Development of purification and test procedures of tumorspecific immunoglobulin fragments for their further diagnostic use  Page description

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Details of project

  
Identifier
48933
Type K
Principal investigator Kotlán, Beatrix
Title in Hungarian Tumorspecifikus ellenanyagfragmens tisztítási és tesztelési eljárásainak kidolgozása a diagnosztikus alkalmazhatóság céljából
Title in English Development of purification and test procedures of tumorspecific immunoglobulin fragments for their further diagnostic use
Panel Molecular and Structural Biology and Biochemistry
Department or equivalent Sebészeti és Molekuláris Patológiai Osztály (National Institute of Oncology)
Participants Ladányi, Andrea
Tímár, József
Tóth, József
Starting date 2005-01-01
Closing date 2009-12-31
Funding (in million HUF) 8.662
FTE (full time equivalent) 2.52
state closed project





 

Final report

  
Results in Hungarian
A szolid tumorokban fellelhető B sejtek kiaknázása volt a cél. A tumort infiltráló B sejtek (TIL-B) tumorspecifikus antigénkötő kapacitását nemzetközileg is elsőként igazoltuk. Az eljárás diagnosztikus célú tumorfelismerő „jelölő molekulákat” eredményez. A tématervi munka sokrétű volt: 1/Új molekuláris biológiai és biotechnológiai módszerek, 2/ preparatív fehérjetisztítási munkák, 3/ összehasonlító DNS elemzések, 4/ különböző ellenanyag – antigén kötődési tesztek, 5/ immun hiszto- és citokémiai jelölési technikák történtek. Eredményeink a célkitűzéseknek megfelelőek. A medulláris emlőkarcinómából nyert egyláncú ellenanyag fragmens tisztítása sikerült. Tumorsejtkultúra lemezek és fagyasztott tumorszövet metszetek a reagensünkkel pozitív reakciót adtak immunfluoreszcencs tesztben konfokális mikroszkópos értékeléssel. Melanomák, kolorektális karcinomák és több emlőtumor típus paraffinos metszetein immunhisztokémiai eljárással is pozitív reakciót kaptunk az új GD3 gangliozid ellenanyaggal. A TIL-B immunglobulin fág megjelenítéses módszer lehetővé tette további érdekes immunglobulin variábilis régiók szelektálását invaziv ductalis emlőcarcinomákban és melanomákban. GD3 gangliozid specifikus ellenanyag fragmensünk a tumorprogresszió és invázió szempontjából fontos struktúrákat szelektíven és megfelelően jelölte a tumorszöveti mintákon. Ezen szialilált glikoszfingolipidek differenciál diagnosztikus értékét alapozta meg a munka, lehetőséget nyitva mikrometastazisok kimutatására is.
Results in English
We aimed to reveal the capacity of B cells present in small amounts in solid tumors. We were internationally the first to prove the characteristic tumorspecific binding capacity of tumor infiltrating B cells (TIL-B). Our startegy helps to develop novel diagnostic tumor recognizing „labelling arrow molecules”. The project work consisted of: 1/ Novel molecular biological and biotechnological techniques, 2/ preparative protein purification processes, 3/ comparative DNA analyses, 4/ different antibody – antigen binding assays, 5/ immunohistochemical and immunocytochemical methods. The results correlated our specific aims. We could purify our medullary breast carcinoma originated antibody fragment. Tumor cell chamber slides and frozen tissue sections gave positive reactions with our reagent in immunofluorescence – confocal laser microscopy. On paraffin embedded tissue sections of melanomas, colorectal carcinomas and some other breast cancer types we got positive reaction with this unique GD3 ganglioside specific antibody. Due to our TIL-B immunglobulin phage display technique further antibody variable region genes were selected in invasive ductal breast carcinomas and melanomas. The GD3 ganglioside specific antibody labelled selectively the structures important for tumorprogression and invasion on tumor tissue specimens. The work supports the differential diagnostic value of these sialylated glycosphingolipides, opening a window towards the detection of tumor micrometastases.
Full text https://www.otka-palyazat.hu/download.php?type=zarobeszamolo&projektid=48933
Decision
Yes





 

List of publications

  
Kotlan, B., Teillaud, J.L., Toth J., McKnight M., Glassy M.: The renaissance of the tumor infiltrating B lymphocytes. „ The proof of principle.”: Novel ganglioside antigen recognizing capacity of B cells in breast, 96th AACR meeting, Abstract book: No5128, p:1211 2005, 2005
Kotlan B, Toth J., McKnight M., Glassy M: Human Antibodies against miscellanous antigens. Weak tumor associated GD3 ganglioside detection on invasive ductal breast carcinomas and other solid tumors, (B56 abstract / Kongresszusi kiadvány / ”Advances in Breast Cancer Research: Genetics, Biology and Clinical Applications”,, 2005
Kotlan B, McKnight M., Teillaud J.L and Glassy M: „A novel approach to obtain unique GD3 and GD2 ganglioside specific antibody fragments for cancer diagnostics.”, A7/ abstract/ Kongresszusi kiadvány, pp70, „Frontiers in Cancer Prevention Research, (Baltimore,, 2005
Kotlan B, Simsa P., Teillaud J.L, Fridman W.H., Toth J., McKnight M., Glassy M.:: Novel ganglioside antigen identified by B cells in human medullary breast carcinomas. The proof of principle concerning the infiltrating B lymphocytes., J. Immunology, 175: 2278 – 2285,, 2005
Simsa P, Teillaud JL, Stott I D, Toth J, Kotlan B: Tumor-infiltrating B cell immunoglobulin variable region gene usage in invasive ductal breast carcinoma., Pathology Oncology Research 11 (2): 92-7,, 2005
Kotlan B, Padanyi A, Reti M, Gyodi E, Rajczy K, Batorfi J, Petranyi Gy, Fulop V:: Alloimmune and autoimmune background in recurrent pregnancy loss – Successful immunotherapy by IVIG., American Journal of Reproductive Immunology Vol 55: 331- 337., 2006
Beatrix Kotlan1,2, Michael McKnight2,3, Jean-Luc Teillaud4, Mark C Glassy2,3: A novel approach to generate unique GD3 Ganglioside Tumorspecific Human Immunoconjugates with apoptotic effects for a potential use in the immunotherapy of solid tumors, Imaging Tecnology in Drug Discovery and Development/2006/ Kongresszusi Kiadvany pp197, 2006
Kotlan, B; Toth, J; McKnight, M; Glassy, MC: Characteristics of tumor gangliosides revealed by B cells infiltrating human breast carcinomas, Human Antibodies [Hum. Antibodies]. Vol. 15, no. 1-2, pp. 9-13. 2006., 2006
Beatrix Kotlan1,3, Jozsef Toth2, Michael McKnight3,4, Mark C Glassy3,4: Generation and functional characteristics of novel single chain Fv antibody fragments specific to unique gangliosides on breast tumors, AACR 97th Annual Meeting/ Washington, Kongresszusi Kiadvany No: 2238, pp528-529, 2006
Beatrix Kotlan1,2, Michael McKnight2, Andrea Ladanyi3, Erzsebet Raso3, Jozsef Toth3, Jozsef Timar3, Mark C Glassy2: B cells on the road to defeat cancer: A novel approach to generate tumorspecific human immunoconjugates from breast carcinomas and melanomas, AACR Frontiers in Cancer Prevention Research /Boston 2006, No: B92 pp161pp, 2006
Beatrix Kotlan1,2, Michael McKnight2, Andrea Ladanyi3, Erzsebet Raso3, Jozsef Toth3, Jozsef Timar3, Mark C Glassy2: A novel approach to generate tumorspecific human immunoconjugates from breast carcinomas and melanomas, International Congress of Immunogenomics and Immunomics 2006/ P-1-44 pp: 302, 2006
Beatrix Kotlan1,2, Michael McKnight2, Andrea Ladanyi3, Erzsebet Raso3, Jozsef Toth3, Jozsef Timar3, Mark C Glassy2: A novel approach to generate tumorspecific human immunoconjugates from breast carcinomas and melanomas, Antibody Engineering/ San Diego/ 2006 poster No: 1C, 2006
Mark C Glassy1,2, Michael McKnight1,2, Beatrix Kotlan2,3, Eric Glassy 4, Keiji Koda5: Cocktails of human anti cancer antibodies show a synergistic effect in nude mouse tumor xenografts, Human Antibodies, 2007
Beatrix Kotlan1,2, Michael McKnight2, Andrea Ladanyi3, Erzsebet Raso3, Jozsef Toth3, Mepur H ravindranath4, Jozsef Timar3, Mark C Glassy2: Novel findings obtained by single chain Fv antibody library establishment from tumor infiltrating B lymphocytes of human breast carcinomas and melanomas, Proceedings of the American Association for Cancer Research Annual Meeting 2007 /Los Angeles April 14-18 2007 Vol 48, No: 3326 pp791, 2007
Beatrix Kotlan1, Agnes Padanyi2, Jozsef Batorfi3, Margit P Buzas3, Katalin Rajczy2, Katalin Miklos4, Peter Simsa1, Julia Németh4, Eva Gyodi2, Marien Reti5, Vilmos Fulop3, Gyozo Petranyi2: Defining alloimmune and autoimmune background in recurrent pregnancy loss leads to successful intravenous immunoglobulin immunotherapy, American Journal of Reproductive Immunology/ISIR June 10-14 2007 / Blackwell, Munksgaard No: ICRI-07-2-97 Vol57 pp472-473, 2007
Kotlan B: Novel findings to establish a new generation of breast cancer diagnostic and therapeutic immunoconjugates., 2008 In: My Fulbright Experience / Reports of Hungarian Grantees Ed by: Nagypal Cs Published by: Bruckner H, pp: 201 – 218 (www.Fulbright.hu/book3/kotlanbeatrix.pdf), 2008
Kotlan B: Role of cellular and humoral immune response in pregnancy failure., In: Questions in highligts in reproductive immunology. Ed: Fülöp V, Semmelwess Kiadó, Budapest 2008. pp. 229 – 237., 2008
Kotlan B: A novel aspect leads to the renaissance of key tumor associated molecules, 6th International Congress on Autoimmunity, Porto, Sept 10 -14, 2008. Előadás, PS17 session, 2008
Kotlan B, Timar J, Glassy MC and Ravindranath MH:: Sensitive detection of overexpressed tumor associated structures on various cancer cell types., 8th International Conference of Anticancer Research , Kos, 17 -22. 2008, Lecture in: Tumorprogression session, 2008
Kotlan B:: The power and promise of immunoglobulin repertoire analysis for cancer therapy., 14th International Congress on Human Antibodies and Hybridomas, New York, 2008. november 12 -14, Cancer I. session / lecture Abstract in: Human Antibodies and Hybridomas, 2008
Kotlan B, Timar J, Ravindranath MH, Glassy MC.: A comparative sequence data analysis and phage display based novel technique reveal key tumor-associated antigens that influence cancer progression., 100th AACR annual meeting, proceedings, Denver 2009 Április 18 -22 No4147 kongresszusi kiadványban, 2009
Kotlan B, Glassy MC. Szentirmay Z and Ravindranath MH:: Key tumor associated molecules revealed by the exploitation of the natural human immune response., Signal and Signal Processing in the Immune System, Balatonoszod, 2009 szeptember 2 -6. Konferenciakiadv., 2009
Kotlan B, Ravindranath MH, Teillaud JL and Glassy MC,: A novel challenge to define tumor associated gangliosides based on investigations of the natural humoral immune response., First International Conference Cancer Immunotherapy and Immunomonitoring Kiev 2009 május 18 -21, Kongresszusi Kiadványban, 2009
Kotlan B, Ravindranath MH, Timar J, Glassy MC,: Can B cells, the small ignored guys hidden in melanoma tissues, put us on the right track for tumor specific antigens?, First International Conference Cancer Immunotherapy and Immunomonitoring Kiev 2009 május 18 -21, Kongresszusi Kiadványban, 2009
Kotlan B, Ravindranath MH, Dobos J, Szentirmay Z, Glassy MC: Tumorasszociált glikoszfingolipid antigének kimutatásának új lehetősége epiteliális és neuroektodermális tumorok esetében., Magyar Onkológusok Társaságának XXVIII. Kongresszusa, Budapest, 2009. november 12 -14. Magyar Onkológia, 2009. 53: suppl pp: 62., 2009
Kotlan B: Tumor infiltráló B limfociták tumor specifikus antigen felismerő képessége., Az Országos Onkológiai Intézet Sebészi és daganatpathológiai Centrum és a Frank Diagnosztika- DAKO Szimpóziuma 2009 dec 4.: Előadás és CD anyag, 2009
Kotlan B, Stroncek DF, Marincola FM: Placing laboratory findings into therapy: a marathon goal that has to be reached! Review, Poliskie Archiwum Medycyny Wewnetrznej119 (9): 586 – 593., 2009
Kotlan B, Stroncek DF, Marincola FM: Intravenous immunoglobulin - based immunotherapy: An arsenal of possibilities for patients and science. Review, Immunotherapy 2009. 1(No6): 995 - 1014, 2009
Kotlan B and Glassy MC: Antibody phage display – a powerful technology that has quickly opened windows into the clinic., In: Antibody Phage Display 2nd ed * ISBN: 9781603273015 Ed: Aitken R. The Humana Press, Inc, Totowa, New Jersey USA, Series Methods Mol Biol. 2009. 562:1-15., 2009



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