Altered expression of TNF-alpha signaling pathway proteins in systemic lupus erythematosus.
OBJECTIVE
To investigate the expression of tumor necrosis factor receptors (TNFR1 and TNFR2) and adapter proteins (TRADD, RIP, and TRAF2) in peripheral blood mononuclear cell (PBMC) subsets from patients with systemic lupus erythematosus (SLE).


METHODS
PBMC were isolated from 45 SLE patients and 25 controls, and stained with labeled antibodies that enabled identification of various T cell, B cell, and monocyte subpopulations. Expression of TNF-related signaling molecules was measured by staining with labeled antibodies either directly or following fixation and permeabilization. Apoptosis was quantified using an anti-active caspase 3 antibody. RNA expression of TNF-related signaling molecules was assessed by quantitative RT-PCR and serum levels of TNF-alpha by ELISA.


RESULTS
SLE patients had increased levels of TNFR1, TNFR2, and TRAF2, together with decreased levels of RIP, on various B, CD4+ T, and CD8+ T cell subsets as compared to controls. This altered expression was seen in both naive and memory subpopulations, and reflected altered staining of the whole population rather than a subset of cells that were activated. The levels of these molecules were not significantly correlated with serum TNF-alpha levels or their RNA expression in whole peripheral blood. TNFR1 and TNFR2 expression was negatively correlated with disease activity. There was no association between the proportion of apoptotic cells in any of the subpopulations and serum TNF-alpha levels or expression of TNF-related signaling molecules.


CONCLUSIONS
Patients with SLE had altered expression of TNF-related signaling molecules, suggesting that there may be an imbalance in TNF-alpha signaling favoring cellular activation as opposed to proapoptotic pathways.
Arthritis Centre of Excellence and Toronto Western Hospital Research Institute, Divisions of Genetics and Development and Population Health, Toronto Western Hospital, Toronto, Ontario, Canada.
Lang-Jing
LJ
Zhu
Carolina
C
Landolt-Marticorena
Timothy
T
Li
Xiao
X
Yang
Xue-Qing
XQ
Yu
Dafna D
DD
Gladman
Murray B
MB
Urowitz
Paul R
PR
Fortin
Joan E
JE
Wither
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics / Q000378:metabolism*; D000328:Adult; D000368:Aged; D017209:Apoptosis / Q000276:immunology; D053420:CRADD Signaling Adaptor Protein / Q000235:genetics / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D016130:Immunophenotyping; D007963:Leukocytes, Mononuclear / Q000276:immunology / Q000378:metabolism*; D008180:Lupus Erythematosus, Systemic / Q000276:immunology / Q000378:metabolism*; D016131:Lymphocyte Subsets / Q000378:metabolism / Q000473:pathology; D008297:Male; D008875:Middle Aged; D047888:Receptors, Tumor Necrosis Factor, Type I / Q000235:genetics / Q000378:metabolism*; D047889:Receptors, Tumor Necrosis Factor, Type II / Q000235:genetics / Q000378:metabolism*; D053199:TNF Receptor-Associated Death Domain Protein / Q000235:genetics / Q000378:metabolism; D055815:Young Adult
D048868:Adaptor Proteins, Signal Transducing; D053420:CRADD Signaling Adaptor Protein; D047888:Receptors, Tumor Necrosis Factor, Type I; D047889:Receptors, Tumor Necrosis Factor, Type II; C446759:TIFA protein, human; D053199:TNF Receptor-Associated Death Domain Protein