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The Dow closes at 10, for the first time. Buddy and Socks the Clinton dogs. Learn more. Our cellular immune system has to cope constantly with foodborne substances that enter the bloodstream postprandially. Here, they may activate leukocytes via specific but yet mostly unknown receptors. Ectopic RNA expression out of gene families of chemosensory receptors, i.
The mRNA expression and the size of subpopulations expressing certain chemosensory receptors varied largely among individual blood samples, suggesting a regulated expression of olfactory and taste receptors in these cells. Moreover, our cellular immune system not only is exposed continuously to endogenous cytokines, hormones, growth factors, and mediators but also to exogenous foodborne chemicals, such as aroma compounds or tastants, which may enter the bloodstream postprandially [ 11 — 13 ]. Thus, it is widely accepted that our cellular immune system responds to certain food ingredients or metabolites [ 5 , 14 — 18 ].
However, their molecular targets on our immune cells remained largely unknown so far. Recently, we have demonstrated in different types of isolated human blood leukocytes the functional expression of members of an olfactory receptor family, the TAARs [ 19 — 21 ], which detect certain biogenic amines with high sensitivity [ 22 , 23 ]. Human and bovine blood leukocytes were purified by use of Ficoll density gradient centrifugation, as described previously [ 22 ]. Total RNA from blood leukocytes was isolated and purified by CsCl gradient centrifugation, as described previously [ 22 ].
Standardised viewing conditions for visual colour matching
Immunocytochemistry for TAAR1 was performed as described previously [ 22 ]. The cell preparation was the same as described previously [ 22 ]. Antibody concentrations were the same as described above for WB. PMN were seeded in the chemotaxis chamber, and live cell imaging of individual cell tracking was monitored under light microscope at 0. Migration data from both gradient conditions, as well as from controls, were collected. Only genes with clear melting curves and single specific product peaks were taken for further data analysis.
Samples that showed irregular melting peaks were excluded from the quantification procedure. Class I OR gene transcripts are expressed in human and bovine blood leukocytes. Data were normalized to an averaged expression of 6 different reference genes, with the lowest value [human, OR56B1 blue ; cow, OR51B4 red ] set as 1.
We next asked how transcript levels are displayed across Class I OR. We picked the cow to investigate OR gene expression in a species with modes of nutrition and digestion different than human but with a high genome homology to human [ 54 ] and a large number of OR genes [ 55 ]. The averaged Cq values ranged from 27 to 31 over all bovine and human chemosensory receptor qPCR experiments. The presence of taste receptors in specific human blood cell types has never been reported. C Upper scale, Human receptors; lower scale, cow receptors.
The cow has 16 functional TAS2R genes [ 57 , 58 ], of which, we identified 12 as putative orthologs [ 58 ] to human genes. We then asked whether different, most abundant receptors from the same chemosensory modality coexpress in the same cells or not. In the olfactory system, typically only 1 OR allele is expressed per olfactory sensory neuron in the nose [ 59 — 61 ]. Whereas flow cytometry would appear to be the method of choice to answer this question for blood cells, the lack of antibodies suitable for life cell staining prevented us from the use of this method. Again, the quality of the antibodies available limited the number of investigated receptors in our study.
For the 3 OR investigated, we observed overlapping signals in the same cells; however, the signals never overlapped completely Fig. The detection methods for the 3 OR were diverse, as the primary antibodies used were labeled directly or required detection by a labeled secondary antibody or labeled streptavidin. Chemosensory receptor protein expression in subpopulations of human blood leukocytes. Horizontal lines indicate median; X, data mean.
Lower and upper blue bars indicate 2nd and 3rd quartiles of data distribution; horizontal lines, median; X, data mean. By taking PMN, which had been fixed and labeled by the same procedures and antibodies as in Fig. Therefore, we refrained from investigating coexpression of TAS2R. We then asked whether in PMN, olfactory and taste receptors coexpress in the same cells. We found that the signals for both subunits partially overlapped in vesicles lining up at the cell surface Fig.
D Merge of signals from B and C, with overlapping signals appearing in yellow. E Overlay of the transmission light picture of the same cell. Shown are percentage and distance of 40 migrated PMN.
Similar results were obtained from 3—6 independent experiments. Furthermore, we asked whether we can induce a cellular function in PMN via their chemosensory receptors. In vivo, during the innate immune response, PMN leave blood vessels and actively migrate along a chemoattractant gradient toward a site of tissue injury or infection [ 67 ]. We found that Do leukocytes express olfactory and taste signaling molecules?
Flegel et al. Here, we show mRNA expression at levels of Their expression levels Fig. C Lactisole at 0.
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We observed chemotactic migration of The concept of expression of olfactory and taste receptors in different primary human blood leukocyte types is new. So far, there have been only 3 reports on mRNA expression of few chemosensory receptors and only in total leukocyte RNA preparations [ 38 , 47 , 48 ]. TAAR, too, may be considered olfactory receptors, at least from experiments with rodents [ 19 , 20 , 73 ].