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Published on: April 1, 2015
Role of perforin-mediated cell apoptosis in murine models of infusion-induced bone marrow failure
Annahita K Sarcon1, Marie J Desierto, Wenjun Zhou
1Hematology Branch, National Heart, Lung, and Blood Institute, National Institutes of Health, Bethesda, MD 20892-1202, USA.
Objective:
To investigate the role of perforin-mediated cell apoptosis in murine models of immune-mediated bone marrow (BM) failure.
Materials And Methods:
We compared C57BL/6J (B6) mice carrying a perforin gene deletion (Prf(-/-)) with wild-type (WT) controls for cellular composition in lymphohematopoietic tissues. Lymph node (LN) cells from Prf(-/-) mice were coincubated with BM cells from B10-H2(b)/LilMcdJ (C.B10) mice in an apoptosis assay in vitro. We then infused Prf(-/-) and WT B6 LN cells into sublethally irradiated C.B10 and CByB6F1 recipients with mismatches at the minor and major histocompatibility loci, respectively, in order to induce BM failure. Cellular composition was analyzed by flow cytometry.
Results:
Prf(-/-) mice showed normal lymphoid cell composition, but Prf(-/-) LN cells had reduced ability to induce C.B10 BM cell apoptosis in vitro. Infusion of 5 to 10 x 10(6) Prf(-/-) LN cells produced obvious BM failure in C.B10 and CByB6F1 recipients; pancytopenia and BM hypocellularity were only slightly less severe than those caused by infusion of 5 x 10(6) WT B6 LN cells. Infused Prf(-/-) LN cells showed less T-cell expansion, normal T-cell activation, and higher proportions of T cells expressing gamma-interferon, tissue necrosis factor-alpha, and Fas ligand CD178, in comparison to infused WT B6 LN cells. Fas expression was equally high in residual BM cells in recipient of both Prf(-/-) and B6 LN cells.
Conclusion:
Perforin deficiency alters T-cell expansion but upregulates T-cell Fas ligand expression. Perforin-mediated cell death appears to play a minor role in mouse models of immune-mediated BM failure.
Insights
Perforin deficiency minimally impacts immune-mediated bone marrow failure in mice, despite altering T-cell responses and upregulating Fas ligand. This suggests perforin plays a minor role in this condition.
Area of Science:
- Immunology
- Hematology
- Cell Biology
Background:
- Immune-mediated bone marrow (BM) failure is a serious condition.
- Perforin is a key molecule in cytotoxic T lymphocyte (CTL) mediated killing.
- The role of perforin in immune-mediated BM failure is not fully understood.
Purpose of the Study:
- To investigate the role of perforin-mediated cell apoptosis in murine models of immune-mediated bone marrow failure.
- To compare the effects of perforin-deficient (Prf(-/-)) and wild-type (WT) mice in inducing BM failure.
Main Methods:
- Compared Prf(-/-) and WT C57BL/6J mice for cellular composition.
- Performed in vitro apoptosis assays using lymph node (LN) cells and BM cells.
- Induced BM failure by infusing Prf(-/-) and WT LN cells into irradiated recipients.
- Analyzed cellular composition by flow cytometry.
Main Results:
- Prf(-/-) mice had normal lymphoid cell composition but reduced ability of LN cells to induce BM cell apoptosis in vitro.
- Infusion of Prf(-/-) LN cells caused BM failure, comparable to WT LN cells.
- Infused Prf(-/-) LN cells showed altered T-cell expansion and activation, with increased Fas ligand expression.
Conclusions:
- Perforin deficiency alters T-cell expansion and upregulates T-cell Fas ligand expression.
- Perforin-mediated cell death plays a minor role in established mouse models of immune-mediated BM failure.
- Further research is needed to elucidate the mechanisms of immune-mediated BM failure.
