The results presented in Figure 3 show which the FPLC peptide fractions induced IFN in autologous PBMC, probably due to the presence of anti-P0 antigen memory T cells, which may have arisen after broken tolerance to self-antigen after an infection, as we have interpreted the data. (immunoglobulin M/protein zero) complexes were pronounced after exposure to protein zero. Naturally processed or synthetic protein zero peptide (194C208)-pulsed TJ2 cells significantly induced interleukin-2 secretion from autologous T cells compared to control antigen-pulsed cells (gene sequencing was carried out as previously described36 and detailed in the KLH populations in C (both panels) were statistically analyzed using Fishers exact test and showed a significantly different response (and was authentic to the patients monoclonal anti-P0 autoreactive cells, and we found that it retained surface BCR with affinity for P0, efficiently mediated BCR-ligand uptake, processed the antigen and presented myelin peptides in MHCII molecules. We show here for the first time that these naturally processed P0 peptides presented by the B-cell clone induce production of the T-cell-specific cytokine IL2 from autologous T cells and production of the Th1 cytokine IFN from both CD4+ and CD8+ autologous T cells. The involvement of T cells in the pathogenesis of PNMGUS has been previously BRD4770 suggested14,16,28C30 and our results presented in this study add further evidence on how these MGUS-derived T helper cells can be triggered by the patients monoclonal B-cell populace that aberrantly presents myelin peptide antigens. The results presented in Physique 3 show that this FPLC peptide fractions induced IFN in autologous PBMC, probably due to the presence of anti-P0 antigen memory T cells, which may have arisen after broken tolerance to self-antigen after an infection, as we have interpreted the data. A transforming event in a self-reactive cell may then lead to growth of an autonomous MGUS-clone. These memory T-cell clones that we observe might be remnants of a disease process that occurred some time previously. A recent study identified P0 BRD4770 as the key CD4+ T-cell antigen in the NOD-B7-2KO autoimmune peripheral neuropathy mouse model.49 NOD-B7-2KO mice deficient of IFN did not develop peripheral neuropathy, suggesting an inflammatory Th1 response to P0 in these mice. The authors also generated a P0-specific TCR transgenic mouse (NOD-POT) and CD4+ T cells from this mouse proliferated when exposed to peripheral nerve lysate and P0. T cells from the NOD-POT mouse exposed to peripheral nerve lysate or P0 also produced IFN and IL17. A contrasting hypothesis was suggested by Horna also enabled the FLJ16239 expanded, fibromodulin-specific T cells to secrete IFN upon recognition of the antigen.51 This CD5+ B-cell-specific antigen-presentation thus enabled the expansion of autologous tumor-specific T cells. Another example illustrating the important role of B cells is usually experimental autoimmune encephalitis, the experimental animal model for multiple sclerosis. Experimental autoimmune encephalitis can be induced by exposing wild-type mice to myelin oligodendrocyte protein, but in mice deficient of B cells, there is no induction, suggesting that B cells have a clear antigen-presenting role in this condition.52 In this study (Physique 2 DCF), we show that myelin peptides are processed in the endosome/lysosome compartment (LAMP-2+) and physically associated with MHC class II (Physique 2 GCI). Non-endocytosed myelin could remain on the surface membrane, which would dim the interpretation. However, based on the data illustrated in Physique 2A, we found that all native surface-bound myelin was associated with surface IgM (none was found in other sites). Secondly, the FPLC fractions analyzed (Physique 3) contained peptides only, no native myelin. Specific anti-peptide reagents/monoclonal antibodies would, however, be helpful in distinguishing between processed and unprocessed myelin. Previous studies have shown Th1 activation and IFN production by PBMC from a PN-MGUS BRD4770 patient after synthetic P0 peptide 194C208 stimulation.16 We confirmed in this study that P0 peptide.