Pregled bibliografske jedinice broj: 107068
Identification of pan B-cell neoplasm markers using a combination of cDNA PCR subtraction and microarray analysis
Identification of pan B-cell neoplasm markers using a combination of cDNA PCR subtraction and microarray analysis // Blood 2001 98(11)
Orlando (FL), Sjedinjene Američke Države, 2001. (predavanje, međunarodna recenzija, sažetak, znanstveni)
CROSBI ID: 107068 Za ispravke kontaktirajte CROSBI podršku putem web obrasca
Naslov
Identification of pan B-cell neoplasm markers using a combination of cDNA PCR subtraction and microarray analysis
Autori
Gaiger, A. ; Ordonez, N. ; Mannion-Henderson, J. ; Carter, L. ; Kusec, Rajko ; Jaksic, Branimir ; Jaeger, U. ; Greinix, H. ; McNeill, P. ; Jeffery, E. ; Reed, S. ; Jelinek, D. ; Greipp, P. ; Persing, D. ; Cheever, M.A. ; Algate, P.
Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni
Izvornik
Blood 2001 98(11)
/ - , 2001
Skup
43th Annual Meeting of the American Society of Hematology
Mjesto i datum
Orlando (FL), Sjedinjene Američke Države, 07.12.2001. - 11.12.2001
Vrsta sudjelovanja
Predavanje
Vrsta recenzije
Međunarodna recenzija
Sažetak
Malignant cells express multiple antigens that can be recognized by the immune system and can be targeted by antigen specific immunotherapies including vaccines, antibodies and T cells. To identify genes that are over-expressed in B-cell malignancies PCR subtracted cDNA libraries in conjunction with microarray screening were used. Ten subtracted libraries (lymphoma n=6, CLL n=2, myeloma n=2) were constructed to enrich for B-cell neoplasm or lymphatic tissue specific cDNA sequences. cDNA pools from lymphomas, B-CLL and multiple myeloma (MM) were subtracted against cDNA pools of related normal hematopoietic tissues or normal non-hematopietic tissues. Subtracted libraries showed enrichment for both B-cell neoplasm and lymphatic tissue specific genes compared to an unsubtracted library. 14, 000 clones were sequenced and analyzed by comparison to sequences in 3 electronic databases: GenBank, huEST and GenSeq. At the time of analysis, 1380 (10%) clones had no significant similarities to known genes and 12, 120 (90%) matched known genes. These 14, 000 cDNA fragments were then analyzed using DNA microarray technology. Genes over-expressed in B-cell malignancies were identified using pairs of fluorescence-labeled cDNA probes synthesized from lymphomas, CLL or MM (n=50) and normal tissue poly A+ RNAs (n=50). Over 1.4 Million hybridization signals were analyzed. Expression patterns of 65 overexpressed genes were confirmed and characterized further by Real Time PCR using a panel of 170 cDNAs comprising of lymphomas, CLL, MM, normal tissues and MACS sorted hematopoietic subpopulations. Expression profiles were compared to known therapeutic antibody targets, CD20, CD52 and CD45. Based on their mRNA expression profile a group of cDNA fragments (Ly1448, Ly1456 and Ly1464) was identified demonstrating expression in normal tissues similar to CD20 but displaying broader coverage in B-cell malignancies. These cDNA fragments are expressed in the majority of B-cell lymphomas, B-CLL and multiple myeloma patient samples and were thus termed Pan B-cell neoplasm markers. So far, potential full length open reading frames of 2 of the 3 cDNA fragments have been identified, encoding for proteins with a predicted transmembrane region. Further characterization of these genes is in progress.
Izvorni jezik
Engleski
Znanstvena područja
Kliničke medicinske znanosti
