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Phenotype and morphology of murine NC-1.1+ natural cytotoxic cells
J H Brien1, Y C Smart, M L Farrelly
1Discipline of Surgical Science, Faculty of Medicine, University of Newcastle, New South Wales, Australia.
Insights
Natural cytotoxic (NC) cells, identified by anti-NC-1.1 mAb, exhibit diverse phenotypes in mice spleen. These NC cells are part of a "horizontal lineage," originating from various lymphohematopoietic cell types.
Area of Science:
- Immunology
- Cell Biology
- Hematology
Background:
- Natural cytotoxic (NC) cells play a role in innate immunity.
- The lineage and differentiation pathways of NC cells remain incompletely understood.
Purpose of the Study:
- To investigate the morphology and phenotype of NC cells.
- To determine the relationship of NC cells to other lymphohematopoietic lineages.
Main Methods:
- Flow cytometry (single and two-color) was used to analyze splenocytes from CBA and (CBA x C57BL/6) F1 mice.
- Anti-NC-1.1 monoclonal antibody (mAb) 1C4 was employed to identify and characterize NC cells.
- Analysis included co-expression of various cell surface markers (Thy-1.2, L3T4, Mac-1, IgM, FcR gamma II, J11d, NK-1.1).
Main Results:
- The anti-NC-1.1 mAb blocked NC cell activity in vitro and in vivo.
- In CBA mice, 6% of splenocytes were NC-1.1+, with granular cells showing the highest proportion (29%).
- NC-1.1+ granular spleen cells co-expressed markers like IgM and FcR gamma II, while small agranular cells did not express T cell markers. In F1 mice, 82% of granular NC-1.1+ cells expressed NK-1.1.
Conclusions:
- NC cells exhibit a heterogeneous phenotype, with granular cells expressing a distinct set of markers.
- The findings support the 'horizontal lineage' theory, suggesting NC cells represent different lymphohematopoietic lineages at specific differentiation stages.
Abstract:
The morphology and phenotype of cells identified by the anti-NC-1.1 mAb, 1C4, were studied in CBA and (CBA x C57BL/6) F1 mice. This mAb blocked splenic natural cytotoxic (NC) cell activity in vitro and in vivo. Single colour flow cytometric analysis showed that 6 +/- 1% of all CBA splenocytes were NC-1.1+, and that granular cells of varying sizes, as defined by their forward and side light scatter, contained the highest proportion of NC-1.1+ cells (29 +/- 7%). When analysed by two colour flow cytometry, the large and the granular NC-1.1+ spleen cells in CBA mice were shown to co-express Thy-1.2 and L3T4 (< or = 33%), Mac-1 (< or = 26%), IgM (< or = 60%) and FcR gamma II and J11d (< or = 100%). In contrast, T cell markers were not co-expressed on the small agranular NC-1.1+ spleen cells. This pattern of marker expression was evident in CBA spleens from 2 days of age. When (CBA x C57BL/6) F1 spleen cells were similarly analysed, 82% of granular NC-1.1+ cells also co-expressed NK-1.1. Single colour analysis of CBA bone marrow, thymus, lymph node and peripheral blood leucocytes revealed that < or = 10% of all cells examined were NC-1.1+ while the most granular cells in these organs were 16-43% NC-1.1+. These results support the 'horizontal lineage' theory that NC cells are cells of different lymphohaemopoietic cell lineages at particular stages of differentiation.
