Barbara Kwiatkowska-Patzer

Oral Administration of Lactococcus lactis Expressing Synthetic Genes of Myelin Antigens in Decreasing Experimental Autoimmune Encephalomyelitis in Rats

Background Multiple sclerosis is a human autoimmunological disease that causes neurodegeneration. One of the potential ways to stop its development is induction of oral tolerance, whose effect lies in decreasing immune response to the fed antigen. It was shown in animal models that administration of specific epitopes of the three main myelin proteins – myelin oligodendrocyte glycoprotein (MOG), myelin basic protein (MBP), and proteolipid protein (PLP) – results in induction of oral tolerance and suppression of disease symptoms. Use of bacterial cells to produce and deliver antigens to gut mucosa seems to be an attractive method for oral tolerance induction in treatment of diseases with autoimmune background. Material/Methods Synthetic genes of MOG35-55, MBP85-97, and PLP139-151 myelin epitopes were generated and cloned in Lactococcus lactis under a CcpA-regulated promoter. The tolerogenic effect of bacterial preparations was tested on experimental autoimmune encephalomyelitis, which is the animal model of MS. EAE was induced in rats by intradermal injection of guinea pig spinal cord homogenate into hind paws. Results Rats were administered preparations containing whole-cell lysates of L. lactis producing myelin antigens using different feeding schemes. Our study demonstrates that 20-fold, but not 4-fold, intragastric administration of autoantigen-expressing L. lactis cells under specific conditions reduces the clinical symptoms of EAE in rats. Conclusions The present study evaluated the use of myelin antigens produced in L. lactis in inhibiting the onset of experimental autoimmune encephalomyelitis in rats. Obtained results indicate that application of such recombinant cells can be an attractive method of oral tolerance induction.

Suppression of Experimental Autoimmune Encephalomyelitis in the Rat by Oral Administration of Spinal Cord Protein Hydrolysate

Multiple sclerosis is an autoimmune disease and the myelin proteins MBP(myelin basic protein), MOG (myelin oligodendrogial glycoprotein), MAG (myelin associated glycoprotein) are the main antigens for the immunologic system (1). Application of the oral toleration method using myelin proteins or their components gives the possibility for a decrease in the number of autoimmunologic reactions. Oral tolerance was first described by Wells in 1911 as a method of prevention of systemic anaphylaxis in guinea pigs by previous feeding of hen’s eggs protein. Long before modern immunology, an observation was made that orally fed antigens could suppress immune response to this antigen. Recently the oral tolerance method has been re-examined in view of the therapy of autoimmune diseases. The mechanism involved in oral tolerance is as follows: clonal deletion, clonal anergy and active suppression. Some studies show that the primary mechanism of oral tolerance is generation of active suppression. Depending on the antigen, the amount fed and the nature of the fragments generated absorbed antigen could induce active suppression or clonal anergy being processed by the gut and absorbed into circulation. Some authors suggested that digestion of the antigen is required for the generation of oral tolerance.

Use of pig spinal cord waste as a valuable source of biologically active substances

Streszczenie — Rdzen kregowy zwierz1t rzeŸnych jest uci1?liwym odpadem przemys3u miesnego. Wchodz1ce w ich sk3ad bia3ka strukturalne mog1 byae u?yte jako potencjalne Ÿrod3o strukturalnie zmodyfikowanych biopolimerow indukuj1cych neurodegeneracyjne przemiany oœrodkowego uk3adu nerwowego ssakow, w tym cz3owieka. Bia3ka prionowe stanowi1 jednak du?e zagro?enie. Zaproponowano zastosowanie procesu enzymatycznego trawienia wieprzowego rdzenia kregowego w celu otrzymywania hydrolizatow zawieraj1cych niskocz1steczkowe peptydy. Peptydy te nie tworz1 trwa3ych struktur trzeciorzedowych, a ich preparaty mog1 byae skutecznie oczyszczone, co pozwala na usuniecie niepo?1danych substancji biologicznych. G3ownym sk3adnikiem rdzenia kregowego ssakow s1 bia3ka mieliny, strukturalne biopolimery izoluj1ce i chroni1ce komorki nerwowe. Du?e podobienstwo sekwencji bia3ek mielinowych wieprzowych i ludzkich pozwala na zastosowanie hydrolizatow wieprzowych rdzeni nerwowych jako preparatow do indukcji tolerancji pokarmowej w stwardnieniu rozsianym. Pilotowe badania na modelach zwierzecych potwierdzaj1 potencjaln1 skutecznoœae terapeutyczn1 otrzymanych preparatow. S3owa kluczowe: wieprzowy rdzen kregowy, cholesterol, bia3ka mieliny, hydrolizaty bia3kowe, tolerancja pokarmowa, stwardnienie rozsiane.