Project description:We used protein arrays to measure IgG autoantibodies associated with Connective Tissue Diseases (CTDs) from retrogenic chimeras Sera was isolated from irradiated Icos-/-;CD45.1 mice reconstituted with 564Igi;Icos-/- bone marrow mixed with Icos-/-;CD45.1 bone marrow and WT bone marrow (BMchim.564) or retrogenic HSCs expressing TCR-A (BMchim.564-Icos.RAG-TCRA), TCR-B (BMchim.564-Icos.RAG-TCRB), or TCR-C (BMchim.564-Icos.RAG-TCRC). 564Igi (564homo) and 564Igi;Icos-/- (564homo.Icos) sera were included as controls.
Project description:We used protein arrays to measure IgG2a autoantibodies associated with Connective Tissue Diseases (CTDs) from retrogenic chimeras Sera was isolated from irradiated Icos-/-;CD45.1 mice reconstituted with 564Igi;Icos-/- bone marrow mixed with Icos-/-;CD45.1 bone marrow and WT bone marrow (BMchim.564) or retrogenic HSCs expressing TCR-A (BMchim.564-Icos.RAG-TCRA), TCR-B (BMchim.564-Icos.RAG-TCRB), or TCR-C (BMchim.564-Icos.RAG-TCRC). 564Igi (564homo) and 564Igi;Icos-/- (564homo.Icos) sera were included as controls.
Project description:We used protein arrays to measure IgG2c autoantibodies associated with Connective Tissue Diseases (CTDs) from retrogenic chimeras Sera was isolated from irradiated Icos-/-;CD45.1 mice reconstituted with 564Igi;Icos-/- bone marrow mixed with Icos-/-;CD45.1 bone marrow and WT bone marrow (BMchim.564) or retrogenic HSCs expressing TCR-A (BMchim.564-Icos.RAG-TCRA), TCR-B (BMchim.564-Icos.RAG-TCRB), or TCR-C (BMchim.564-Icos.RAG-TCRC). 564Igi (564homo) and 564Igi;Icos-/- (564homo.Icos) sera were included as controls.
Project description:T-cell based therapies have shown remarkable efficacy in multiple myeloma (MM), yet the disease remains largely incurable. Here, we investigated the constant domains of the immunoglobulin heavy chain (IgH) as novel targets for therapeutic T-cell receptors (TCRs), after confirming high and homogeneous IGH expression in >95% of MM patients. MM cells secrete excessive monoclonal immunoglobulins (M-proteins) that drive complications but are inaccessible to CAR T-cell or antibody targeting. Peptides from IgA and IgG constant regions were eluted from HLA-A*02:01, and reactive TCRs were isolated from healthy donors using allo-HLA-A*02:01 presentation to circumvent self-tolerance. T cells engineered with two TCRs specific for IgA or IgG passed a stringent multi-tier safety screen and selectively eliminated MM cells from 20 HLA-A*02:01+ patients secreting the relevant IgH in vitro. In vivo, IgA-TCR T cells eradicated IgA+HLA-A*02:01+ MM cells in xenograft models and reduced circulating IgA in humanized PBMC mice. These findings establish immunoglobulin constant domains as viable TCR targets in MM, potentially making ~40% of patients of European descent eligible for TCR T cell therapy, and extension to additional HLA alleles could further broaden eligibility. The approach may also be applicable to lymphoma and antibody-mediated autoimmune diseases.
Project description:This dataset contains peptide identifications derived from immunoprecipitated IgG isolated from mouse serum. Serum samples were obtained from 10 month old Vλ2 315m⁺/⁻ TCR TG⁺/⁻ mice, representing the experimental cohort used to study Ig repertoire composition. A recombinant human IgG3 (hIgG3) expressing mouse VH+VL of interest was processed in parallel and used as a positive control.
Project description:The carboxy-terminal domain (CTD) of the largest subunit of RNA Polymerase II (RNAPII) consists of multiple tandem repeats of the heptapeptide consensus Y1-S2-P3-T4-S5-P6-S7. RNAPII CTD is intrinsically disordered and has been shown to promote liquid-liquid phase-separation (LLPS) of RNAPII in vivo. However, understanding the precise role of the conserved heptad residues in LLPS has been hampered by the lack of direct characterization of the biochemical properties of the CTD. Here, we generated a systematic array of RNAPII CTD variants to unravel the sequence-encoded molecular grammar underlying LLPS of the human CTD.
Project description:Sera were acquired from the Sjogren's International Collaborative Clinical Alliance (SICCA) biorepository and were assayed for IgG autoantibodies. The autoantigen array was performed to identify the specific autoantibodies that were enriched in pSS patients.