Cytokines specificity

Once activated, T cells undergo clonal expansion (they multiply) under the influence of cytokines, specifically IL-2. These steps elicit an anti-donor T cell response that results in graft destruction. 

A single cell suspension of LNC is prepared under aseptic conditions and cultured for various periods of time. The production by LNC of type lcytokines (such as IFN-y and IL-12) and type 2 cytokines (such as IL-4, IL-5, IL-10 and IL-13) is measured using cytokine specific ELIS As or cytokine microarrays. It is possible to measure the spontaneous production of IL-5, IL-10, IL-12, IL-13 and IFN-y by LNC without restimulation in vitro. However, IL-4 appears to be produced in comparatively small amounts and to induce detectable levels of this cytokine an additional stimulus is required concanavalin A (con A), a T lymphocyte mitogen. Culture of LNC derived from mice treated with chemical respiratory sensitizer with con A will stimulate the production of detectable levels of IL-4. Treatment with the same mitogen of LNC derived from naive or vehicle-treated mice fails to induce measurable IL-4 secretion [86],... 

DUBl, 2, 2A mouse unknown cytokine specific growth regulators... 

Grotzinger, J., Kurapkat, G., Wollmer, A., Kalai, M., and Rosejohn, S. (1997). The family of the IL-6-type cytokines Specificity and promiscuity of the receptor complexes. Proteins 27, 96-109. 

Drumm K, Buhl R, Kienast K. 1999. Additional N02 exposure induces a decrease in cytokine specific mRNA expression and cytokine release of particle and fibre exposed human alveolar macrophages. Eur J Med Res 4 59-66. 

Figure 7.3 Production by draining lymph node cells of IL-4, IL-10 and IFN-yfollcwing repeated topical exposure of mice to TMA or DNCB. Groups of BALB/c strain mice (n=10) received 50 pi of 1 per cent DNCB or 10 per cent TMA (both in AOO) bilaterally on both shaved flanks. Five days later this treatment was repeated. After a further 5 days, 25 pi of chemical was applied to the dorsum of both ears daily for 3 consecutive days. One day following the final exposure mice were killed and draining auricular lymph nodes excised and pooled for each experimental group. A single cell suspension of lymph node cells was prepared and cultured in the presence (for mitogen-inducible IL-4 production), or absence (for the measurement of spontaneous IL-10 and IFN-y secretion) of 2 pg. ml 1 concanavalin A. Culture was terminated after various periods and the concentrations of IL-4, IL-10 and IFN-y measured in supernatants by cytokine-specific ELISA. In each case cytokine concentrations are recorded as mean values in ng. mf1. Standard errors are shewn when greater than 0.3 ng. mf1. A, IL-4 B, IL-10 and C, IFN-y.

Two types of soluble binding protein have been described, nonspecific serum proteins and cytokine-specific soluble receptor proteins. The major nonspecific serum protein capable of binding cytokines appears to be a2-macroglobuIin (B57, B58, Mil). A number of soluble cytokine inhibitors related to the relevant receptor have been described. The soluble form of the IL-2R a chain has been found in the serum of apparently normal individuals and is increased in the serum of individuals with inflammatory diseases such as rheumatoid arthritis (W31). Similar molecules have been described for IL-1, IL-6, IFNy, and IL-7 (F7, N14, S62). The mechanism of release has not been properly established but appears to require proteolytic cleavage of the membrane-bound receptor. Soluble inhibitors for two other cytokines, IL-4 and TNF, appear to be derived by alternative RNA splicing sites that give rise to receptors lacking a transmembrane sequence and that are secreted (M50, S22). 

Specific and nonspecific inhibitors have now been described for many cytokines. Specific inhibitors such as the IL-lra (H4) or soluble receptor components shed from cells may be inhibitory in bioassays (F7). IL-1 and IL-6 in plasma may be bound to carrier proteins such as aj-macroglobulin (B57, Mil). In addition circulating autoantibodies to cytokines have been found in normal individuals. 

These multi-analyte platforms (e.g., Luminex) consist of a benchtop flow cytometer/analyzer for the detection of cytokines captured onto microspheres ( beads ) with unique fluorescent intensities. The beads are covalently coupled to cytokine-specific antibodies so that cytokines present in biological fluids can be captured when mixed with the desired assortment of cytokine-specific beads. Detection antibodies carry the reporter molecule, phycoerythrin (PE), so that fluorescent signal is proportional to the amount of cytokine present in... 

Figure 9 RT-PCR analysis of cytokine mRNA expression in the spleens of DBA/2 mice immunized with casein/CKA. Total RNA was extracted from pooled spleens of S DHA/2 mice or CVE-administercd mice 3 weeks after immunized using RNAzol and reverse-transcribed, and the cDNA concentration was adjusted by co-amplification of two fold serially diluted cDNA and constant amounts of control plasmid pMCQ with [5-actin specific primers. Cytokine expression patterns of IL-12, IFNy, IL-6, and 11.-10 were determined by PCR using murine cytokine-specific primers. The PCR products were separated in 2 % agarose gel and visualized by ethidium bromide staining. Relative quantification of the amounts of the RT-PCR products was performed using a Computing Densitometer and NIH Image software,...

The effect of PDT on wound healing was tested in full-thickness incisional wounds in 24 hairless Sprague Dawly rats [114]. Cytokines, specifically TGF-beta, are believed to be instrumental in sustaining the fibrotic process, which leads to scarring. Rats were injected with 0.25 or 0.5 mg kg" BPD-MA or 5 or 10 mg kg CASP, 3 and 24 h prior to irradiation with light (1-20 J cm ), respectively. However, there was no apparent influence of PDT on either the rate or final appearance of wound healing. 

Thompson RD, Noble KE, Larbi KY, Dewar A, Duncan GS, Mak TW, Nourshargh S Platelet-endothelial cell adhesion molecule-1 (PECAM-l)-deficient mice demonstrate a transient and cytokine-specific role for PECAM-1 in leukocyte migration throu the perivascular basement membrane. Blood 2001 97 1854-1860. 

Zalcman, S., Green-Johnson, J.M., Murray, L., Nance, D.M., Dyck, D., Anisman, H., and Greenberg, A.H. 1994. Cytokine-specific central monoamine alterations induced by interleukin-1, -2 and -6. Brain Res. 643, 40-49. 

---