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Colostrinin: a Proline-Rich Polypeptide Complex of Potential Therapeutic Interest

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Colostrinin: a Proline-Rich Polypeptide Complex of Potential Therapeutic Interest Cell. Mol. Biol. 59 (1): 4-11 Cellular & Molecular Biology Published on November 3, 2013 http://www.cellmolbiol.com DOI 10.1170/T942 Copyright © 2013. All rights reserved. COLOSTRININ: A PROLINE-RICH POLYPEPTIDE COMPLEX OF POTENTIAL THERAPEUTIC INTEREST M. JANUSZ AND A. ZABŁOCKA Department of Immunochemistry, Ludwik Hirszfeld Institute of Immunology and Experimental Therapy, Polish Academy of Sciences, Rudolfa Weigla 12, 53-114 Wrocław, Poland Abstract Article information A proline-rich polypeptide complex (PRP) subsequently known as ColostrininTM was found for the first time in ovine colostrum Received on February 12, 2011 as a fraction accompanying colostral IgG2. Subsequently, similar polypeptides were found in human, bovine and caprine colostrum. PRP is a complex of peptides of molecular masses from 500 to 3000 Da. It contains 25% proline residues and 40% Accepted on January 30 ,2012 hydrophobic amino acids. It is not species specific, and is active bothin vivo and in vitro. PRP possesses immunoregulatory properties, including effects on humoral and cellular immune responses, shows regulatory activity in Th1 and Th2 cytokine Corresponding author induction, and has the ability to inhibit the overproduction of reactive oxygen species and nitric oxide. PRP has also shown Tel: +48-71 337 11 72 psychotropic properties. Both immunoregulatory and psychotropic properties suggest potential clinical use of PRP for Fax: +48-71 337 13 82 neurodegenerative disorders. Beneficial effects of PRP/Colostrinin in the case of Alzheimer’s disease were shown in double- E-mail:[email protected] blind placebo-controlled trials, in long-term open-label studies and in multicenter clinical trials. A very important property of PRP/Colostrinin and one of its components, a nonapeptide (NP), is the prevention of Aβ aggregation and the disruption of aggregates already formed. Moreover, PRP has been found to modulate neurite outgrowth, suppress uncontrolled activation of cells, and reduce 4-HNE-mediated cellular damage. Biological response modifying activity of PRP/Colostrinin can play an important role in its use in the treatment of Alzheimer’s disease and suggests its application beyond neurodegenerative disorders. Key words: Proline-rich polypeptide complex (PRP)/Colostrinin/(CLN)TM, immunomodulatory properties, cytokine induction, reactive oxygen species and nitric oxide, Alzheimer’s disease. INTRODUCTION nomodulatory substances was shown for the first time in our laboratory during studies on ovine and bovine serum Proline-rich proteins and peptides are widespread in and colostral IgG. We have found that ovine colostral IgG2 microorganisms, plants and animals. They can influence is contaminated with a peptide with high content of proline many biological functions such as regulation of cell and residues (30, 41). Attempts to obtain anti-ovine IgG2 an- tissue processes and signal and regulatory protein interac- tibodies by immunization with peptide-contaminated im- tions. The most important are the peptides with potential munoglobulins were unsuccessful. However, IgG2 sepa- therapeutic significance, i.e. showing antiviral, antibac- rated from peptide, named proline-rich polypeptide (PRP), terial, antitumor, or immunoregulatory activity. Peptides was a good immunogen. This indicated that PRP might with immunotropic activity, especially constituents of co- have immunotropic activity. Proline-rich polypeptide, sub- lostrum and milk, seem to be of great importance. sequently called ColostrininTM (CLN), was also found in Colostrum and milk – the first mammalian nourishment human, bovine, and caprine colostrum (48, 57). The yield – are the richest reservoir of constituents of great impor- of proline-rich polypeptide is dependent on time after de- tance in newborn development. They contain protective livery. The level of PRP isolated by the method of Janusz and supporting factors. Immunoglobulins are a reservoir et al. (31) is the highest, between 400 and 800 mg/l, in of immunity transferred from the mother to the newborn. sheep colostrum collected within the first 6 hours. PRP Immunity can be transferred not only by immunoglobu- is also present in colostrum collected several days after lins. Besides immunoglobulins, another constituent of delivery and in milk but at a much lower level. PRP is a milk, lactoferrin, has a protective function, exhibiting an- complex of low-molecular-weight peptides, ranging from tibacterial, antifungal, antiviral, antiparasitic, and antitu- 500 to 3000 Da. Proline residues in PRP comprise up to mor activities. Supporting factors such as growth factors, 25%. It contains a high proportion of hydrophobic amino nucleotides, vitamins, and minerals stimulate the growth acids (40%), a low percentage of glycine, and no alanine, and regeneration of cartilage, nerve, bone, and muscle, and arginine or cysteine residues. PRP is not phosphorylated balance blood sugar. Colostrum and milk are rich in cyto- or glycosylated. Circular dichroism spectra in water and kines, lymphokines and peptides which may stimulate the in 50% (v/v) trifluoroethanol suggest block sequences of neonate immune system and play a regulatory role. proline residues forming helices of poly-proline II. PRP A possibility that colostrum could be a source of immu- is susceptible to proteases but collagenase resistant. It is a 4 M. JANUSZ And A. ZABŁOCKA/ Colostrinin-potential therapeutic interest. weak immunogen. In PRP complex peptides with homolo- intraperitoneally into NZB mice significantly lowered the gy to β-casein and its hypothetical homolog and to annexin incidence of a positive Coombs reaction and prolonged the have been found. Peptides with no significant homology to mean age of mice. From a pool of precursor cells, PRP any specific protein in the current GenBank database have may induce suppressor cells controlling the development also been identified (31, 36). When analyzing gene expres- of hemolytic anemia (76). Proline-rich polypeptide com- sion changes it was found that Colostrinin elicited highly plex is the first protein of mammalian origin that functions complex and multiphasic changes in the gene expression as a B cell-trophic mitogen. PRP induces resting mouse B profile of treated cells (63). Proline-rich polypeptide com- cells into and supported their progression through the cell plexes of both ovine and bovine origin are not species spe- cycle at frequencies comparable to those seen for bacterial cific. They are active in mice, rats, chickens, and humans. lipopolysaccharide (35). Immunoregulatory activity simi- Two possible biological functions of PRP were consid- lar to whole PRP complex was reflected by a nonapeptide ered. The first was that the polypeptide could prevent the fragment, VESYVPLFP (NP), isolated from chymotryptic proteolysis of colostral immunoglobulins in the digestive digestion of PRP and obtained by chemical synthesis (32, tract of newborns and the second that polypeptide com- 59). plex could act as an immunomodulator. It was shown that the presence of PRP did not prevent proteolysis of ovine PROLINE-RICH POLYPEPTIDE COMPLEX IN- colostral IgG1 and IgG2 by trypsin, pepsin, and papain. FLUENCES INNATE IMMUNITY The polypeptide did not act as an inhibitor of proteolytic enzymes with other substrates, either (31). Innate (natural) immunity refers to the basic resistance to disease that a species possesses. It is the first line of de- IMMUNOTROPIC EFFECTS OF PRP fense against infections. The reactions of innate immunity include, among others, phagocytosis, the production and As shown in a mouse model, PRP complex exerted ef- activity of cytokines, chemokines, adhesion molecules, fects on the humoral and cellular immune response, both acute phase proteins, reactive oxygen species (ROS) and in vivo and in vitro. The effect depended strongly on the reactive nitrogen species (NOS). It was shown that PRP intensity of the response and not on the time of administra- complex did not induce the production of nitric oxide tion (67). PRP induces differentiation of cells exhibiting (NO) in mice; it was able to inhibit the release of NO in- helper or suppressive function from glass-nonadherent and duced by LPS by about 30-50%. This effect was accompa- glass-adherent precursor cells, respectively. The fact that nied by lower iNOS protein expression in peritoneal cells. PRP may generate two opposite activities from the pool In the liver sections of mice treated with PRP 6 h after of thymocytes explains its immunoregulatory effect (42, LPS application, the number of iNOS-positive cells was 78, 79. It is suggested that the activity of PRP could be significantly reduced (71,74). Proline-rich polypeptide mediated by prostaglandins (67). Proline-rich polypeptide complex, and at a lesser intensity its nonapeptide constitu- complex reversibly converts sensitivity of thymocytes to ent (NP), show both an inductive and regulatory effect on cortisone and activity in the GvH reaction (77). PRP act- cytokine and reactive oxygen species release in human ing on cortisone-sensitive (CS) or cortisone-resistant (CR) peripheral blood leukocytes and whole blood cultures. In thymocytes changes the number of cells forming autolo- cultures of human whole blood cells, PRP in concentra- gous rosettes in mice (66). tions of 1-100 μg was able to induce the secretion of both PRP is able to reduce binding of peanut agglutinin Th1, i.e. IFN and TNF-α, and Th2 cytokines, i.e., IL-6 and (PNA) to murine PNAhigh and to increase the binding of IL-10, involved in the cellular
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