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PubMed1990

Tumor immunophenotype: comparison between primary neoplasm and its metastases.

Monoclonal antibodies (MAb) are firmly established diagnostic adjuvants both in vitro and in vivo. Their potential for immunotherapy is highly promising. Antigenic heterogeneity of cells within the same tumor is a well known phenomenon; however, no large-scale studies are available to ascertain to what degree metastases maintain the immunophenotype of the primary tumors. For that purpose, we studied 54 commonly epithelial malignancies using immunohistochemistry (IHC) with a panel of seven frequently used MAb recognizing a gamut of membrane and cytoplasmic antigens (AE-1, CAM 5.2, B72.3, MC10, anti-carcinoembryonic antigen (anti-CEA), epithelial membrane antigen (EMA), and human milk fat globule (HMFG)). The number of metastases per primary tumor ranged between 1 and 26, with a total of 344 tissues studied. Metastases were located in regional and distal lymph nodes as well as in a diversity of organs (pancreas, adrenal, colon, spleen, soft tissues, etc). Only those cases in which all the tissues were obtained from a single surgical procedure and, therefore, uniformly fixed and processed, were selected. All the metastases from three cases (5.5%) were found to express one or two antigens not present on their primary. In no case did all metastases from a positive primary become negative for one MAb. Twelve cases (18.5%) showed modifications of the phenotype in one or more metastases. This study demonstrates that a broad phenotypic variation does not follow when tumors metastasize, and that it is, therefore, safe to foretell the metastatic immunophenotype based upon that of the primary tumor.

Antibodies, Monoclonal↗
PubMed1988

Application of a T cell receptor antibody beta F1 for immunophenotypic analysis of malignant lymphomas.

One hundred sixty-five non-Hodgkin's lymphomas (101 B, 63 T, one histiocytic) were immunostained with an antibody (beta F1) reactive with a common framework determinant on the beta-subunit of the T cell receptor (TCR). beta F1 stained T lymphomas exclusively, including 53% of peripheral T cell lymphomas but only 33% of T lymphoblastic lymphomas. When expression of beta F1 and CD3 were considered together, 4 types of T lymphoma were delineated: 1) beta F1+CD3+; 2) beta F1+CD3-; 3) beta F1-CD3+, and 4) beta F1-CD3-. The first represented lymphomas with classical T immunophenotype. The second might represent T lymphomas with aberrant loss of CD3 expression. The third might represent T lymphomas with a putative second TCR or cases with an immature phenotype expressing cytoplasmic CD3 only. The fourth type included cases that may be derived from natural killer cells instead of T cells, cases of T lymphoma with aberrant loss of both beta F1 and CD3, and some cases of immature T cell (lymphoblastic) lymphoma. beta F1-CD3- lymphomas exhibited a remarkable predilection for the nasal region. beta F1 is useful in studying T cell lymphomas and distinguishing a novel immunophenotype frequently expressed by nasal lymphomas.

Antibodies, Monoclonal↗
PubMed1989

Stability of multiple antigen receptor gene rearrangements and immunophenotype in Hodgkin's disease-derived cell line L428 and variant subline L428KSA.

The Hodgkin's disease (HD) derived cell line L428 and a phorbol ester-selected subline L428KSA, which have been independently passaged in tissue culture for several years, were studied for possible antigen receptor gene and immunophenotypic differences. Multiple but identical alterations of these genes were found, including: the deletion of one and rearrangement of the other immunoglobulin (Ig) heavy chain allele; the rearrangement of one kappa and one lambda light chain allele; and the rearrangement of one T cell receptor (TCR) beta allele. Restriction mapping of the Ig heavy chain locus indicated that rearrangement of the retained allele produced a JH-C gamma 4 fusion product by an isotype switch mechanism. The 14q+ chromosome [t(14q32;?)] present in both cell cultures derived either from translocation 5' (telomeric) to the rearranged JH allele or 3' (centromeric) to the deleted Ig heavy chain allele and did not involve detectable rearrangement of the c-myc, bcl 1, or bcl 2 oncogenes. No differences in the immunophenotype were found between the L428 and L428KSA cells: both expressed leukocyte activation antigens and some determinants associated with myelomonocytic cells but no lymphoid markers. It is postulated that these phenotypic characteristics derived from secondary genetic events/differentiative reprogramming which produced extinction of primary lymphoid characters, including terminal deoxynucleotidyl transferase (TdT) essential to generation of the Ig and TCR gene rearrangements, and expression of an incomplete set of myelomonocytic markers.

Blotting, Southern↗
PubMed1989

Relationship between immunophenotype and the development of second primary neoplasms in patients with non-Hodgkin's lymphoma.

We reviewed the records of 107 patients with non-Hodgkin's lymphoma (NHL) to evaluate the relation between second primary neoplasms and the NHL immunophenotype. The incidence of second primary neoplasms was 3.7%. There were one case of hepatocellular carcinoma and 3 cases of gastric adenocarcinoma including one patient who had a history of metachronous malignant lymphomas. Three patients had B cell lymphoma with monoclonal IgM kappa phenotype, and one patient had follicular mixed cell type lymphoma with serum monoclonal IgM kappa. The dominant immunophenotype of B cell lymphomas in Japanese patients is IgM lambda. We believe that the association of the uncommon phenotype of IgM kappa with second primary neoplasms, especially gastric cancer, reflects an underlying genetic predisposition. NHL patients with IgM kappa phenotype should be evaluated carefully for second primary neoplasms.

Adenocarcinoma↗
PubMed1989

Membrane antigen analysis in the diagnosis of lymphoid leukemias and lymphomas. Differential diagnosis, prognosis as related to immunophenotype, and recommendations for testing.

The immunophenotypic features of lymphoid leukemias and non-Hodgkin's lymphomas are reviewed. Emphasis is placed on the more recent literature, particularly the insights provided by monoclonal antibodies specific for T and B lymphocytes, and the contributions of flow cytometry. Characteristic immunophenotypes for T- and B-lineage acute lymphoblastic leukemia, the non-Hodgkin's lymphomas, chronic B- and T-cell lymphoproliferative disorders, and multiple myeloma are described. The importance of this information in the diagnostic process and in providing prognostic information concerning disease progression is considered. Limitations associated with monoclonal antibody reagents and flow technology are noted. Representative panels of monoclonal antibodies as used in the author's laboratory for characterizing these disorders are presented and discussed in light of the author's experience.

Antigens, Neoplasm↗
PubMed1989

Immunophenotyping of malignant lymphomas--a clinico-pathological approach.

The use of monoclonal antibody especially in immunophenotyping of lymphomas is firmly established in clinical practice. This has helped in refining the classification of non-Hodgkin's lymphomas and the updated Kiel Classification is now much favoured by the Europeans. Nonetheless, clinicians by and large preferred the National Cancer Institute Working Formulation Classification. This article explores the potential usage of monoclonal antibodies reactive to the so-called Clusters of Differentiation antigens on lymphoid cells and the role of a clinical pathologist in achieving an accurate clinical diagnosis. Such immunophenotyping is only complementary to the morphological diagnosis of lymphomas.

Antibodies, Monoclonal↗
PubMed1989

Immunophenotypes of Reed-Sternberg cells: a study of 19 cases of Hodgkin's disease in plastic-embedded sections.

The immunophenotype of Reed-Sternberg (RS) cells in Hodgkin's disease (HD) has not been clearly defined, partly owing to difficulties in studying RS cells in cell suspensions or identifying them with certainty in frozen sections. We studied the immunophenotype of RS cells with a recently developed plastic section immunohistochemical technique on acetone-fixed tissues that affords superior morphological detail while preserving a wide variety of lymphoid differentiation antigens. Nineteen cases of HD [16 nodular sclerosing (NS), 2 mixed cellularity (MC), and 1 lymphocyte depleted (LD)] were embedded in plastic and stained for pan-B, pan-T, and various T-subset markers, as well as leukocyte common antigen (CD45), interleukin-2 (IL-2) receptor (CD25), and RS cell markers CD15 and CD30. RS cells were positive for CD45, CD15, CD30, and CD25, except for 3 cases (2 NS, 1 MC) that were CD15 negative and 2 cases (NS) that were CD45 negative. In 10 cases (NS), RS cells were positive for at least two pan-T-cell markers and CD4; pan-B cell markers were uniformly negative. RS cells in 6 cases (3 NS, 2 MC, 1 LD) were positive for at least one T-cell marker (CD2) and one B-cell marker (CD22). Two cases of NSHD showed no T- or B-cell marking. These data provide further evidence that RS cells in some cases of NSHD have T-cell phenotypes and that RS cells are not homogeneous in their immunoreactivity.

Antibodies, Monoclonal↗
PubMed1989

Immunophenotypic and genotypic characterization of diffuse mixed non-Hodgkin's lymphomas.

Diffuse, mixed small and large cell lymphomas (DML) are a heterogeneous group of non-Hodgkin's lymphomas. There are only a few published immunophenotypic and/or genotypic studies of DML, and they include a small number of cases. It is unclear whether these neoplasms are monoclonal, oligoclonal, or perhaps of dual lineage. Using monoclonal antibodies (UCHL1, MB2, MT1, LN1, LN2, and L26) that are effective in paraffin-embedded, B5-fixed tissue, 13 cases of DML were studied. This method allowed for improved correlation between cell morphology and immunophenotype compared with frozen section immunohistology. In addition, Southern blotting/DNA hybridization was used to identify directly the lineage of the neoplastic cell population. In each case a population of large B lymphocytes was demonstrated by immunohistology. In six of the cases, a single class of immunoglobulin light chains was detected by frozen section immunohistology, cytospin immunocytology, or both supporting the hypothesis that the B lymphocytes are monoclonal. In almost all the cases (12 of 13), the small cell population consisted predominantly of T lymphocytes. Immunoglobulin gene rearrangements were detected in seven cases, but no T cell receptor gene rearrangements were detected. It was concluded that DML are monoclonal lymphomas of B cell lineage with a non-neoplastic T cell component.

Blotting, Southern↗
PubMed1988

Cytochemical, cytogenetic, immunophenotypic and tumorigenic characterization of two hairy cell lines.

Two cell lines (EH and HK) were derived from two patients with hairy cell leukemia (HCL). Both patients exhibited a clinical picture characteristic of HCL, including splenomegaly, cytopenias, and tartrate-resistant acid phosphatase (TRAP)-positive "hairy" lymphocytes in blood and marrow. EH and HK were demonstrably of B lineage, as judged by cytochemistry and immunophenotype, including expression of B1, B2, and LEU-12 antigens and of monoclonal surface immunoglobulins (slgs). Monoclonality was confirmed by clonal karyotype abnormality demonstrated in cell line HK. HCL parentage suggested by cytochemistry and electron microscopy was confirmed by the immunophenotypic observation that HCL lines expressed antigens alpha S-HCL1, alpha S-HCL3, and cCLLa. While alpha S-HCL1 and alpha S-HCL3 are nonspecific, their co-expression is characteristic of HCL cells. The cCLLa is a novel 69-kd membrane HCL-associated polypeptide antigen not shared by circulating normal T or B lymphocytes nor by malignant cells from unrelated lymphoid or nonlymphoid malignancies. The doubling time of EH and HK was 24 and 36 hours, respectively. While HK included a small subset of Epstein-Barr virus (EBV) nuclear antigen-positive cells, EH cells were homogeneously negative for the presence of this antigen. Both cell lines were consistently implantable in irradiation-preconditioned immunodeficient mice giving rise to primary tumors and widespread metastasis.

Animals↗
PubMed1988

Immunophenotypic and cytogenetic analysis of Molt-3 and Molt-4: human T-lymphoid cell lines with rearrangement of chromosome 7.

Molt-3 and Molt-4 are T-cell lines originally derived in 1971 from a patient with T-cell acute lymphoblastic leukemia. An unusual T-cell antigen receptor gamma-chain gene (T-gamma) rearrangement detected by Southern blot analysis of Molt-4 prompted an in-depth study of the immunophenotype and karyotype of both cell lines. Molt-3 and Molt-4 had immunophenotypic characteristics of thymocytes with expression of CD1 and CD5. Both cell lines had a hypertetraploid karyotype with two rearranged no. 7 chromosomes: 2der(7)t(7;7)(p15;q11). The presence of a break in chromosome band 7p15 suggested the involvement of T gamma. We cloned the rearranged BamHI fragments spanning the known T-gamma constant and joining regions. Comparison with germline clones of T gamma did not suggest any of the clones included a breakpoint region. Thus the 7p15 chromosomal abnormality in Molt-3 and Molt-4 is not associated with the currently described joining and constant regions of T gamma.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1986

Cytogenetic and immunophenotypic analysis of cell lines established from patients with T cell leukemia/lymphoma.

Cell lines were established from five patients with T cell malignancies. Two patients had T cell lymphoblastic lymphoma (T-LL), whereas three patients had T cell acute lymphoblastic leukemia (T-ALL). Both T-LL cell lines expressed cell surface antigens characteristic of midthymocytes (Leu 2, 3, 6+). One T-ALL cell line also expressed this immunophenotype, one expressed suppressor/cytotoxic antigens (Leu 2+; Leu 3, 6-), and one expressed antigens of a mature but uncommitted T cell (Leu 4+; Leu 2, 3, 6-). Cytogenetic analysis showed that each cell line had 46 chromosomes with pseudodiploidy. The three T-ALL cell lines had only a few chromosome changes; one cell line had one deletion, another had two deletions, and the third had a translocation and two deletions (including loss of part of 9p). In comparison, both T-LL cell lines had complex chromosome changes, including most notably a rearrangement of band 14q11.2. The immunophenotypes and chromosome breakpoints showed patterns of interlock between the T-LL and T-ALL cell lines because common abnormalities occurred at six distinct chromosome sites. Cell lines with limited and specific chromosomal abnormalities are important because they can provide the basic material for molecular genetic studies that could elucidate the genetic mechanisms involved in neoplasia.

Adolescent↗
PubMed1988

The aberrancy of immunophenotype and immunoglobulin status as indicators of prognosis in B cell diffuse large cell lymphoma.

To assess the prognostic significance of the immunophenotype in diffuse large cell lymphoma (DLCL), 105 DLCL patients were studied between 1978 and 1987 using a panel of 40 monoclonal antibodies applied to frozen tissue. Eighty-three patients were found to have B cell phenotypes, and 20 patients had T cell phenotypes. Focusing on markers relevant to clinical outcome among B cell LCL showed that lack of expression of the pan B antigens Leu14 and Leu16 were correlated with decreased survival (Leu14, P = 0.01; Leu16, P = 0.06; log-rank). HLA-DR activity also showed that lack of expression of this antigen correlated with poor survival (P = 0.004, log-rank). Kappa light chain immunoglobulin lack of expression showed predictive value for decreased survival as well (P = 0.005, log-rank). Multivariate analyses of known clinically important variables and the immune phenotypes confirm that the loss of HLA-DR and B cell aberrancy are independent factors predicting a poor clinical outcome. Losing some B activation/kappa antigens appears to be a broad biologic phenomenon linking surface antigen lack of expression with decreased survival. This suggests that aberrancy of immunophenotype and immunoglobulin status are key predictors of survival in B-LCL.

Antigens, Surface↗
PubMed1985

Immunophenotypic differences between dermatopathic lymphadenopathy and lymph node involvement in mycosis fungoides.

The authors applied a battery of monoclonal antibodies to T cells and other hematolymphoid cells on frozen tissue sections of lymph nodes with involvement by MF and DL, both with and without a history of MF. All 13 lymph nodes showing histologic involvement with MF showed immunophenotypic abnormalities. All of these cases showed significant loss of Leu-9 expression, and 10 cases showed significant loss of Leu-8 expression. In addition, occasional cases showed loss of the pan-T-cell markers Leu-1, 4, and 5. All cases of DL of histologic grade LN-0 or 1, with or without a history of MF, showed a predominance of T helper cells in paracortical regions without evidence of immunophenotypic abnormalities. Three of the 4 cases of DL of histologic grade LN-2 or 3 showed significant loss of Leu-8 and/or Leu-9 expression identified by a decrease in the ratio of paracortical Leu-8/Leu-4 or Leu-9/Leu-4-expressing cells, all cases with a history of possible or definite MF. These results raise the possibility that the immunologic methods employed may be able to identify cases of DL with early involvement by MF.

Antibodies, Monoclonal↗
PubMed1988

Nuclear contour irregularity correlates with Leu-9-, Leu-8- cells in benign lymphoid infiltrates of skin. An ultrastructural morphometric and quantitative immunophenotypic analysis suggesting the normal T-cell counterpart to the malignant mycosis fungoides/Sézary cell.

The purpose of the present study was to determine if a nonmalignant skin-associated T-cell might exhibit nuclear irregularity and represent the normal counterpart to the malignant Sézary T-cell found in mycosis fungoides (MF) and the Sézary syndrome (SS). Punch skin biopsies from ten benign lymphoid infiltrates of skin were subjected to ultrastructural morphometric analysis and quantitative immunohistochemistry (on serial frozen sections) using a battery of monoclonal antibodies. Form factor (FF) (4 pi A/p2) was used to assess nuclear contour irregularity. The lymphoid infiltrates were morphometrically heterogeneous (range of FF values: 0.482-0.704). Immunophenotypically, there was marked variation in host response to the variety of antigens presented. Linear regression analysis was used to determine if nuclear contour irregularity showed any statistical relationship to immunophenotypically defined lymphocyte subpopulations. It was determined that nuclear contour irregularity (mean FF values) did not correlate with the presence of any surface antigen tested. This included the antigens Leu-2a, Leu-3a + b, Leu-4, Leu-6, Leu-7, Leu-8, Leu-9, Leu-14, and Ia. There was, however, significant correlation of increased nuclear contour irregularity with the presence of Leu-9- and Leu-8- cells (r = 0.7 and 0.6, respectively), lymphocyte subsets reportedly deficient in MF and the SS. This finding leads to the speculation that the Leu-9-, Leu-8- reactive T-cell with an irregular nucleus might represent the normal counterpart to the malignant clonally expanded T-cell found in MF and the SS. It was also determined that helper to suppressor ratios varied 68-fold from 0.2 to 13.5 among these ten benign lymphoid infiltrates of skin. This finding underscores the futility of using helper to suppressor ratios as a diagnostic tool in defining T-cell malignancies.

Adult↗
PubMed1988

Immunophenotyping in the diagnosis and classification of acute lymphoblastic leukemia.

The identification of ALL immunophenotypes with distinctive clinical features and prognostic significance indicates the importance of these studies in the evaluation of ALL patients for both clinical and research purposes. For differential diagnosis, the expression of pan-B-cell or pan-T-cell lymphoid antigens and the absence of myeloid/monocyte antigens represent the most useful markers for distinguishing ALL from AML. However, the increasing appreciation of large numbers of patients with clinically significant mixed lymphoid-myeloid phenotypes suggests that rigid classification of acute leukemias into exclusive lymphoid and myeloid categories may be somewhat artificial. In adult ALL, patients with My+ phenotypes (B+sIg-T-My+ and B-sIg-T-My+) have a lower incidence of complete remission and shorter survival times than do patients with My- marker profiles. Preliminary studies in childhood ALL also suggest a correlation between myeloid antigen expression and poor prognostic factors. In addition, children with sIg+ "B-cell," cIg+ "pre-B-cell," and T-cell ALL phenotypes have shorter disease-free survival times than do patients with more common "early pre-B" (B+cIg-sIg-T-) marker profiles. Application of immunologic markers in concert with cytogenetic and gene rearrangement studies has led to the identification of novel subgroups of leukemia with distinct clinical characteristics. Future studies incorporating a multiparameter diagnostic approach including immunophenotyping, gene rearrangement studies, and karyotypic analyses should further our understanding of the heterogeneity of acute leukemias, guide the development of new therapeutic strategies, and provide for more clinically relevant classification of these disorders.

Antibodies, Monoclonal↗
PubMed1987

Classification of acute myeloid leukemias--a comparison of FAB and immunophenotyping.

A large number of monoclonal antibodies (McAbs) directed against components on myeloid (granulocytic/monocytic) cells have been generated. Individual McAbs were identified which are selectively reactive with antigenic determinants expressed by myeloid cells at specific stages of differentiation in a lineage-restricted fashion. The composite phenotype obtained by a combination of antimyeloid McAbs allows for a precise definition of the normal or malignant cell type under investigation. Cell binding studies on normal and leukemic cells and the biochemical characterization of the antigens provided the basis for a grouping of those antimyeloid McAbs into clusters of differentiation (CD). The reactivity patterns of CD11, CD13, CD14, CD15, and CD33 McAbs and the characteristics of the respective antigens are reviewed. These CD McAbs distinguish leukemic cells of myeloid from those of lymphoid origin. The monocytic nature of AML cells can be recognized by CD14 McAbs, whereas the other CD McAbs react with both monocytic and nonmonocytic types of acute myeloid leukemia. The expression of these differentiation antigens is not concordant with the morphological-cytochemical French-American-British (FAB) classification of leukemia; nevertheless, tendencies for agreement are apparent. If used in combination, FAB typing and immunophenotyping could provide complementary information. Their potential use for mapping of myeloid differentiation and for cell type recognition in leukemia phenotyping demonstrates the utility of antimyeloid CD McAbs for biological or clinical investigations. The diagnostic value of antimyeloid McAbs is enhanced if the reagents are included in a panel of McAbs standardized for routine immunophenotyping.

Antibodies, Monoclonal↗
PubMedDate not supplied

A practical approach to immunophenotyping of lymphomas. Comparison of immunohistologic and immunocytologic techniques.

Comparison of immunohistologic (IH) and immunocytologic (IC) techniques was conducted on 86 specimens. Forty-seven B-cell, five T-cell, and two null-cell lymphomas were identified by IH as well as 16 cases of lymphoid hyperplasia. The results of IC were generally identical to those of IH except for two T-cell and two B-cell lymphomas. The diagnosis of T-cell lymphoma was a major problem for IC because of the presence of normal T-cell count and/or normal helper/suppressor ratio in these cases. Twenty-one percent of the B-cell lymphomas failed to express surface immunoglobulin but did express B1 and HLA-DR antigens. Such a discrepancy was not demonstrated in cases of lymphoid hyperplasia, thus serving as a useful criterion in the diagnosis of B-cell lymphoma. While combined IH and IC should be used for immunophenotyping in large medical centers, IH is recommended for community hospitals. The identification of kappa, lambda, B1 and T11 (Leu 5) antigens in frozen sections with the immunoperoxidase technique should be sufficient to phenotype most lymphoproliferative diseases. The criteria for immunophenotyping of lymphomas are discussed.

Antibodies, Monoclonal↗
PubMed1987

Intermediate lymphocytic lymphoma: immunophenotypic and cytogenetic findings.

A detailed immunohistologic and cytogenetic analysis of 12 cases of intermediate lymphocytic lymphoma was performed. The characteristic immunophenotype of intermediate lymphocytic lymphoma was: surface IgM and IgD+, BA1+, B1+, BA2-, B2-, B4+, Leu 14+, Leu 1+, HLA-DR+, and common acute lymphocytic leukemia associated (CALLA) antigen negative. Clonal chromosome abnormalities were identified in ten cases, with structural or numerical abnormalities of chromosomes 11 or 12 in nine cases. Five cases had structural abnormalities involving the long arm of chromosome 11; three of these had translocations with chromosome 14 at band q32. Three cases had trisomy 12, and one case had a translocation involving the long arm of chromosome 12. The tenth case had a translocation involving the long arms of chromosomes 7 and 9. These characteristic immunophenotypic and cytogenetic findings suggest a close lineage relationship between intermediate lymphocytic lymphoma and small lymphocytic (well differentiated) lymphoma/chronic lymphocytic leukemia. Their differing clinical, cytologic, and architectural features suggest, however, that intermediate lymphocytic lymphoma should be considered a separate category of lymphocytic lymphoma in the International Working Formulation.

Biopsy↗