36 research outputs found
Targeted panel sequencing in the routine diagnosis of mature T- and NK-cell lymphomas: report of 128 cases from two German reference centers
Diagnosing any of the more than 30 types of T-cell lymphomas is considered a challenging task for many pathologists and currently requires morphological expertise as well as the integration of clinical data, immunophenotype, flow cytometry and clonality analyses. Even considering all available information, some margin of doubt might remain using the current diagnostic procedures. In recent times, the genetic landscape of most T-cell lymphomas has been elucidated, showing a number of diagnostically relevant mutations. In addition, recent data indicate that some of these genetic alterations might bear prognostic and predictive value. Extensive genetic analyses, such as whole exome or large panel sequencing are still expensive and time consuming, therefore limiting their application in routine diagnostic. We therefore devoted our effort to develop a lean approach for genetic analysis of T-cell lymphomas, focusing on maximum efficiency rather than exhaustively covering all possible targets. Here we report the results generated with our small amplicon-based panel that could be used routinely on paraffin-embedded and even decalcified samples, on a single sample basis in parallel with other NGS-panels used in our routine diagnostic lab, in a relatively short time and with limited costs. We tested 128 available samples from two German reference centers as part of our routine work up (among which 116 T-cell lymphomas), which is the largest routine diagnostic series reported to date. Our results showed that this assay had a very high rate of technical success (97%) and could detect mutations in the majority (79%) of tested T-cell lymphoma samples
Imatinib Treatment Induces CD5+ B Lymphocytes and IgM Natural Antibodies with Anti-Leukemic Reactivity in Patients with Chronic Myelogenous Leukemia
Imatinib mesylate is a first line treatment of Chronic Myelogenous Leukemia and of a rare form of gastrointestinal stromal cancer, where the response to the drug is also linked to the immune system activation with production of antineoplastic cytokines. In this study, forty patients in the chronic phase of disease, treated with imatinib mesylate, were analyzed. Bone marrow aspirates were drawn at diagnosis, after 3, 6, 12, 18 months for haematological, cytofluorimetric, cytogenetic, biomolecular evaluation and cytokine measurement. Responder and non responder patients were defined according to the European LeukemiaNet recommendations. In responder patients (nâ=â32), the percentage of bone marrow CD20+CD5+sIgM+ lymphocytes, and the plasma levels of IgM, were significantly higher, at 3 months and up to 9 months, than in non responders. These IgM reacted with O-linked sugars expressed by leukemic cells and could induce tumor cell apoptosis. In responeĂŹder patients the stromal-derived factor-1 and the B-lymphocyte-activating factor of the tumor necrosis factor family significantly raised in the bone marrow after imatinib administration, together with the bone morphogenetic proteins-2 and â7. All patients with high number of CD20+CD5+sIgM+ cells and high stromal-derived factor-1 and B lymphocyte activating factor levels, underwent complete cytogenetic and/or molecular remission by 12 months. We propose that CD20+CD5+sIgM+ lymphocytes producing anti-carbohydrate antibodies with anti-tumor activity, might contribute to the response to imatinib treatment. As in multivariate analysis bone marrow CD20+CD5+sIgM+ cells and stromal-derived factor-1 and B-lymphocyte-activating factor levels were significantly related to cytogenetical and molecular changes, they might contribute to the definition of the pharmacological response
The âearly birdsâ: natural IgM antibodies and immune surveillance
Precancerous epithelial lesions are sites of
uncontrolled cellular proliferation generated by
irreversible genetic alterations. Not all of those lesions
progress to invasive cancer, some may even regress, but
the early detection of abnormal cells can be crucial for
patient survival. Immune surveillance mechanisms are
responsible for the removal of transformed cells and
antibodies play an important role in these immune
processes. In the past, analysis of the immunoglobuline
repertoire has focused mainly on xenoimmunizations or
the investigation of cancer patient immunity. The human
hybridoma technology (Trioma technique) offers the
unique possibility to study the humoral immunity of
healthy people. Using this technique a series of tumorbinding
antibodies could be isolated which all have
several features in common: they are germ-line coded
IgM antibodies, they predominantly bind to
carbohydrates on post-transcriptionally modified
antigens, they induce apoptosis and, most importantly,
they detect not only malignant cells but also precursor
stages. These data demonstrate that the body has a
comprehensive defense system against malignant cells
based on the production of natural antibodies
Natural IgM antibodies, the ignored weapons in tumour immunity
During its lifetime each multi-cellular
organism is permanently exposed to infectious agents
and transformed cells. Without an early recognition and
a rapid elimination system, there would be no
development and no life. The innate or natural immunity,
seems to be more important for the detection of
âforeignâ cells and particles than has been thought. Even
if not every transformed cell has the ability and potency
for malignant behaviour, the important question is not,
why malignant cells arise, but instead, why malignancy
occurs so infrequently. We have shown in a recent paper,
by using the human hybridoma technology, that tumour
immunity is not induced by malignant cells, but instead
the result of innate immunity and that natural IgM
antibodies play an important role in immunosurveillance
mechanisms against transformed cells in humans
(BrÀndlein et al., 2003b). In this review typical features
of natural IgM antibodies are discussed and tumourspecific
reactivities and different apoptotic functions on
epithelial cancer cells are illustrated
Natural IgM antibodies: from parias to parvenus
Over the years, natural IgM antibodies were
considered as the parias among the immune competent
molecules. Their characteristic properties, like low
affinity, cross-reactivity and pentameric structure, were
assessed as difficult and nebulous. Today, mainly based
on the persistent work of a few researchers and the key
discoveries on innate immunity, natural IgM antibodies
are âback on stageâ. Their important role in the immune
response against invasive particles, modified selfcomponents
and altered cells is accepted. All the so far
negatively judged features have to be seen in a different
light, e.g. low affinity seems to be good for function and
does not exclude specificity, cross-reactivity is no longer
judged as unspecific, but instead as a very economic way
of immune recognition and the pentameric structure is
important for binding capacity and functional activities.
In addition, with the use of natural IgM antibodies, a
new field of tumour-specific targets has been
encountered, the carbo-neo-epitopes, which are
commonly found on post-transscriptionally modified
membrane receptors. Having understood the typical
features of natural IgM antibodies, their renaissance
opens a new area of cancer therapeutics and diagnostics