Expression of receptors for interleukin 4 and interleukin 7 on human T cells.
Human recombinant interleukin 4 (IL-4) and interleukin 7 (IL-7) have been modified with biotin-N-hydroxysuccinimide and used to examine the expression of human IL-4 and IL-7 receptors (R) on activated peripheral blood T cells by flow cytometry. Freshly isolated T cells expressed only a low level of IL-4R which remained unchanged when cells were cultured in the absence of stimuli. In the presence of IL-4, IL-7, phytohemagglutinin A (PHA) or immobilized CD3 monoclonal antibody the intensity of biotinylated IL-4 staining increased approximately twofold on the majority of cells. A combination of mitogen with either IL-4 or IL-7 caused a considerable increase in IL-4 receptor expression over that seen in the presence of mitogen alone. IL-2 alone failed to induce IL-4R although it was able to cause a significant increase in receptor expression on T cells co-cultured with PHA or CD3. Freshly isolated T cells expressed high levels of IL-7R, as determined by biotinylated IL-7 binding and flow cytometry, which did not change significantly with culture in medium alone. Stimulation with PHA, Concanavalin A (Con A) or CD3 had little effect on the intensity of staining. In contrast, activation with phorbol ester resulted in a decrease in IL-7R expression. Similarly, in the presence of IL-4 or IL-7, but not IL-2, the intensity of staining with biotinylated IL-7 was lowered. Analysis of purified T-cell populations showed that IL-7R were present, and IL-4R could be induced, on both CD4+ and CD8+ populations. Analysis of IL-4 receptor expression by this flow cytometric technique was supported by results from 125I-labeled IL-4 binding and by Northern blot analysis of mRNA levels. Taken together, the results of these studies show that the use of biotinylated cytokines and flow cytometry provides a very sensitive method with which to study the expression and regulation of cytokine receptors.
Armitage
R J
RJ
Immunex Corporation, Seattle, WA.
Ziegler
S F
SF
Beckmann
M P
MP
Idzerda
R L
RL
Park
L S
LS
Fanslow
W C
WC
D000818:Animals; D015704:CD4 Antigens; D016827:CD8 Antigens; D006801:Humans; D066298:In Vitro Techniques; D015847:Interleukin-4 / Q000378:metabolism; D015851:Interleukin-7 / Q000378:metabolism; D010835:Phytohemagglutinins / Q000276:immunology; D012333:RNA, Messenger / Q000096:biosynthesis; D051381:Rats; D011971:Receptors, Immunologic / Q000378:metabolism*; D019948:Receptors, Interleukin-4; D020395:Receptors, Interleukin-7; D011975:Receptors, Mitogen / Q000378:metabolism*; D016176:T-Lymphocyte Subsets / Q000276:immunology; D013601:T-Lymphocytes / Q000276:immunology*
D015704:CD4 Antigens; D016827:CD8 Antigens; D015851:Interleukin-7; D010835:Phytohemagglutinins; D012333:RNA, Messenger; D011971:Receptors, Immunologic; D019948:Receptors, Interleukin-4; D020395:Receptors, Interleukin-7; D011975:Receptors, Mitogen; D015847:Interleukin-4