The role of the P53 gene and the P53 protein in non-Hodgkin malignant lymphomas
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Study design Scope This paper aimed to highlight the stages of Chronic Lymphocytic Leukemia type of B (CLL-B), which did not meet the standard treatment criteria for malignant hematological diseases due to mutations in the P-53 gene, with progression to Diffuse Large Lymphoma. Method The frequency of p-53 protein expression in 85 patients diagnosed with CLL was analyzed by the Enzyme-Linked Immune-Absorbent Assay (ELISA) technique to investigate the relationship of this protein to the stage of the disease, as well as the impact on response to treatment and survival. Cell extracts 10³×10³/L in 100 μl lysis buffer were applied to ELISA plates coated with PAb 240 capture antibody. Results The frequency of increased positivity of the protein with the modified structure, the isoform, p-53 in type CLL-B, was 17% in the 85 cases initially included in the study. The percentage of p-53 protein isoforms, positive above normal value, with very high values, 50, 60, respectively 140 µg / dl was found in the percentage of 3.5% with the transformation of CLL into non-Hodgkin's malignant lymphomas, (NHL) type Diffuse Large Lymphoma, (DLL) or in Mantle Lymphoma, from Richter syndrome. Conclusions In the context of a heterogeneous condition such as LL-B, this cheap and safe method, ELISA seems to provide a useful prognostic tool capable of identifying patients who can be considered candidates for therapeutic, targeted, personalized strategies. Primary study design Observational; The cases were classified as CLL with> 5000 lymphocytes in absolute value, present at the cytological examination of the blood smear, from the peripheral blood or LLC with less than 10% prolymphocytes based on the peripheral blood smears May-Grunwald Giemsa, stained. Cohort study: After analyzing the 85 LLC samples, in different stages of disease evolution, starting with stage zero (stay and watch) and up to stage IV, 20 patients were selected, eligible for this study, to be investigated for the detection of p-53 protein isoforms responsible for resistance to oncological treatments of the disease with Rituximab, Cyclophosphamide, Doxorubicin hydrochloride, Vincristine sulfate (Oncovin), and Prednisone, (R-CHOP), after 2 cycles of relapses, representing a group of 16 men and 4 women aged 39-85 years. Trial setting; The presence of protein isoforms in the studied group was calculated as a percentage of 17%, and the un-favorable evolution and transformation of Chronic Lymphocytic Leukemia in the stages studied in Diffuse Large Lymphoma was calculated to be in a percentage of 3.5% of the studied cases, as in the meta-analyzes from the specialized literature, Diffuse Large lymphoma is considered a rare disease.
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Protocol/serial number: Dr. AURELIAN UDRISTIOIU; Web of Science Researcher ID: I-3687-2019; UEFISCDI ID (UEF-iD): U-1900-063Y4656 . Genomic analysis showed that in most patients, Richter syndrome is derived from the initial predominant CLL clone through a linear path of genetic evolution involving the development of additional genetic lesions in extra poetic tissues by migration of bone marrow stem cells including bone marrow events. activation of the MYC oncogene and disruption of the functionality of the P-53 gene. Finally, Richter Syndrome has been described in some patients who received the BCL2 Venetoclax or Ibrutinib inhibitor although the exact prevalence and characteristics of this transformation remain to be determined Non-Hodgkin malignant lymphomas. Intervention The antibody is suitable for the techniques: ICC / IF and ELISA. Research antibody PAb 1620 has been reported to be specific for the conformation of the normal p-53 protein, (Cook and Milner 1990) and PAb 240 antibodies bind specifically to denatured p-53 protein, i.x. p53 in the "mutant" conformation. Complete physical examination: In patients diagnosed with CLL-B, symptoms such as frequent cough, night sweats, and retrosternal pain were evaluated. Clinical examination and ultrasound revealed lymphadenopathy and/or splenomegaly, with an enlarged spleen. Laboratory examinations: Hemogram with 5 Diff and cytological examination of the blood smear on the peripheral blood in the May Grunwald-Giemsa staining, and bone marrow puncture, BM with medullary forcing. The cases were classified as CLL with >5000 lymphocytes in absolute value, present at the cytological examination of the blood smear, from the peripheral blood or LLC with less than 10% prolymphocytes based on the peripheral blood smears May-Grunwald Giemsa, stained. Immunophenotyping: The diagnosis of CLL was confirmed by immune phenotyping. All samples that entered the study were lymphocytes with positive CD19⁺, CD20⁺⁻, CD5⁺ and CD23⁺ cell receptors. The CD38+ receptor was considered positive if the distinct lymphocytes of the population showed a higher intensity of staining than the granulocytes in the sample and was associated with the presence of protein ZAP-70. A sandwich ELISA colorimetric quantitative method was used for direct detection of the p53 isoform protein, the product of gene p53: Specificity: human p53 protein (aa20-25); Format: Purified product: Monoclonal antibody clone: Isotype DO-1: IgG2a. The antibody is suitable for the techniques: ICC / IF and ELISA. The research antibody PAb 1620 has been reported to be specific for the conformation of the normal p-53 protein, and PAb 240 antibodies bind specifically to denatured p-53 protein. Compatible sample types: cell culture supernatants, plasma, serum; solid support: 96-well microplate; firm: Ray Biotech Life, Inc.
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- UEFISCDI ID (UEF-iDGrant ID: U-1900-063Y4656