Colostrinin: a proline-rich polypeptide (PRP) complex isolated from ovine colostrum for treatment of Alzheimer's disease. A double-blind, placebo-controlled study.

A proline-rich polypeptide (PRP) complex, subsequently called Colostrinin, was isolated from ovine colostrum. The complex showed immunomodulatory properties in mice, rats, and chickens, inducing maturation and differentiation of thymocytes. It was recently found that Colostrinin is a cytokine-like factor that acts as an inducer of interferon gamma (IFN-gamma) and other cytokines in human peripheral blood and cord blood leukocyte cultures and has psycho-immuno-enhancing activity in volunteers. These observations prompted us to study the effect of Colostrinin on patients with Alzheimer's disease (AD). Forty six AD patients were divided into 3 groups and randomly assigned to receive orally either Colostrinin (100 microg per tablet, every second day), commercially available bioorganic selenium (100 microg selenium per tablet, every second day) or placebo tablets. One cycle of the treatment lasted 3 weeks and was separated from the next cycle by a 2 week hiatus. Each patient received 10 cycles of treatment during the year of the clinical trial. Outcomes were assessed by psychiatrists blinded to the treatment assignment. Eight of the 15 AD patients treated with Colostrinin improved and in the 7 others the disease had stabilized. In contrast, none of the 31 patients from the selenium or placebo groups with similar mild or moderate AD improved. The administration of selenium promoted stabilization in 13 of the 15 patients, whereas in the placebo group only 8 of the 16 patients were stabilized at the 12 month trials end-evaluation. Colostrinin was found to be a remarkably safe drug. Mild and transient effects were anxiety, stimulation, insomnia, and tiredness. The results obtained showed that oral administration of Colostrinin improves the outcome of AD patients with mild to moderate dementia. The results are very encouraging and deserve further research.

Antifungal activity of 2-(4-chlorophenyl)-1,2-benzisoselenazol-3(2H)-one, the analog of Ebselen.

The antimicrobial activity of 8 selenoorganic compounds (3-benzisoselenazoles, 1-benzisoselenazolone oxide and 4-disaryl diselenides) was investigated. It was found that selenoorganic compounds from benzisoselenazolone group suppress growth of some fungi and bacteria. The growth of Saccharomyces cerevisiae sigma 127-8b and Candida albicans 258 strains was strongly inhibited by the 2-(4-chlorophenyl)-1,2-benzisoselenazol-3(2H)-one. Also Ebselen and 2-acetyl-1,2-benzisoselenazol-3(2H)-one caused strong inhibition of Saccharomyces cerevisiae sigma 127-8b growth but a lower effect was observed in assays with Candida sp. strains. Benzisoselenazolones were also found to have antibacterial activities. They significantly reduced the growth of Gram-negative Escherichia coli K-12 ROW and Gram-positive Staphylococcus aureus 209P (Oxford) bacteria strains.

Development and disappearance of tolerance to induction of interferon and tumor necrosis factor response in athletes treated with natural immunostimulant.

The effect of nonspecific immunostimulation was examined in 15 basketball players subjected to extensive physical effort. The Tolpa* Torf Preparation (TTP*), a natural immunostimulating drug, was applied orally, one 5 mg tablet daily, in two 21-day cycles, separated by 2-week hiatus. Blood samples were collected 4 times, after each of two TTP* cycles and after the first and second hiatus. Whole blood assay was used to determine the spontaneous and induced production of interferon (IFN) and tumor necrosis factor (TNF). The levels of the cytokines were measured by microbioassays. TTP* stimulated synthesis of IFN and TNF in the whole blood cultures. However, after the oral administraton of TTP* for 3 weeks the leukocytes of the athletes developed hyporeactivity to IFN induction by TTP* and to a lesser extent to another "superinducer"--a mixture of phytohemagglutinin and bacterial lipopolysaccharide. The hyporeactivity state disappeared spontaneously within 2 weeks. In contrast, the tolerance to TNF induction did not develop during the TTP* administration. The increase of immunoglobulins, mainly of IgM and IgG classes and an acute phase protein--alpha1-antitrypsin, was observed at the late phase of the treatment. We suggest that the cytokine levels may be early markers for immunoprophylaxis. Furthermore, high production of IFN and TNF may be associated with extensive physical effort.

[Selenium in health and disease]. / Selen w zdrowiu i chorobie.

Selenium deficiency occurs in many regions of the world including Poland. It leads to human body dysfunctions and diseases among which Keshan disease is the most known. Biologically active selenium is usually incorporated into proteins as selenocysteine, which is the most useful in nutritional supplementation. Selenium addition is important since many viruses are more pathogenic when deficiency of this element occurs.

Tumor-associated antigens are cytokine inducers and hyporeactivity factors to the immune system.

We investigated possible mechanisms leading to the inhibition of the immune system in people with chronic disorders. Tumor cell produce protein released into the circulation, such as tumor associated antigens, may play an important role in processes preceding paralysis of the immune system. To test this hypothesis the following tumor associated antigens were used: AFP, OFP, CA-125, CA-50 and CA-19-9. Their role was assessed by modulating cytokine production in cord blood lymphocytes and peripheral white blood cells obtained from grown population of patients treated with colostrinin, an cytokine inducer. PHA, LPS and colostrinin were used as positive control in those essays. Each antigen tested individually induced IFN, TNF alpha and IL-6 in dose dependent fashion. None of the tested cytokines were spontaneously released by the cells. Data generated from these experiments indicated that tumor associated antigens are inducing type 1 cytokines in similar fashion as LPS or colostrinin. However, lymphocytes taken from patients undergoing therapy with colostrinin revealed progressive loss capability to produce type 1 cytokines as they did in case of colostrinin. The loss of the capability to respond to antigen may represent phenomenon leading to immune tolerance.

Modulation of cytokine production by a selenoorganic compound (AE-22) in hyperreactive or hyporeactive bronchoalveolar leukocytes of asthmatics or lung cancer patients.

We have found that many synthetic selenoorganic compounds, including ebselen, have immunotropic activity. These studies were designed to assess the effect of the analog of ebselen bis[2-pyridyl (2-carbamoyl) phenyl]diselenide (AE-22) on human leukocytes that may express various activation states. The cells were obtained from bronchoalveolar lavage (BAL) cells of patients with various inflammatory lung diseases. The AE-22-treated BAL cells from patients with bronchial asthma (n = 6) and with small cell lung cancer (SCLC) (n = 6) were compared with these in the peripheral blood leukocytes (PBL) from the same donors. The control group comprised 5 patients who underwent diagnostic examination and were free of any cancer or concomitant diseases. Secretion of TNF-alpha, IL-6, and IFN-gamma was considered as a marker of BAL or PBL cell activation. Different response of the cells and various effects of AE-22 were observed in relation to the origin and functional state of leukocytes. It was established that AE-22 can induce TNF-alpha, IL-6, and IFN-gamma in a dose-dependent manner in BAL cells and PBL isolated from healthy individuals. However, BAL cells were found to be less reactive than PBL as cytokine producers. In contrast, AE-22 had no effect on BAL cells obtained from patients with lung cancer, which were found to be hyporeactive to phytohemagglutinin and bacterial lipopolysaccharide and did not produce TNF-alpha, IL-6, or IFN spontaneously. The spontaneous release of cytokines by BAL cells from bronchial asthma patients, but not by PBL from the same individuals, was significantly (p < 0.01) higher than that from the cultures of healthy control subjects. The high secretion of cytokines by the locally activated BAL cells was significantly (p < 0.01) reduced after administration of AE-22. The results suggest that AE-22 has immunomodulatory activity. AE-22 can downregulate the hyporeactive BAL cells from asthmatics, but it appears to be inactive in BAL cells of cancer patients who can tolerate the cytokine inducers.

Coincidence between spontaneous release of interferon and tumor necrosis factor by colostral leukocytes and the production of a colostrinine by human mammary gland after normal delivery.

We have tentatively identified colostrinines as novel cytokines produced by the mammary gland after delivery and detectable in colostrum. The primary colostrinine, the proline-rich polypeptide, was isolated from ovine colostrum in 1974. It is generally understood that the various factors present in colostrum play a pivotal role in transmitting of passive or active immunity from mother to child. We have found previously that both ovine and human colostrinines are inducers of interferon (IFN) gamma and other cytokines. In this paper, we reported that the leukocytes isolated from human colostrum donated by healthy mothers at 1-9 days after delivery, produced IFNs and tumor necrosis factors (TFNs) spontaneously. The release of IFNs and TNFs coincided with production of a colostrinine that has been isolated from the human colostrum samples and partially characterized. Our results suggest that the maximum production of colostrinine occurs 3 days after delivery. The tolerance (hyporeactivity) of the colostral leukocytes to IFN inducers and the modulation of the TNF response may be the late effects of the colostrinine release.

Classification of cytokines according to the receptor code.

A simple classification of cytokines is presented. It is based on voluminous information available from published papers of many outstanding researchers of the cytokines and their receptors and/or summarized by experts in this field. The cytokines are divided into 7 families according to their receptor code. Such arrangement of apparently different cytokines may explain the biological significance of pleiotropy and redundancy. Furthermore, it may stimulate the development of the comprehensive classification of cytokines together with classical hormones, neuromediators, neuromodulators and related substances that participate in complex biological communication system.

Seleno-organic compounds as immunostimulants: an approach to the structure-activity relationship.

Our studies on the seleno-organic compounds were focused at their activities as the modest cytokine inducers in human peripheral blood leukocyte cultures. Our bioassays used in the screening methods were based on the quantitative determinations of mainly two types of cytokines: interferons (IFNs) and tumor necrosis factors (TNFs). More recently we have found that several of the compounds have direct immunotropic actions in vitro and in vivo, in mice and in chickens. The paper summarizes the data related to the cytokine-inducing activity of 65 seleno-organic compounds divided into 4 groups according to their chemical structures. The reference compound was ebselen, the well known experimental drug with various biological activities. Approximately 50% of the compounds were found to be active in our bioassays. The selected compounds induced also IL-6 and GM-CSF. Their activities were clearly correlated with defined chemical structures as well as with the presence of selenium. We suggest that some of the selected by us compounds, other than ebselen, are interesting as immunostimulants and potential antiviral agents and cytokine inducers active in humans.

[Successful therapy of the psychiatric patient with catatonic paranoidal syndrome and monitoring of induced cytokine production]. / Skuteczne leczenie chorej z zespolem katatoniczno-paranoidalnym i monitorowaniem wytwarzania cytokin.

The paper reports the case of a 42-year-old woman with a diagnosed catatonic-paranoidal syndrome and a noted dysfunction as regards lymphocyte subpopulation and the production of some cytokins: Tumor Necrosis Factor (TNF) and interferons (IFN). As the first phase of the treatment had proved unsuccessful and there appeared symptoms hinting at the possibility of a subclinical inflammatory process in the brain, parallel to psychiatric therapy, treatment was instituted in the system of successive three-week cycles with Tolpa's Peat Extract (PTT), Selenium and Zinc on a natural yeast basis (Selenium and Zinc preparations) as well as with encortone. After a month of treatment a complete clinical remission was achieved as well as normalization as regards all the tested immunological markers.

Colostrinine: a proline-rich polypeptide from ovine colostrum is a modest cytokine inducer in human leukocytes.

A proline-rich polypeptide (PRP), now named colostrinine, molecular weight 18,000, was isolated from ovine colostrum and characterized by Janusz, Lisowski et al. The nonapeptide (NP) which is an active fragment of PRP was obtained by chemical synthesis. In mice, PRP has many regulatory effects on the humoral and cellular immune response. The present paper describes PRP as a cytokine inducer. PRP at concentration of 1-100 micrograms/ml was found to induce production of interferon (IFN) and tumor necrosis factor (TNF) in human peripheral blood leukocytes and in whole blood cultures. The effects were dose related. The identified till now cytokines induced by PRP were IFN-gamma and TNF-alpha but many other cytokines may be stimulated also. NP was considerably less active as the cytokine inducer than the natural PRP. Two volunteers given orally once daily for two to three weeks 100 or 200 micrograms PRP in tablets were found to develop the tolerance of IFN induction and had the modified TNF response. Furthermore, the PRP-treated volunteers showed signs of psycho-stimulation. Taken together our observations suggest that ovine PRP is active in humans and may have therapeutic value as an immunostimulant and/or neurotropic cytokine.

Effect of benzisoselenazolones and organic diselenides on graft versus host reaction and immunoglobulins levels in chickens.

Two week old chickens were treated once daily for 5 days with AE8--1-pyridyl-1,2-benzisoselenazol-3(2H)-one, AE22--bis-2-(N-phenyle-carboxamido) 1 pyridyl diselenide and AE31--bis(phenylo-diselenide with R3 = CONHC18H37). Their whole blood alone or blood mixed with thymus cells were used to generate graft versus host (GvH) reaction in 15 day old chicken embryos. The treatment of the chickens with the compounds stimulated the GvH reaction modifying activity of the donor cells as measured by increase of the spleen weight of the recipient chicken embryos. On the other hand, treatment with these compounds inhibited the IgG or IgM production in chickens immunized with human albumin.

Immunotropic activities of benzisoselenazolones and organic diselenides in mice.

We have investigated the immunotropic effects of 23 seleno-organic compounds (8 benzisoselenazolones, 3 benzisoselenazolone oxides and 12 organic diselenides). All of the compounds increased the rosette formation of sheep red blood cells (SRBC) with spleen cells obtained from thymectomized C53BL/6 mice and incubated in vitro in the presence of imuran. Furthermore, 16 of the compounds were also assayed in vitro in the hydrocortisone test performed with C57BL/6 mouse thymocytes. It was found that all of them significantly protected the cells against hydrocortisone induced cytotoxicity. Also in the Jerne's assay, performed in 129Ao/Boy mice pretreated in vivo with 3 selected compounds 5 days before immunization with SRBC, the stimulation of plaque forming cells (PFC) was observed. Only one compound (AE22, an analog of piroxicam) was found to be inhibitory in this assay. In contrast, in the graft versus host (GvH) assay performed in hybrid mice the donor lymphoid cells obtained from C57BL/6 mice pretreated with 9 selected seleno-organic compounds, suppressed the GvH reaction in the recipient hybrid mice. Thus, in all of the immunotropic assays except the GvH reaction in adult mice, the seleno-organic compounds were found to have immunostimulating activities.

Interferon responses in schizophrenia and major depressive disorders.

The spontaneous and induced interferon (IFN) production in whole blood cultures was examined in 45 psychiatric inpatients and in 65 normal controls. Among inpatients there were 32 who were chronic schizophrenics (14 women, 18 men) and 13 who were severely depressed (11 women, 2 men). The analysis of the pooled results of assays in the heterogeneous population showed that leukocytes of the psychiatric patients produced significantly lower levels of IFN after stimulation with virus (NDV), lipopolysaccharide (LPS), and IFN spontaneously released without the inducers that control cells. In contrast, there was no difference between the psychiatric patients and controls in IFN response to phytohemagglutinin and phorbol myristate acetate (PHA + PMA). The results apparently confirmed observations made by Moises et al (1985) and Katila et al (1989). We have also tested our hypothesis that the statistics may mask the individual pattern of IFN response related to the specific psychiatric diagnosis, however. In fact, in the group of chronic schizophrenics we have found either high or low responders to all IFN inducers (NDV, PHA + PMA and LPS). Furthermore, the patients with high IFN response had dominant positive symptoms of schizophrenia (delusions, hallucinations, bizarre behavior and thought disorder). Whereas, in the patients with low IFN response the negative symptoms prevailed (asociality or withdrawal, flat affect, attention impairment, abolition or apathy). In plasma samples of schizophrenics, factors were detected that transferred a hypersensitivity to the IFN inducers to normal donor leukocytes. For instance, in leukocytes cultured in the presence of plasma from schizophrenics, there were 71% of high IFN responders after stimulation with NDV, versus 26% of high IFN responders in the presence of plasma from normal controls. We suggest that the factors may belong to the class of opioid peptides, which interact with the production of cytokines including IFNs.

Interferon and tumor necrosis factor responses of HIV+ patients as markers for monitoring of the AIDS progression.

The blood samples taken from 31 HIV+ and AIDS patients were used to study interferon (IFN) and tumor necrosis factor (TNF) responses. The IFN and TNF levels in plasma were determined. In the whole blood assay (whole blood diluted 1:10 with culture medium) Newcastle disease virus (NDV) and phytohemagglutinin (PHA) were used as cytokine inducers. Blood leukocytes of HIV+ patients produced significantly less IFN-alpha after NDV stimulation than the cells of healthy (HIV-) individuals. On the other hand, the production of IFN-gamma in response to PHA was impaired only in AIDS patients with stage CDC IV and CD4+ cell number < 200/microliters. These patients had also increased IFN levels in plasma. Particularly, the high level of IFN in plasma was frequently detected in patients with progressing AIDS with CD4+ cell number < 50/microliters. This type of IFN was identified as a mixture of acid-labile and acid-stable IFN-alpha. The IFN responses of HIV+ patients may be considered as markers for monitoring progression of AIDS and therapy.

Elevated release of tumor necrosis factor-alpha and interferon-gamma by bronchoalveolar leukocytes from patients with bronchial asthma.

Bronchoalveolar lavage (BAL) leukocyte-secreted cytokines are considered to be important mediators of the inflammatory and allergic reactions in the lung. This study examines quantitative changes in the level of tumor necrosis factor-alpha (TNF alpha) and interferon-gamma (IFN gamma) production in BAL cell cultures derived from patients (n = 11) with bronchial asthma. The secretion of TNF alpha and IFN gamma was determined in intact (unstimulated) and phytohemagglutinin/phorbol myristate acetate (PHA + PMA)-stimulated BAL leukocyte cultures and compared with that in control cultures. In all patients studied, the background and PHA + PMA-induced secretion of TNF alpha and IFN gamma was significantly (p < 0.001) higher than that in parallel control cultures. In contrast to BAL cell preparations, the capacity of TNF alpha and IFN gamma secretion by patients' peripheral blood mononuclear cells (PBMC) did not differ from that of control subjects. High spontaneous release of TNF alpha and IFN gamma by patients' BAL leukocytes, but not PBMC, suggest that in the pathophysiology of bronchial asthma, these cytokines may act as local pathogenic agents in the lung.

Production of cytokines by mouse peritoneal cells treated with Tolpa Torf Preparation (TTP): dependence on age and strain of mice.

Production of two cytokines: Interferon beta (IFN-beta) and tumor necrosis factor alpha (TNF-alpha) by freshly isolated resident mouse peritoneal cells (RPC) treated with immunostimulating drug Tolpa* Torf Preparation (TTP) was investigated. Nontoxic concentrations of TTP (10 and 100 micrograms/ml) potentiated the cytokine production by RPC of BALB/c mice. The observed effects were dose-dependent. The optimal cytokine-inducing concentration of TTP was 100 micrograms/ml. The effect of TTP depended on the age of mice. The high potentiation (16-fold) of IFN-beta and TNF-alpha production by TTP was observed when 5 and 8 week-old mice were used. However, in RPC isolated from one year old mice, only two (fold) potentiation of IFN and TNF was observed. The activities of the same commercial preparations of TTP were compared in the RPC cultures isolated from mice of the three different strains: BALB/c, C3H/HeJ and NZB. The significant stimulation (16-32-fold) of TNF and IFN was observed in the cells isolated from BALB/c mice. In contrast, only slight stimulation of IFN and TNF (1-3-fold) was observed in cells of NZB mice. Peritoneal cells of C3H mice did not respond to stimulation with TTP.

Tolpa Torf Preparation (TTP) induces interferon and tumor necrosis factor production in human peripheral blood leukocytes.

Tolpa Torf Preparation (TTP) is a natural immunomodulating drug registered in Poland for use in humans. TTP is a biologically active low molecular weight fraction of an extract from peat containing organic substances, primary bound sugars, amino-acids, uronic and huminic acids and mineral salts. The toxicity of TTP is remarkably low, eg. cytotoxicity (CD50) for human peripheral blood leukocytes (PBL) is 1-9 mg/ml. We have discovered that TTP is an interferon (IFN) and tumor necrosis factor (TNF) inducer in human PBL. The IFN and TNF response of the PBL cultures was dose dependent. The optimal concentration of TTP for IFN or TNF response was 10-100 micrograms/ml. The cytokines stimulated by TTP were IFN-gamma, IFN-alpha and TNF-alpha. Ten commercial batches of TTP have been found to be active as cytokines inducers although variations in their activities were observed. On the other hand, 8 batches of TTP rejected by the producer because of the inadequate immunostimulating activity determined in mice, were found to be significantly less active than the commercial preparations. Over 115 buffy coats from the individual blood donors were used to prepare PBL cultures for this study. Approximately 20% of the PBL cultures were unresponsive to TTP. The IFN and TNF response of PBL to other inducers: phytohemagglutinin (PHA) or lipopolysaccharides (LPS) also varied. Whereas only 7% of PBL could not be stimulated by PHA, as much as 20-50% of PBL failed to produce IFN or TNF when treated with LPS. We suggest that TTP may have clinically useful activities connected with the capacity of stimulation of IFNs and TNF production.

A method to assess the immunomodulating effects of the Tolpa Torf Preparation (TTP) by measuring the hyporeactivity to interferon induction and tumor necrosis factor response.

Tolpa Torf Preparation (TTP) is an immunomodulator which was found to be interferon (IFN) and tumor necrosis factor (TNF) inducer in human peripheral blood leukocytes (PBL). PBL cultures prepared from the blood of healthy volunteers taking 5 mg TTP tablet daily loose ability to respond to IFN induction by TTP, phytohemagglutinin (PHA) or lipopolysaccharide (LPS). The phenomenon of hyporeactivity or tolerance is characteristic of every IFN inducer described so far. In the TTP treated volunteers the tolerance developed during three weeks and it lasted one to two weeks. In comparison with the tolerance to the IFN induction the hyporeactivity to TNF induction developed slowly and it was only partial. Because the detection of the tolerance to IFN induction is a very sensitive indicator of TTP activity in vivo in patients taking the drug orally, we suggest that it may be applied to assess the mode of action, dosage and schedule of administration of the IFN-inducing immunomodulator.

Human lymphoid target cells for the cytokine-inducing seleno-organic compounds.

Several seleno-organic compounds including ebselen are known as antiinflammatory and antioxidant agents. They also have glutathione peroxidase-like activity and are inhibitors of leukotrienes and prostaglandins. We have recently discovered that these drugs are inducers of cytokines, mainly interferon gamma (IFN-gamma) and tumor necrosis factor alpha (TNF-alpha) and mitogenic interleukins in human peripheral blood leukocytes (PBL) but not in the mouse or rat lymphoid cells. We described a production of IFN-gamma and TNF-alpha by various subsets of PBL stimulated with 2-phenyl-1,2-benzisoselenazol-3(2H)-one (ebselen) or bis [2-(N-phenyl-carbamoyl)]phenyl diselenide. IFN-gamma was produced mainly by E-rosette positive lymphocytes. However, the presence of monocytes was required for the optimal production of IFN-gamma. Also soluble mediators released by monocytes enhanced IFN-gamma synthesis. On the other hand, TNF-alpha was produced mainly by the adherent monocytes. Its synthesis was enhanced by the addition of T or B lymphocytes or conditioned medium from the culture of the stimulated lymphocytes. The relative concentrations of the subsets of lymphocytes or monocytes was important for the maximum production of both IFN-gamma and TNF-alpha. High concentration of lymphocytes inhibited the cytokine production.