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Assays for the Specific Growth Rate and Cell-binding Ability of Rotavirus
Published on: January 28, 2019
Induction of Cell Death in the Human Acute Lymphoblastic Leukemia Cell Line Reh by Infection with Rotavirus Isolate
Rafael Guerrero1, Carlos Guerrero1, Orlando Acosta1
1Department of Physiological Sciences, Faculty of Medicine, Universidad Nacional de Colombia, Carrera 30 No. 45-03 Bloque 47, Ciudad Universitaria, Bogotá 111321, Colombia.
Abstract:
Cancer is a major health problem that poses a great challenge to health care systems worldwide. Tools for cancer treatment have rapidly advanced in recent years, resulting in therapeutic strategies which are alternative and complementary to conventional treatment. To identify the cell surface receptors used by a tumor cell-adapted rotavirus and the cell death markers induced by its infection, we use Wt1-5, a rotavirus isolate recently adapted to tumor cells, to infect the human acute lymphoblastic leukemia cell line, Reh. The expression of cell surface receptors used by Wt1-5 was determined using flow cytometry and an antibody blocking assay to test for their implication in virus infection. Viral antigens and cell death markers induced by rotavirus infection were followed by flow cytometric analysis. The present study showed that rotavirus Wt1-5 was able to use cell surface proteins such as heat shock proteins (HSPs) 90, 70, 60 and 40, Hsc70, PDI and integrin β3. Rotavirus Wt1-5 induced cytotoxic effects including changes in cell membrane permeability, alteration of mitochondrial membrane potential, DNA fragmentation and activation of cell death signaling. Wt1-5 deserves to be further studied as a candidate oncolytic agent due to its ability to induce apoptosis in lymphoblastic leukemia-derived cells.
Insights
A rotavirus adapted to tumor cells, Wt1-5, uses cell surface proteins like heat shock proteins (HSPs) and integrin β3 to infect leukemia cells. This virus induces apoptosis, showing potential as an oncolytic agent.
Area of Science:
- Oncology
- Virology
- Cell Biology
Background:
- Cancer presents a significant global health challenge, driving the search for novel therapeutic strategies.
- Advances in cancer treatment include complementary and alternative approaches beyond conventional methods.
- Tumor-cell adapted viruses are being explored as potential oncolytic agents.
Purpose of the Study:
- To identify cell surface receptors utilized by a tumor-adapted rotavirus (Wt1-5).
- To determine the cell death markers induced by Wt1-5 infection in leukemia cells.
- To evaluate Wt1-5 as a potential oncolytic agent.
Main Methods:
- Infection of human acute lymphoblastic leukemia (Reh) cells with Wt1-5 rotavirus.
- Flow cytometry and antibody blocking assays to identify cell surface receptors.
- Analysis of viral antigens and cell death markers (e.g., membrane permeability, mitochondrial potential, DNA fragmentation) via flow cytometry.
Main Results:
- Rotavirus Wt1-5 utilizes cell surface proteins including heat shock proteins (HSPs 90, 70, 60, 40), Hsc70, PDI, and integrin β3 for infection.
- Wt1-5 infection induced significant cytotoxic effects in Reh cells.
- Observed effects included altered membrane permeability, mitochondrial membrane potential, DNA fragmentation, and activated cell death signaling.
Conclusions:
- Rotavirus Wt1-5 effectively infects leukemia cells by engaging specific cell surface proteins.
- The virus induces apoptosis, a programmed cell death pathway, in lymphoblastic leukemia cells.
- Wt1-5 demonstrates promise as a candidate oncolytic agent for treating leukemia.
