Loss of Fas (CD95/APO-1) regulatory function is an important step in early MALT-type lymphoma development.

Fas (CD95, APO-1) mutations were found in autoimmune diseases and some lymphomas, suggesting impairment of Fas-mediated cell death signaling that may cause tumor development. Because mucosa-associated lymphoid tissue (MALT)-type lymphoma B cells recognize autoantigens and proliferate in response to antigen and T cell-mediated signals, it is suggestive that autoreactive B cell lymphoma precursor cells may have escaped the Fas-mediated checkpoint that normally operates in healthy individuals. Using different biochemical, molecular, and functional approaches, we analyzed the Fas signaling in malignant B cells from seven MALT-type lymphomas that were additionally characterized for the t(11;18)(q21;q21) and four gastric diffuse large B cell lymphomas (DLBL). All DLBLs and three of seven MALT-type lymphomas were resistant to Fas-mediated apoptosis in vitro. Moreover, four of five MALT-type lymphomas analyzed and one of three DLBLs analyzed showed mutations in Fas mRNA transcripts but no loss of heterozygosity in the Fas promotor region. Alternative mechanisms of resistance to apoptosis, such as decreased expression of Fas or production of soluble Fas were not operative. Therefore, it is suggestive that a subgroup of MALT-type lymphoma B cells, irrespective of t(11;18)(q21;q21), escape the censoring Fas pathway by mutating and inactivating Fas. This identifies a key regulatory step in early MALT-type lymphomagenesis.

[Impaired CD95-(Fas,APO-1-)apoptosis regulation is a progression factor in early MALT-type lymphoma genesis]. / Eine gestörte CD95-(Fas,APO-1-)vermittelte Apoptoseregulation ist ein Progressionsfaktor in der frühen MALT-Typ-Lymphomgenese.

BACKGROUND: MALT-type lymphomas are B cell tumors arising for so far unknown reasons on the background of chronic inflammation, e.g. Helicobacter pylori-associated gastritis. T cells are supposed to have a positive impact on tumor growth, but fail in their control function. We therefore examined the interaction of T cells with malignant B cells in vitro and focused on T cell control which normally operates by CD95L/CD95-mediated apoptosis. PATIENTS AND METHODS: Malignant B cells were isolated from tumor tissues of 7 patients with low-grade MALT-type lymphoma and 4 patients with DLBL and cocultured in vitro with activated T cells. Normal memory B cells were used as control. We developed a T/B coculture assay for investigation of CD95L/CD95-mediated apoptosis. The influence of T cells on CD95 expression and survival of B cells was measured by FACS analysis. RESULTS: Activated T cells induced CD95 expression and thus enhanced sensitivity to CD95L-mediated apoptosis in normal memory B cells. However, malignant B cells from 3 out of 7 low-grade MALT-type lymphomas and all 4 gastric DLBLs resisted apoptosis, although the cells showed enhanced CD95 expression. CONCLUSION: Resistance to CD95L/CD95-mediated apoptosis allows malignant B cells from MALT-type lymphoma to escape T cell control and leads to prolonged survival. This phenomenon acts as a progression factor in early lymphomagenesis.

Tumor biology of mucosa-associated lymphoid tissue lymphomas.

Extranodal lymphomas arising at mucosal sites exhibit clinicopathological features that suggest a closer relationship of these tumors to the structure and function of mucosa-associated lymphoid tissue (MALT) than to lymph nodes. The factors that induce MALT in these tissues are operative in early MALT lymphoma development and the progressive independence on T-cell help defines late stages of MALT lymphoma genesis.

Expression of costimulatory molecules in low-grade mucosa-associated lymphoid tissue-type lymphomas in vivo.

B-cell lymphomas of mucosa-associated lymphoid tissue (MALT) type develop against a background of chronic inflammation and have functional autoantigen receptors. Because they respond to environmental factors in vivo, the expression of costimulatory molecules, which play a key role in the differentiation of normal B-lymphocytes and in T-/B-cell interaction, may be critical in early MALT-type lymphoma pathogenesis until further chromosomal aberration leads to progression. We found a high number of tumor-infiltrating T-lymphocytes (TITLs) in all low-grade MALT-type lymphomas. The TITLs in low-grade lymphomas were activated and expressed a memory and immunocompetent phenotype. Reverse transcriptase-polymerase chain reaction analyses and immunohistochemistry confirmed the presence of CD40-ligand and Fas-ligand in 80% of low-grade lymphomas. In contrast to the TITLs, the tumor B cells did not express CD40-ligand or Fas-ligand in vivo or in vitro. Moreover, the cytokine profile in vivo suggested a Th2/Th3-weighted profile (interleukin-10, interleukin-13, transforming growth factor beta(1)) rather than Th1-weighted (interferon-gamma, interleukin-2). By interphase fluorescence in situ hybridization analysis the translocation t(11;18)(q21;q21) was found in four of nine (44%) cases studied. Interestingly, there was a four times higher proliferation and survival rate of purified t(11;18)-positive tumor B cells in vitro, although there were no significant profile differences from the TITLs in vivo. The finding of essential costimulating molecules in low-grade MALT-type lymphomas in vivo indicates a locally directed cognate T-/B-cell interaction. Consequently, a potentially equipped inflammatory background may not only determine the fate of autoreactive B-cells, but is also crucial to lymphoma maintenance and progression.

Oligoclonal expansions of T-cell repertoire in gastric mucosa associated lymphoid tissue type B-cell lymphoma and adjacent gastritis.

Local stimulation by Helicobacter pylori (HP), autoantigen, and a concurrent T-cell-mediated stimulation of B cells are believed to play an important role in gastric mucosa-associated lymphoid tissue (MALT) type B cell lymphomagenesis. Many autoimmune diseases have shown to lead to a skewed T-cell repertoire with autoantigen specific expansions and deletions. Characterization of lymphoma and gastritis areas of seven gastrectomy specimens using a T-cell receptor beta variable chain (TCR betaV) family-specific reverse transcriptase (RT)-polymerase chain reaction (PCR) assay and fluorescence-activated cell sorter (FACS) analysis revealed a local chronic and acute activation of T cells in lymphoma and an oligoclonal T-cell repertoire in gastritis and in lymphoma, partially sharing the same clones. Local activation and a partial identity suggest that an antigenic challenge caused by a common local pathogen may still continue to take place in MALT type lymphoma as in gastritis, consistent with the view that gastritis may be a precursor lesion of MALT type lymphoma. Expansions that were found only in one of the compartments suggest that also an immune hyperstimulation may contribute to the T-cell repertoire, possibly because of certain tissue antigens. Deletions of TCR betaV families found only in gastritis underline the view that autoantigen may play an important role in its pathogenesis.

[MALT-type B-cell lymphomas escape fas-mediated apoptosis]. / MALT-Typ B-Zell Lymphome entkommen der FAS-vermittelten Apoptosekontrolle.

Important prognostic factors in primary MALT-lymphomas are stage and grading since the localized low-grade lymphoma may be cured with antibiotic treatment that may result in lymphoma regression as exemplified in Helicobacter eradication (H.p.) in gastric low-grade MALT-lymphomas. For obscure reasons, around 20% of low-grade MALT-lymphomas do not respond to eradication therapy or contain in 25-33% high-grade components as a possible consequence of tumor cell transformation. Given that MALT-lymphoma B-cells are autoimmune in nature and proliferate in response to antigen and T-cell-mediated signals, it is suggestive that autoreactive B-cell lymphoma precursors generated during chronic inflammation escaped the tolerance checkpoint. This mechanism normally operates in healthy individuals through CD40-L/FAS-L expressing activated T-cells and is fundamental in the deletion of harmful B-cells. Analyzing the role of FAS/FAS-L mediated apoptosis in lymphomagenesis, we purified B- and T-cells from low- and high-grade MALT B-cell tumors and tonsillar memory B-cells as control. T-cells were stimulated in vitro to express CD40-L and FAS-L using anti-CD3 antibodies and were subsequently cocultivated with B-cells. Apoptosis was 3 times increased in normal memory B-cells and, to a lesser extent, in a subgroup of low-grade MALT-lymphomas as compared with the spontaneous apoptosis without T-cell coculture. In striking contrast, all primary high-grade MALT-type lymphomas showed increased survival but were resistant to FAS-mediated apoptosis. Furthermore, 40% of low-grade MALT-type lymphomas were resistant to apoptosis and showed mutations in the FAS-gene. The conclusion that self-tolerant high-grade but also a subgroup of low-grade MALT-lymphoma B-cells escaped censoring by the FAS pathway may therefore identify a novel regulatory step in early MALT-lymphomagenesis.