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Gonzalez-Martin, A.

Publications and source records attributed to Gonzalez-Martin, A..

3 recordsLinked to original sources

Regulation of B cell tolerance and autoimmunity by miR-130b

Breakdown of B cell tolerance is a central feature of autoimmune diseases, yet the molecular mechanisms underlying the female predominance of these diseases remain unclear. Here, we identified miR-130b as an essential component of a miRNA network, defined by the GUGCA seed motif, that regulates central B cell tolerance. Elevated miR-130b levels impaired tolerance by promoting the survival of immature B cells upon B cell antigen receptor engagement. This network converged on the estrogen receptor pathway through downregulation of Esr1 and Pten. Genetic ablation of Esr1 in immature B cells was sufficient to compromise central B cell tolerance in females, but not in males, revealing an unrecognized role for ER in this process. In males, Esr1 deficiency reduced bone marrow B cell numbers, whereas in females, B cell numbers were preserved, with a greater proportion of cells displaying lower surface CD19 levels. Transcriptomic analysis revealed sex-specific genetic programs characterized by defective B cell development and skewed immunoglobulin gene usage in males, and increased PI3K-AKT signaling in females. In females, this molecular rewiring compensated for developmental defects, but also attenuated central tolerance mechanisms, allowing the escape of autoreactive B cells to the periphery. In patients with multiple sclerosis, elevated levels of miR-130b in circulating vesicles correlated with more severe disease, including increased formation of new demyelinating lesions, cognitive decline, and neurodegeneration. Together, our findings identify a sex-specific ER-dependent checkpoint in B cell tolerance controlled by a seed-driven miRNA network, providing a mechanistic framework for female predisposition to autoimmunity.

immunology↗

Vaccine Elicitation of HIV Broadly Neutralizing Antibodies from Engineered B cells

HIV broadly neutralizing antibodies (bnAbs) can suppress viremia and protect against infection1. However, their elicitation is made difficult by low frequencies of appropriate precursor B cell receptors and the complex maturation pathways required to generate bnAbs from these precursors2. Antibody genes can be engineered into B cells for expression as both a functional receptor on cell surfaces and as secreted antibody3-5. Here, we show that HIV bnAb-engineered primary mouse B cells can be adoptively transferred and vaccinated in immunocompetent wild-type animals resulting in the expansion of durable bnAb memory and long-lived plasma cells. Somatic hypermutation after immunization indicated that engineered cells have the capacity to respond to an evolving pathogen. These results encourage further exploration of engineered B cell vaccines as a strategy for durable elicitation of HIV bnAbs to protect against infection and as a contributor to a functional cure.

immunology↗

Reprogramming the antigen specificity of B cells using genome-editing technologies

We have developed a method to introduce novel paratopes into the human antibody repertoire by modifying the immunoglobulin genes of mature B cells directly using genome editing technologies. We used CRISPR-Cas9 in a homology directed repair strategy, to replace the heavy chain (HC) variable region in B cell lines with that from an HIV broadly neutralizing antibody, PG9. Our strategy is designed to function in cells that have undergone VDJ recombination using any combination of variable (V), diversity (D) and joining (J) genes. The modified locus expresses PG9 HC which pairs with native light chains resulting in the cell surface expression of HIV specific B cell receptors (BCRs). Endogenous activation-induced cytidine deaminase (AID) in engineered cells allowed for Ig class switching and generated BCR variants with improved anti-HIV neutralizing activity. Thus, BCRs engineered in this way retain the genetic flexibility normally required for affinity maturation during adaptive immune responses.

immunology↗