Nonlymphoid Malignancies Research Articles

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.

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.

Angioimmunoblastic lymphadenopathy with malignant transformation during long-term remission of ocular melanoma

A 52-year-old woman presented with angioimmunoblastic lymphadenopathy five years after undergoing enucleation for ocular melanoma. She was first treated by corticotherapy and azathioprine and had two successive remissions over an 18-month period. Lymph node biopsy performed when lymphadenopathy relapsed for the second time showed focal malignant transformation of angioimmunoblastic lymphadenopathy. Intensive polychemotherapy induced a complete remission. Three months after completion of chemotherapy, the patient presented with hepatic metastases from her melanoma and died shortly thereafter. This is believed to be the first documented association between angioimmunoblastic lymphadenopathy and melanoma. Issues regarding the relationship between angioimmunoblastic lymphadenopathy and nonlymphoid malignancies are raised.

Spurious Detection of Terminal Deoxynucleotidyl Transferase in Phytohemagglutinin-Stimulated Lymphocytes

Expression of terminal transferase (TdT) is believed to be restricted to primitive lymphoid cells; recently, however, indirect immunofluorescent (IF) assays have been used to demonstrate the apparent presence of TdT in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes and in various nonlymphoid malignancies. Using an IF assay, we found that a heteroantiserum to TdT reacted with cultured and PHA-stimulated human peripheral blood mononuclear cells, but we were unable to confirm the presence of TdT in these cells using immunoblotting and biochemical assays. We conclude that the IF results are spurious and most likely represent recognition by the heteroantiserum of inducible protein(s) other than TdT.

Spurious detection of terminal deoxynucleotidyl transferase in phytohemagglutinin-stimulated lymphocytes

Expression of terminal transferase (TdT) is believed to be restricted to primitive lymphoid cells; recently, however, indirect immunofluorescent (IF) assays have been used to demonstrate the apparent presence of TdT in phytohemagglutinin (PHA)-stimulated peripheral blood lymphocytes and in various nonlymphoid malignancies. Using an IF assay, we found that a heteroantiserum to TdT reacted with cultured and PHA-stimulated human peripheral blood mononuclear cells, but we were unable to confirm the presence of TdT in these cells using immunoblotting and biochemical assays. We conclude that the IF results are spurious and most likely represent recognition by the heteroantiserum of inducible protein(s) other than TdT.

T cell receptor and immunoglobulin gene rearrangements in acute myeloblastic leukemia.

The organization and expression of the beta chain of T cell antigen receptor gene (beta-TCR) and Ig H and L chain genes were analyzed by Southern blot technique in 24 patients with a diagnosis of acute myeloblastic leukemia (AML). Rearrangements of the beta-TCR genes were seen in DNA samples from 3 of the 24 patients. One of these three patients also showed rearrangement of the Ig H chain gene. RNA samples from all three patients expressed a beta-TCR gene transcript on dot blot analysis. However, on Northern blot analysis, one patient expressed an incomplete 1.0 kb transcript and no Ig H chain mRNA, despite a rearranged configuration. The karyotypes of two of these patients showed abnormalities involving chromosome 7. Rearrangements of T cell antigen receptor genes may occur in nonlymphoid malignancy, and is consistent with the concept of lineage infidelity in AML.

11 - Laboratory Investigation of Non-lymphoid Malignancy

Abnormalities of genetic expression associated with malignant transformation cause a number of phenotypic changes including the production of substances by cancer cells which differ quantitatively or qualitatively from the products of normal cells. These ‘cancer markers’ or tumour associated antigens may be used as objective parameters in screening, diagnosis, staging, histopathological evaluation and monitoring treatment of non-lymphoid neoplasia. Specific and sensitive tests for alphafetoprotein, carcinoembryonic antigen, human chorionic gonadotropin and a number of other cancer markers have been developed. Established tests rely upon polyclonal heterologous antisera in radioimmunoassay or enzyme-linked immunoassay. Theoretical advantages of monoclonal antibodies in this setting have yet to be realized. Monoclonal antibodies do however show practical advantages over extensively absorbed polyclonal antisera in the histopathological evaluation of solid tumours and further progress with these reagents is expected. Studies of host-tumour immunological interactions suggest that future routine investigations of non-lymphoid neoplasia may include assessments of autochthonous reactivity.

Prospective study of natural cytotoxicity in peripheral blood of patients with nonlymphoid solid malignancies

Natural killer (NK) cells have been implicated as an initial immunosurveillance mechanism for carcinogenesis in humans. Work in the murine system as well as the findings of depressed NK activity in patients with advanced malignancies and the discovery of increased incidences of cancer in humans congenitally deficient in NK ability have supported this. Few prospective studies have demonstrated a prognostic change in NK activity with respect to malignant disease course. In 32 healthy donors, NK activity against K562 was determined. No race or sex difference existed with respect to NK cell function. Esophageal (5), bronchogenic (3), breast (3), cervical (3), and endometrial (1) cancer patients who had received no prior chemotherapy were compared to controls. All patients subsequently received radiotherapy. Prior to such treatment NK activity could not be associated with stage of malignancy. Of the 15 patients studied, 11 were sequentially followed. Five of eight patients with stable or improving clinical courses as assessed by weight and Karnofsky scores were found to have increasing NK activity. Two of three patients with poor clinical courses presented with subnormal killing which never rose to normal while the third declined to subnormal before expiring. Esophageal, cervical, and endometrial carcinoma patients all presented with low or subnormal NK activity. Of these, only cervical and endometrial cancer patients exhibited an increase to normal levels.

Skin-associated lymphoid tissues (SALT): origins and functions.

The skin has an unusual set of immunologic requirements. It is confronted by a specialized set of pathogenic organisms and environmental chemicals that represent a distinctive spectrum of antigenic specificities. Skin is subjected to physicochemical stresses such as irradiation with ultraviolet light that alter dramatically its immunologic properties. It is proposed that nature has provided skin with a unique collection of lymphoid cells, reticular cells, and organized lymphoid organs to deal with these special demands. Evidence in favor of the existence of skin-associated lymphoid tissues (SALT) includes (1) the cutaneous microenvironment is capable on its own of accepting, processing, and presenting nominal antigen; (2) strategically located peripheral lymph nodes are able to accept immunogenic signals derived from skin; (3) subsets of T lymphocytes display differential affinity for skin and its associated peripheral nodes; and (4) acquisition of this affinity by T cells is determined at least in part by differentiation signals received in situ from resident cutaneous cells. Responsibility for the establishment and integration of SALT rests with keratinocytes, Langerhans cells, and immunocompetent lymphocytes, each of which contributes uniquely to the synthesis. Together they provide skin with immune surveillance that effectively prejudices against the development of cutaneous neoplasms and persistent infection with intracellular pathogens. In patients who have been under long-term immunosuppressive therapy, the large majority of nonlymphoid malignancies arise within the skin, rather than other types of tissues. These data suggest that immune surveillance, once thought to be an immune defense operative in all somatic tissues, is a specialized immune function dedicated to the skin and mediated by SALT.

Malignant Lymphoma, T-cell Type: A Distinct Morphologic Variant with Large Multilobated Nuclei, with a Report of Four Cases

Clinical, morphologic, and immunologic features in four cases of a cytologically distinctive variant of T-cell lymphoma of large multilobated lymphoid (“histiocytic”) cell type are described. Clinical features included weight loss, fever, and night sweats. The ages of the patients ranged from 42 to 63 years. An extranodal manifestation and distribution of disease was observed in three of the four patients, with a predilection for skin and subcutaneous tissue, bone, gonads, and the central nervous system. Lymphadenopathy was present in all cases but represented the major clinical finding in only one. Splenomegaly was not observed. Histologically, the lymphomatous infiltrate was characterized by large lymphoid cells with markedly irregular, multilobated or hypersegmented nuclei, with relatively fine chromatin, and small to inconspicuous nucleoli. Neoplastic lymphoid cells were identified as T-cell type on the basis of spontaneous rosette formation with sheep erythrocytes (E rosettes). Cytochemically, these cells demonstrated strong punctate cytoplasmic acid phosphatase activity, and α -naphtyl butyrate esterase activity (punctate or dot-like staining pattern). The unusual cytologic features of this variant of malignant lymphoma differentiate it from other types of non-Hodgkin’s lymphomas, and from non-lymphoid malignancies, which it may simulate histologically and clinically.

Combined T and B Lymphocyte Marker Test in Lymphoproliferative Disorders

The combined T and B lymphocyte marker test of Mendes et al (1974) has been developed to provide an easy, reproducible test for clinical practice. It has been used to study subpopulations of peripheral lymphocytes in patients with lymphoid malignancies. Significant increases in the numbers of lymphocytes which bind both sheep erythrocytes (E) and the fixed, third component of complement (C3) have been found using this test, in the peripheral blood of all untreated lymphosarcoma (L/SA) patients and T cell acute lymphoblastic leukaemia (ALL) patients. Such increases were not found in blood from patients with chronic lymphatic leukaemia (CLL), other non-lymphoid malignancies, null-cell ALL or normal individuals. These mixed E and C3 receptor bearing cells may be Fc receptor positive and represent a separate subpopulation of lymphocytes.

Qualitative changes in serum immunoglobulins in patients with malignant and other conditions

Sera from patients suffering from malignant lymphoma, non-lymphoid malignancies and non-malignant conditions were examined by isoelectric focusing in thin layer polyacrylamide gels. Three distinct qualitative changes in immunoglobulin pattern were observed and designated Type I (monoclonal), Type II (oligoclonal) and Type III (pluriclonal), 95/354 patients (27%) suffering from malignant lymphoma and 11/71 patients (15%) suffering from non-lymphoid malignancies showed a qualitative change in their immunoglobulin pattern. A control group of healthy laboratory workers and normal donors failed to show this abnormality. Non-malignant conditions showing the abnormality included autoimmune diseases, primary and drug induced immune deficiency disorders and alcoholic cirrhosis. Many patients showed the abnormal pattern at diagnosis and prior to the commencement of therapy. However, in other patients the pattern appeared, sometimes transiently, during the course of the diseases. In these patients therapy could not be excluded as the cause of the abnormality. It is proposed that the common feature in all these patients is an immune deficiency state due either to the disease process itself or the effect of therapy. In many instances the abnormality was not detectable by conventional methods (electrophoresis, Immunoelectrophoresis or quantitation of immunoglobulins by radial immunodiffusion). It is suggested that more sensitive techniques (e.g., isoelectric focusing) should be employed in the investigation of immune deficiency disorders and other diseases leading to changes in immune competence.