Takao Shibata

Addition of rituximab to cyclophosphamide, doxorubicin, vincristine, and prednisolone therapy has a high risk of developing interstitial pneumonia in patients with non-Hodgkin lymphoma

There are a few reports suggesting that rituximab (RTX) might be a risk for interstitial pneumonitis (IP). We also experienced such patients in the era of RTX. Here, we reviewed all the patients with non-Hodgkin lymphoma who were treated with RTX-CHOP-like regimen (R-CHOP) to determine the risk of developing IP. One of 59 (1.7%) patients who received CHOP alone and 8 of 129 (6.2%) patients who were treated with R-CHOP experienced IP (p = 0.28). Furthermore, three of eight patients who have had IP during R-CHOP were confirmed having Pneumocystis jirovecii pneumonia (PCP). PCP occurred during the fourth, sixth, and seventh cycle of chemotherapy, respectively. Among the patients treated by R-CHOP, 3 of 32 (9%) patients whose lymphocyte counts were <1000/μL before chemotherapy developed PCP, while 70 patients whose lymphocyte counts were >1000/μL did not (p = 0.03). In four of eight patients, IP occurred during the administration of granulocyte-colony stimulating factor. RTX seems to have a certain risk to induce IP including PCP. Patients with lymphoma who were treated by R-CHOP regimen, might be considered as PCP prophylactic, especially if the number of lymphocytes is low at the beginning of chemotherapy.

CLASSICAL HODGKIN AND REED-STERNBERG CELLS DEMONSTRATE A NON-CLONAL IMMATURE B LYMPHOID LINEAGE: EVIDENCE FROM A SINGLE CELL ASSAY AND IN SITU HYBRIDIZATION

Hodgkin and Reed‐Sternberg (H & RS) cells are generally considered to be the neoplastic cells of Hodgkins disease (HD), however such cells are only found in a minority of the lesions. Recently in a few studies on HD, the clonality of H & RS cells was examined, using a single‐cell polymerase chain reaction (PCR) examination. To clarify the lineage and clonality of H & RS cells, we performed single cell PCR and in situ hybridization (ISH), and nine cases of classical HD were thus studied. By ISH, the immunoglobulin J chain, and the |gk and |gl light chain were rarely expressed in the H & RS cells, however, no T‐cell markers could be detected. The expression of the recombination activating genes (RAG‐1, 2) could be determined in the H & RS cells. We isolated CD30+ H & RS cells, CD3+ T cells and CD20+ B cells from suspended materials using a mechanical sorter. We performed single cell PCR in a sorted individual cell, to amplify the complementarity determining region of the Ig heavy chain (IgH) gene and T‐cell receptor γ chain (TCR γ) gene. In all cases, TCR γ could be frequently amplified in the T cells, but was only rarely amplified in the H & RS and B cells. In contrast, the IgH was frequently amplified in the H & RS and B cells, but not in the T cells. In addition, the PCR production of the H & RS cells all showed different lengths. The results therefore support the polyclonal nature and immature B lymphoid cell origin of H & RS cells.

Weekly Paclitaxel in the Treatment of Advanced or Metastatic Breast Cancer Previously Treated or Not Treated with Docetaxel: A Phase II Study

Background: Paclitaxel has been approved for 3-weekly administration in Japan. Recent reports suggest that weekly paclitaxel can achieve a higher tumor response and lower toxicity. Methods: This study was designed to investigate the usefulness and tolerability of weekly paclitaxel by 1-hour infusion in patients with metastatic breast cancer who were previously treated with docetaxel or other anticancer agents. Results: Thirty-five patients were enrolled. The overall response rate was 41.2% (14/34, 95% confidence interval: 24.6–59.3%). The median time to progression and the median survival time were 218.5 and 624 days, respectively. One patient developed dyspnea, probably induced by a hypersensitivity reaction. The most common hematological toxicities were leukopenia and neutropenia, although no patients developed grade 4 leukopenia or neutropenia and G-CSF support was not required. Conclusions: Weekly paclitaxel could be safely administered and achieved a relatively high response rate. Weekly paclitaxel would be a good candidate second-line therapy for recurrent or advanced breast cancer.

Clonal Analysis of Hodgkin's Disease Shows Absence of TCR/Ig Gene Rearrangement, Compared with T-Cell-Rich B-Cell Lymphoma and Incipient Adult T-Cell Leukemia/Lymphoma

To better characterize the clonality and pathogenesis of Hodgkins disease (HD), we used polymerase chain reaction (PCR) and Southern blot to analyze the rearrangement of immunoglobulin (Ig) and T-cell receptor (TCR) genes, the bcl-2 oncogene, and the Epstein-Barr virus (EBV) genotype. In situ hybridization studies of EBV were also done. Twenty-six cases of HD were compared with 15 cases of non-specific lymphadenitis, 7 with incipient adult T-cell leukemia/lymphoma (ATLL), and 4 T-cell rich B-cell lymphomas (TRBL), all of which histologically resembled HD. EBV genes were detected in 20 of 26 HD patients (77%) and in 7 of 15 patients with non-specific lymphadenitis (47%), 5 of 7 with incipient ATLL (71%), and 1 of 4 with TRBL (25%). In contrast to specimens of non-specific lymphadenitis, TRBL, and incipient ATLL, only one EBV genotype was evident in the specimens of HD. EBV latent membrane protein (LMP) was detected immunologically in 16 of 26 HD patients (62%), one of four TRBL (25%) and one of seven incipient ATLL (14%), but it was not evident in non-specific lymphadenitis. The LMP positive cases showed amplified EBV genomes. Only one of the 26 cases of HD had a bcl-2 gene rearrangement by PCR, but this was not seen in any other disease. The bcl-2 protein was detected immunologically in seven of the 26 HD patients (27%) and in one of the seven incipient ATLL cases (14%). EBV has been reported to upregulate bcl-2 expression, but in this study the presence of bcl-2 protein did not correlate with the presence of the t(14;18) translocation or EBV-LMP. All TRBLs showed rearrangement of the immunoglobulin genes by PCR and/or Southern blot, and the giant cells were of B-cell type. All incipient ATLLs displayed rearrangement of the TCR genes, and the giant cells were of T-cell origin. In seven of 26 HD cases, the giant cells were weakly stained with T-cell antibodies, in another seven positive with B-cell antibodies and in 18 instances polyclonally positive for both kappa and lambda. However, PCR and Southern blot displayed only two cases of TCR gene rearrangement, while two others had very weak rearrangements of immunoglobulin gene positive only by PCR. Thus the T and B-cell genotype did not correlate with the T and B-cell phenotype recorded in these cases. The absence of Ig and TCR gene rearrangements seems to be common in HD, compared with in TRBL and incipient ATLL.

Rearrangement of immunoglobulin heavy and light chains and VH family in thyroid and salivary gland lymphomas.

It is often difficult to differentiate extranodal marginal zone B‐cell lymphoma of mucosa‐associated lymphoid tissue (MALT lymphoma) from non‐neoplastic inflammatory conditions. Demonstration of clonal lymphoid proliferation by molecular procedures is important for accurate diagnosis. We examined the clonal population of B‐cell lymphomas in nine cases of thyroid and two cases of salivary gland B‐cell lymphoma using semi‐nested polymerase chain reaction (PCR)‐based assay for IgH gene arrangement and reverse transcription (RT)‐PCR single‐strand conformation polymorphism (SSCP) for the detection of IgL gene rearrangement. Clonality was evident in nine out of 11 cases of B‐cell lymphomas examined by PCR, and in six of eight cases by RT‐PCR SSCP. In addition, analysis of VH families was performed in eight cases. Although VH3 family was frequently used, each case demonstrated the VH4, VH5 or VH6 family. It is possible that the normal counterpart of thyroid or salivary gland lymphoma might be different from peripheral blood B lymphocytes, which usually use VH3 family. Our results indicate that although no clonality was noted in one case by both PCR and SSCP, these molecular methods are useful as supplementary diagnostic tests for both thyroid and salivary gland lymphomas.

Biased VH family usage in primary gastric B cell lymphoma of mucosa‐associated lymphoid tissue‐type and the selected Vβ expression of T cells infiltrating into the lymphoma cells

Seven cases of primary gastric low‐grade B cell lymphoma of mucosa‐assoclated lymphoid tissue (MALT) type, two cases of high‐grade B cell lymphoma with a low‐grade component and three cases of pure high‐grade lymphoma were selected for the current study. The Ig VH gene use of lymphoma cells and the Vβ repertoires of infiltrating T cells were Investigated. The VH gene analysis showed multiple VH family usage In 12 cases, but the MALT‐type lymphoma cell usage was found to be biased for the families that have a low number of VH genes (VHIV and V). Another analysis of lymphoma‐lnfiltrating T cells showed restricted expressions of the Vβ repertoire in all seven low‐grade cases and three high‐grade cases. In those 10 cases, a considerable number of CD4‐postttve T cells Infiltrated Into lymphoma cells and RAG‐1 was also prominently expressed. Based on these findings, It was thus assumed that the normal counterpart of gastric B cell lymphoma of MALT type is different from the conventional B cell lymphoma, and the restricted expression of Vβ repertoires Is therefore considered to be a characteristic finding in low‐grade B cell lymphomas of MALT type as well as in a proportion of high‐grade lymphomas (the so called ‘high‐grade lymphoma of MALT type’).

Limited TCR Vβ Usage of Infiltrating T cells in Synovial Tissues from Patients with HTLV-I Associated Arthropathy

Human T cell lymphotropic virus type-I (HTLV-I) is the etiologic agent of adult T cell leukemia/lymphoma and recently has also been suggested to be involved in chronic arthritis. The synovia of patients with rheumatoid arthritis (RA) contains activated T lymphocytes, with a restricted expression of T cell receptor (TCR) variable (V) beta gene segments. To characterize the T-cell populations of RA among HTLV-I carriers and noncarriers, we performed the immunohistochemical staining of CD4 and CDB, as well as a reverse transcription polymerase chain reaction (RT-PCR) to estimate the proportion of TCR beta RNA containing any particular V elements on the synovial specimens. In all but one HTLV-I carrier, the proviral DNA and/or RNA expression of HTLV-I was detected in the synovium. The CD4-positive cells proliferated markedly in the HTLV-I carriers compared with the noncarriers. In contrast to mononuclear cells in the peripheral blood, synovial T cells expressed only a few V beta transcripts, and no definite difference was observed between the carriers and the noncarriers. These results suggest that a common major antigen associated with the pathogenesis of RA may thus selectively interact with the V beta component of the TCR. Using RT-PCR, we studied the expression of the recombination-activating gene-1 (RAG-1), which was used in the V(D)J recombination of immunoglobulin and TCR genes. In all cases, RAG-1 was transcripted. The results supported the possibility that the extrathymic development of the selected TCR V beta T cells occurred in the synovia.