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J Immunol Methods. 2006 Apr 20;311(1-2):47-56. Epub 2006 Feb 21.

Novel two-parameter flow cytometry (MIL4/SSC followed by MIL4/CT7) allows for identification of five fractions of guinea pig leukocytes in peripheral blood and lymphoid organs.

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1
AIDS Research Center, National Institute of Infectious Diseases, Tokyo, 162-8640, Japan.

Abstract

Though the guinea pig has been an extremely useful animal model for a variety of diseases, the tools necessary to undertake a full-scale immunological analysis of the guinea pig have been lacking. For instance, traditional two-parameter forward/side scatter (FSC/SSC) flow cytometry, though effective in human and other animal models, is unable to adequately identify the distinct fractions of guinea pig peripheral blood leukocytes (PBL). We introduce here a new flow cytometric technique (MIL4/SSC followed by MIL4/CT7) which redresses this lack by identifying and characterizing five distinct fractions of PBL: neutrophils, lymphocytes, monocytes, eosinophils plus basophils, and the novel MIL4(-)SSC(large)CT7(high) population. The MIL4(-)SSC(large)CT7(high) cells possess cytoplasmic inclusion bodies of variable size that were positive for periodic acid Schiff (PAS). Their cell surface stained positive for the helper/inducer lymphocyte markers, T cell markers, CD45, Thy-1, asialo GM1 and FcR, but negative for B cell markers, such as membrane-type IgM, CD8 and MHC class II. The novel flow cytometric technique also allowed us to establish that the five leukocyte fractions were found in PBL, splenocytes, thymocytes and lymph node cells. Cells which were positive for inclusion bodies comprised 16.6% of splenocytes, 9.9% of PBL and 4.3% of liver cells, but were comparatively rare in lymph node cells, thymocytes, and BM cells. The novel flow cytometric technique introduced here will allow a better understanding of the response of each type of guinea pig leukocyte and thereby shed light on the diseases with which they are associated.

PMID:
16533513
DOI:
10.1016/j.jim.2006.01.010
[Indexed for MEDLINE]

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