The mo abs were used with two primary aims: To evaluate their role in distinguishing acute myeloid leukaemia (AML) from ALL

The mo abs were used with two primary aims: To evaluate their role in distinguishing acute myeloid leukaemia (AML) from ALL. To determine the cells of lineage in cases of ALL and classify them either as B-cell lineage (precursor B-cells) or T-cell lineage. == Material and Methods == Thirty five cases of acute leukaemia diagnosed on peripheral blood smear and bone marrow examination were included in this study. phosphatase antialkaline phosphatase technique served as a reliable and convenient method for immunophenotyping of leukaemias. KEY WORDS:Immunophenotyping, Surface markers, Leukemia == Introduction == Leukaemias, till recently, were recognized and classified by morphologic and cytochemical criteria [1,2]. Recent improvements in immunology have led to important insights into the stages of human lymphocyte and granulocyte differentiation [3]. These stages can be precisely defined using monoclonal antibodies [mo abdominal muscles] enabling a more accurate classification of leukaemias [3,4]. The established protocols of immunophenotyping acute leukaemias, especially the acute lymphoblastic leukaemias (ALL), are resource rigorous and beyond the reach of most of the developing countries. In this study, a modified protocol was evaluated for immunophenotyping acute leukaemias. The mo abdominal muscles were used with two main aims: To evaluate their role in distinguishing acute myeloid leukaemia (AML) from ALL. To determine the cells of lineage in cases of ALL and classify them either as B-cell lineage (precursor B-cells) or T-cell lineage. == Material and Methods == Thirty five cases of acute leukaemia diagnosed on peripheral blood smear and bone marrow examination were included in this study. There were 20 cases of ALL, 14 cases of AML and one case of acute undifferentiated leukaemia. The classification of leukaemia was based on the criteria laid by the French-American-British (FAB) cooperative study group [1,5]. Cytochemical staining such as myeloperoxidase (MPO), Sudan Black B (SBB) were usually utilised. Immunophenotyping was carried out Bupivacaine HCl on routinely prepared blood and bone marrow smears using the alkaline phosphatase antialkaline phosphatase (APAAP) technique [6,7]. The protocol followed was : Prepare blood and bone marrow smears. Air dry for 218 hours. Fix in acetone : methanol for 90 seconds. Transfer directly to Tris buffer saline (TBS). Leave in TBS for Bupivacaine HCl 15 minutes. Add suitably diluted main mouse mo ab and incubate in a moist chamber at room heat for 30 minutes. Tap off antibody and place slide in TBS for 5 minutes. Add anti-mouse-immunoglobulins. Incubate for 30 minutes. Tap off antibody and place slide in TBS for 5 minutes. Add APAAP complex and incubate for 30 minutes. Tap off APAAP complex. Place slide in TBS for 5 minutes. Add alkaline phosphatase substrate and incubate for 30 minutes. Wash in TBS and then with tap water. Counterstain with haematoxylin and mount in an aqueous mounting medium. All incubation was carried out in room heat in a moist chamber. Positive and negative controls (e.g. normal peripheral blood smear, known case of leukaemia) were used for comparison. The following mo abs were used : CD 2, CD 7, CD 10, CD 13, CD 14, CD 19, CD 20, and CD 33. The B-cell antigen expression was defined as CD 19 positivity. CD 20 was used as an additional B-cell marker. The T-cell antigen expression was CD 2 and CD 7 positivity. The cells of B and T lineages were by definition unfavorable for myeloid cytochemical markers and mo abdominal muscles CD 13 and CD 33. CD 14 was used as a monocytic marker. The common ALL antigen (CALLA) was defined as CD 10 positivity. A positive reaction was seen as cell having reddish staining of an intensity greater than that seen in the background. The criterion for surface marker positivity was its expression in at least 20% of the leukaemic blast cell populace [8]. Electron microscopy was carried out for detection of MPO in the case diagnosed as acute undifferentiated leukaemia using the method of Fraham and Karnovsky [9]. The cases were treated using standard chemotherapeutic regimens and their Bupivacaine HCl end result was correlated with the type of leukaemia. == Results == There were 20 cases of ALL. Amongst the ALL, there were 9 of FAB L1 and 11 of FAB L2 subtypes. After Rabbit Polyclonal to ARFGAP3 immunophenotyping, the cases of ALL were placed in three groups (Table 1). == TABLE 1. == Immunophenotypes of ALL CD 10 positivity was seen in 12 out of 20 cases of ALL. In child years ALL, 10 out of 12 cases reacted with CD 10 and were mostly FAB L1 subtype (Table 2). The adult group showed mostly FAB L2 subtype and a lesser CD 10 positivity (Table 3). CD 19 expression was seen in all cases of precursor B-cell ALL. CD 20 was seen in 6 out of the 20 cases. Bupivacaine HCl == TABLE 2. == FAB and immunophenotypes of child years ALL and their response to therapy == TABLE 3. == FAB and immunophenotypes of adult cases of ALL and their response to therapy Remission by 30 days Remission in 90 days, disease free for 1 year. The CALLA positive.