Precursor B-lineage acute lymphoblastic leukemia patients with aberrant natural killer cell and T cell - lineage antigen expression: experience from a tertiary cancer care center

Abstract Introduction Flow cytometric immunophenotyping (FCI) plays a major role in diagnosing hematologic malignancies. In patients diagnosed with precursor B-lineage acute lymphoblastic leukemia (B-ALL), expression of certain non-lineage/cross lineage antigens is of prognostic and cytogenetic relevance. There is a paucity of studies that have comprehensively analyzed the clinical and laboratory profiles of B-ALL patients showing aberrant T/natural killer (NK) cell antigen expression. Materials and methods This is a prospective study where 152 consecutive B-ALL patients were analyzed for aberrant expression of T/NK cell antigens (CD1a, CD5, CD4, CD7, CD8 and CD56) by FCI. The clinical and laboratory profile of these T/NK-cell antigen-expressing B-ALL patients was statistically analyzed against conventional B-ALL patients. Results In our B-ALL cohort, CD5, CD7 and CD56 expression were observed in one, six and nine patients, respectively. CD56-expressing B-ALL patients were predominantly children (89%) and presented as standard clinical risk (p = 0.010) disease with frequent ETV6-RUNX1 fusion (p = 0.021) positivity. On the contrary, CD7-expressing B-ALL patients were adolescent-young adult/adult-age skewed (83%) and had an adverse cytogenetic profile (p = 0.001), especially for the frequent presence of BCR-ABL1 fusion (p = 0.004) and KMT2A rearrangement (p = 0.045). CD7-expressing B-ALL patients had inferior event-free survival (p = 0.040) than their CD56-expressing counterparts, but there was no significant difference in the overall survival (p = 0.317). Conclusion In comparison to conventional B-ALL patients, there are significant differences in the age, cytogenetic profile and event-free survival of T/NK-cell antigen-expressing B-ALL patients.

Myeloid/natural killer cell precursor and myeloid/natural killer cell acute leukemia / 中国实验血液学杂志

With the popularity of flow cytometry, the classification of leukemia become more detailed. Myeloid/natural killer cell precursor acute leukemia and myeloid/natural killer cell acute leukemias are generally recognized as two kinds of rare leukemias and have poor prognosis. The cells expressed both myeloid and lymphatic antigens in these two leukemia and can not be diagnosed by morphology. The only basis to make a definite diagnosis is their unique Immunophenotyping. The role of CD7 and CD56 in these two leukemia are compelling, in the other hand, as the progress of cell differentiation research, there are many new awareness of NK cell differentiation. In this article, the biological origin, clinical manifestation, diagnosis, treatment and the role of CD7 and CD56 in these two leukemia are briefly summarized.

Loss of pan-T cell antigens CD2, CD3, CD5 and CD7 in Kikuchi's disease / 中华病理学杂志

<p><b>OBJECTIVE</b>To study the possible loss of pan-T cell antigens CD2, CD3, CD5 and CD7 in Kikuchi's disease and to evaluate the role of T cell antigen loss in distinguishing benign from malignant T-cell lymphoid lesions.</p><p><b>METHODS</b>Formalin-fixed and paraffin-embedded tissues of 33 cases of Kikuchi's disease and 15 cases of reactive lymphoid hyperplasia were studied by EliVision immunohistochemical staining for CD2, CD3, CD5 and CD7.</p><p><b>RESULTS</b>Twenty-four of the 33 (72.7%) cases of Kikuchi's disease lost one or more of the pan-T cell antigens, including the loss of CD5 only (13 cases), CD7 only (1 case), CD2 only (1 case), CD2 and CD7 (2 cases), CD5 and CD7 (4 cases), CD2 and CD5 (2 cases), and CD2, CD7 and CD5 (1 case). Amongst these cases, the commonest antigen loss was CD5 (20 cases, 60.6%), followed by CD7 (8 cases, 24.2%) and CD2 (6 cases, 18.2%). Compared with the xanthomatous subtype of Kikuchi's disease, the loss of antigens was more commonly seen in the proliferative and necrotizing subtypes. Analysis of follow-up data showed that the loss of antigens in Kikuchi's disease was not significantly associated with the prognosis. In reactive lymphoid hyperplasia, the expression of CD2, CD3, CD5 and CD7 was seen in all cases with similar intensity, with no obvious pan-T cell antigen loss.</p><p><b>CONCLUSION</b>Loss of one or more pan-T cell antigens in Kikuchi's disease is demonstrated in present study, suggesting that the immunophenotypic pattern is not unique in T cell lymphoma.</p>

Significance of TCR gene clonal rearrangement analysis in diagnosis of mycosis fungoides / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To investigate the significance of detecting TCR gene clonal rearrangement in the diagnosis of mycosis fungoides (MF) and to optimize the primers used for detecting the TCR gene clonal rearrangement with PCR in paraffin embedded tissues of MF.</p><p><b>METHODS</b>Nineteen cases of MF were enrolled into the study. A panel of 10 antibodies were used for immunophenotypic analysis and polymerase chain reaction for TCR-γ and TCR-β gene rearrangement detection in this study.</p><p><b>RESULTS</b>TCR gene clonal rearrangements were detected in all 19 cases, in which 84.2% cases (16/19) had TCR-γ gene clonal rearrangements. The positive rates of the primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-γ were 47.4%, 78.9% and 31.6%, respectively. The positive rate of V(2-5)/V(8-12)/JGT(1) was statistically significantly higher than that of T(VG)/T(JX) and BIOMED-2-TCR-γ (P < 0.05). No TCR gene clonal rearrangement was detected using the primers V(γ11)/V(γ101)/Jγ12 and V(γ11)/V(γ101)/J(p12). TCR-β gene clonal rearrangement was detected in 31.6% (6/19) cases.</p><p><b>CONCLUSIONS</b>TCR gene clonal rearrangement analysis is a useful tool in the diagnosis of MF and TCR-γ gene is a good target gene for the detection. The primers T(VG)/T(JX), V(2-5)/V(8-12)/JGT(1) and BIOMED-2-TCR-γ can be used in clinicopathologic detection for TCR gene clonal rearrangement and V(2-5)/V(8-12)/JGT(1) may be the first choice.</p>

Significance of myeloid antigen expression in precursor T lymphoblastic lymphoma / 癌症

<p><b>BACKGROUND AND OBJECTIVE</b>Precursor T lymphoblastic lymphoma (T-LBL) is a highly aggressive lymphoma. Myeloid antigen expression was found in some of the patients, and its clinical significance is worth studying. This study was to compare the clinical features, short-term efficacy and survival of T-LBL patients with or without myeloid antigen expression so as to evaluate its prognostic significance.</p><p><b>METHODS</b>Forty-five T-LBL patients, with a median age of 14 years, were treated at Sun Yet-sen University Cancer Center between January 2000 and July 2008. These patients were divided into myeloid antigen-positive group (My(+) group) and myeloid antigen-negative group (My(-) group) based on the flow cytometric (FCM) analysis in bone marrow or pleural fluid. Myeloid antigen expression and its correlation with the short-term efficacy and overall survival were assessed in the two groups.</p><p><b>RESULTS</b>There were 18 patients (40.0%) in the My(+) group and 27 (60.0%) in the My(-) group. The myeloid antigen expression was negatively correlated with the initial level of lactate dehydrogenase (LDH), but not with other clinical features. The remission rate was lower in the My(+) group than in the My(-) group (38.8% vs. 70.3%, P = 0.028). The 2-year overall survival rate was lower in the My(+) group than in the My(-) group (51.9% vs. 78.7%, P = 0.036). By age subgroup analysis, there were no differences in response and survival rate among children and adolescents with or without myeloid antigen expression. But the remission rate and the 2-year overall survival rate were significantly lower in adult patients with myeloid antigen expression than in patients without it. Univariate and multivariate analysis demonstrated that age and myeloid antigen expression were adverse prognostic factors.</p><p><b>CONCLUSION</b>Myeloid antigen expression is a predictor of a poor response to chemotherapy, and adverse prognostic factor in adult T-LBL, but not in children with T-LBL.</p>

Clinicopathologic study of 128 cases of T-lymphoblastic lymphoma/leukemia / 中华病理学杂志

<p><b>OBJECTIVE</b>To clarify clinical and morphological features and immunophenotype of T lymphoblastic lymphoma/leukaemia (T-LBL/ALL) and to further improve the knowledge and diagnostic accuracy for T-ALL/LBL.</p><p><b>METHODS</b>128 cases of T-LBL/ALL were analyzed for the clinical features, morphology, immunophenotype and TCR gene rearrangement using routine eosin and haematoxylin stain, immunohistochemistry and polymerase chain reaction combining with the clinical findings.</p><p><b>RESULTS</b>In 128 cases of T-LBL/ALL, there were 94 male and 34 female. The ratio of male/female was 2.8:1. The age of patients ranged from 4 to 88 years, with an average of 27 years and a median of 22 years. Lymph nodes and extranodal areas were involved in 58/128 and 27/128 cases of T-LBL/ALL, respectively. The other 43 cases had involvement of both nodal and extranodal areas. Cervical node and mediastinum were involved in 74 cases and 43 cases, respectively. Diffuse growth pattern of tumor cells was predominant. Nodular growth pattern was seen only in a few cases. Most cases composed of small to medium-sized lymphoblasts, and other 7 cases showed a composition of large lymphoblasts. Tumor cells expressed TdT in 121/128 (94.5%) cases, CD34 in 48/98 (49.0%) cases, CD3 in 78/108 (72.2%) cases, CD7 in 104/108 (96.3%) cases, CD43 in 56/63 (88.9%) cases, CD79a in 5/70 (7.1%) cases, CD10 in 25/76 (32.9%) cases, CD99 in 58/60 (96.7%) cases and Pax-5 in 4/91(4.4%) cases. All of the cases were negative for MPO. A follow up data, ranging from 1 to 53 months, was obtained in 51/128 (39.8%) patients. The over all survival rate was 68.6% and the median survival time was 12 months. Under a similar condition of carrying a positive staining result on CD3 in tumor cells, there was a statistically significant difference between patients in the group of over 30 of age and that with the age ranging from 11 to 30. Patients associating with a CD10 positive staining of tumor cells showed also a shorter survival period. In addition, there were 4 out of 5 cases showing the presence of TCR gene rearrangement.</p><p><b>CONCLUSIONS</b>T-LBL/ALL are aggressive in behavior, associating mainly with enlarged cervical lymph nodes and masses in the mediastinum, occurring predominantly in children and young adults. Although small to medium-sized tumor cells with diffuse pattern were found in most cases, however, large-sized tumor cells and nodular pattern could also be obtained in a few cases. Immunohistochemistry staining particularly adoption of CD7, Pax-5, TdT, CD34 and Ki-67 stainings in combination are helpful of making a diagnosis for T-LBL/ALL. Analysis of TCR gene rearrangement will be helpful for the diagnosis of a few difficult cases.</p>

Expression of 5 genes in CD7 positive acute myeloid leukemia stem/progenitor cells from bone marrow / 中国实验血液学杂志

This study was aimed to investigate abca5, mdr-1, kdr, dapk and irf-1 expressions in leukemia stem/progenitor cells (LSC) from CD7 positive acute myeloid leukemia, the expression of these 5 genes in mononuclear cells (MNC) from 15 normal bone marrow (NBM) and 16 AML patients bone marrow (AML BM) specimen were detected by real-time quantitative PCR (RQ-PCR). CD34(+)CD38(+) progenitor cells and CD34(+)CD38(-)Lin(-) stem cells were sorted by flow cytometry (FCM) from the MNCs of 10 NBM and 21 AML BM specimen. These 5 gene expressions in the sorted cells were detected by small amount cell RQ-PCR. The results showed that these 5 genes above mentioned all expressed in NBM-MNC, in which the expression levels of irf-1 and dapk were highest with the relative expression levels 4.08 and 3.86, the expression levels of abca 5 and mdr-1 were in the middle with the relative expression 0.49 and 0.84 respectively, the kdr expression was lowest with the relative expression level 0.02. In CD34(+)CD38(+) progenitor cells, the expression level of kdr increased dramatically (p < 0.05) while irf-1 and dapk dramatically decreased (p < 0.05). There was no obvious change of expression in the rest 2 genes. In CD34(+)CD38(-) stem cells the expression level of these 5 genes all increased nearly 2 times as much as that in CD34(+)CD38(+) progenitor cells, but kdr increased 3 times as much, and the increase of kdr and irf-1 expressions was of statistical significance (p < 0.05). Compared with the NBM, expression levels of 5 genes in AML-MNC decreased, and out of them abca 5, mdr-1, kdr and dapk were decreased most remarkably (p < 0.05). Comparison between AML CD34(+)CD38(+) cells and AML MNC showed that the expression level of irf-1 and dapk were decreased dramatically (p < 0.05) while the rest 3 genes increased their expression with statistical significance (p < 0.05). The expression levels of these 5 genes were higher in CD34(+)CD38(-) cells than those in CD34(+)CD38(+) stem cells, and the increase of kdr and irf-1 expressions showed statistical difference (p < 0.05). These 5 genes expression levels were all higher than those in CD34(+)CD38(+) cells whether in AML CD34(+)CD38(-)CD7(+) cells or CD34(+)CD38(-)CD7(-) cells. The increase of kdr expression in CD34(+)CD38(-)CD7(+) cells as well as kdr and irf-1 expressions in CD34(+)CD38(-)CD7(-) cells were all of statistical significance (p < 0.05). In conclusion the expression level of kdr in NBM was highest in stem cells while dapk and irf-1 were highest in differentiated cells. The expression levels of these 5 genes in CD34(+)CD38(-)Lin(-) stem cells were higher than those in CD34(+)CD38(+) progenitor cells. The gene expressions in AML CD34(+)CD38(-)CD7(+) cells and CD34(+)CD38(-)CD7(-) cells are in accordance with the characteristics of stem cells.

Frequent CD7 Antigen Loss in Aggressive Natural Killer-Cell Leukemia: A Useful Diagnostic Marker / 대한진단검사의학회지

BACKGROUND: Aggressive natural killer-cell leukemia (ANKL) is a rare neoplasm characterized by systemic proliferation of NK cells. However, the differential diagnosis of NK lymphoproliferative disorders is difficult because of the absence of a distinct diagnostic hallmark. Therefore, to identify diagnostic markers for ANKL, we analyzed the clinical data and laboratory findings obtained for 20 patients with ANKL. METHODS: From January 2000 to July 2007, 20 patients were diagnosed with ANKL on the basis of bone marrow studies. We retrospectively analyzed the clinical features and laboratory findings, including the complete blood count, Epstein-Barr virus status, immunophenotype, and the cytogenetic results. RESULTS: The subjects included 6 women and 14 men (median age, 44 yr; range, 2-70 yr). Cytogenetic studies were performed in 18 patients, and karyotypic abnormalities were observed in 9 patients (50%). None of the cytogenetic abnormalities were constantly observed in all the patients. However, 6q abnormalities were observed in 4 patients (4/18, 22%). The immunophenotype of the leukemic NK-cells was cytoplasmic CD3+, surface CD3-, CD16/56+, CD2+, and CD5-. Notably, the CD7 antigen was absent in 10 patients (50%). When the CD7 loss was combined with cytogenetic abnormalities, clonal markers could be identified in 75% of the ANKL cases. CONCLUSIONS: The CD7 antigen loss was frequently observed in our series of ANKL patients. In conjunction with the cytogenetic findings, this characteristic immunophenotypic finding can serve as a reliable marker for the timely diagnosis of ANKL. Therefore, immunophenotypic analysis of CD7 expression should be included in the diagnosis of NK cell neoplasms.

Clinical and laboratory features of patients with CD34(+) acute promyelocytic leukemia / 中华肿瘤杂志

<p><b>OBJECTIVE</b>To explore the expression of CD34 in patients with acute promyelocytic leukemia (APL) and investigate the clinical and laboratory features of CD34(+) APL patients.</p><p><b>METHODS</b>262 APL patients diagnosed by chromosome analysis and/or fusion gene examination in the last five years were retrospectively analyzed in this study. To survey the expression of CD34 in those patients, all the cases were divided into two groups (CD34(+) APL vs. CD34(-) APL). The clinical features including age, gender, abnormal values of the peripheral hemogram before treatment, the complete remission (CR) rate and the incidence of DIC and laboratory data such as the results of morphology, immunology, cytogenetics and molecular biology (MICM) between those two groups were compared.</p><p><b>RESULTS</b>Of the 262 APL patients, 38 (14.5%) cases were positive for CD34 expression. There were no statistically significant differences between CD34(+) APL and CD34(-) APL groups in gender and age (P > 0.05). Before treatment, the median level of WBC in CD34(+) APL was 25.92 x 10(9)/L, which was significantly higher than that of CD34(-) APL (5.3 x 10(9)/L, P < 0.05). CD34(+) APL by morphology classification were mostly of the subtypes M3b and M3v (65.8%), while these subtypes in CD34(-) APL (40.3%) were significantly less (P < 0.01). There were no statistically significant differences between the two groups compared in respect of complete remission (CR) rate and the incidence of DIC (P > 0.05). The expression level of CD34 in APL had correlation to the expression level of CD2, CD7 and CD117; the latter three phenotypes in CD34(+) APL were significantly higher than those in CD34(-) APL (P < 0.01). No significant difference was found between those two groups by chromosome analysis, but there was more PML-RAR-alpha transcript short form in CD34(+) APL than that in CD34(-) APL (P < 0.05).</p><p><b>CONCLUSION</b>CD34(+) acute promyelocytic leukemia is a unique subtype of APL with different biological characteristics.</p>

CD7 expression in differentiating mycosis fungoides from benign cutaneous lymphocytic infiltrates

Diagnosis of the early phase of mycosis fungoides [MF] is sometimes difficult. Loss of CD7 expression is considered a distinguishing characteristic of MF. The aim of this study was to determine the range of CD7 expression in MF and compare the results with benign inflammatory dermatosis and equivocal cases of possible MF. During a period of 30 months, we examined 15 patients with MF, 12 patients suspicious for MF, and 15 patients with benign inflammatory dermatosis. The slides stained by H andE were reviewed by two pathologists. Immunostaining was done for CD43, CD3, CD5, CD7, and CD20 on paraffin embedded tissues. All the patients in MF group showed absence of CD7 expression in epidermotropic mycosis cells. Compared with benign inflammatory dermatosis, patients with MF had significantly lower CD7 expression in the dermal infiltrate [P< 0.0001]. In patients with MF, the mean CD7 was significantly lower than CD43, CD3, and CD5 [P=0.001]. The mean CD7 count of parapsoriasis was significantly higher than MF [P= 0.01]. The mean CD7 count of parapsoriasis was significantly lower than benign inflammatory dermatosis [P=0.016]. The lowest mean CD7 counts were found in patch stage of MF. Low CD7 expression < 10% lymphocytes had sensitivity and positive predictive values of 75% and 100% and specificity and negative predictive values of 100% and 83.3% for the diagnosis of patch stage of MF. Minimal expression of CD7 is a specific finding for patch stage of MF. Benign inflammatory dermatosis can also show low expression of this marker, but rarely matches that of patch stage of MF

The immunophenotypic analysis of CD7+ and (or) CD56+ acute myeloid leukemic stem cells and its applications in minimal residual disease detection / 中华血液学杂志

<p><b>OBJECTIVE</b>To explore the immunophenotypic characteristics of CD7 and/or CD56 positive acute myeloid leukemic stem cells, and the relationship between minimal residual disease (MRD) and the leukemic stem cells (LSC).</p><p><b>METHODS</b>The immunophenotype of leukemia cells from 51 CD34+ CD38+ CD7+ and/or CD34+ CD38+ CD56+ acute myeloid leukemia (AML) patients (exclude M3) at diagnosis was analyzed by using 4 - 6 panels of 4 color antibodies, and cells from 28 normal bone marrow (NBM) samples were served as control. The expression of CD7 and CD56 in the CD34+ CD38+ subpopulation was used as a leukemic cell marker for monitoring MRD of 53 samples from 26 CD7+ and (or) CD56+ patients.</p><p><b>RESULTS</b>In CD7+ and/or CD56+ AML patients at diagnosis, the average positivity of CD7 in CD34+ CD38+ subpopulation and CD34+ CD38- Lin- stem cells subpopulation was (77.39 +/- 20.71)% and (44.57 +/- 22.70)%, and that of CD56 was (56.71 +/- 32.56)% and (33.51 +/- 29.64)%, respectively, all significantly higher than that of NBM (P < 0.01 and P < 0.05). Compared with that of NBM, the expression of CD90 in AML patients was significantly lower in the CD34+ CD38- Lin- subpopulation (P < 0.01), the expression of CD123 was significantly higher than NBM (P < 0.01), and the expression of CD117 was no significant difference (P > 0.05). In follow up of CD7+ and (or) CD56+ patients, the expression rate of CD7 and (or) CD56 in the CD34+ CD38- Lin- subpopulation MRD+ group was significantly higher than that in the MRD- group. The actual rate for CD7 was 71% (15/21) and 16% (4/25) (P = 0.001), and its relative rate was 81% (17/ 21) and 24% (6/25) (P = 0.000), respectively. The actual rate of CD56 is 100% (4/4) and 12% (3/25) (P = 0.001), and its relative rate was 75% (3/4) and 20% (5/25) (P = 0.031), respectively. A high CD7+ CD34+ CD38- Lin- subpopulation frequency at diagnosis in CD7+ AML patients predicted a high frequency of positive MRD in later detection (P < 0.05).</p><p><b>CONCLUSIONS</b>CD7 and CD56 are expressed on the stem cells in CD7+ and/or CD56+ AMLs and a high frequency of CD7 and CD56 in the CD34+ CD38- Lin- stem cell subpopulation predicts a high frequency of positive MRD in later detection.</p>

Clinical study on prognosis of acute leukemia subtypes Ly + AML and My + ALL / 中国实验血液学杂志

The purpose of this study was to investigate the prognosis of acute myelogenous leukemia (AML), acute lymphoblastic leukemia (ALL), lymphoid antigen-positive acute myeloid leukemia (Ly + AML), myeloid antigen-positive acute leukemia (My + ALL) and biphenotypic acute leukemia (BAL). Immunophenotyping was performed on medullary specimens of 197 acute leukemia (AL) patients by using three-color flow cytometry analysis and CD45/SSC gating. The scoring systems proposed by EGIL was adopted to classify the AL patients into five groups: 43 of ALL, 53 of AML, 53 of My + ALL, 39 of Ly + AML and 9 of BAL patients. The results showed that in Ly + AML, CD7 was the most common (53.8%) as compared to other lymphoid markers, however, in My + ALL CD13 was the most common (47.2%) as compared to other myeloid markers. Compared with Ly + AML, My + ALL had higher incidences of enlargement of liver, spleen and lymphnodes significantly (P<0.05). As for the case numbers of WBC counts > 100 x 10(9)/L, the positive rate of CD34 and the complete remission rate there was no obvious difference between groups of Ly + AML and My + ALL (P>0.05). As for incidences of enlargement of liver, spleen and lymphnodes, the case numbers of WBC counts > 100 x 10(9)/L, the positive rate of CD34 and complete remission rate, no obvious difference was found between ALL and My + ALL (P>0.05). Compared with AML, Ly + AML had lower complete remission rate significantly (P<0.05). As for incidences of enlargement of liver, spleen and lymphnodes, the case numbers of WBC counts > 100 x 10(9)/L and the positive rate of CD34, no obvious difference was found between AML and Ly + AML (P>0.05). Compared with Ly + AML and My + ALL, BAL showed no significant difference in complete remission rate (P>0.05) because the number of BAL patients was too small. It is concluded that since Ly + AML has lymphoid markers, and the prognosis of Ly + AML is worse than AML, the clinical therapy for Ly + AML should contain both AML and ALL. Though My + ALL had myeloid markers, no significant difference was found between My + ALL and ALL, it might be supposed that their therapy could be the same.

Immunologic characteristics and prognosis of acute myeloid leukemia M1 / 中国实验血液学杂志

The study was aimed to investigate the immunological characteristics and prognosis of acute myeloid leukemia (AML) M(1) and to find the main points in immunology to differentiate AML M(1) from M(2), and M(1) from ALL (proB, preB, T). Immunophenotyping was performed in 41 AML M(1) patients by three-color flow cytometry analysis using CD45/SSC gating, meanwhile the cytogenetic analysis was performed in 17 patients. 51 newly diagnosed AML M(2) patients and 58 newly diagnosed ALL patients were used as control at the same time. The results showed that the positive rate of CD33 in M(1) was 100%, which was high in sensitivity, but low in specificity; the positive rate of CD11b, CD15, MPO, CD117 in M(1) were significantly lower than that in M(2) (p < 0.05); the positive rate of T-lineage antigen in Ly + AML M(1) was higher than that in M(2) (p < 0.05); compared with ALL ProB, M(1) had high expression of HLA-DR, simultaneously myeloid antigen CD13, CD15, CD33, CD117, MPO and T-lineage antigen CD4, CD7 were all highly expressed (p < 0.05); compared with ALL PreB, M(1) had high expression of HLA-DR, CD34, meanwhile myeloid antigen CD13, CD15, CD33, CD117, MPO and T-lineage antigen CD4, CD5 were all highly expressed (p < 0.05); as compared with T-ALL, the early-phase antigen HLA-DR, CD34, myeloid antigen CD13, CD15, CD33, CD117, MPO of M(1) were all significantly highly expressed (p < 0.05). In M(1), the complete remission (CR) rate in patients with CD7 positive had no statistical difference from that in patients with CD7 negative (p > 0.05); the CR rate of patients with CD34 positive had no statistical difference from that of patients with CD34 negative (p > 0.05); CR rate in M(1) was lower than that in M(2) (p < 0.05), time to reach CR was longer, the incidence of hyperleukocytic acute leukemia was higher (p < 0.05), CR rate in hyperleukocytic acute leukemia was lower (p < 0.05). It is concluded that the myeloid antigen CD33, CD13 in M(1) are highly expressed, early-phase antigen HLA-DR in M(1) is also highly expressed, but the myeloid antigen CD11b, CD15, MPO, CD117 in M(1) are lowly expressed, T-lineage antigen CD4, CD7 in M(1) are highly expressed in the meantime. There is no definite characteristic marker in immunology to differentiate M(1) from M(2), but as the positive rate of CD11b, CD15, MPO, CD117 in M(1) are significantly lower than that of M(2), CD11b, CD15, MPO, CD117 can be used as reference indicators to differentiate M(1) from M(2). AML M(1), ALL ProB, ALL PreB and T-ALL, which are difficult to differentiate in morphology can be well seperated through the analysis of immunological phenotype. CD117 is mainly expressed in AML, which is useful for the differentiation diagnosis between AML and ALL. The prognosis of M(1) is worse than that of M(2).

Immunologic classification used in typing of 68 cases of acute leukemias / 中国实验血液学杂志

To evaluate the significance of immunologic classification for typing of acute leukemia (AL). 68 cases of AL were classified by morphologic and immunologic typings. The results showed that the consistency rate was 94.1% between morphology and immunology, and 4 morphologic misdiagnosed cases were corrected by immunology; CD13 and CD33 were special myeloid lineage-associated antigens; AML-M(3) was often CD34 low-expressed and HLA-DR-negative; CD14 was often expressed in AML-M(4) and M(5); lymphoid lineage-associated antigens (CD7) were easily found in ANLL, and myeloid lineage-associated antigens were also found in ALL. In conclusion, immunologic classification can improve the accuracy in acute leukemia diagnosis. The diagnosis of some special AL, such as acute unidentified leukemia (AUL), AML-M(0) and so on, must rely on immunologic classification.

Study of caspase-3 level in relation to prognostic risk factors in children with acute lymphoblastic leukemia

Apoptosis refers to a biochemically regulated process of automated cell death mediated through a highly organized network of interacting proteases. Malignant transformation of hematopoietic progenitors into leukemic cells results in defects in the cell cycle regulation; including defects in apoptosis. Multiple mechanisms may account for why leukemic cells become resistant to chemotherapy. One common cause includes defects in the cell inherent programmed cell death [apoptosis]. This study was conduced on 20 children with newly diagnosed ALL and 10 controls. Our aim was to evaluate the level of caspase-3 among them and relate it to prognostic factors. It can be concluded that caspase-3 level after induction therapy proved to be a significant predictor of remission stage especially for B-ALL. CD10 positive cases also showed a significant difference between caspase-3 level before and after induction therapy. The same was demonstrated with low risk cases. We can also conclude that failure of caspase-3 level to increase after induction is associated with poor prognosis

CD34+ antigen expression relating to prognosis in acute myeloid leukemia / 中国实验血液学杂志

To explore CD34(+) antigen expression in new diagnosed acute myeloid leukemia (AML) and analyze the prognosis for CD34(+) AML patients, the expression of antigen CD34 in 238 AML patients was detected by indirect immunofluorescence assay. The results showed that CD34 in 92 out of the 238 patients (38.7%) were positive, there was relationship between the CD34(+) expression and FAB subtypes (M(0), M(1)), and no CD34(+) expression was observed in M(3) subtypes. The complete remission rate of CD34(+) AML patients was 32%, which was lower than that of CD34(-) AML (61%). The lymphoid-associated antigen (CD7) was significantly increased in CD34(+) AML patients, compared with CD34(-) patients (P < 0.05). It is concluded that CD34(+) AML patients show poor prognosis and lower CR rate. The detection of CD34 expression is of some value in predicting prognosis in AML.

Evaluation of CD[7] T-cell deletion marker for diagnosis of mycosis fungoides

Clinical diagnosis of mycosis fungoides [MF] in its early stages can be difficult and requires biopsy confirmation of disease. Even with histological evaluation, early-stage MF is still difficult to distinguish from various benign inflammatory dermatoses [BID]. Recent attempts to enhance the diagnostic sensitivity and specificity have mainly focused on lymphocyte Immunophenotyping and genotyping [1, 2]. Our purposes in this study were clinical and histopathologic assessment of MF patients in addition to evaluate the diagnostic value of Immunophenotyping with special reference to CD7 deletion. The study was carried on 19 MF patients and 16 cases of BID as a control group. They were subjected to full clinical examination, routine laboratory investigations and chest x- ray. Skin biopsies were obtained from all MF cases and BID control group. The paraffin embedded skin specimens were stained with hematoxylin-eosin stain [H and E] and immunostains for CD3, CD4, CD8 and CD7. According to Smoller's [3] histological criteria, the diagnosis of MF was confirmed; showing various pathologic patterns including granulomatous MF in one case. Immunophenotyping results revealed that T-cells in all MF and BID specimens were predominantly expressed CD3+, CD4+ and negatively stained by CD8. Conversely, the mean CD7+ count as a percentage of total T-cells in MF specimens [16.8%] was significantly lower than those of BID [61.7%]. The sensitivity and specificity of CD7 deletion [expressed by <50% of T-cells] for diagnosis of MF was 89.5% and 93.75% respectively. CD4/CD8 ratios greater than 2:1 and 10:1 were noticed in 73.7% and 31.3% of MF biopsies. CD4/CD8 ratios more than 10:1 were specific for MF while ratios greater than 2:1 were also found in 15.8% of BID. We can conclude that MF may manifest a variety of histological forms. Expressions of conventional T-cell markers including CD3, CD4 and CD8- do not generally distinguish benign T-cell infiltrate from MF, however, deletion of CD7 [<50%] has demonstrated considerable usefulness in the diagnosis of MF [89.5%] and was occasionally found in BID [6.25%]

Study on the immunophenotypes of bone marrow cells from patients with myelodysplastic syndromes and its clinical implications / 中国实验血液学杂志

The phenotypes of the bone marrow cells in various subtypes of myelodysplastic syndromes (MDS) and its clinical implication were explored. The antigen expression of a panel of antigens expressed in marrow cells from 30 patients with subtypes of MDS was assayed by alkaline phosphatase anti-alkaline phosphatase method. The results showed that the expression of myeloid antigens appeared abnormality, CD13 and CD33, found on granulocyte and macrophage precursors, increased, and CD15 decreased. There were no significant changes for monocytic antigen CD14 and lymphoid antigens CD7 and CD10. CD34 was increased in RAEB/RAEB-t and was not increased in RA/RAS patients. CD71, expressed by erythroblast and proliferative cells, was higher in all subtypes of MDS than that in control group. It is suggested that the bone marrow cells from MDS patients showed abnormality of more than two series of immunophenotypes, detection of immunophenotype in MDS cells might be contributed to the diagnosis and predicting prognosis.

Flow cytometric immunophenotyping in fine-needle aspiration of lymph nodes

Fine-needle aspiration (FNA) of lymph nodes has been regarded as a useful method in the diagnosis of lymphadenopathy. However, this procedure has been shown to be of limited value in the diagnosis of low or intermediate grade malignant lymphomas in some studies. Immunophenotyping is an essential adjunct to cytomorphology for the diagnosis of lymphoma by FNA. Immunophenotyping using flow cytometry (FCM) is rapid, objective and reliable. Using FCM, multiparametric analysis of 33 FNA materials from lymph nodes was performed and profiles of surface markers of lymphoid cells were assessed. In reactive hyperplasia, patterns of cell surface markers were quite variable, but disclosed polyclonality. Most of the B-cell lymphomas showed immunophenotypes for B-cell lineages with their kappa: lambda or lambda: kappa ratio being over 3:1. In T-cell lymphomas, T-cell surface markers were predominantly expressed as well. In conclusion, our results suggest that immunophenotyping of lymph node aspirates is a valuable diagnostic adjunct for lymphoproliferative disorders, particularly in B-cell lymphomas because immunophenotyping can be easily and adequately performed by FCM.