acid component of the eluate was shown by urea-polyacrylamide gel electrophoresis to contain five polynucleotides. Two of the five, estimated to contain 40 and 60 nucleosides, had antigenic activity. The other three polynucleotides which had more than 77 nucleosides had no antigenic activity. No modified nucleosides were found in these ribonucleic acid molecules. Even in this highly purified ribonucleoprotein antigen, Sm antigen was detected by immunodiffusion. This evidence indicated that there are some molecular associations between ribonucleoprotein and Sm antigens as has previously been suggested.

Amino Acids↗

Thymostimulin therapy in melanoma patients: correlation of immunologic effects with clinical course.

Thirty-two nonmetastatic melanoma patients with low T-lymphocyte values were treated with a thymic extract, thymostimulin (TS) (8 patients), DTIC (8 patients), or surgery alone (16 patients). In the 8 patients receiving TS, active E-rosette (T-Ea) and total E-rosette (T-Et) counts rose to normal levels and there was a significant rise in IgM and IgD receptors. Six out of eight patients treated with TS showed no evidence of metastases after 34 months, while 7/8 patients on DTIC and 13/16 patients on surgery alone developed metastases. Twenty metastatic patients with low T-lymphocyte values received either DTIC plus TS or DTIC alone. Total lymphocyte, T-Ea, and T-Et counts did not increase in either group nor was there a significant difference between the group on DTIC plus TS and the group on DTIC alone. The survival rate of patients on DTIC plus TS did not differ significantly from that on DTIC alone.

Adult↗

In vivo effects of thymustimulin on hematopoiesis of mice treated with cyclophosphamide.

This study reports on the in vivo effects of thymustimulin (TST), a thymic extract, on the hematopoiesis of mice treated with cyclophosphamide (CTX). Peripheral blood counts and both bone marrow pluripotent (spleen colony-forming units, CFU-S) and committed (granulocyte-macrophage colony-forming units, CFU-GM; erythroid burst-forming units, BFU-E) hematopoietic progenitor cells were assayed by in vitro methods. Administration of CTX alone was associated with severe hemotoxicity, which was followed by a gradual recovery of hematopoiesis. Hematotoxic effect of CTX was less pronounced when TST was administered in association with CTX. All the studied parameters were higher in TST + CTX-treated animals than those in CTX-treated animals, especially after Day 5 from the beginning of treatment for blood leukocytes, bone marrow cell counts, and packed red cell volume, and at Day 10 or 15 for CFU-S, CFU-GM, and BFU-E. These findings suggest that, in this experimental model, the simultaneous administration of TST reduces the myelosuppressive activity of CTX in vivo.

Animals↗

Nonspecific immunomodulation influences the course and location of Cryptosporidium parvum infection in neonatal BALB/c mice.

The influence of nonspecific immunomodulation with Thymomodulin (a calf thymic extract with immunomodulatory activity) and hydrocortisone on the course and location of Cryptosporidium parvum infection in neonatal BALB/c mice (infected with 10(6) or 10(5) oocysts on day 7 of life) was studied using scanning electron microscopy of the inner surface of different parts of intestine. Daily peroral treatment of suckling mice with 20 mg/kg/day of Thymomodulin for 5 days before inoculation resulted in an earlier peak and earlier termination of cryptosporidial infection when compared with control infected mice. On the other hand, peroral administration of 25 mg/kg of hydrocortisone every second day led to the persistence of cryptosporidial infection in the ileum of immunosuppressed mice until the end of observation (day 15 post infection), whereas only transient infection was observed in the intestine of control infected mice. The location of infection was also altered in hydrocortisone--treated mice--the severe infection was observed in more proximal parts of the intestine (anterior and middle jejunum), whereas no cryptosporidia were found in these parts of the intestine in nontreated infected mice.

Adjuvants, Immunologic↗

In vivo hemopoietic activity of thymic extract 'Thymustimulin' in aged healthy humans.

The in vivo effects of the administration of Thymustimulin (TST), a bovine thymic extract, on the hemopoiesis of healthy elderly subjects were evaluated. Five healthy individuals over 70 years of age were treated with TST for one month. Before and at the end of TST treatment, peripheral blood cell counts were performed and the in vitro growth of peripheral blood erythroid (BFU-E) and granulocyte-macrophage (CFU-GM) progenitor cells was assessed, together with the determination of T-lymphocyte subpopulations. Our results indicate that in elderly subjects, TST significantly (p less than 0.05) enhances the number of BFU-E; furthermore, a slight increase in the hemoglobin levels was observed. BFU-E positively correlated with CD3+ T-lymphocytes (r = 0.84; p less than 0.02), which significantly (p less than 0.01) increased after TST administration. No differences were found after TST treatment in the other parameters considered. TST seems able to exert a stimulatory effect on the erythropoiesis of elderly subjects, probably by its effects on T-lymphocyte functions.

Aged↗

Effect of thymic extract 'thymostimulin' on the incidence of infections and myelotoxicity during adjuvant chemotherapy for breast cancer.

A clinical trial was made to evaluate the effectiveness of thymostimulin (TST), a bovine thymic extract, in preventing infections in breast cancer patients treated with polychemotherapy. Fifty-one women treated with adjuvant CMF were randomly divided into two groups: 25 patients received TST plus chemotherapy and 26 patients were subjected to chemotherapy only. Periodic clinical and laboratory checks were performed for at least three months to monitor the occurrence of infections and variations in immunological parameters. Infections (mostly cystitis, conjunctivitis and stomatomucositis) were observed in 37% of the TST treated patients and in 77% of the controls. (p less than 0.005). Serum immunoglobulin concentrations in the two groups were not significantly different, while quantitative analysis of peripheral T lymphocyte subsets showed a higher incidence of decreased T4/T8 ratio in the control group (p = 0.055). Finally, there was a significantly lower incidence of myelotoxicity in the TST treated patients (p less than 0.0005). In conclusion, thymostimulin seems effective in preventing some of the commonest side effects of cancer chemotherapy.

Adult↗

The purification, characterization and amino acid analysis of nuclear ribonuclear protein and Sm antigens reacting with human autoimmune sera.

'Monospecific' antibodies directed against nuclear ribonucleoprotein (nRNP) and Smith (Sm) antigens from the sera of patients with mixed connective tissue disease (MCTD) and systemic lupus erythematosus (SLE) were used to isolate and purify the respective antigens from calf thymus nuclear extracts. The antibodies were allowed to interact with the nuclear antigens and the resulting complexes were purified by Protein-A-Sepharose and poly-U-Sepharose chromatography. When the Protein-A-Sepharose antigen-antibody complex was sequentially eluted with 20% and 60% ethylene glycol, 2 fractions could be identified, one containing antigenically active Sm and the other antigenically active Sm and nRNP. The nRNP fraction contained 5 major proteins with mol. wts ranging from 38,000 to 150,000. The Sm antigen had a mol. wt of 68,000 and was noted to 'co-purify' with the nRNP antigen. Amino acid analysis of the purified proteins demonstrated a high content of glycine, serine and acidic amino acid residues similar to previously described core proteins of heterogeneous nuclear RNP (hnRNP), a precursor to mammalian messenger RNA. Analysis of the RNA moiety following nuclease phosphorylase reactions demonstrated the presence of a RNA polynucleotide with a 'capped' structure at the 5' terminus, a feature consistent with the concept that the nRNP antigen is part of the hnRNP complex.

Amino Acids↗

Oral thymic extract for chronic hepatitis C in patients previously treated with interferon. A randomized, double-blind, placebo-controlled trial.

BACKGROUND: Hepatitis C is an important cause of chronic liver disease. It is claimed that Complete Thymic Formula, an over-the-counter herbal dietary supplement, is beneficial for patients with hepatitis C. OBJECTIVE: To evaluate the efficacy and safety of Complete Thymic Formula. DESIGN: Randomized, double-blind, placebo-controlled trial. SETTING: Tertiary referral center. PATIENTS: 38 patients with hepatitis C who did not respond to or were intolerant of interferon therapy. INTERVENTION: Complete Thymic Formula for 3 to 6 months or placebo for 3 months. MEASUREMENTS: Serial measurements of hepatitis C virus (HCV) RNA titers. RESULTS: No differences were noted at 3 months between the placebo group (n = 13) and the treatment group (n = 19) in mean HCV RNA titers (4.06 +/- 1.52 x 10(6) copies/mL compared with 3.48 +/- 1.92 x 10(6) copies/mL; P > 0.2). The 19 patients who completed 6 months of treatment with Complete Thymic Formula remained positive for HCV, and their mean HCV RNA titers were similar at 6 months and at baseline (2.78 +/- 1.96 x 10(6) copies/mL compared with 3.12 +/- 1.94 x 10(6) copies/mL; P > 0.2). CONCLUSIONS: Complete Thymic Formula did not benefit patients who had previously received interferon therapy. Patients should be advised about use of this over-the-counter compound.

Adult↗

In vivo effects of thymic hormones (Imunotim) on immunologic parameters of patients with secondary immunodeficiency diseases.

In previous papers we revealed the immuno-restorative capacity of thymic hormones both in vitro and in vivo. This paper is concerned with the investigation in a phase I trial of the effects of a thymic extract (Imunotim) prepared in the Cantacuzino Institute, Bucharest upon patients with immunodeficiencies manifest as recurrent or persistent infections resistant to classical treatments. Twenty five testings were performed on 23 patients with secondary immunodepressions: carcinomas with immunodepressions following the acute phase of the disease or cytoreductive treatments (10 cases), allergic and autoimmune conditions (3 cases) and frequent, recurrent infections with prolonged evolution. In 73.9% of cases (17/23) after administration of Imunotim per os the immunologic parameters showed a tendency to return to normal values. In patients with low pre-therapy values the following were noticed: a remarkable increase of the number of lymphocytes from 1317.2 +/- 506.40 to 1961.1 +/- 899.11 (p < 0.01), of PMN from 1401.7 +/- 444.21 to 2651.0 +/- 755.31 (p < 0.01), of the absolute number of B lymphocytes from 192.0 +/- 79.00 to 444.8 +/- 299.75 (p < 0.05); the rise in the rate and absolute number of total T lymphocytes (628.2 +/- 192.81, 1041.0 +/- 441.84, respectively; p < 0.05), T helper (from 29.84 +/- 12.75% to 44.14 +/- 18.76%, respectively from 421.6 +/- 127.29 to 1058.7 +/- 411.05; p < 0.01 respectively p < 0.02) and the rise of the absolute number of T suppressor/cytotoxic lymphocytes from 92.8 +/- 97.11 to 368.3 +/- 368.53 (p < 0.05). In some patients with lymphocytosis and adenopathies the absolute lymphocyte number and the maturation of the lymphocyte populations and subpopulations in blood showed a decline. Individual variations could be noticed even within one and the some primary affection.

Adjuvants, Immunologic↗