Neutrophils accumulate around Trichinella spiralis larvae encysted in skeletal muscle cells of the host. The magnitude of the neutrophil infiltration follows a pattern relating to the stage of infection of T.spiralis. This study was performed to determine if there are factors chemotactic for neutrophils present in the sera of mice infected with T.spiralis, and if present, to compare the chemotactic potential of sera from several time points during the infection. Female MRL++ mice hosted the T.spiralis infection and provided neutrophils for all experiments. The chemotactic potentials of sera were tested by placing 150 ul of serum collected at zero, four, 11, or 28 days following initial infection of the mouse with T.spiralis, into the bottom well of a modified Boyden chamber called a deep well chemotaxis chamber. Next a PVP-free polycarbonate micropore membrane with a pore size of 5 um was placed over the bottom well. The top chamber was then fastened to the bottom well and filled with a neutrophil suspension which was obtained by gradient layer centrifugation using the NIM • 2 step gradient reagent system. A cover glass was placed over the opening of the top well and the entire apparatus was incubated at 37°C under 5% C02 in a humidified incubator for four hours. Following the incubation cells which had migrated through to the bottom well were counted using a hemocytometer and the membranes were stained with Diff Quick. The results demonstrate that there are factors present in the sera of mice infected with T.spiralis that elicit the chemotactic response of neutrophils in vivo. This work also demonstrates that sera from mice infected 11 and 28 days are significantly more chemotactic for neutrophils than are sera from uninfected mice and mice infected four days. These findings correlated with the histologic appearance of the infected skeletal muscle at four, 11 and 28 days post infection as determined by other members of our laboratory. The chemokines IL-8 and KC, and other chemotactic factors such as C5a are discussed as potential mediators of the neutrophil chemotaxis found in this experiment.
| Identifer | oai:union.ndltd.org:pdx.edu/oai:pdxscholar.library.pdx.edu:open_access_etds-6138 |
| Date | 05 July 1995 |
| Creators | Youngman, Sara Lynn |
| Publisher | PDXScholar |
| Source Sets | Portland State University |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | Dissertations and Theses |
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