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The Inducible Defense System: Antibody Molecules and Antigen-Antibody Reactions
Published in Julius P. Kreier, Infection, Resistance, and Immunity, 2022
In addition to the antibody diversity resulting from “mixing and matching” of different DNA sequences, other modifications may occur during the recombination process. For example, during the rearrangement process when the DNA is cleaved and religa ted, two nucleotides from a codon of one V segment of DNA may be spliced next to one nucleotide of a codon from a J segment. As a result, an entirely new triplet may be produced and a different amino acid is incorporated into the chain. In addition, the enzyme terminal deoxynucleotidyl transferase (TdT) is able to add nucleotides randomly to the H chain between the time the DNA is broken at the V, D, and J segments, and the DNA is religated. This process also adds a significant amount of diversity to the recombinant CDR3 region. It is estimated that the human immune system can make over a billion different antibodies by randomly rearranging fewer than 200 segments of DNA. As a consequence, the immune system is able to respond to new pathogens it has never before encountered.
Defective Tumor Suppressor Gene Replacement and Oncogene Inactivation for the Treatment of Cancer
Published in Eric Wickstrom, Clinical Trials of Genetic Therapy with Antisense DNA and DNA Vectors, 2020
Thus, all patients, with the exception of one who did not complete the treatment protocol (patient 9), showed some evidence of gene transfer. Six of the seven patients also showed increased terminal deoxynucleotidyl transferase biotin dUTP nick end-labeling (TUNEL) staining in posttreatment versus pretreatment biopsies, indicating apoptosis (programmed cell death) was occurring in cancer cells (Table 2).
Hairlessness and Metabolic Compensations
Published in Miroslav Holub, Immunology of Nude Mice, 2020
Spleen cells from 1-month-old nude mice which have been inoculated with Moloney murine leukemia virus after birth do develop lymphomas upon transfer to syngeneic recipients (400 R-irradiated and with opposite sex to permit identification of donor cells).48 Spleen cells taken from euthymic donors (CBA, BALB/c) after the same leukemogenic treatment produce lymphomas in the recipients faster than nu/nu spleen cells and arise from at least two T-cell subsets, one terminal deoxynucleotidyl transferase positive, the other 20 β-hydroxysteroid dehydrogenase positive. The BALB/c nu/nu spleen cells were negative for these enzymes and had to mature into the enzyme-positive cells in the thymus-bearing recipient.48
Protective effect of dendropanoxide against cadmium-induced hepatotoxicity via anti-inflammatory activities in Sprague-Dawley rats
Published in Toxicology Mechanisms and Methods, 2023
Sreevarsha Gali, Swati Sharma, Amit Kundu, Eunah Lee, Joo Hee Han, Joo Kyung Shin, Ji Soo Choi, So Young Kyung, Jae-Sung Kim, Hyung Sik Kim
Cadmium chloride (CdCl2) and silymarin were purchased from Sigma-Aldrich (St. Louis, MO, USA). Enzyme-linked immunosorbent assay (ELISA) kits for analyzing aspartate aminotransferase (AST), alanine aminotransferase (ALT), alkaline phosphatase (ALP), γ-GGT: gamma-glutamyl transpeptidase, triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL), total cholesterol, interleukin-1β (IL-1β), and interleukin-10 (IL-10) levels were purchased from Abcam (Cambridge, UK). A tumor necrosis factor-alpha (TNF-α) kit was purchased from Invitrogen (Thermo Fisher Scientific, MA, USA). Interleukin-6 (IL-6) kit was purchased from Abbkine, Inc. (International Biomedicine Park, Wuhan, China). Superoxide dismutase (SOD), catalase (CAT), and malondialdehyde (MDA) assay kits were purchased from Abcam (Cambridge, UK) and glutathione (GSH) from Thermo Fisher Scientific. Primary antibodies against IL-1β, TNF-α, IL-6, IL-10, heme oxygenase-1 (HO-1), nuclear respiratory factor 2 (NRF2), Kelch-like ECH-associated protein 1 (Keap 1), BCL2 associated X protein (Bax), B-cell lymphoma protein 2 (Bcl-2), cleaved caspase 3, caspase 3, and β-actin were purchased from Abcam (Cambridge, MA, USA). Horseradish peroxidase (HRP)-conjugated secondary antibodies were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Bicinchoninic acid (BCA) was purchased from Thermo Fisher Scientific. The terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) test assay kit was obtained from Abcam (Cambridge, UK).
A novel polymerase β inhibitor from phage displayed peptide library augments the anti-tumour effects of temozolomide on colorectal cancer
Published in Journal of Chemotherapy, 2022
Lihong Qin, Mao Huiwen, Jianguo Wang, Yuanyaun Wang, Salman A. Khan, Ying Zhang, Hong Qiu, Longwei Jiang, Lingfeng He, Yan Zhang, Shaochang Jia
Moreover, we performed a terminal deoxytransferase-mediated dUTP biotin nick-end labelling (TUNEL) assay to examine cell apoptosis. Accumulation of unrepaired DNA damage will consequently activate the intrinsic pathway of apoptosis, which can be detected by TUNEL (TdT-mediated dUTP Nick-End Labeling) assay. TUNEL assay uses terminal deoxynucleotidyl transferase (TdT) to catalyze the incorporation of deoxynucleotides at the free 3′-hydroxyl ends of fragmented DNA. The deoxynucleotides are then labelled for detection of the degree of DNA fragmentation. As shown in Figure 4C, the ratio of apoptotic cells were 2-fold higher in combination group than that of single-treated group. The representative micrograph of γH2AX foci detection and TUNEL assay were shown in Figure 4D,E respectively. The synergistic effect of TMZ and 10 D was analyzed with the isobologram method, as shown in Figure S3. These above results demonstrated that 10 D in combination with TMZ efficiently induce cytotoxicity in SW480 cancer cells.
Epicutaneous Administration of 17β-Estradiol Induces Langerhans Cells Depletion
Published in Immunological Investigations, 2022
Katia Jarquín-Yáñez, Miguel Ángel Herrera-Enriquez, Cristina Lemini, Edith Melendez-Moreno, Paulina Villena-López, Maria Estela Ávila, Beatriz Hernández-Téllez, Gabriela Piñón-Zárate, Enrique Agustin Sampedro-Carrillo, Andrés Eliú Castell-Rodríguez
The solvents and reagents were of analytical reagent grade and were used without further purification. Na2EDTA and paraformaldehyde were obtained from Merck (Darmstadt, Germany). The phosphate-buffered saline (PBS) solution was prepared according to standard procedures. Anti- MHCII antibodies were obtained from Neomarker Antibodies (CA, USA). Antibodies against CD11c, CD207, CD40, and CD86 were obtained from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Biotinylated goat anti-mouse streptavidin/peroxidase-conjugated IgG was obtained from ICN Biomedicals (Costa Mesa, CA, USA). The streptavidin/peroxidase staining kit (Histostain Plus Kit) was obtained from Zymed (San Francisco, CA, USA). 17β-Estradiol (1,3,5(10)-estratrien-3,17ß-diol) was a gift from Syntex (Cd. Mex., México). 3-Amino-ethyl carbazole, dimethylformamide, diaminobenzidine (DAB), and 1-fluoro-2,4-dinitrobenzene (DNFB) were obtained from Sigma–Aldrich (MO, USA). The terminal deoxynucleotidyl transferase (TdT)-mediated dUTP nick end-labeling (TUNEL) assay kit was obtained from Promega (WI, USA).