SUMMARYCoeliac disease is apparently a T cell-mediated disease, precipitated in the proximal small intestine of susceptible individuals by gluten. Preferential presentation of gluten peptides most probably takes place in coeliac mucosa by the disease-associated HLA-DQ2 and -DQ8 molecules. In peripheral blood, however, both HLA-DR, -DQ and -DP-restricted T cell responses to gluten have been observed. We examined gluten-specific T cell clones (TCC) derived from peripheral blood for cytokine production to see if their profiles were related to the HLA restriction or the disease state of the donors. As previously found for mucosal TCC, the main product was interferon-gamma (IFN-), often with additional IL-4, IL-5, IL-6, IL-10, tumour necrosis factor, and transforming growth factor-beta. Regardless of restriction element or disease state, gluten-reactive TCC from peripheral blood therefore seem to secrete cytokines compatible with a Th0 profile.

Coeliac disease (CD) is probably caused by an abnormal immune response towards wheat gliadin in the small intestine. We found that gliadin-specific T cells from the small intestinal mucosa of HLA-DQ2 positive CD patients were almost exclusively restricted by the disease-associated DQ2 molecule. In the peripheral blood of CD patients, a large proportion of gliadin-specific T cells were found to be restricted by DQ molecules, including DQ2, but many were instead restricted by DR or DP molecules of the patient. We have now investigated gliadin-specific T cell responses in peripheral blood from healthy individuals. Four of 20 persons tested had strong in vitro responses and were used as donors for gliadin-specific T cell clones. We found gliadin-specific T cells restricted by the CD-associated DQ2 molecule in peripheral blood for two of these four individuals. It is the presence of such T cells also in the small intestinal mucosa which seems typical of CD.

Work on t h e paper chromatography of glycosides and aglycones from Digitalis purpwra and Digitalis lanata has already been published (BBR-HEIM SVEXDSEN and BRISEID JENSEN 1950). The method described there has been developed further and used in coiinection with a work on the separation and fluorimetric determination of gitoxigenin, gitoxin and purpureaglycoside B (BRISEID JENSEN 1052 b). I n elaborating the method consideration was only given in the earlier work to the possible presence of the hitherto known cardioactive substances in the digitalis leaf, namely purpureaglycoside A, digitoxin, digitoxigenin and the three aforementioned B-series substances. However, further paper chromatographic investigations of ''pure" substances, pharmaceutical preparations and drug extracts show that the glycoside mixture is considerably more complex : several new glycosides can be detected, and a couple of them seem to occur in significant quantities. A satisfactory separation of the sometimes highly complex glycoside mixtures requires further refinement of the paper chromatographic technique. In the present paper are described the increase of separation effected by combining the previously used liquid system (chloroform-methanol-water) and the system chloroformbenzene-forrnamide as well as new and more discriminating developers.Chromatography on forrnamitle-impregnated filter paper was introduced by ZAFFAROM, BURTON and KEUTMANN (1960) for rorticosteroitls. The method has subsequently been developed and largcly usc~cl by SCHINDLER and REICHSTEI~ (1 951) in invt,stigatioris of strophanthus glyrosides. I t has also becn used by NEHER and WETTSTEIN ( 1 952) in the chromatography of a number of slightly polar steroids, arid by HEFTMANN anti LEVANT (19.52) in the chromatography of varioiis cardiac glyc*osides, thcir acetates and thcir aglycones.In the previous work on paper chromatography of digitalis glycosides or digitaloid substances the following reagents have been i i s e d for localizing the individual substances : trtchloroacotic acid (B~ZRHEIM SVENDSEN and BRISEID JENSEN 1950 ; BRISEID JENSEN 1952 b ; HEFVMANN and LEVANT 1952), hydrochloric acid vapor (MESNARD anti DEVZE 1950; HASALL 1951), alkaline m-dinitrobcnzene (JAMINET 1950; SCHINDLER and REICHSTEIN 1951). amnioniacal nitrate of silver (HEFTMANN and LEVANT 1952).1 Art8 ~iharma~ologira vol. 9, fasc. 2.

The stability of adrenaline in some injections has been investigated by chemical assays based on determinations of adrenochrome and of adrenolutine, while the rat blood pressure method and the rat uterus inhibition method served as biological control procedures. The colorimetric method, in which adrenaline is oxidised to adrenochrome by means of potassium ferricyanide in acid solution, proved unable to detect any deterioration of adrenaline in procaine and adrenaline injections. The corresponding fluorimetric method, in which the adrenochrome formed is rearranged to adrenolutine by addition of strong alkali, gave results which agreed well with the biological results. In injections, adjusted to pH 3 with hydrochloric acid, adrenaline was stable for at least 20 months at 4" in solutions heated at 120" for 20 min., at 100" for 20 min. or unheated. Replacement of the hydrochloric acid by sodium metabisulphite (pH 3.6) prevented the discoloration. In injections of procaine and adrenaline, adjusted to pH 3 with hydrochloric acid, the adrenaline content decreased over the 20 month period, and was most pronounced in the solutions that had been heated before storage. Sodium metabisulphite significantly increased the stability of adrenaline in these injections and also prevented any colour formation.

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