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Identification and propagation of a putative immunosuppressive orphan parvovirus in cloned T cells
M D McKisic1, D W Lancki, G Otto
1Department of Pathology, University of Chicago, IL 60637.
Abstract:
A putative parvovirus related to minute virus of mice (MVM), but distinct from MVM-prototype and MVM-immunosuppressive, was identified, using serologic techniques and Southern blot analysis, in maintenance cultures of established T cell clones. This putative viral agent resulted in a lytic infection of cloned L3 cytotoxic T cells but was unable to produce a productive infection in BHK.21 or EL-4(G) cells. Moreover, maintenance cultures of several distinct subsets of cloned T cells apparently contaminated with this putative viral agent contained poorly growing cells and erythrocyte aggregates. The aggregation of mouse erythrocytes appeared to be a reliable indicator of infection with this putative virus and may be related to the ability of this agent to agglutinate mouse erythrocytes. This putative virus also was found to inhibit the proliferative response of certain cloned T cells to IL-2 and Ag. Viremic mice and secondary MLC supernatant were identified as two potential sources of contamination and represent ways of propagating this agent in vitro. The finding that this agent interferes with the ability of T cell clones to thrive and, therefore has the potential to alter immune responses, emphasizes the importance of identifying and excluding parvoviral infections in cultures of murine T lymphocytes.
Insights
A novel parvovirus distinct from minute virus of mice (MVM) infects T cell clones, causing cell death and erythrocyte aggregation. This viral contamination impacts T cell proliferation and immune responses in murine lymphocyte cultures.
Area of Science:
- Virology
- Immunology
- Cell Biology
Background:
- Murine T cell cultures are essential for immunological research.
- Contamination by adventitious viruses can compromise experimental results.
- Minute virus of mice (MVM) is a known parvovirus affecting murine models.
Purpose of the Study:
- To identify and characterize a novel parvovirus in T cell cultures.
- To investigate the effects of this parvovirus on T cell function.
- To establish methods for detecting and preventing viral contamination in T cell cultures.
Main Methods:
- Serologic techniques and Southern blot analysis were used for viral identification.
- Infection assays were performed on various cell lines (L3, BHK.21, EL-4(G)).
- Erythrocyte aggregation assays and T cell proliferation assays (IL-2 and Ag response) were conducted.
Main Results:
- A parvovirus, distinct from known MVM strains, was identified in T cell clones.
- The virus caused lytic infection in L3 cytotoxic T cells but not in BHK.21 or EL-4(G) cells.
- Infected cultures showed poor cell growth, erythrocyte aggregates, and inhibited T cell proliferation.
Conclusions:
- A novel parvovirus contaminates murine T cell cultures, impacting cell viability and function.
- Erythrocyte aggregation serves as a potential indicator of parvoviral infection.
- Preventing parvoviral contamination is crucial for maintaining the integrity of T cell-based immunological studies.