Five new binary vectors have been constructed which have the following features: (1) different plant selectable markers including neomycin phosphotransferase (nptII), hygromycin phosphotransferase (hpt), dihydrofolate reductase (dhfr), phosphinothricin acetyl transferase (bar), and bleomycin resistance (ble); (2) selectable markers are located near the T-DNA left border and; (3) selectable marker and beta-glucuronidase (uidA) reporter genes are divergently organized for efficient expression, and can easily be removed or replaced as needed.

With the binary vectors pBIG and pBIB it is possible to: i) exchange the plant selectable marker/promoter fusion and the reporter gene, ii) use one vector and Agrobacterium strain system, thus reducing variables associated with using different vectors and strains, iii) allow for divergent transcription of the selectable marker gene and of promoter/reporter gene fusions.The Hyg (4), the neomycin-phosphotransferase (NPT II) gene from pBI 101 (1), and a modified hygromycin B phosphotransferase (HPT) gene was taken from pGDW 11 (5). Subsequently, the GUS-gene was exchanged by the pUC 19 polylinker (Hind lI/Sst I) without a Bam HI site to create pBIB-KAN and pBIB-HYG, respectively.The vectors have been tested by leaf disk transformation of Nicotiana tabacum cv. Petit Havana SRI and were effective in producing resistant plants which produce shoots and roots on MSmedium containing the appropriate antibiotics (kanamycin sulfate 200 Ag/ml; hygromycin B 45 /tg/ml).

SununaryInterleukin 10 (IL-10) is released during the induction phase of contact sensitivity and was shown in prior functional studies to convert epidermal Langerhans cells (LC) from potent inducers of primary immune responses to specifically tolerizing cells in vitro. To investigate whether IL-10 also subserves the function of a tolerizing agent in vivo ears of BALB/c or C3H mice were injected intradermally with 1-2 pg of recombinant mouse (rm)IL-10 8 h before epicutaneous application of 3% trinitrochlorobenzene (TNCB; a contact allergen). As a control, mice were injected with phosphate-buffered saline or IL-10 plus neutralizing amounts of anti-IL-10 mAb. 5 d later, mice were challenged with 1% TNCB on contralateral ears and ear swelling response was measured 24 h later. Whereas control-treated mice showed a normal ear swelling response to epicutaneous challenge (A mm -2 = 25 _+ 5), ear swelling response of IL-10-treated animals was significantly inhibited (A mm -2 = 3 _+ 2). Coinjection of IL-10-specific mAb together with traiL-10 completely abrogated this effect. To differentiate between a state of nonresponsiveness and induction of tolerance by IL-10, mice initially treated with IL-10 and TNCB were resensitized with 3% TNCB in the absence of any treatment after 14 d of rest (group 1). Again mice were challenged 5 d later and ear swelling responses were tested. Whereas control mice treated with allergen alone (group 2) showed a good swelling response (Amm -2 = 28 _+ 6), IL-10-treated mice (group 1) showed a minimal response towards application of allergen (A mm -2 = 4 + 2). To show that anergy induction by IL-10 was antigen-specific, mice initially treated with IL-IO plus TNCB were exposed to 0.5% dinitrofluorobenzene (DNFB) 14 d later (group 1). After challenge with 0.1% DNFB, IL-10-treated mice showed an ear swelling response (A mm -2 = 13 -+ 3; group 1) similar to that of control mice only sensitized with DNFB (A mm -2 = 14 _+ 3; group 3). In an attempt to show the induction of antigen-specific tolerance in these mice in vitro, regional lymph nodes of mice initially treated with TNCB plus IL-10 (group 1) and control-treated mice (groups 2 and 3) were prepared and cultured in the presence of TNBS, dinitrobenzene sulfonate (DNBS), or medium to measure antigen-specific proliferation. Lymph node cells from animals sequentially treated with IL-10 plus TNCB and afterwards DNFB in the absence of IL-10 (group 1) showed a low, but significant proliferation (,,,15,000 cpm) to DNBS, but only background proliferation (•3,000 cpm) to TNBS, or medium. In contrast, lymph node cells from animals treated with TNCB or DNFB in the absence of IL-10 (groups 2 and 3) proliferated to the sensitizing agent, but not control allergens. To elucidate the mechanism of action of IL-10, the epidermal cytokine pattern was analyzed on the mRNA level after injection of IL-10 or controls and application of allergen. Injection of IL-10 (but not controls) significantly impeded the induction of proinflammatory cytokines IL-lfl, tumor necrosis ...

Summary Epicutaneous patch testing is the diagnostic standard for the detection of allergic contact dermatitis. The present guidelines are aimed at residents and board‐certified physicians in the fields of dermatology and allergology as well as other medical specialties involved in establishing the indication for patch testing and its execution in patients with contact dermatitis and other forms of delayed‐type hypersensitivity. The target audience also includes other health care providers and insurance funds. Based on a systematic literature search and a formal consensus process (S3), the guidelines were developed by dermatologists in collaboration with pediatricians, occupational medicine physicians, nursing staff as well as patient representatives. The systematic methodological approach and appraisal of evidence upon which the recommendations are based are outlined in a separate method report that also contains evidence tables. The guidelines address general aspects of patch testing as well as medicolegal issues. The recommendations given relate to topics such as the indication for patch testing, informed patient consent, as well as the choice of test substances, test chambers and test site, duration of exposure, reading times and interpretation of test reactions. Furthermore, recommendations are provided with respect to endogenous and exogenous factors, specific patient groups (children, pregnant women, immunosuppressed individuals) as well as possible risks and adverse events associated with patch testing using contact allergens. Note: This publication is part 1 of the short version of the S3 guidelines for “Epicutaneous patch testing using contact allergens and drugs” (registry no. 013 – 018; date: March 20, 2019; valid until December 31, 2021). Part 2 of the short version will be published in the next issue. The long version of these guidelines can be accessed at http://www.awmf.org. The method report is available as online publication (https://www.awmf.org/leitlinien/detail/ll/013-018.html) and contains the evidence tables in its appendix.

SummaryEpicutaneous patch testing is the diagnostic standard for the detection of allergic contact dermatitis. The present guidelines are aimed at residents and board‐certified physicians in the fields of dermatology and allergology as well as other medical specialties involved in establishing the indication for patch testing and its execution in patients with contact dermatitis and other forms of delayed‐type hypersensitivity. The target audience also includes other health care providers and insurance funds.Based on a systematic literature search and a formal consensus process (S3), the guidelines were developed by dermatologists in collaboration with pediatricians, occupational medicine physicians, nursing staff as well as patient representatives. The systematic methodological approach and appraisal of evidence upon which the recommendations are based are outlined in a separate method report that also contains evidence tables.The guidelines address general aspects of patch testing as well as medicolegal issues. The recommendations given relate to topics such as the indication for patch testing, informed patient consent, as well as the choice of test substances, test chambers and test site, duration of exposure, reading times and interpretation of test reactions. Furthermore, recommendations are provided with respect to endogenous and exogenous factors, specific patient groups (children, pregnant women, immunosuppressed individuals) as well as possible risks and adverse events associated with patch testing using contact allergens.