Hairy roots derived from the infection of a plant by Rhizobium rhizogenes (previously referred to as Agrobacterium rhizogenes) bacteria, can be obtained from a wide variety of plants and allow the production of highly diverse molecules. Hairy roots are able to produce and secrete complex active glycoproteins from a large spectrum of organisms. They are also adequate to express plant natural biosynthesis pathways required to produce specialized metabolites and can benefit from the new genetic tools available to facilitate an optimized production of tailor-made molecules. This adaptability has positioned hairy root platforms as major biotechnological tools. Researchers and industries have contributed to their advancement, which represents new alternatives from classical systems to produce complex molecules. Now these expression systems are ready to be used by different industries like pharmaceutical, cosmetics, and food sectors due to the development of fully controlled large-scale bioreactors. This review aims to describe the evolution of hairy root generation and culture methods and to highlight the possibilities offered by hairy roots in terms of feasibility and perspectives.
†These authors contributed equally to this work.Most kidney transplant recipients who discontinue immunosuppression reject their graft. Nevertheless, a small number do not, suggesting that allogeneic tolerance state (referred to operational tolerance) is achievable in humans. So far, however, the rarity of such patients has limited their study. Because operational tolerance could be linked to anergy, ignorance or to an active regulatory mechanism, we analyzed the blood T-cell repertoire usage of these patients. We report on comparison of T-cell selection in drug-free operationally tolerant kidney recipients (or with minimal immunosuppression), recipients with stable graft function, chronic rejection and healthy individuals. The blood T cells of operationally tolerant patients display two major characteristics: an unexpected strongly altered T-cell receptor (TCR) Vb usage and high TCR transcript accumulation in selected T cells. The cytokine transcriptional patterns of sorted T cells with altered TCR usage show no accumulation of cytokine transcripts (IL10, IL2, IL13, IFN-c ), suggesting a state of hyporesponsiveness in these patients. Identification of such a potential surrogate pattern of operational tolerance in transplant recipients under life-long immunosuppression may provide a new basis and rationale for exploration of tolerance state. However, these data obtained in a limited number of patients require further confirmation on larger series.
A collection of 332 PVY isolates, originating from potato crops, was characterized in order to seek the newly described PVY variants, PVYNW and PVYZ. Serologically, 284 samples were found to contain only PVYO, 36 samples to contain only PVYN, whereas 12 samples were doubly infected by both PVYO and PVYN. All 36 isolates, serologically PVYN‐typed and 16 isolates serologically PVYO‐typed, induced vein necrosis on tobacco. The latter 16 isolates reacted like a Polish standard PVYNW isolate and are probably new members of that PVY subgroup; in the panel of samples, they originated from potato cultivars never previously reported as infected by PVYNW and their high incidence probably indicates that they are more widespread than commonly accepted. Indexing of 108 PVYO isolates on three potato cultivars (Desiree, Eersteling and Maris Bard) carrying hypersensitivity resistance genes to PVYO, PVYC and PVYZ, respectively, revealed the presence of various pathotypes. Two isolates reacted as two standard isolates, designated PVYZE, originating from Spain and previously suspected to be PVYZ members; these isolates were able to overcome the hypersensitivity genes in the three potato cultivars and consequently were members of a newly recognized pathotype. Another isolate evoked the reactions expected after infection with PVYZ. Surprisingly, PVYC was only found in one sample that was doubly infected by PVYO.
Chronic rejection represents a major cause of long-term kidney graft loss. T cells that are predominant in long-term rejected kidney allografts (35 ± 10% of area infiltrate) may thus be instrumental in this phenomenom, which is likely to be dependant on the indirect pathway of allorecognition only. We have analyzed the variations in T cell repertoire usage of the Vβ chain at the complementary determining region 3 (CDR3) level in 18 human kidney grafts lost due to chronic rejection. We observed a strongly biased intragraft TCR Vβ usage for the majority of Vβ families and also a very high percentage (55%) of Vβ families exhibiting common and oligoclonal Vβ-Cβ rearrangements in the grafts of patients with chronic rejection associated with superimposed histologically acute lesions. Furthermore, Vβ8 and Vβ23 families exhibited common and oligoclonal Vβ-Jβ rearrangements in 4 of 18 patients (22%). Several CDR3 amino acid sequences were found for the common and oligoclonal Vβ8-Jβ1.4 rearrangement. Quantitative PCR showed that biased Vβ transcripts were also overexpressed in chronically rejected kidneys with superimposed acute lesions. In contrast, T lymphocytes infiltrating rejected allografts with chronic rejection only showed an unaltered Gaussian-type CDR3 length distribution. This pattern suggests that late graft failure associated with histological lesions restricted to Banff-defined chronic rejection does not involve T cell-mediated injury. Thus, our observation suggests that a limited number of determinants stimulates the recipient immune system in long-term allograft failure. The possibility of a local response against viral or parenchymatous cell-derived determinants is discussed.
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