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Blood, 1 June 2001, Vol. 97, No. 11, pp. 3621-3627
PHAGOCYTES
CD44 ligation on peripheral blood polymorphonuclear cells induces
interleukin-6 production
Giuseppe Sconocchia,
Laura Campagnano,
Domenico Adorno,
Angela Iacona,
Nella Y. Cococcetta,
Vittorio Boffo,
Sergio Amadori, and
Carlo U. Casciani
From the Institute of Tissue Typing and Dialysis, CNR;
and the Department of Surgery and the Department of Biopathology,
University "Tor Vergata," Rome, Italy.
Polymorphonuclear cells (PMNs) contribute to the initiation and
progression of the immune response by mediating cytotoxicity, phagocytosis, and cytokine secretion. Because CD44 serves as a cytotoxic-triggering molecule on PMNs, it was hypothesized that it
could also trigger cytokine production. In this study, the effect of
anti-CD44 antibodies on interleukin-6 (IL-6) production in human PMNs
was assessed. By using a reverse transcriptase-polymerase chain
reaction, it was shown that PMNs stimulated with a mouse monoclonal or
a rabbit polyclonal F(ab)2 anti-CD44 transcribe IL-6
messenger RNA. A similar effect was obtained when an anti-CD44 antibody
was replaced with hyaluronic acid (HA). Kinetic studies showed that
anti-CD44 and HA induced IL-6 gene transcription, initiated 3 hours
after stimulation, peaked between 12 and 24 hours, and disappeared
after 48 hours. Analogous results were achieved when secreted IL-6
protein was measured by enzyme-linked immunosorbent assay in the PMN
culture supernatants. To characterize which metabolic pathways
regulated CD44-dependent IL-6 production in PMNs, an RNA polymerase
inhibitor, actinomycin D, and 2 protein kinase inhibitors, such as
genistein and staurosporine, were tested. Actinomycin D and genistein
blocked IL-6 production, whereas staurosporine did not, suggesting that
CD44-dependent IL-6 production requires gene transcription and tyrosine
kinase activity. Furthermore, the relationship between CD44 and
cytokines that affect PMN function, including interferon  (IFN)
and IL-2, was investigated. Without CD44 cross-linking, IFN did not
trigger IL-6 production. However, on CD44 cross-linking, IFN
produced a strong synergistic effect on IL-6 syntheses in human PMNs.
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