Human T-cell Leukemia-lymphoma Virus Research Articles

Establishment of an Interleukin 2-Dependent Human T Cell Line From a Patient With T Cell Chronic Lymphocytic Leukemia Who Is Not Infected With Human T Cell Leukemia/Lymphoma Virus

AbstractWe established an interleukin 2 (IL-2)-dependent human T cell line, Kit 225, from a patient with T cell chronic lymphocytic leukemia (T-CLL) with OKT3+, –T4+, –T8– phenotype. Southern blot analysis showed that Kit 225 is not infected with human T cell leukemia/lymphoma virus (HTLV) type I or II, and is probably derived from the major clone in the fresh leukemic cells. Kit 225 cells express a large amount of IL 2 receptors constitutively and their growth is absolutely dependent on IL 2. No other stimuli, such as lectins or antigens, are required for maintaining the responsiveness to IL 2. As abnormal IL 2 receptor expression was also seen originally in the fresh leukemic cells, the establishment of this cell line with IL 2 suggests that IL 2-mediated T cell proliferation is involved in the leukemogenesis of some cases of HTLV-negative T-CLL.

Non-Hodgkin lymphoma in Jamaica and its relation to adult T-cell leukemia-lymphoma.

Of 95 patients consecutively diagnosed with non-Hodgkin lymphoma, 52 (55%) had antibodies to human T-cell leukemia-lymphoma virus, type I. Antibody positivity was strongly associated with skin involvement, leukemia, and hypercalcemia (p less than 0.02). Two patients had systemic opportunistic infections. Neither meningeal nor lung infiltration was detected, and lymph node infiltration was diffuse in all patients. Of 36 patients who received immunophenotypic classifications, 30 had diseases that affected the T-cell system, and the cells of all tested patients with these diseases showed the helper/inducer (T4) phenotype. Twenty-seven of these thirty-six patients were found to have adult T-cell leukemia-lymphoma, and of the 27, 24 had antibodies to HTLV-I. The median duration of survival in patients with adult T-cell leukemia-lymphoma was 17 weeks, but a subgroup of 9 patients had indolent courses and a median survival of 81 weeks, which suggests that the disease has differing expression with courses that range from smoldering and indolent to acute and rapidly fatal. Hypercalcemia was the most important prognostic determinant of adult T-cell leukemia-lymphoma.

The isolation of human T-cell leukemia lymphoma virus I.

The isolation of human T-cell leukemia lymphoma virus I.

T lymphomas associated with human T-cell leukemia-lymphoma virus may show phenotypic and functional differences from adult T-cell leukemias

We report phenotypic and functional characterization of lymph node cells from 27 patients with T lymphoma from the Nagasaki district of Japan. Nagasaki is one of several areas where adult T-cell leukemia is endemic, and sera from 18 of 23 patients examined (78.3%) were positive for antibody to adult T-cell leukemia-associated antigen. The majority of cases (24 of 27) showed the Leu-2a −, 3a +, MASO36c − phenotype similar to adult T-cell leukemia, but seemed to be subdivived into four groups according to the presence or absence of Leu-1 antigen and the receptor for sheep erythrocytes (SRBC). Eleven cases had both Leu-1 antigen and the receptor for SRBC (E-RFC +, Leu-1 +, 2a −, 3a +), but 6 cases lacked Leu-1 antigen (E-RFC +, Leu-1 −, 2a −, 3a +), 4 cases lacked receptors for SRBC (E-RFC −, Leu-1 +, 2a −, 3a +), and 3 cases lacked both of these markers (E-RFC −, Leu-1 −, 2a −, 3a +). The effects of lymph node T cells on pokeweed mitogen (PWM)-induced normal B-cell differentiation was studied in 10 cases. Lymph node T cells of 5 cases showed helper activity without any suppressor activity, and 2 cases showed suppressor activity with almost no detectable helper activity, similar to adult T-cell leukemia cells. Two other cases lacked particular functional effects. These results suggest the possibility that Leu-3a + T-cell lymphoma may be functionally subdivided into at least two types, a helper type and a suppressor (adult T-cell leukemia) type.

Human T cell leukemia/lymphoma virus I infection and subsequent cloning of normal human B cells. Direct responsiveness of cloned cells to recombinant interleukin 2 by differentiation in the absence of enhanced proliferation.

A human T cell leukemia/lymphoma virus (HTLV)-I-infected B cell clone expressed Tac antigen on its cell surface and responded to recombinant interleukin 2 (IL-2) by increased production of IgM without any increase in proliferation. Anti-Tac antibody completely inhibited the IL-2-induced differentiation of this HTLV-I-infected B cell clone. This study demonstrates that HTLV-I can directly infect normal mature human B cells, and that the Tac antigen, which may be induced by infection with HTLV-I, is the functional receptor for IL-2-induced B cell differentiation. The availability of such cell lines and clones should provide useful tools to delineate precisely the differentiation step in the human B cell cycle.

Human T cell leukemia/lymphoma virus-infected antigen-specific T cell clones: indiscriminant helper function and lymphokine production.

The ability of human T cell leukemia/lymphoma virus (HTLV)-I to alter the function of infected T lymphocytes was examined directly by investigating the properties of an antigen-specific T cell clone before and after transformation with HTLV-I. Following infection, the T4 antigen-specific clone manifested a tenfold increase in its surface interleukin 2 (IL 2) receptor (Tac) density and acquired the viral determinants p19, p24, and 4D12 not present in the uninfected clone. Prior to infection, the T cell clone responded to antigen stimulation in the presence of histocompatible antigen-presenting cells with proliferation and secretion of multiple lymphokines, including IL 2, B cell growth factor (BCGF), B cell differentiation factor (BCDF), and interferon-gamma (IFN-gamma). Following infection, the T cell clone both proliferated and produced constitutively three of these lymphokines (BCGF, BCDF, and IFN-gamma) in the absence of accessory cells or antigen. Co-cultivation with any accessory cells regardless of histocompatibility resulted in increased proliferation and lymphokine production. IL 2 production by the HTLV-I-transformed cell, however, could not be detected. Similarly, the uninfected clone was able to provide B cell help for Ig production only when stimulated with both histocompatible cells and antigen. In contrast, the infected cell provided T cell help to B cells in an unregulated manner, independent of antigen or histocompatibility. Thus, functions such as the induction of proliferation, B cell help, and lymphokine production, which are finely regulated in uninfected antigen-specific T cell clones, became indiscriminant after HTLV-I infection.

HTLV-I Antibodies in Childhood Leukemia

To the Editor.— There have been reports of adults with leukemia and lymphoma with serum antibodies to human T-cell leukemia-lymphoma virus (HTLV-I), the agent linked to adult T-cell leukemia-lymphoma.1-3This disease has been identified in specific geographic regions: Japan, the Caribbean basin, and in the southeastern United States.2In these studies, patients with non-T-cell type did not have seropositive antibodies for HTLV-I, although normal persons did have some antibody titers in the endemic area.4All these studies were performed on adult patients. We explored the possibility that HTLV-I could be associated with acute leukemia in children. Materials and Methods.— Serum specimens were obtained from 92 children with acute leukemia at the time of initial diagnosis when evaluated at the Division of Pediatric Hematology-Oncology at Emory University, Atlanta. Patients were all younger than 21 years at the time of diagnosis. Ninety-two patients had acute lymphocytic leukemia, of which

Racial and other characteristics of human T cell leukemia/lymphoma (HTLV-I) and AIDS (HTLV-III) in Trinidad.

Adult T cell leukaemia/lymphoma was first recognised as a clinical entity in southwest Japan. Subsequently the Caribbean has been found to be another area where the disease is endemic, and sporadic cases have been identified in different parts of the world. The human T cell leukaemia/lymphoma virus (HTLV-I) is causally related to adult T cell leukaemia/lymphoma. A subgroup of HTLV, designated HTLV-III, has recently been isolated from many patients with the acquired immunodeficiency syndrome (AIDS) and preAIDS, and there is now evidence that this variant is the primary cause of AIDS. This is the first report from Trinidad to describe 12 cases of adult T cell leukaemia/lymphoma and 14 of AIDS. All were in patients of African descent. No cases were seen in subjects of East Indian descent, who, like those of African descent, comprise as much as 40% of the population. West Indians of African descent may have increased susceptibility to infection with both HTLV-I and HTLV-III.

Burkitt cell acute lymphoblastic leukemia with partial expression of T-cell markers and subclonal chromosome abnormalities in a man with acquired immunodeficiency syndrome

A 45-year-old white male, bisexual, with a 2-year history of acquired immunodeficiency syndrome (AIDS) prodrome, developed a Burkitt cell-like acute lymphoblastic leukemia (ALL). Marker studies of marrow blasts show an unusual and possibly unique pattern, in that an unequivocal monoclonal B cell leukemia, having K-IgM with HLA-DR and B cell subset antigen (BA-1) expression, was superimposed with a mature suppressor T cell marker profile (pan-T, mature T, and suppressor/cytotoxic T antigens). The leukemic blasts were totally negative for terminal deoxynucleotidyl transferase (TdT), human T cell leukemia-lymphoma virus (HTLV) p19 antigen, and other immunoglobulin isotypes. Chromosome analysis of marrow cells disclosed that 70% of the cells had 47,XY,+12,t(8;14)(q24;q32) chromosome complement, and 30% of the cells had a 47,XY,+12,dup1q+(q22–31),t(8;14)(q24;q32). The consistent finding of the specific chromosome rearrangement ( 8 14 translocation) in all abnormal cells suggests that the cells were derived from a common precursor. With regard to the partial T cell marker expression, the significance of these markers in B cell leukemia is unclear, as is their relation to the additional chromosome abnormalities that apparently developed in the process of clonal evolution.

A monoclonal antibody specific for a 52,000-molecular-weight human T-cell leukemia virus-associated glycoprotein expressed by infected cells.

A monoclonal antibody, designated HT462, is described which is specific for an antigen expressed in human T-cell leukemia/lymphoma virus (HTLV) preparations and by HTLV-infected cells. In indirect immunofluorescence assays, the antigen was detected on the surface of both HTLV-transformed producer and nonproducer cells, including cells infected in vitro with either HTLV subgroup I (HTLV-I) or HTLV-II. Normal human peripheral blood lymphocytes stimulated with phytohemagglutinin, cord blood T cells cultured with T-cell growth factor, and a variety of HTLV-negative T- and B-cell lines all lacked HT462 antigen expression. The HT462 antigen is a 52,000-molecular-weight glycoprotein, as shown by Western blotting procedures and treatment of viral preparations with neuraminidase, endoglycosidase F, and trypsin. The unglycosylated molecule is approximately 42,000 daltons. That the antigen is virus associated was demonstrated by its banding at the density of HTLV in gradients of metrizamide and by its concomitant synthesis with HTLV gag proteins after short-term culture of primary HTLV-positive leukemic cells. Differential expression of the HT462 antigen and HTLV gag-pol gene products was observed. In one case, low HT462 expression was correlated with the known inability of the particular cell line to produce syncytia in vitro. The properties of the HT462 antigen are most consistent with it being a gene product of the HTLV px region or else a cellular antigen specifically induced after viral infection. We cannot rule out, however, that the antigen is a variant cleavage product of the env gene. The monoclonal HT462 will be useful in further definition of the proteins and functions encoded by the env-px genetic sequence and in studying the biological properties of HTLV-transformed cells. Furthermore, the monoclonal, by recognizing HTLV-transformed nonproducers, will allow a greater spectrum of virus-infected cells to be detected.

HTLV-I antibodies associated with cutaneous T cell lymphoma in Denmark.

The first isolation and characterization of a human retrovirus was done by Gallo and co-workers from patients with aggressive cases of adult T cell leukemia/lymphoma in the United States [15, 16]. This virus, named human T cell leukemia/lymphoma virus (HTLV-I) is a T cell-tropic retrovirus [8, 9], initially found sporadically among United States cases of adult T cell leukemias and lymphomas (ATLL) with cutaneous manifestations resembling aggressive variants of mycosis fungoides. Subsequently, HTLV-I was specifically linked to adult T cell malignancies in Japan and the Caribbean [8]. A second human retrovirus, HTLV-II, which is related to but distinct from all previous HTLV-I isolates, was identified and isolated from a patient with hairy cell leukemia [12]. Recently, a third retrovirus, HTLV-III has been isolated and is a probable etiologic agent in the acquired immune deficiency Syndrome (AIDS) [4–7].

A possible association between HTLV-I and B-cell chronic lymphocytic leukemia in Jamaica.

Cell surface markers were determined in 10 patients with chronic lymphocytic leukemia (CLL) in Jamaica, an area endemic for the human T-cell leukemia/lymphoma virus (HTLV-I) and with a high positivity for HTLV-I antibody titers in CLL patients. The results demonstrated that the predominant cell phenotype in all patients was of B-cell origin. When surveyed for HTLV-I antibody, 3 out of 5 patients with B-CLL from this series were found positive. A possible association between HTLV-I and B-CLL is discussed.

Retroviruses and human cancer: evaluation of T-lymphocyte transformation by human T-cell leukemia-lymphoma virus.

(1985). Basic Science Review: Retroviruses and Human Cancer: Evaluation of T-Lymphocyte Transformation by Human T-Cell Leukemia-Lymphoma Virus. Cancer Investigation: Vol. 3, No. 2, pp. 145-160.

Molecular studies of human T cell leukemia/lymphotropic retroviruses.

Human T cell leukemia/lymphoma virus (HTLV), a name originally used to designate a retrovirus associated with a form of mature T cell malignancy, namely adult T cell leukemia (ATL), now stands for a family of human retroviruses isolated in nature, which preferentially infect a subset of T lymphocytes [20]. In addition to their common cell tropism, viruses of this family share other biological, physiochemical, and biochemical properties. However, one major difference in the biological properties of these viruses is directly reflected in their different disease spectrums and transforming capabilities. The virus associated with ATL, renamed HTLV-I, transforms T cells efficiently in culture. A second virus, HTLV-II, which was obtained initially from culture T cells of a patient with hairy cell leukemia, also has the capacity to transform T cells in vitro. A third virus subgroup, HTLV-III, lacks transforming activity but is instead highly cytopathic. HTLV-III has been unambiguously shown to be the etiological agent of the acquired immunodeficiency Syndrome (AIDS) [4, 9, 11, 12].

Human T-cell leukemia/lymphoma viruses: clinical and epidemiologic features.

The discovery of the human T-cell leukemia/lymphoma virus (HTLV) family of retroviruses represents the culmination of a search that dates back more than 50 years, stimulated chiefly by the fact that leukemias and lymphomas in animals are often caused by retroviruses (Poiesz etal. 1980; Gallo and Wong-Stall 1982). Since the first virus isolation of HTLV in 1979, more than 100 isolates of human retroviruses, representing three distinet classes, have been reported from many laboratories in the world (Popovic etal. 1983b; Haynes etal. 1983; Miyoshi etal. 1981; Kalyanaraman etal. 1982b; Gallo etal. 1984). All of these isolates have an affinity to infect mature T cells and are, therefore, said to be T-lymphotrophic. The three classes of HTLV are termed HTLV-I, HTLV-II and HTLV-III. HTLV-I has been most thoroughly charac- terized from a molecular and clinical perspective and has been found to be associated with certain forms of mature T-cell leukemia/lymphoma, the description of which will be a major focus of this review (Blatnner et al. 1982; Gallo et al. 1983a; Blayney et al. 1983c). HTLV-II has more recently been isolated, and its relationship to human disease is unclear, although isolates have been obtained from a single patient with T hairy cell leukemia and from some patients with the acquired immune deficiency syndrome (Kalyanaraman et al. 1982b; Hahn et al., in press). In the latter case, it is likely that these isolations represented passenger viral infections, given the recent discovery of HTLV-III as the etiological agent for the acquired immune deficiency syndrome (AIDS) (Gallo et al. 1984). The molecular features of this class of viruses are reviewed elsewhere in this volume.KeywordsAcquire Immune Deficiency SyndromeBovine Leukemia VirusHairy Cell LeukemiaAcquire Immune Deficiency Syndrome PatientAcquire Immune Deficiency Syndrome CaseThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Decline of natural cytotoxicity of human lymphocytes following infection with human T-cell leukemia/lymphoma virus (HTLV)

Cell-mediated natural cytotoxicity (CMNC) of fresh or long-term cultured lymphocytes collected from HTLV-positive patients or infected in vitro with the virus, was tested against K562 target cells. Severe depression of reactivity was found in fresh lymphocytes of three patients with advanced disease, in 12 in vitro established T-cell malignant lines, and two HTLV-infected cord blood (C5/MJ and C91/PL) lines. Moreover, all (eight) HTLV-1 infected cell lines listed showed a significant inhibition of CMNC of peripheral blood lymphocytes of healthy donors. Whether virus infection promotes the outgrowth of pre-existing suppressor cells and/or produce changes of the T-lymphocyte function is unknown.

Presence of antibodies to human lymphoma-leukemia virus (HTLV-I) in Germans with symptoms of the acquired immunodeficiency syndrome (AIDS).

Sera from German patients exhibiting symptoms compatible with the acquired immune deficiency syndrome (AIDS) or the lymphadenopathy syndrome (LAS) were assayed for antibodies against human T-cell lymphoma/leukemia virus (HTLV-I)-related antigens by enzyme immunoassay, indirect immunofluorescence, and radioimmunoprecipitation. Antibodies against HTLV were detected in 3 out of 31 sera.

Retrovirus-induced expression of interleukin 2 receptors on cells of human B-cell lineage.

We provide direct evidence for interleukin 2 receptor (IL2R) induction by human T-cell leukemia/lymphoma virus (HTLV) in human B-cell lines. A lymphoblastoid B-cell line (LCL-Ter) was established by Epstein-Barr virus-induced transformation of peripheral blood lymphocytes derived from a healthy HTLV carrier and cloned in vitro. HTLV gp21 and/or p19 antigens were detected in eight LCL-Ter clones, all of which also expressed the IL2R antigens Tac and Hiei (defined by monoclonal antibodies). However, five other LCL-Ter clones, which were negative for the HTLV antigens, did not express the IL2R antigens. Furthermore, when the IL2R-negative B-cell line LCL-Kan, derived form a normal donor, was cocultured with HTLV-producer cells, three HTLV-carrying clones were obtained and found to constitutively express IL2R. IL2R induced by HTLV on these B-cell lines bound recombinant interleukin 2 and were similar in apparent molecular mass (approximately equal to 60 kDa) to the IL2R of peripheral blood lymphocytes stimulated with phytohemagglutinin.

Efficient transformation of previously activated and dividing T lymphocytes by human T cell leukemia-lymphoma virus

Modifying previously reported techniques, we attempted to increase the efficiency of human T cell leukemia-lymphoma virus (HTLV) transformation of human T lymphocytes. Lethally irradiated donor cells (DCs) were cultured with target mononuclear cells (TMCs). DCs included ten HTLV+ T cell lines with varying degrees of virus expression or seven cell lines that do not express HTLV. TMCs were prepared from 20 cord and 16 adult peripheral blood samples, including eight patients with acquired immunodeficiency syndrome (AIDS). TMCs were either added directly to the DCs or were first stimulated with phytohemagglutinin (PHA) (5 micrograms/mL) and grown in T cell growth factor (TCGF) prior to exposure to DCs. The presence of integrated HTLV proviral DNA in the transformed cells was determined by dot blot hybridization, utilizing a cloned probe to the HTLV-I genome. HTLV production by transformed TMCs was assessed for HTLV p19, reverse transcriptase, and virus particles. No transformation occurred with T cell donor lines that do not express HTLV. Low virus expressor DCs could only, with rare exception, transform preactivated TMCs. High-titer virus-producing DCs could transform activated and nonactivated cord blood cells and activated adult TMCs. Only MT-2 could routinely transform nonactivated normal adult and activated AIDS TMCs. HUT 102 B2 could transform only one activated AIDS sample, the cells of which initially expressed HTLV-like proteins and virions. Transformed cell lines contained subsets of mature T lymphocytes with variable HTLV expression. Prior activation and culture of the T lymphocytes increases the probability and rate of transformation by HTLV, allowing for biologic detection of low HTLV- producing cells and for in vitro expansion of T lymphocyte subsets from selected patients.

Efficient Transformation of Previously Activated and Dividing T Lymphocytes by Human T Cell Leukemia-Lymphoma Virus

Modifying previously reported techniques, we attempted to increase the efficiency of human T cell leukemia-lymphoma virus (HTLV) transformation of human T lymphocytes. Lethally irradiated donor cells (DCs) were cultured with target mononuclear cells (TMCs). DCs included ten HTLV+ T cell lines with varying degrees of virus expression or seven cell lines that do not express HTLV. TMCs were prepared from 20 cord and 16 adult peripheral blood samples, including eight patients with acquired immunodeficiency syndrome (AIDS). TMCs were either added directly to the DCs or were first stimulated with phytohemagglutinin (PHA) (5 micrograms/mL) and grown in T cell growth factor (TCGF) prior to exposure to DCs. The presence of integrated HTLV proviral DNA in the transformed cells was determined by dot blot hybridization, utilizing a cloned probe to the HTLV-I genome. HTLV production by transformed TMCs was assessed for HTLV p19, reverse transcriptase, and virus particles. No transformation occurred with T cell donor lines that do not express HTLV. Low virus expressor DCs could only, with rare exception, transform preactivated TMCs. High-titer virus-producing DCs could transform activated and nonactivated cord blood cells and activated adult TMCs. Only MT-2 could routinely transform nonactivated normal adult and activated AIDS TMCs. HUT 102 B2 could transform only one activated AIDS sample, the cells of which initially expressed HTLV-like proteins and virions. Transformed cell lines contained subsets of mature T lymphocytes with variable HTLV expression. Prior activation and culture of the T lymphocytes increases the probability and rate of transformation by HTLV, allowing for biologic detection of low HTLV-producing cells and for in vitro expansion of T lymphocyte subsets from selected patients.