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001-es BibID:	BIBFORM054365
Első szerző:	Kappelmayer János (laboratóriumi szakorvos)
Cím:	Flow cytometric detection of intracellular myeloperoxidase, CD3 and CD79a : interaction between monoclonal antibody clones, fluorochromes and sample preparation protocols / Janos Kappelmayer, Jan W. Gratama, Eva Karaszi, Pablo Menendez, Juana Ciudad, Rosana Rivas, Alberto Orfao
Dátum:	2000
ISSN:	0022-1759
Megjegyzések:	Detection of intracellular myeloperoxidase (MPO), CD79a and CD3 has become the most specific tool for the assignmentof myeloid, B- and T-lymphoid lineages in acute leukemias. In order to establish the best combination of monoclonalantibody reagent and sample preparation technique for the intracellular detection of these three markers, we compared sixdifferent cell fixation?permeabilization kits (Cytofix/CytopermE, Fix and PermE, IntraprepE, IntrastainE, PermeacyteEand PermeafixE) using 12 fluorochrome conjugates derived from seven monoclonal antibody (mAb) clones. A total of 21samples corresponding to normal peripheral blood (n54), normal bone marrow (n53), acute myeloblastic leukemia (AML,n56), precursor B-acute lymphoblastic leukemia (ALL, n56) and T-ALL (n52) cases, were analysed in two centers. Allfixation / permeabilization methods resulted in decreased side scatter and mostly increased forward scatter as compared toerythrocyte-lyse-washed and 1% paraformaldehyde fixed samples. The autofluorescence levels of the leukocyte populationswas only significantly increased with use of the Cytofix/CytopermE kit and mildly with the other techniques. In addition,non-specific staining increased significantly for combinations of any anti-MPO mAb with the Cytofix/CytopermE kit andfor the CD3 clone S4.1 combined with any intracellular method. Anti-MPO antibodies gave a stronger fluorescence signalwhen conjugated to PE than when coupled to FITC. In conclusion, MPO-7-PE, UCHT-1-PE (CD3) and any HM57-PEconjugate (CD79a) in combination with Fix and PermE, IntraprepE, IntrastainE or PermeafixE, provided specific stainingof the respective markers in sufficient intensities. Thus, combined selection of fixation / permeabilization kits and monoclonalantibody reagents against CD3, CD79a and MPO is required for obtaining optimal cytoplasmic detection of these antigens.Ó 2000 Elsevier Science B.V. All rights reserved.
Tárgyszavak:	Orvostudományok Klinikai orvostudományok idegen nyelvű folyóiratközlemény külföldi lapban
Megjelenés:	Journal of Immunological Methods. - 242 : 1-2 (2000), p. 53-65. -
További szerzők:	Gratama, Jan Willem Karászi Éva Menendez, Pablo Ciudad, Juana Rivas, Rosana Orfao, Alberto
Internet cím:	Intézményi repozitóriumban (DEA) tárolt változat
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