Roles of calpain-calpastatin system (CCS) in human T cell activation.
The immune response is determined by the speed of the T cell reaction to antigens assured by a state of readiness for proliferation and cytokine secretion. Proliferation, apoptosis and motion of many cell types are controlled by cytoplasmic proteases - µ- and m-calpain - and their inhibitor calpastatin, together forming the "calpain-calpastatin system" (CCS), assumed to modify their targets only upon activation-dependent cytoplasmic Ca2+ increase. Contrastingly to this notion, using quantitative real time PCR and semiquantitative flow cytometry respectively, we show here that the CCS genes are constitutively expressed, and that both calpains are constitutively active in resting, circulating human CD4+ and CD8+ lymphocytes. Furthermore, we demonstrate that calpain inhibition in the resting T cells prevents them from proliferation in vitro and greatly reduces secretion of multiple cytokines. The mechanistic reason for these effects of calpain inhibition on T cell functions might be the demonstrated significant reduction of the expression of active (phosphorylated) upstream signalling molecules, including the phospholipase C gamma, p56Lck and NFκB, in the inhibitor-treated cells. Thus, we propose that the constitutive, self-regulatory calpain-calpastatin system activity in resting human T cells is a necessary, controlling element of their readiness for complex and effective response to antigenic challenge.
Department of Pathophysiology, Medical University of Gdańsk, Gdańsk, Poland.
Anna
A
Mikosik
Department of Pathophysiology, Medical University of Gdańsk, Gdańsk, Poland.
Aleksandra
A
Jasiulewicz
Department of Pathology and Experimental Rheumatology, Medical University of Gdańsk, Gdańsk, Poland.
Agnieszka
A
Daca
Department of Pathology and Experimental Rheumatology, Medical University of Gdańsk, Gdańsk, Poland.
Izabella
I
Henc
Department of Pathophysiology, Medical University of Gdańsk, Gdańsk, Poland.
Joanna E
JE
Frąckowiak
Department of Pathology and Experimental Rheumatology, Medical University of Gdańsk, Gdańsk, Poland.
Katarzyna
K
Ruckemann-Dziurdzińska
Department of Clinical and Social Gerontology, Medical University of Gdańsk, Gdańsk, Poland.
Jerzy
J
Foerster
Research Center on Ageing, University of Sherbrooke, Sherbrooke, Quebec, Canada.
Aurelie
A
Le Page
Department of Pathology and Experimental Rheumatology, Medical University of Gdańsk, Gdańsk, Poland.
Ewa
E
Bryl
Research Center on Ageing, University of Sherbrooke, Sherbrooke, Quebec, Canada.
Tamas
T
Fulop
Department of Pathophysiology, Medical University of Gdańsk, Gdańsk, Poland.
Jacek M
JM
Witkowski
D015703:Antigens, CD / Q000378:metabolism; D017209:Apoptosis / Q000187:drug effects / Q000235:genetics; D002135:Calcium-Binding Proteins / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology; D002154:Calpain / Q000235:genetics / Q000378:metabolism*; D002470:Cell Survival / Q000187:drug effects / Q000235:genetics; D016207:Cytokines / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D008213:Lymphocyte Activation / Q000187:drug effects / Q000276:immunology*; D010766:Phosphorylation; D016192:Resting Phase, Cell Cycle / Q000235:genetics; D015398:Signal Transduction; D016176:T-Lymphocyte Subsets / Q000276:immunology / Q000378:metabolism; D013601:T-Lymphocytes / Q000187:drug effects / Q000276:immunology* / Q000378:metabolism*
D015703:Antigens, CD; D002135:Calcium-Binding Proteins; D016207:Cytokines; C033668:calpastatin; D002154:Calpain
Immune response; Immunity; Immunology and Microbiology Section; T cells; calpain; calpastatin; cytokines; proliferation