The N-terminus domain of the a2 isoform of vacuolar ATPase can regulate interleukin-1beta production from mononuclear cells in co-culture with JEG-3 choriocarcinoma cells.
OBJECTIVE
a2V-ATPase is the a2 isoform of vacuolar ATPase and is expressed in human trophoblast cells. a2V-ATPase resides as a 70-kDa molecule in intracellular vesicles. Upon cell stimulation, it migrates to the surface as a 50-kDa molecule, after a 20-kDa portion [N-terminus domain of the a2V-ATPase (a2NTD)] is cleaved and secreted to the extracellular environment. Previous studies showed that a2NTD-regulated cytokine production from stimulated T cells. The aim of this study was to determine if a2NTD can regulate cytokine production from immune cells that were in contact with JEG-3 cells.


METHODS
Peripheral blood mononuclear cells (PBMC) from females were co-cultured with JEG-3 cells in the presence or absence of a2NTD, and supernatants were analyzed by enzyme-linked immunosorbent assay for interleukin (IL)-1beta. Additionally, PBMC cultured with JEG-3 cells, in the presence or absence of a2NTD, were analyzed for cytokine gene expression by gene arrays.


RESULTS
There was an increased secretion of IL-1beta and a decrease in type I and II IL-1 receptors (IL1RA and IL-1R2) gene expression in PBMC that were co-cultured with JEG-3 cells in the presence of a2NTD.


CONCLUSIONS
These data suggest a role for a2NTD in the regulation of IL-1beta pro-inflammatory cytokine production at the fetal-maternal interface.
Ntrivalas
Evangelos
E
Clinical Immunology Laboratory, Rosalind Franklin University of Medicine and Science, 3333 Green Bay Road, N. Chicago, IL 60064, USA.
Gilman-Sachs
Alice
A
Kwak-Kim
Joanne
J
Beaman
Kenneth
K
D000906:Antibodies / Q000494:pharmacology; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002822:Choriocarcinoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D018920:Coculture Techniques; D005260:Female; D015870:Gene Expression / Q000187:drug effects; D006801:Humans; D053583:Interleukin-1beta / Q000378:metabolism*; D042541:Intracellular Space / Q000378:metabolism; D007527:Isoenzymes / Q000378:metabolism; D007963:Leukocytes, Mononuclear / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D008954:Models, Biological; D011247:Pregnancy; D021122:Protein Subunits / Q000378:metabolism; D053573:Receptors, Interleukin-1 Type I / Q000235:genetics; D053574:Receptors, Interleukin-1 Type II / Q000235:genetics; D011994:Recombinant Proteins / Q000276:immunology / Q000494:pharmacology; D014594:Uterine Neoplasms / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D025262:Vacuolar Proton-Translocating ATPases / Q000737:chemistry / Q000378:metabolism*
D000906:Antibodies; D053583:Interleukin-1beta; D007527:Isoenzymes; D021122:Protein Subunits; D053573:Receptors, Interleukin-1 Type I; D053574:Receptors, Interleukin-1 Type II; D011994:Recombinant Proteins; D025262:Vacuolar Proton-Translocating ATPases