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Using this tool to survey and quantify the expression selecting of self-pMHCII, we have shown unequivocal proof that a known CD4+ selecting ligand can be presented on both positive and negative selecting thymic APCs.
However, CD4loCD8lo thymocytes were able to initiate CD8 differentiation upon release from the negative selecting environment by transfer to in vitro systems.
Because negative selection of immature thymocytes remove high affinity TCR specific for self-antigens [7], we expected that this should leave a "hole" in the T cell repertoire around negative selecting self-antigens.
Hence, if an infected cell presents a nonself antigen that is highly similar to a negative selecting self-antigen, then this foreign antigen might not be matched by any available TCR which could provide an explanation for why around half of foreign pMHC do not generate a T cell response [3], [4], [21].
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To purify T cells, splenocytes from naïve C57BL/6 mice were negative selected by exclusion of NK1.1+, Gr-1+, CD11b+, B220+ cells.
All neutrophils are negative selected from the peripheral blood.
Therefore, most of the proteins encoded by these negative selected HGT genes appear to be functional.
Conversely, 217 significant negative selected sites (dN-dS < 0) were identified.
All neutrophils are negative selected by the EasySep™ Human Neutrophil Enrichment Kit (Stemcell) from a polymorphonuclear cell-rich fraction of peripheral blood.
The region between FHA to PINc domains included few negative selected sites which were mostly located near the functional domains.
Since the library used in the NSH method is made from potentially full-length cDNAs, each negative selected has a good chance of containing a full-length cDNA.
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