Giant Septic Lymphadenitis with Marked Gas Formation Caused by Bacteroides fragilis in a Patient with Adult T-cell Leukemia/lymphoma.

Adult T-cell leukemia/lymphoma (ATL) sometimes causes opportunistic infections. A 53-year-old woman with systemic lymphadenopathies was diagnosed with ATL by inguinal lymph node biopsies and underwent oral chemotherapy. Two months later, high grade fever, lower abdominal pain and lymphadenopathy recurred. Computed tomography revealed the presence of lymphadenopathy with marked gas formation in the pelvic lesion. Blood cultures were suggestive of septic lymphadenitis by Bacteroides fragilis (BF). This represents the first demonstration of giant lymphadenitis with gas formation caused by BF in a patient with ATL. Notably, septic lymphadenitis is pivotal in the differential diagnosis of systemic lymphadenopathy in ATL.

Invasive rhino-maxillary mucormycosis diagnosed before HSCT.

We report a case of successful allogeneic hematopoietic stem cell transplantation (HSCT) with full myeloablative conditioning in a patient with pre-existing invasive mucormycosis. The mucormycosis involved the maxilla, the nasal septum, and the hard palate. Sustained antifungal therapy and aggressive surgery both before and after HSCT were required.

A novel adult T cell leukemia-derived cell line (SALT-3) susceptible to human immunodeficiency virus type 1 infection.

A novel human T cell line (SALT-3) was established from the pleural effusion of a patient with adult T cell leukemia (ATL) of lymphoma type. SALT-3 showed atypical T cell markers such as CD1-CD2-CD3-CD4+CD5+CD7+CD8-CD19-CD20-CD25+HLA-DR+. T cell receptor alpha/beta and gamma/delta were undetectable. Human T cell lymphotropic virus type 1 (HTLV-I) particles were seen on SALT-3 cells by electron microscopic analysis. HTLV-I gag p19, proviral DNA and mRNA of HTLV-I genes were also detected in the cells. Chromosome analysis showed abnormal karyotypes as 47, XY, partial trisomy of No.3 chromosome, and trisomy of No. 7 chromosome. Furthermore, SALT-3 were susceptible to the infection of human immunodeficiency virus type 1 (HIV-1) and the cells were rapidly killed after HIV-1 infection. This newly established HTLV-I-infected human T cell line would be a useful tool to study biological activities of atypical type of ATL cells and to examine the cytotoxic effects of HIV-1 and it's modulators.

[Detection of HTLV-1 provirus DNA in T-cell neoplasms].

Using polymerase chain reaction (PCR) amplification and Southern hybridization, gag, pol, env and pX region genes of HTLV-1 provirus were detected in T-cell malignancy such as adult T-cell leukemia/lymphoma (ATLL), mycosis fungoides in leukemic phase (MF) and, CD8-leukemia. The gag, pol, and/or env regions, were not amplified in some cases of ATLL, which was considered to be induced by mutation, but not deletion. However, the pX-1 and pX-2 regions could be amplified in all cases examined. As it is suggested that the pX gene plays an important role in leukemogenesis, the mutation may not occur in this region gene. Interestingly, the pX-2 was amplified in the cases with MF and CD8-leukemia as well. The amplified products were hybridized to the HTLV-1 pX sequence, even though the products contained DNA fragments with a size larger than expected as well as those of an expected size. These results indicated the possibility that the virus having sequence identical with HTLV-1 pX is integrated in the tumor cells of MF and CD8 leukemia.

Retroviruses and cutaneous T-cell lymphoma.

Since the discovery of the first human retrovirus, HTLV-I, and its etiologic role in ATL, the search for a retrovirus and its role in the development and progression of CTCL has been vigorously pursued and debated. Current studies in CTCL have evaluated serum antibodies to retroviral proteins, electron microscopy to identify viruslike particles, and Southern blot analysis and PCR amplification to detect proviral DNA sequences. There have been inconsistent findings within and between a variety of studies, emphasizing the need for critical evaluation of experimental methods and their potential shortcomings. Several interesting observations have included (1) serologic evidence of HTLV-I infection in a small subset of CTCL patients, (2) cloning of a deleted HTLV-I proviral genome from a B-cell line established from the peripheral blood of a CTCL patient, (3) detection of retrovirus in Langerhans cells and B cells, and (4) molecular evidence for the presence of an HTLV-I-like retrovirus. By viewing CTCL as a model of tumor progression, mechanisms by which retroviruses play a role in the development and progression of CTCL are facilitated. Future studies will need to correlate the detection of proviral sequences and the nature of a retroviral infection with specific cell types and stage of disease and determine if these findings demonstrate a causal role in CTCL or a secondary phenomenon due to CTCL-associated immunosuppression. It is likely that new data will be reported between the writing of this article and the time of publication; however, the currently available data reviewed in this article do not provide conclusive evidence that retroviruses play a primary etiologic role in CTCL.

[HTLV-I negative adult T cell leukemia; a case report of acute type].

We described a case of adult T cell leukemia (ATL) not associated with human T-cell leukemia virus type I (HTLV-I), a clinical entity that was first reported by Shimoyama et al. A 79-year-old male was admitted with anorexia and fever in October, 1989. Physical examination revealed marked hepatosplenomegaly and superficial lymphadenopathies. Hematological examination revealed marked leukocytosis (136,300/microliters) with abnormal lymphoid cells showing highly lobulated nuclei. Hypercalcemia (11.2 mg/dl) and elevation of lactic dehydrogenase were also recognized. Surface marker analysis showed that the abnormal lymphoid cells in the peripheral blood were positive for CD2 and CD4 but negative for CD8. Southern blot analysis of the DNA from peripheral blood leukemic cells revealed monoclonal rearrangement of T-cell receptor beta-chain gene. The clinical and hematological findings of the patient were compatible with those of acute type ATL, however, serum anti-HTLV-I antibody was negative and HTLV-I proviral DNA was not detected in the leukemic cells by Southern blot analysis. Furthermore, the polymerase chain reaction showed no integration of the HTLV-I proviral DNA in the leukemic cells.

Adult T cell leukaemia-lymphoma.

Adult T cell leukaemia-lymphoma (ATL) was first discovered and reported in Japan, where it has a high incidence in the south-west region. The first human retrovirus HTLV-I (human T-cell lymphotropic virus type I) is considered to be related to its aetiology. In ATL endemic areas, HTLV-I carriers form a fairly high percentage of the population, even among healthy individuals. ATL shows diverse clinical features. It can be divided into four subtypes: acute, chronic, smouldering and lymphoma type. ATL cells originate from the CD4-positive subset of peripheral T cells; they show a characteristic notch in the nucleus and a tendency to lobulation. ATL resists chemotherapy, and patients with acute and lymphoma types have a fairly poor prognosis. A definite diagnosis of ATL is made by documenting the presence of HTLV-I proviral DNA in the DNA of tumour cells. HTLV-I infection is caused by transmission of live lymphocytes via three routes (from mother to child, from males to females, and by transfusion). Familial occurrence of ATL is frequently seen. HTLV-I infection is seen in other countries, but its incidence is highest in Japan. Infection with HTLV-I is a direct cause of ATL. Furthermore, infection with this virus can indirectly cause many other diseases via the induction of immunodeficiency, such as chronic lung disease, opportunistic lung infection, cancer of other organs, monoclonal gammopathy, chronic renal failure, strongyloidiasis, non-specific dermatomycosis, HTLV-I-associated lymphadenitis, HTLV-I uveitis and HTLV-I-associated myelopathy-tropical spastic paraparesis (HAM/TSP).

Adult T-cell leukemia/lymphoma revealed by a surgically cured cardiac valve lymphomatous involvement in an Iranian woman: clinical, immunopathological and viromolecular studies.

A 60-year-old woman from the town of Mashhad in northeastern Iran developed cardiac failure due to aortic and mitral regurgitations which needed cardiac valve replacement. Histopathological study of the valves revealed a T-cell non-Hodgkin's lymphoma. Blood examination showed leukemic features with 32% of abnormal white blood cells. Human T-cell leukemia/lymphoma virus type I (HTLV-I) antibodies were present in the serum and the specific env HTLV-I sequences were detected in the DNA extracted from the valves and peripheral blood mononuclear cells (PBMC) using polymerase chain reaction technique. Clonal integration of two HTLV-I copies was found in both the valves and PBMC DNA, thus the diagnosis of adult T-cell leukemia/lymphoma (ATL) was established. In contrast to the acute life-threatening cardiac localization, our case met the diagnostic criteria of chronic ATL, this was confirmed by favorable evolution without chemotherapy during the 24 months after diagnosis. According to our knowledge, this is the first report of an isolated lymphomatous cardiac valve involvement, without other cardiac abnormalities. It seems important to underline that the patient originated from Iran where endemicity of HTLV-I has only recently been discovered.

Detection of human T-cell lymphotropic virus type-1 proviral DNA in the saliva of an adult T-cell leukaemia/lymphoma patient using the polymerase chain reaction.

We report a case of adult T-cell leukaemia/lymphoma (ATLL), in whom the polymerase chain reaction (PCR) on genomic DNA from saliva demonstrated the monoclonal integration of human T-cell lymphotropic virus type-1 (HTLV-1) proviral DNA in lymphocytes in the saliva. These results provided evidence of the possibility of saliva-borne transmission of HTLV-1.

Epstein-Barr virus in adult T-cell leukemia/lymphoma.

Adult T-cell leukemia/lymphoma (ATLL) is a well-known human T-cell lymphotropic virus type-1-related disease. We studied Epstein-Barr virus (EBV) in the tumor cells of ATLL, to investigate the etiological significance of double infection with these viruses. We used polymerase chain reaction and EBV-encoded small RNA-1 in situ hybridization to investigate the presence of EBV and immunohistochemistry to detect EBV-related oncoproteins, such as EBV-determined nuclear antigen-2 and latent membrane protein. Polymerase chain reaction performed on DNA of frozen specimens from 96 cases of ATLL revealed that the tumor tissue from 21 cases contained EBV DNA. EBV-encoded small RNA-1 in situ hybridization performed on the paraffin sections of the polymerase chain reaction-positive cases indicated EBV in the nuclei of ATLL tumor cells in 16 cases, nine of which were in the pleomorphic nuclei. Latent membrane protein was also detected in the cytoplasm of ATLL tumor cells in 15 cases, and EBV nuclear antigen-2 was observed in the nuclei of ATLL tumor cells in 11 cases. We conclude that EBV was present within tumor cells in about 17% of cases with ATLL and expressed EBV oncoprotein in the tumor cells. It is hypothesized that EBV and human T-cell lymphotropic virus-1 may infect the same T cells in early life and may play a role in the oncogenesis of ATLL.

Hematopoietic progenitor cells from patients with adult T-cell leukemia-lymphoma are not infected with human T-cell leukemia virus type 1.

The in vivo host range of human T-cell leukemia virus type 1 (HTLV-1) has not been definitively established. To determine if hematopoietic stem cells from patients with adult T-cell leukemia-lymphoma (ATL) are infected with HTLV-1, we used a clonogenic progenitor assay followed by the polymerase chain reaction for the detection of HTLV-1 DNA. In vitro growth characteristics of myeloid (CFU-GM) and erythroid (BFU-E) progenitor cells among nonadherent T-cell-depleted bone marrow (BM) mononuclear cells (NA-T-MNCs) from 10 patients with ATL was not significantly different from those of HTLV-1-seronegative controls (P = .20); numbers of colonies per 1 x 10(5) NA-T-MNCs were 34.9 +/- 7.6 for CFU-GM and 39.0 +/- 12.5 for BFU-E in patients with ATL, whereas those were 32.1 +/- 9.5 for CFU-GM and 41.4 +/- 12.7 for BFU-E in normal controls. HTLV-1 DNA was not detected in individual colonies formed by CD34+ cells from any of the patients. Similarly HTLV-1 DNA was not detected in 1 x 10(3) CD34+ cells sorted on a fluorescence-activated cell sorter (FACS) from six patients with ATL studied. In contrast, HTLV-1 DNA was detected in BM mononuclear cells from all patients. These observations clearly indicate that hematopoietic progenitor cells from patients with ATL are normal in their colony-forming capacity and that CD34+ cells from patients with ATL are not infected with HTLV-1 in vivo.

In vivo infection of human T-cell leukemia virus type I in non-T cells.

Using three different procedures, immunomagnetic isolation, FACS and panning, each of the T4 cell, T8 cell, monocyte, and B cell population was fractionated from peripheral blood mononuclear cells (PBMC) obtained from 10 human T cell leukemia virus type I (HTLV-I)-associated myelopathy (HAM)/tropical spastic paraparesis (TSP) cases, 3 adult T cell leukemia and lymphoma (ATLL) cases and 9 HTLV-I-infected healthy carriers. DNA from each fraction was then subjected to the quantitative polymerase chain reaction. Although T4 cells possessed the highest copy number of HTLV-I in general, we also detected a significant amount of HTLV-I provirus in T8 cells, monocytes, and B cells. To confirm virus infection of the non-T cell populations, especially the monocyte fraction, we performed further experiments and the following results were obtained: (1) HTLV-I tax RNA expression was confirmed simultaneously in the RNA of monocytes that were negative for T cell receptor beta chain (TCR beta) expression in vivo. (2) We cultured these monocytes for a short period of time and detected HTLV-I antigen expression in non-specific esterase-positive monocyte/macrophage cells. Our data indicate that HTLV-I has a broad host range in vivo and that monocytes or cells of monocyte lineage such as tissue macrophages might comprise a virus reservoir in vivo.

Effects of fixation and varying target length on the sensitivity of polymerase chain reaction for detection of human T-cell leukemia virus type I proviral DNA in formalin-fixed tissue sections.

In this study, the fixation condition most suitable for maintaining the sensitivity of the polymerase chain reaction (PCR) was investigated by using the alpha-tubulin gene sequence, and the PCR procedure most effective for detecting human T-cell leukemia virus type I (HTLV-I) proviral sequences in fixed, embedded tissues of adult T-cell leukemia patients was explored. First, the sensitivity of the PCR targeting a 286-bp alpha-tubulin sequence was studied in tissue sections fixed in several fixatives for various periods at 25 or 4 degrees C. For histological examination, fixation with 10% buffered formalin at a lower temperature for a shorter period was found to be preferable to retain the sensitivity. And the HTLV-I sequence was detected in only 7 of 18 specimens (38.9%) when the 374-bp sequence of the gag region was targeted, but the rate increased to 77.8% (14 of 18 specimens) when the length of the target sequence was reduced to 120 bp within the same region. Therefore, the one-round PCR targeting a shorter sequence is preferable for application of PCR to archival fixed tissue specimens, the fixation condition of which may not be ideal for DNA preservation.

Detection of human T-cell lymphotropic virus type 1 infection by the polymerase chain reaction using dried blood specimens on filter papers.

A simple method for detection of proviral DNA sequences of human T-cell lymphotropic virus type 1 (HTLV-1) was developed using dried blood specimens on filter papers. The whole blood was blotted onto the Guthrie paper. After the blood has dried, the blotted paper was punched out into small discs. The discs were then boiled to prepare the template for PCR (filter paper-PCR method). The filter paper-PCR method detected even a single HTLV-1-infected cell in three discs. The sensitivity of the filter paper-PCR method was equivalent to that of the method in which DNA was extracted with phenol and used as the template for PCR (DNA extraction-PCR method). In addition, DNA in the blotted filter paper was still utilizable as the template after the storage at 25 degrees C for at least 7 wk. A total of 53 clinical specimens from 30 seropositive and 23 seronegative individuals who were screened by particle agglutination (PA) test were analysed for HTLV-1 DNA by both PCR methods. Of 30 PA-positive specimens, 28 were also positive for HTLV-1 antibody by Western blot (WB) analysis, but two were indeterminate. The twenty eight WB-positive and one of the two indeterminate specimens were positive for HTLV-1 proviral DNA by both PCR methods. Of 23 PA-negative specimens, 22 were negative for HTLV-1 proviral DNA by both PCR methods. However, one PA-negative specimen was positive by both PCR methods. This patient was a 16-mth-old infant who was born to an HTLV-1 carrier mother and fed thereafter without her breast milk. In comparison to DNA extraction-PCR method, the sensitivity and specificity of the filter paper-PCR method was 100%, respectively.

Atypical human T-cell lymphotropic virus type-I-associated T-cell lymphoma in a low-prevalence Alaska Native population. Implications for disease surveillance.

An atypical case of adult T-cell leukemia/lymphoma (ATL) associated with human T-cell lymphotropic virus type I (HTLV-I) occurred in a 46-year-old Inupiat Eskimo man with no behavioral risk factors for HTLV-I infection. The case was characterized by lack of atypical circulating lymphocytes, hypercalcemia, and opportunistic infections; and by complete remission of the initial renal parenchymal lymphoma. The lymphoma cells had a helper T-cell (CD4) immunophenotype. Serum antibodies to HTLV I/II, detected by Western immunoblot, were identified in specimens collected 31 months before the onset of illness, at the time of diagnosis, and up to 37 months later, shortly before the patient's death. Polymerase chain reaction was used to identify HTLV-I DNA in peripheral blood mononuclear cells and in lymphoma in involved skin. Clinicians should be alert to sporadic cases of both atypical and classic ATL, even in populations in which the prevalence of HTLV-I infection is low.

Rheumatic manifestation of human leukemia virus infection.

Rheumatic disorders associated with retroviruses are described in this article. A recent study of human T-cell leukemia virus type-I (HTLV-I) revealed that it appeared to be associated with the pathogenesis of several immune disorders such as myelopathy, broncho-pneumopathy, Sjogren's syndrome, and arthropathy. HTLV-I-associated arthropathy (HAAP) shows remarkable synovial proliferation with nuclear convoluted T-cell infiltration in both synovium and synovial fluid. Synovial cells obtained from HAAP patient-integrated HTLV-I proviral DNA and also expressed mRNA for HTLV-1 tax gene; moreover, HTLV-1 integrated synovial cell clones expressed a high level of mRNA for several oncogene and growth factors compared with HTLV-I non-integrated clones. These findings suggest that HTLV-I is the first exogenous retrovirus that contributes to synovial proliferation with immune disorders in humans.

Comparative analysis of HTLV-I promoter activities reveals no disease-linked pattern of expression.

To investigate the possibility of an association between the type of pathology caused by HTLV-I and the activity of its promoter, we compared the levels of transcription obtained with six LTRs isolated from patients with two different HTLV-I-related diseases: ATL and TSP/HAM. The patients came from different geographical endemic areas. The LTR region was amplified by polymerase chain reaction (PCR) from the DNA of uncultured peripheral blood lymphocytes, and directly cloned upstream of the luciferase reporter gene. Constructs were tested by a transient transfection assay in a variety of cell lines. Although the activities of these LTRs were statistically different in some of the cell lines tested, no correlation could be demonstrated between the promoter activity and the nature of the disease. Thus, the data suggest that the LTR is not a major determinant of the nature of the disease associated with the infection by HTLV-I.