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PubMedDate not supplied

The immunophenotype of blood and cerebrospinal fluid mononuclear cells in dogs.

Inflammatory neurologic diseases are common in dogs, but establishing a definitive diagnosis often is difficult. Nucleated cell number and type in cerebrospinal fluid (CSF) rarely are suggestive of an etiologic agent. We speculated that CSF leukocyte immunophenotyping would be a useful adjunct in the investigation of canine inflammatory neurologic diseases by yielding more specific etiologic information. The goals of this study were to establish the feasibility of flow cytometric evaluation of individual canine CSF samples and to identify the cell distribution in healthy dogs. The mononuclear cell populations of paired blood and CSF samples from 23 healthy dogs were characterized by labeling of cells with antibodies against CD4, CD8alpha, CD21, and CD14 molecules and by flow cytometric analysis of their expression. The mean proportion of CD4+ and CD21+ cells was significantly higher in blood than in the CSF (P < .002 and P < .001, respectively). In contrast, the mean proportion of CD14+ and CD8a+ cells was not significantly different between blood and CSF (P = .5 and p = .9, respectively). These findings demonstrate differences in the distribution and function of mononuclear cells in the circulating venous and subarachnoid compartments in the dog.

Animals↗
PubMed1991

An improved method of alkaline phosphatase--antialkaline phosphatase immunophenotyping using microwave irradiation.

The application of microwave irradiation from a domestic microwave oven to the alkaline phosphatase-antialkaline phosphatase immunophenotyping has resulted in substantial time saving of the procedure. This has been achieved without compromising the quality of the final preparation. It can be carried out on peripheral blood smears, bone marrow smears and cytocentrifuge preparations on fresh slides, slides that have been stored at -20 degrees C wrapped in aluminium foil and unfixed smears left at room temperature for up to 8 days.

Alkaline Phosphatase↗
PubMedDate not supplied

Immunophenotyping pediatric leukemias in Kelantan, Malaysia.

This study reviewed the immunophenotyping results of children with acute leukemia in Kelantan, Malaysia. In the 3.5-year period (January 1994 to June 1997), 45 cases were identified. All children were under the age of 12 years and the predominant ethnic group was Malay. Thirty-six cases (80%) were acute lymphoblastic leukemia (ALL) and 9 cases (20%) were acute myeloblastic leukemia (AML). Of the ALL cases, 3% were of B-cell and 22% of T-cell origin, and 96% of the B-lineage ALL were CD10 positive. All the AML cases expressed CD33 and 78% were positive for CD13. The incidence of mixed-lineage leukemias was 13.8% for My+ ALL and 11.1% for Ly+ AML.

Bone Marrow Cells↗
PubMed2000

Aberrant immunophenotypes detected by flow cytometry in acute lymphoblastic leukemia.

The present study was designed to analyse the proportion of ALL patients in which the phenotypic detection of minimal residual disease (MRD) is feasible, based on the presence of aberrant phenotypes: lineage infidelity, asynchronous expression, overexpression and ectopic phenotype. For this purpose we have prospectively investigated the phenotype of blast cells from 25 patients at diagnosis using a large panel of monoclonal antibodies by multiparametric flow cytometry. The mean age was 23.3 +/- 17.3 with 10 children and 15 adults. 14 patients were classified as L1, 9 L2 and 2 L3 according to the FAB classification. 17 cases were B-lineage ALL and 8 T-ALL. 23 out of 25 cases (92%) included in this study displayed phenotypic aberrations at diagnosis (15 out of 17 cases of B-lineage ALL and all T-ALL patients). 76% of patients displayed two or more than two aberrancies. The phenotypic aberrations were lineage infidelity, found in 12 patients, asynchronous antigen expression detected in 17 patients, antigen overexpression in 4 patients and ectopic phenotype in 7 patients. In summary our results show that when a large panel of MoAbs is used for the immunophenotypical characterization of ALL, most patients display aberrant phenotypes, the coexistence of more than two aberrant antigen expressions being frequently detected. These results suggest that the use of immunological methods for the detection of MRD in ALL based on the existence of aberrant phenotypes could be of great help for the follow-up of patients in complete remission.

Adolescent↗
PubMedDate not supplied

Immunophenotype changes and loss of CD52 expression in two patients with relapsed T-cell prolymphocytic leukaemia.

T-cell prolymphocytic leukaemia (T-PLL) is an aggressive disease often resistant to conventional chemotherapy. Long lasting remissions with the monoclonal antibody CAMPATH-1H (anti-CD52) have been documented. We describe two unusual T-PLL patients treated successfully first with CAMPATH-1H in whom, at the time of relapse, the cells underwent a phenotypic switch with loss of CD52 expression. In one of them, cytogenetic analysis demonstrated the same chromosome abnormalities in the cells at diagnosis and relapse. The reasons for the immunophenotypic changes are unknown but it is likely that loss of CD52 antigen expression contributed to the resistance to CAMPATH-1H in one of the patients when re-treated.

Adult↗
PubMed1994

Peroxidase expression in acute unclassified leukemias: ultrastructural studies in combination with immunophenotyping.

The lineage affinity of 61 cases of acute unclassified leukemias (AUL) was reevaluated by ultrastructural analysis of peroxidase expression (POEM) in combination with immunophenotyping. In 24 cases a significant proportion of the blasts displayed ultrastructural myeloperoxidase (UMPO) and in another 3 cases platelet peroxidase (UPPO) suggesting the allocation of these leukemias to the myeloid or megakaryoblastic lineage. No significant correlation between myeloid surface marker expression and POEM positivity could be detected, while the presence of the CD 19 or CD 24 antigen significantly correlated with POEM negativity. The detection of lymphoid markers on POEM+ blasts indicates, that these leukemias might be derived from bipotential progenitors, which have retained their lymphoid antigens during myeloid differentiation. In one case a bilineage leukemia with a POEM+/CD 19- and a POEM-/CD 19+ population could be identified by immunoelectron microscopical studies (IEM). The remaining 34 AUL cases were POEM negative. Combined data suggest that these cases probably derive from early lymphoid progenitors. Taken together, AUL are heterogenous in presentation and in their cellular origin including a major portion of cases with an unequivocal myeloid differentiation detectable by ultrastructural analysis of myeloperoxidase expression.

Acute Disease↗
PubMed1994

Quality assessment for immunophenotyping of leukaemias and lymphomas: the Dutch experience. The SIHON Study Group.

Sending material at regular intervals to laboratories that perform immunophenotyping of leukaemias and lymphomas showed a large variety in the percentage of positive cells between the different laboratories but an agreement in diagnosis of more than 90%. Each laboratory used his own methods but antibody panels and guide-lines for interpretation were standardised. Studies are in progress to answer the question if standardisation in flow cytometry will result in more comparable results.

Flow Cytometry↗
PubMed1995

Leukemic transformation of primary myelofibrosis: immunophenotype, genotype and growth characteristics of blast cells.

Blast cells from six patients with leukemic transformation of primary myelofibrosis (PMF) were studied by morphology, immunophenotype and genotype as well as response to hematopoietic growth factors. The majority of the patients showed granulocytic or granulo-monocytic blasts, and only one had T lymphoid-monocytic blasts. None of the patients showed rearrangement of Ig or TCR genes, or the existence of the bcr-abl fused gene. A prominent growth response to GM-CSF and IL-3 was evident in all of the patients examined in liquid as well as semisolid cultures. The response to G-CSF was observed in four of the six patients in suspension culture, and in two of three patients in the clonogenic assay. Stem cell factor (SCF) was a weak growth stimulant, however the combination of this factor with GM-CSF or IL-3 was synergistically stimulatory. These results suggest that leukemic transformation of PMF occurs mainly at the level of myeloid stem cell, and that GM-CSF, IL-3, G-CSF and SCF are major growth factors for the blast cells in these cases.

Aged↗
PubMed1998

PreB1 (CD10-) acute lymphoblastic leukemia: immunophenotypic and genomic characteristics, clinical features and outcome in 38 adults and 26 children. The Groupe dEtude Immunologique des Leucémies.

The less differentiated stage (CD10-) of B-lineage acute lymphoblastic leukaemia (ALL) described as preB1-ALL in the GEIL nomenclature, accounts for less than 10% of ALL. It is classically considered to be associated with translocation (4;11)(q21;q23), and to have a poor prognosis. We report an extensive immunophenotypic, genomic and clinical study of a series of 64 preB-1 ALL patients, representing 6.3% of a cohort of consecutive ALLs. The engagement of preB1-ALL cells in the B-lineage was confirmed by their B-lineage score, equal to or higher than 2. In addition, more than 90% of the cases tested showed rearranged IGH genes. Translocation (4;11) was the most frequent karyotypic anomaly seen, but only accounted for 24% of the preB1-ALL cases tested. Expression of the myeloid associated antigen CD15 was also found with high incidence in this subset. Clinical and biological features at presentation showed more significant differences between preB1- and T-ALL than between preB1- and preB2-ALL (CD10+). However, outcome characteristics of the 22 children with preB1-ALL confirmed the worse prognosis of this entity.

Adolescent↗
PubMedDate not supplied

An algorithm for leukaemia immunophenotype pattern recognition.

Since leukaemia-specific leucocyte antigen has not been identified to date, the immunological diagnosis of leukaemia is achieved through the application of a wide set of monoclonal antibodies specific for surface markers on leukaemic cells. Thus, the interpretation of leukaemia immunophenotype seems to be a mathematically determined comparison of 'what we found' and 'what we know' about it. The objective of this study was to establish an algorithm for transformation of empirical rules into mathematical values to achieve proper decisions. Recognition of leukaemia phenotype was performed by comparison of phenotyping data with reference data, followed by scoring of such comparisons. Systematic scoring resulted in the formation of new numerical variables allocated to each state, whereas a most significant variable was described as a complex measure of compatibility. A system of recognized states was described by mathematical variables measuring the confidence of information systems, i.e. maximal, total and relative entropy. The entire algorithm was derived by matrix algebra and coded in a high-level program language. The list of the states recognized appeared to be especially helpful in differential diagnosis, occasionally pointing to states that had not been in the scientist's mind at the start of the analysis.

Algorithms↗
PubMedDate not supplied

Composition of the inflammatory infiltrate in pediatric penile lichen sclerosus et atrophicus (balanitis xerotica obliterans): a prospective, comparative immunophenotyping study.

Dermatopathological evaluation of pediatric preputial inflammatory disease rarely allows for specific diagnosis other than pediatric penile lichen sclerosus et atrophicus (balanitis xerotica obliterans, LSA/BXO). A prospective immunopathological study was performed on 20 consecutive, unselected, clinically and histopathologically confirmed LSA/BXO cases to determine the relative presence of T and B lymphocytes. There were seven cases with early stages of disease, eight with florid disease, and five with later stages of disease. Two ritual circumcision specimens and 12 specimens with non-LSA/BXO balanitis, collected during the same period, were used as controls. The infiltrate in LSA/BXO patients was wholly composed of T cells (positive with UCLH-1 antibody) in all cases. B cells (positive with L-26 antibody) were found only focally in small, discreet, easily recognizable (follicular or early follicle-like) aggregates, positioned slightly deeper than the band-like infiltrate of T cells. T cells were inconspicuous in 9 of the 12 control specimens. In the three other controls, T cells were much more obvious and these patients showed clinical features possibly suggestive of LSA/BXO in early, prediagnosable phases of development. We conclude that limited immunophenotyping may be a useful adjunct to diagnosis in pediatric cases in which only limited tissue is available or the disease may be more difficult to classify with confidence.

Adolescent↗
PubMed1991

Peripheral T-cell lymphomas--clinicopathologic and immunophenotypic analysis of 25 cases.

The clinicopathologic and immunophenotypic findings of 25 cases of peripheral T-cell lymphoma in Korea were analysed. Seventeen cases (68%) of the 25 T-cell lymphomas presented in the extranodal sites including the nasal mucosa, tonsil, oral cavity, skin and rarely bone, mediastinum and breast. Immunologic studies showed that 12 cases (48%) of the lymphomas were of T-helper phenotype, 5 cases (20%) were of cytotoxic/suppressor phenotype, 1 case (4%) expressed both helper and cytotoxic/suppressor markers, and 7 cases (28%) lacked detectable markers for subsets. Histologically, fourteen cases (56%) showed histologic features suggestive of peripheral T-cell lymphoma. The more frequently seen histologic types by Working Formulation (WF) included large cell type and immunoblastic type. Classification by WF was straightforward in most cases of large cell, immunoblastic type. However, with some cases of small cell, large cell and mixed types, there were problems fitting the morphology seen into the WF category. We hope that the establishment of a world wide immunologic and clinicopathologic classification for peripheral T-cell lymphoma will be made in the near future.

Adolescent↗
PubMed1992

Clinicopathologic and immunophenotypic study of non-Hodgkin's lymphoma in Korea. Lymphoreticular Study Group of the Korean Society of Pathologists.

This study sponsored by the Lymphoreticular Study Group of the Korean Society of Pathologists was carried out to provide nationwide data about the histopathologic-immunophenotypic features of malignant lymphomas in Korea. Two hundred and ninety Non-Hodgkin's lymphoma (NHL) among 312 malignant lymphomas collected from three representative areas in Korea were histologically reclassified. Two hundred and fifty three cases were immunohistochemically studied. T-cell lymphoma comprised 35.2% of NHL in this study and showed a quite comparable incidence to that of Japan and China, but it was much higher than in Western countries. A very low prevalence rate of the follicular variety (4.0%) and a higher propensity of primary extranodal involvement (60%) are additional characteristics of NHL in Korea. The most common histologic subtype of B cell lymphoma was diffuse large cell type, whereas the most common subtype of T cell lymphoma was diffuse mixed small and large cell type.

Adolescent↗
PubMed1992

A clinicopathologic study on the diffuse malignant lymphoma--a morphologic and immunophenotypic analysis in 62 patients at Harbor-UCLA Medical Center.

In order to compare the prognoses of patients with diffuse malignant lymphomas on the basis of histology and immunophenotypes, we retrospectively studied 62 cases of diffuse lymphoma arising in lymph nodes. We also evaluated the reactivity patterns of monoclonal antibodies (MoAb) LN1, LN2 and LN3 to determine the criteria for making a differential diagnosis in B cell lymphomas. The immunologic phenotypes were determined by the avidin biotin peroxidase complex method, using frozen or paraffin fixed tissues. The majority (66.3%) were B cell with the remaining 20.9% being T cell and 12.9% were non-B, non-T cell lineage. Immunological heterogeneity was found especially in the mixed small and large cell and the immunoblastic lymphomas. There was no significant difference between B- and T-cell lymphomas with respect to survival and death (P > 0.05). Histologically 79% (49/62) of the lymphoma was large cell and 21% (13/62), small cell lymphoma. There was a difference in prognosis between low, intermediate and high-grade of lymphomas. However there were no significant differences among the subtypes of the diffuse aggressive lymphomas. Factors associated with poor prognosis were advanced stages (P < 0.025) and histology of the malignant lymphomas. MoAb LN1, LN2 and LN3 gave positive staining in 83.3%, 91.7% and 60% of B cell lymphomas, respectively. The most common phenotypic pattern in B cell lymphomas was LN1+, LN2+, LN3+/-, suggestive of follicular center cell origin. As a panel, phenotypic patterns of MoAb LN1, LN2 and LN3 may be useful in differentiation of follicular center cell lymphoma from others.

Adolescent↗
PubMed2000

Differential diagnosis of malignant lymphomas and related disorders by specific pattern of expression of immunophenotypic markers revealed by multiparameter flow cytometry (Review).

The introduction of monoclonal antibodies (mAbs) for cell immunophenotyping and use of flow cytometry with the progressively improving software for multivariate analyses have revolutionized the diagnosis and influenced the classification of hematologic neoplasms. In this review we focus on the practical application of flow cytometry in the diagnosis and classification of malignant lymphomas and related lymphoproliferative disorders with special emphasis on differential diagnosis. A general approach to the utilization of flow cytometry (FC) in hematopathology with an algorithm to diagnose the most common neoplasms is presented. We discuss precursor B-cell neoplasms, mature B-cell neoplasms (SLL/CLL, mantle cell lymphoma, marginal zone lymphoma, hairy cell leukemia, diffuse large B-cell lymphoma, plasma cell dyscrasias and lymphomas with plasmacytic differentiation), precursor T-lymphoblastic leukemia and mature (peripheral) T-cell neoplasms, including T-SLL/PLL, anaplastic cell lymphomas and large granular cell leukemia/lymphoma. The text is accompanied by characteristic FC scatterplots of the discussed entities.

Antibodies, Monoclonal↗
PubMed2000

Utility of flow cytometry immunophenotyping for the diagnosis and classification of lymphoma in community hospital clinical needle aspiration/biopsies.

CONTEXT: Flow cytometry immunophenotyping (FC) of needle aspiration/biopsy (NAB) samples has been reported to be useful for the diagnosis and classification of lymphoma in university and cancer center-based settings. Nevertheless, there is no agreement on the utility of these methods. OBJECTIVE: To further define the utility of adjunctive FC of clinical NAB for the diagnosis and classification of lymphoma, and to determine if this approach is practicable in a routine clinical practice setting. SETTING: A community-based hospital. METHODS: Clinical NABs were submitted for adjunctive FC between June 1996 and September 1999 if initial smears were suspicious for lymphoma. Smears and cell block or needle core tissues were routinely processed and paraffin-section immunostains were performed if indicated. The final diagnosis was determined by correlating clinical and pathologic data, and the revised European-American classification criteria were used to subtype lymphomas. RESULTS: Needle aspiration/biopsies from 60 different patients were submitted for FC. Final diagnoses were lymphoma (n = 38), other neoplasm (n = 15), benign (n = 6), or insufficient (n = 1). For 38 lymphomas (20 primary, 18 recurrent), patients ranged in age from 32 to 86 years (mean, 62 years); samples were obtained from the retroperitoneum (n = 11), lymph node (n = 9), abdomen (n = 8), mediastinum (n = 6), or other site (n = 4); and lymphoma subtypes were indolent B-cell (n = 20; 2 small lymphocytic, 14 follicle center, 4 not subtyped), aggressive B-cell (n = 14; 3 mantle cell, 10 large cell, 1 not subtyped), B-cell not further specified (n = 2), or Hodgkin disease (n = 2). For the diagnosis of these lymphomas, FC was necessary in 20 cases, useful in 14 cases, not useful in 2 cases, and misleading in 2 cases. Thirty-two of 36 lymphoma patients with follow-up data received antitumor therapy based on the results of NAB plus FC. CONCLUSIONS: Adjunctive FC of NABs is potentially practicable in a community hospital, is necessary or useful for the diagnosis and subtyping of most B-cell lymphomas, and can help direct lymphoma therapy. Repeated NAB or surgical biopsy is necessary for diagnosis or treatment in some cases.

Adult↗
PubMed2001

Goblet cell carcinoids of the appendix: immunophenotype and ultrastructural study.

BACKGROUND: Goblet cell carcinoids of the appendix are rare neoplasms with uncertain biological behavior. OBJECTIVE: The aims of our study were to evaluate the immunophenotype of this neoplasm with cell cycle/cell proliferation markers and to understand their histogenesis with ultrastructural analysis using conventional carcinoids as a frame of reference. METHODS: Clinical data and archival pathologic material of all goblet cell carcinoids of the appendix recorded by the Saskatchewan Cancer Registry between 1970 and 1998 were reviewed and evaluated by light microscopy, histochemistry, immunohistochemistry, and electron microscopy. RESULTS: Seven cases of goblet cell carcinoids were identified among 110 cases of conventional carcinoids of the appendix. Histopathology revealed widespread infiltration of the periappendiceal fat in all cases, with extensive perineural invasion. The cells stained strongly positive for mucicarmine, periodic acid-Schiff, periodic acid-Schiff diastase, Alcian blue, cytokeratin, and carcinoembryonic antigen. Most cases were positive for synaptophysin. Increased expression of cell proliferation markers and cell cycle markers was observed. Expression of p53 was strong in one case. Electron microscopy demonstrated the presence of mucinous vacuoles of varying sizes and occasional membrane-bound neuroendocrine granules. CONCLUSIONS: Goblet cell carcinoids of the appendix arise from a pluripotent cell with divergent neuroendocrine and mucinous differentiation. These neoplasms are widely invasive; they demonstrate a high cellular proliferation rate and dysregulation of the cell cycle with up-regulation of cyclin D1 and p21, and down-regulation of p16. Complete removal of the tumor is recommended because of the unpredictable biological behavior of this tumor, which includes delayed local recurrences and lung metastases.

Adult↗
PubMed1992

Immunophenotypical characterization of inflammatory cellular infiltrates in tinea.

In order to elucidate the still poorly understood pathogenetic pathways of acute tinea, the inflammatory cellular infiltrates in this infection were analyzed. Lesional punch biopsies were cryostat-sectioned and stained with monoclonal antibodies for immunophenotypization of T cells, B cells, macrophages and activation markers. For each antibody the positively stained inflammatory cells in the dermis and in the epidermis were quantified separately. Most of the dermal mononuclear cells in acute tinea were identified as T helper lymphocytes of the memory type. Furthermore, considerable amounts of Langerhans' cells and macrophages were found, but virtually no B cells. A high proportion of cells expressed markers of activation. Within the epidermis, accumulations of Langerhans' cells and LeuM5+ dendritic macrophages were detected near fungal element. In view of the otherwise rather similar cellular infiltrates in acute tinea and different non-infectious dermatoses, acute tinea may be particularly suitable to study the functional relationship of Langerhans cells and LeuM5+ macrophages.

Acute Disease↗