Biomedical subjects

J Cossman

Publications and source records attributed to J Cossman.

At least 91 records · Page 5Linked to original sources

PubMed1985

The t(14;18) chromosome translocations involved in B-cell neoplasms result from mistakes in VDJ joining.

In this study, the joining sequences between chromosomes 14 and 18 on the 14q+ chromosomes of a patient with pre-B-cell leukemia and four patients with follicular lymphoma carrying a t(14;18) chromosome translocation were analyzed. In each case, the involved segment of chromosome 18 has recombined with the immunoglobulin heavy-chain joining segment (JH) on chromosome 14. The sites of the recombination on chromosome 14 are located close to the 5' end of the involved JH segment, where the diversity (D) regions are rearranged with the JH segments in the production of active heavy-chain genes. As extraneous nucleotides (N regions) were observed at joining sites and specific signal-like sequences were detected on chromosome 18 in close proximity to the breakpoints, it is concluded that the t(14;18) chromosome translocation is the result of a mistake during the process of VDJ joining at the pre-B-cell stage of differentiation. The putative recombinase joins separated DNA segments on two different chromosomes instead of joining separated segments on the same chromosome, causing a t(14;18) chromosome translocation in the involved B cells.

B-Lymphocytes↗

PubMed1985

Spontaneous alteration of idiotype in a monoclonal B-cell lymphoma. Escape from detection by anti-idiotype.

The surface immunoglobulin of B-cell neoplasms provides a specific point of attack for potential antibody therapy. The capacity of anti-idiotype antibody to home to the target neoplasm requires that the idiotype be unique and that it be expressed by every cell in the neoplastic clone. We describe the evolution of an altered idiotype in a follicular lymphoma that resulted in escape from laboratory detection by monoclonal anti-idiotype antibody. This was not due to the emergence of a second (biclonal) lymphoma, since all the neoplastic cells were otherwise identical both phenotypically and genotypically, as determined by flow cytometry and genomic DNA (Southern blot) hybridization, respectively. All cells expressed the same B-cell immunotype and bore a constant amount of IgMk. The demonstration of a single configuration of immunoglobulin-gene DNA confirmed monoclonality and established that the change in idiotype was not a result of new gene rearrangements but was more likely due to somatic mutation of the variable region--a process presumed to occur naturally in B cells. These data demonstrate the lability of idiotype expression and define a mechanism by which B-cell neoplasms may become unresponsive to anti-idiotype therapy.

Adult↗

PubMed1985

Involvement of the bcl-2 gene in human follicular lymphoma.

Recombinant DNA probes were cloned for the areas flanking the breakpoint on chromosome 18 in cells from a patient with acute lymphocytic leukemia of the B-cell type; cells of this line carry the t(14;18) chromosomal translocation. Two of the probes detected DNA rearrangements in approximately 60 percent of the cases of follicular lymphoma screened. In follicular lymphoma, most of the breakpoints in band q21 of chromosome 18 were clustered within a short stretch of DNA, approximately 2.1 kilobases in length. Chromosome 18-specific DNA probes for the areas flanking the breakpoints also detected RNA transcripts 6 kilobases in length in various cell types. The gene coding for these transcript (the bcl-2 gene) seems to be interrupted in most cases of follicular lymphomas carrying the t(14;18) chromosomal translocation.

B-Lymphocytes↗

PubMed1985

Significant non-S-phase DNA synthesis visualized by flow cytometry in activated and in malignant human lymphoid cells.

The development of a monoclonal antibody to the deoxynucleoside bromodeoxyuridine (BrdU), combined with two parameter flow cytometry, has allowed us to examine large numbers of cells for non-S-phase DNA synthesis. Three human lymphoid cell populations were studied to determine the level of deoxynucleoside (dN) incorporation as a function of DNA content. In each population, non-S-phase DNA synthesis was observed. In a rapidly growing human T-lymphoblastoid cell line (CCRF-CEM), 53% of dN incorporation occurred in G0/G1 plus G2 + M. In chronic lymphocytic leukemia (CLL) cells stimulated with tetradecanoylphorbol acetate (TPA), 45% of the observed burst in thymidine incorporation was found to be localized to G0/G1 cells. Non-S-phase incorporation was not, however, limited to neoplastic cells. Normal human peripheral blood B cells treated with the Cowan strain of Staphylococcus aureus (CSA) undergo a transient burst in thymidine incorporation, but do not go on to divide in the absence of other stimuli. Flow-cytometric analysis showed that 80% of this CSA-stimulated dN incorporation was into G0/G1 cells. These data are consistent with a more dynamic state of DNA synthesis than usually envisioned. Furthermore, the data show that although thymidine incorporation levels are related to incorporation of dN into DNA, they can be unrelated to cell proliferation.

Antibodies, Monoclonal↗

PubMed1985

Production and characterization of a monoclonal antibody that binds Reed-Sternberg cells.

A murine monoclonal antibody, termed HeFi-1, was produced after immunization with the L428 Hodgkin's disease tissue culture cell line. HeFi-1 selectively stained only the Reed-Sternberg or Hodgkin's cells in 18 of 18 cases of Hodgkin's disease, including the nodular sclerosis, mixed cellularity, and lymphocyte-depleted histologic subtypes. HeFi-1 did not stain any cells in normal lung, brain, salivary gland, thyroid, gall bladder, pancreas, liver, testis, breast, endometrium, or kidney. Rare large cells at the edge of the lymphoid follicles were stained in normal tonsil, colon, and hyperplastic thymus. There was no staining of any cells in 14 cases of B cell non-Hodgkin's lymphoma; however, the malignant cells in three of 11 cases of non-Hodgkin's lymphoma which appeared to express T cell markers were also stained with HeFi-1. Tissue culture cell lines including the T cell acute lymphocytic leukemia lines MOLT4 and CEM, the histiocytic cell line U-937, and the amniotic cell line WISH were not stained. Seven Epstein Barr virus (EBV)-positive lymphoblastoid cell lines were stained with HeFi-1, but there was no staining of three EBV+ African Burkitt's lymphoma cell lines or three EBV- American Burkitt's cell lines. HeFi-1 did not block the ability of the L428 cells to stimulate a mixed lymphocyte reaction or function as accessory cells for mitogen-induced human T cell proliferative responses. Modulation of the HeFi-1 cell surface antigen on the L428 cells was not observed. HeFi-1 specifically immunoprecipitated a cell surface protein of approximately 120,000 daltons from both the L428 and EBV+ lymphoblastoid cell lines. HeFi-1 monoclonal antibody should prove useful not only in the diagnosis, staging, and potential therapy of Hodgkin's disease, but also for determining the cell of origin of the Reed-Sternberg cell.

Animals↗

PubMed1985

Induction of immunoglobulin secretion in follicular non-Hodgkin's lymphomas: role of immunoregulatory T cells.

B cell neoplasms are clonal expansions of B lymphocytes thought to be frozen at various points along the normal B cell differentiation pathway. We studied cell suspensions from lymph nodes involved by follicular (nodular) non-Hodgkin's lymphoma to determine the capacity of the malignant B cells to secrete immunoglobulin (Ig). Neoplastic B cells from all 14 follicular lymphomas secreted monoclonal immunoglobulin in culture when appropriate signals were provided. In most cases, maximal Ig secretion occurred when autologous T cells were removed by E rosette depletion, replaced with allogeneic normal T cells, and the cultures were exposed to 12-O-tetradecanoylphorbol-13-acetate. Autologous T cells exerted a suppressor effect on Ig secretion in 8/8 cases studied, diminishing the response of the malignant B cells to allogeneic T cells. This suppressor effect did not correlate with the percentage of cells staining with anti-Leu-2a or with "helper-suppressor" (Leu-3a-Leu-2a) ratios of the lymph node T cells. Our findings demonstrate that the arrested differentiation of most follicular lymphomas is reversible and implicate a T cell-mediated host immunoregulatory mechanism affecting Ig secretion in vivo. An additional contribution of our results is the demonstration of a cell culture system for synthesis of sufficient monoclonal Ig for use as an immunogen in production of anti-idiotype antibodies.

Antibody Formation↗

PubMed1985

Lymphokine-induced phagocytosis in angiocentric immunoproliferative lesions (AIL) and malignant lymphoma arising in AIL.

A factor that augmented the phagocytosis of IgG-coated ox red blood cells by the human monocyte/macrophage line U937 was identified in cell culture supernatants from two of two patients with angiocentric peripheral T cell lymphomas, three of three patients with angiocentric immunoproliferative lesions that were not frankly malignant, and one of two patients with T lymphoblastic malignancies. The factor was not present in supernatants derived from 14 nonangiocentric peripheral T cell lymphomas of other histologic types nor in ten cases of B cell lymphoma and two cases of Hodgkin's disease. A similar factor was present in the supernatants of concanavalin A (Con A)-stimulated normal peripheral blood mononuclear cells and in the supernatants of IL-2-dependent T cell lines derived from normal peripheral blood. The factor had an apparent mol wt of greater than 50,000 daltons, was heat labile (100 degrees C for two minutes), and stable at pH 2.0. Its stimulation of phagocytosis was independent of any increase in number of Fc receptors. Thus, this factor is probably not gamma-interferon. This factor may play a pathogenetic role in the hemophagocytic syndromes associated with certain T cell malignancies and immunodeficient states.

Animals↗

PubMed1985

Antigen presentation by Hodgkin's disease cells.

The L428 tumor cell line is a long-term tissue culture of Reed-Sternberg cells which was derived from the pleural effusion of a patient with Hodgkin's disease. The L428 cells express all known cell surface antigens, cytochemical staining, and cytologic features of freshly explanted Reed-Sternberg cells. In addition to the previously described HLA-DR cell surface antigens, the L428 cells are now demonstrated to express both DS and SB alloantigens. Thus, the L428 cells express all of the known subclasses of the human immune response genes that are located in the major histocompatibility complex. Furthermore, the L428 cells are capable of presenting soluble antigen to T cells in a genetically restricted fashion. T cell lines were established from normal donors previously immunized with tetanus toxoid. The T cells utilized were incapable of tetanus toxoid-induced proliferation unless antigen-presenting cells were added to the cultures. However, T cells from the two normal donors, which like the L428 cells expressed HLA-DR 5, demonstrated significant proliferative responses when cultured with tetanus toxoid and L428 cells. No proliferative response was observed when the L428 cells were used as antigen-presenting cells for a DR (4,-), DR (2,-) or DR (1,7) T cell line. The tetanus toxoid dose-response curve was similar regardless of whether autologous mononuclear leukocytes or L428 cells were used as antigen-presenting cells. The T cell proliferation induced by soluble antigen was also blocked by anti-HLA-DR antibody. Thus, functionally, Hodgkin's disease may be classified as a tumor of antigen-presenting cells.

Antibodies, Monoclonal↗

PubMed1984

Immunologic phenotypes of diffuse, aggressive, non-Hodgkin's lymphomas. Correlation with clinical features.

The immunologic phenotypes of 59 cases of diffuse, aggressive, non-Hodgkin's lymphomas were determined using a battery of immunologic and cytochemical techniques. Included were cases of diffuse, large cell "histiocytic," mixed cell, undifferentiated non-Burkitt's. Burkitt's lymphoma, lymphoblastic lymphoma, and mycosis fungoides/Sézary's syndrome were excluded from this study since these are distinct clinicopathologic entities with well-recognized immunologic phenotypes. The immunotype could be determined in 57/59 (97%) cases tested: 31 of 59 cases (53%) were B-cell type, 25 of 59 (42%) were peripheral T-cell type, and one was true histiocytic. Two cases had no detectable markers and were called "null cell." This relatively high frequency of peripheral T-cell lymphomas in an American series previously has not been observed and may be a result of progressive improvements in immunologic techniques. Monoclonal anti-T cell antibody staining was performed in 11 T-cell cases and corroborated the findings using spontaneous E-rosette formation. Eight of the T-cell lymphomas had a helper cell phenotype whereas one had a suppressor cell phenotype and two could not be subclassified. All B-cell lymphomas in this series possessed monoclonal surface immunoglobulin detected by direct immunofluorescence of viable cells. Enzyme cytochemistry profiles only partially correlated with immunotype and were not believed to be helpful in the determination of specific phenotypes. There were no significant differences between the B-cell and T-cell diffuse aggressive lymphomas with respect to sex, constitutional symptoms, stages, sites of extranodal involvement, complete remission rate, or survival when they were studied prior to the initiation of aggressive therapy. Although immunotyping can be successfully performed in essentially all cases of diffuse, aggressive non-Hodgkin's lymphomas, to date, the authors have been unable to demonstrate that immunotype alone has an independent prognostic effect.

B-Lymphocytes↗

PubMed1984

Acute megakaryoblastic leukemia in childhood.

Routine morphologic and cytochemical study of the leukemic cells of a 13-month-old child did not permit definitive determination of either a lymphoid or a myeloid cell origin. However, ultrastructural cytochemical analysis revealed platelet peroxidase (PPO) reactivity. The malignant cells expressed PPO in both unfixed and fixed preparations. PPO was observed in the perinuclear space and reticulum but not in the Golgi saccules. In a 1% glutaraldehyde, 1% formaldehyde solution known to inhibit PPO but not myeloperoxidase, no reaction product was observed. The demonstration of PPO activity in these undifferentiated cells established their megakaryoblastic lineage. Further studies including DNA flow cytometry confirmed that these cells were megakaryoblasts. This study demonstrates that megakaryoblastic malignancy may occur as an acute leukemia of childhood and emphasizes the value of ultrastructural cytochemistry in cases of acute leukemia which defy routine classification. Abnormalities of chromosome 21 were present in our patient as in four previous cases of megakaryoblastic leukemia in childhood. The nonrandomness of chromosome 21 involvement in this disease should therefore be considered.

Acute Disease↗

PubMed1984

Functional properties of antigen-specific T cells infected by human T-cell leukemia-lymphoma virus (HTLV-I).

Tetanus-toxoid specific helper-inducer T-cell clones, which had been infected and transformed by human T-cell leukemia-lymphoma virus (HTLV-I), were obtained from an antigen-specific human T cell line by using a limiting dilution technique in the presence of the virus. These HTLV-I-infected T-cell clones proliferated specifically in response to soluble tetanus toxoid but, unlike normal T cells, they could do so in the absence of accessory cells. The HTLV-I-infected T-cell clones did not present the antigen to autologous antigen-specific T cells that were not infected with HTLV-I. The capacity of helper-inducer T cells to retain antigen-specific reactivity after infection by HTLV-I, while losing the normal T-cell requirement for accessory cells, has clinical and theoretical implications.

Antigens, Surface↗

PubMed1984

The pathologic spectrum of adult T-cell leukemia/lymphoma in the United States. Human T-cell leukemia/lymphoma virus-associated lymphoid malignancies.

The human T-cell leukemia/lymphoma virus (HTLV) is a novel Type-C retrovirus isolated from patients with post-thymic T-cell malignancies. Thirteen patients diagnosed in the United States were identified as having antibodies to HTLV and a typical clinicopathologic syndrome characteristic of adult T-cell leukemia/lymphoma as described in Japan. The most characteristic diagnostic feature in our series was the presence of highly pleomorphic and lobated lymphoid cells in the peripheral blood. Also notable was acid phosphatase activity which was partially tartrate-resistant in the neoplastic cells. The pathologic spectrum of the associated lymphomas was broad and encompassed several diffuse histologic subtypes in the Rappaport classification, the working formulation, and the classification of the Japanese lymphoma study group. However, differences in survival could not be correlated with differences in histologic subtype. All patients presented with Ann Arbor Stage IV lymphoma. Other common clinical features were generalized lymphadenopathy, hepatosplenomegaly, skin and peripheral blood involvement, hypercalcemia, and lytic bone lesions. The clinical course was aggressive with a median survival of 9 months. In two-third of patients with cutaneous involvement, epidermal infiltration resembling Pautrier microabscesses was observed. However, most cases can be readily distinguished from mycosis fungoides/Sézary syndrome on clinical and epidemiologic grounds. The presence of HTLV-antibodies in patients with lymphoid malignancies appears to define a distinct clinicopathologic syndrome which tends to occur in geographic clusters. Adult T-cell leukemia/lymphoma is favored as the diagnostic term for this clinicopathologic entity.

Adult↗

PubMed1984

In vitro enhancement of immunoglobulin gene expression in chronic lymphocytic leukemia.

B cell chronic lymphocytic leukemia (CLL) cells appear to be arrested in their differentiation so that little immunoglobulin is secreted in most cases. To determine their capacity for further differentiation we stimulated cells from a series of 10 cases of CLL with a phorbol ester and assayed for production of immunoglobulin protein, accumulation of immunoglobulin mRNA, and alterations in cell surface markers. We found that cells from all cases were induced to secret monoclonal immunoglobulin of the same heavy and light chain type as the surface membrane immunoglobulin type. Immunoglobulin secretion was preceded by a rapid increase in the levels of mRNA coding for IgM, predominantly the secretory form, mu s-mRNA, rather than the membrane form, mu m-mRNA. A similar selection of mu s- over mu m-mRNA is known to occur in plasma cells by a mechanism of differential processing of mRNA from a single mu-chain gene. Except for a decline in the expression of surface IgD, cell surface determinants remained unaffected both in terms of the percentage of positive cells and the relative number of sites per cell. In contrast to previous studies, these results indicate that CLL cells consistently retain the capacity to further differentiate toward plasma cells and secrete immunoglobulin. The immunoglobulin secretion is mediated, at least in part, by a developmentally regulated increment in mu s-mRNA.

B-Lymphocytes↗

PubMed1984

Low-grade lymphomas. Expression of developmentally regulated B-cell antigens.

A series of low-grade B-cell lymphomas was analyzed for a battery of immunologic determinants by flow cytometry and immunohistochemistry. Histologically distinctive subclasses of these lymphomas, well-differentiated lymphocytic (WDL), intermediately differentiated lymphocytic (IDL), and follicular center cell (FCC) lymphoma, were found to be readily distinguishable by their expression of immunologic determinants that are known to be developmentally regulated in normal B cells. Although all cases expressed monoclonal surface immunoglobulin (sIg), HLA-DR, and the surface membrane proteins recognized by antibodies B1 (p32) and BA1, staining with other monoclonal antibodies revealed unique immunologic phenotypes for each subclass: WDL p65 (Leu 1)+, p24 (BA2)-; IDL p65+, p24+; FCC p65-, p24-. Additionally, the fluorescence intensities (number of determinants per cell) obtained for sIg, BA-1, and B1, but not HLA-DR, were significantly different among the three lymphoma subclasses. The relative fluorescence intensities of each of these three markers followed the same pattern: FCC greater than IDL greater than WDL. Taken together, these distinguishing features suggest that low-grade B-cell lymphomas represent arrested, and possibly sequential, stages of B-cell differentiation.

Antibodies, Monoclonal↗

PubMed1984

Suppression of immunoglobulin synthesis by lymphocyte subpopulations in patients with Crohn's disease.

In previous studies, patients with mild or inactive Crohn's disease were found to have increased suppressor T-cell activity. To further characterize suppressor T cells in Crohn's disease, studies were carried out with the use of monoclonal antibodies. Excessive suppressor activity was eliminated by removal of OKT 8+ lymphocytes by complement-mediated lysis. However, the percentage of OKT 8+ (or Leu 2a+) cells and the ratio of OKT 4+ to OKT 8+ (or Leu 3a+ to Leu 2a+) cells were not significantly different from normal. Although the subgroup of patients with increased suppression of immunoglobulin synthesis had a significantly lower mean Leu 3a to Leu 2a ratio than that of normal subjects, in the whole group of Crohn's patients studied, neither the percentage of Leu 2a+ cells nor the ratio of Leu 3a+ to Leu 2a+ cells correlated with excessive suppression of immunoglobulin synthesis. A subpopulation of Leu 2a+ lymphocytes reactive with the monoclonal antibody HNK-1 (Leu 2a+ HNK-1+) was increased in patients with Crohn's disease. Furthermore, elimination of HNK-1-reactive lymphocytes by complement-mediated lysis diminished the excessive suppressor cell function in patients with Crohn's disease. The percentage of Leu 2a+ HNK-1+ lymphocytes correlated significantly with the suppression of pokeweed mitogen-stimulated immunoglobulin synthesis in vitro. Thus, patients with mild Crohn's disease have an increased suppressor cell activity in vitro which correlates with the presence of a subset of lymphocytes that have an HNK-1+ Leu-2a+ phenotype.

Adolescent↗

PubMed1983

Immunoglobulin-gene rearrangements as unique clonal markers in human lymphoid neoplasms.

Immunoglobulin genes in their germ-line form are separated DNA subsegments that must be joined by means of recombinations during B-cell development. Individual immunoglobulin-gene rearrangements are specific for a given B cell and its progeny. We show that the detection of such gene rearrangements by Southern hybridization provides a sensitive marker for both clonality and B-cell lineage within lymphoid tissues lacking expression of definitive surface phenotypes. We have used these genetic markers in three ways: to establish a diagnosis of lymphoma in a neoplastic disorder of uncertain cell type, to show that some lymphomas that were previously classified as being of T-cell type in fact contain monoclonal B cells, and to detect clonal B-cell populations within lymphomatous tissues of uncertain immunotype and within an atypical lymphofollicular hyperplasia having no other clonal surface markers. These sensitive and unique indicators of clonality located directly at the DNA level are capable of providing insights into the cellular origin, early detection, and natural history of neoplasia.

B-Lymphocytes↗

PubMed1983

Lymphoid blast crises of chronic myelogenous leukemia represent stages in the development of B-cell precursors.

The origin and stage of differentiation of the blast-crisis cells in chronic myelogenous leukemia have remained uncertain. Because immunoglobulin heavy-chain and light-chain genes must undergo a DNA rearrangement during B-cell development but rarely do so in human non-B-cell lineages, we examined these genes in 18 episodes of chronic myelogenous leukemia. In eight of nine episodes of lymphoid blast crisis, heavy-chain genes were rearranged, and in three, rearrangements in light-chain genes were also present. In contrast, cells from chronic myeloid, myeloid blast, and erythroid-like phases retained germ-like immunoglobulin genes. The observed phenotypic markers and gene configurations revealed that most lymphoid blast crises represent stages of development of B-cell precursors. In two separate episodes of lymphoid crisis, cells from a single patient possessed identical heavy-chain but different light-chain-gene configurations. Thus, the precursor cells that monoclonally expand to produce a lymphoid crisis are capable of immunoglobulin-gene rearrangements and represent discrete steps in early B-cell maturation.

Antigens, Surface↗

PubMed1983

Polymorphonuclear neutrophils express the common acute lymphoblastic leukemia antigen.

Monoclonal antibodies J5, VIL-A1, and BA-3, known to react with the common acute lymphoblastic leukemia antigen (CALLA) were found to specifically stain normal human polymorphonuclear neutrophils (PMN). The antigen detected on PMN had a molecular weight (95,000-110,000 mol wt) close to that of CALLA (95,000-100,000 mol wt) and thus these surface membrane antigens are likely related, if not identical. The fluorescent staining intensity of PMN is comparable to that of CALLA-positive leukemic cells and the presence of PMN in patient samples could potentially produce false-positive results in diagnosis.

Adult↗