The effect of antigen stimulation on alpha(4), beta(1) and beta(7) chain integrin expression and function in CD4+ cells.
BACKGROUND
The alpha(4) integrin, as alpha(4)beta(1) (VLA-4) or alpha(4)beta(7), is critical for T cell migration and proliferation, although its functional modulation remains poorly understood. We hypothesized that increased receptor density, based on new receptor chain synthesis, was one such mechanism. We examined the surface receptor density of the alpha(4) and beta(1) chains on CD4+CD45RO+ cells, and the mRNA expression of these and the beta(7) chain in response to allergen and nonallergen antigen stimulation.


METHODS
Flow-cytometric analyses for CD49d, CD29, and CD45RO were performed on T cell lines specific for timothy, tetanus, and Candida from atopic and nonatopic donors. RNA was extracted from cells sorted to select CD4+/CD49d-positive cells before and after stimulation. Equivalent amounts of cDNA for beta-actin, alpha(4), beta(1) and beta(7) were used in PCR, and the products were quantified using phosphoimaging.


RESULTS
CD49d expression is heterogeneous on T cell lines and is upregulated by antigen stimulation on CD4+ T cells. The surface expression on CD4+CD45RO+ timothy allergen or tetanus toxoid T cell lines is at least double that found on CD45RO- cells. Antigen stimulation upregulated CD49d expression on the CD4+CD45RO+ subpopulation of both cell lines although it was not as significant as in the case of all CD4+ T cells. CD29 surface expression behaves similarly. Candida had no effect on CD49d or CD29. Messenger RNA expression for the alpha(4) chain (CD49d) is significantly upregulated 48 h following the addition of timothy or tetanus. beta(7) chain expression also rises significantly on both cell lines. beta(1) chain expression increases, but not significantly.


CONCLUSIONS
The surface expression of the CD49d is heterogeneous and much higher on CD4+CD45RO+ cells than on CD4+RO- T cells. The CD49d integrin chain on CD4+ T cells is upregulated following antigen exposure. However, the CD4+CD45RO+ subpopulation is only partially responsible for this increase suggesting other T cells to have this receptor expression upregulated. CD29 expression behaves similarly. Messenger RNA expression increases coordinately for alpha(4), beta(7), and not significantly for beta(1) in these cells. These observations provide a potential mechanism for the selective accumulation of T cells at sites of inflammation, and suggest an important point of intervention for allergic and inflammatory disease.
Tarkowski
M
M
Institute of Occupational Medicine, Department of Occupational Diseases, Lodz, Poland.
Pacheco
K A
KA
Rosenwasser
L J
LJ
D000485:Allergens / Q000276:immunology; D000938:Antigen-Presenting Cells / Q000502:physiology; D015703:Antigens, CD / Q000096:biosynthesis / Q000235:genetics; D015139:Blotting, Southern; D015496:CD4-Positive T-Lymphocytes / Q000378:metabolism*; D017931:DNA Primers / Q000737:chemistry; D005434:Flow Cytometry; D006801:Humans; D039441:Integrin alpha4; D039041:Integrin alpha4beta1; D019012:Integrin beta1 / Q000096:biosynthesis / Q000235:genetics*; D016023:Integrins / Q000096:biosynthesis / Q000235:genetics*; D017493:Leukocyte Common Antigens / Q000378:metabolism; D008213:Lymphocyte Activation; D012333:RNA, Messenger / Q000096:biosynthesis*; D016201:Receptors, Lymphocyte Homing / Q000096:biosynthesis / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation
D000485:Allergens; D015703:Antigens, CD; D017931:DNA Primers; D039041:Integrin alpha4beta1; D019012:Integrin beta1; D016023:Integrins; D012333:RNA, Messenger; D016201:Receptors, Lymphocyte Homing; C071777:integrin alpha4beta7; D039441:Integrin alpha4; D017493:Leukocyte Common Antigens