Systemic and Local Expression of Perforin in Lymphocyte Subsets in Acute and Chronic Rheumatoid Arthritis

GORDAN GULAN, JAGODA RAVLIC-GULAN, NATASA STRBO, VLATKA SOTOSEK, BORIS NEMEC, DAMIR MATOVINOVIC, DUSAN RUBINIC, ECKHARD R. PODACK, and DANIEL RUKAVINA

ABSTRACT.

Objective. To investigate the role of the cytolytic action mediated by perforin in the course of rheumatoid arthritis (RA), we studied the immunophenotypic characteristics of lymphocytes containing perforin in peripheral blood (systemic level), in synovial fluid (SF), and in the synovial membrane (local level) in patients during the acute or chronic phase of RA. Cells from patients with osteoarthritis were used as controls.

Methods. Flow cytometry was used for simultaneous detection of intracellular (perforin) and cell surface antigens. Mean fluorescence intensity (MFI) was a measure of the mean perforin content per cell. Immunocytochemical staining was used to visualize perforin in the cytoplasmic compartment of cells.

Results. In acute RA highly significant changes in perforin expression were found in all compartments (peripheral blood, SF, and synovial membrane): (1) increase of percentage of total perforin positive cells; (2) increase of both subsets of cytolytic cells, T (CD8+P+) and NK (CD56+P+) cells; (3) increase in the frequency of perforin positive cells in CD8+ and CD56+ cell populations; and (4) the highest content of perforin/cell (MFI values) in all compartments, except in the synovial membrane.

Conclusion. Perforin positive cells may participate in the acute phase of RA by maintaining and perpetuating inflammation and contributing to tissue destruction. (J Rheumatol 2003;30:660-70)

Key Indexing Terms:

RHEUMATOID ARTHRITIS
LYMPHOCYTE MEDIATED CYTOTOXICITY
BLOOD
SYNOVIAL TISSUE
PERFORIN

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From the Department of Orthopaedic Surgery and Department of Physiology and Immunology, Faculty of Medicine, University of Rijeka, Rijeka, Croatia; and Department of Microbiology and Immunology, University of Miami, Miami, Florida, USA.

Supported by Grant 006201 from the Ministry of Science and Technology of the Republic of Croatia; Dr. Podack is supported by National Institutes of Health grant CA39201 and AI/DK 49829.

G. Gulan, MD, DSc, Senior Assistant of Orthopaedics; B. Nemec, MD, DSc, Professor of Orthopaedics; D. Matovinovic, MD, DSc, Professor of Orthopaedics; D. Rubinic, MD, Department of Orthopaedic Surgery; J. Ravlic-Gulan, MD, DSc, Assistant Professor of Immunology and Physiology; N. Strbo, MD, Assistant of Immunology and Physiology; V. Sotosek, MD, Research Assistant; D. Rukavina, MD, DSc, Professor of Immunology and Physiology, Department of Physiology and Immunology, Faculty of Medicine, University of Rijeka; E.R. Podack, MD, PhD, Professor, Department of Microbiology and Immunology, University of Miami.

Address reprint requests to Dr. D. Rukavina, Department of Physiology and Immunology, Faculty of Medicine, University of Rijeka, B. Branchetta 20, 51000 Rijeka, Croatia. E-mail: daniel@medri.hr

Submitted February 19, 2002; revision accepted September 27, 2002.

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