Mentoring Each Other: Creating a community of practice for aspiring and current library managers

Integrating formal management training into library school curriculum has been a topic of discussion for years, with varying progress. And in our experience, librarians are rarely given formal training on the job before becoming managers. We need to find ways to support new managers, helping them to acquire skills to handle the interpersonal, economic, and political challenges. Human Resources departments can provide valuable training in policy and procedures of the organization, but libraries have unique management challenges that may not exist elsewhere on campus. Librarians may be asked to manage faculty, staff, and student workers. They may need to arrange for coverage for service points, as well as work as an academic department. They work on projects with people from all parts of campus. Sometimes only other librarians understand the different dynamics of our situation. But when you’re a manager, it can be awkward to discuss specific management concerns with colleagues in your organization. Meeting with librarians from other institutions provides a comfortable level of anonymity

Design of immunogens to elicit broadly neutralizing antibodies against HIV targeting the CD4 binding site

A vaccine which is effective against the HIV virus is considered to be the best solution to the ongoing global HIV/AIDS epidemic. In the past thirty years, numerous attempts to develop an effective vaccine have been made with little or no success, due, in large part, to the high mutability of the virus. More recent studies showed that a vaccine able to elicit broadly neutralizing antibodies (bnAbs), that is, antibodies that can neutralize a high fraction of global virus variants, has promise to protect against HIV. Such a vaccine has been proposed to involve at least three separate stages: First, activate the appropriate precursor B cells; second, shepherd affinity maturation along pathways toward bnAbs; and, third, polish the Ab response to bind with high affinity to diverse HIV envelopes (Env). This final stage may require immunization with a mixture of Envs. In this paper, we set up a framework based on theory and modeling to design optimal panels of antigens to use in such a mixture. The designed antigens are characterized experimentally and are shown to be stable and to be recognized by known HIV antibodies

Differences in the Binding Affinity of an HIV-1 V2 Apex-Specific Antibody for the SIVsmm/mac Envelope Glycoprotein Uncouple Antibody-Dependent Cellular Cytotoxicity from Neutralization

Here we show that PGT145, a potent broadly neutralizing antibody to HIV-1, directs the lysis of SIV-infected cells by antibody-dependent cellular cytotoxicity but does not neutralize SIV infectivity. This represents the first instance of cross-reactivity of an HIV-1 Env-specific antibody with SIVsmm/mac Env and reveals that antibody binding affinity can differentiate sensitivity to ADCC from neutralization.As a consequence of their independent evolutionary origins in apes and Old World monkeys, human immunodeficiency virus type 1 (HIV-1) and simian immunodeficiency viruses of the SIVsmm/mac lineage express phylogenetically and antigenically distinct envelope glycoproteins. Thus, HIV-1 Env-specific antibodies do not typically cross-react with the Env proteins of SIVsmm/mac isolates. Here we show that PGT145, a broadly neutralizing antibody to a quaternary epitope at the V2 apex of HIV-1 Env, directs the lysis of SIVsmm/mac-infected cells by antibody-dependent cellular cytotoxicity (ADCC) but does not neutralize SIVsmm/mac infectivity. Amino acid substitutions in the V2 loop of SIVmac239 corresponding to the epitope for PGT145 in HIV-1 Env modulate sensitivity to this antibody. Whereas a substitution in a conserved N-linked glycosylation site (N171Q) eliminates sensitivity to ADCC, a lysine-to-serine substitution in this region (K180S) increases ADCC and renders the virus susceptible to neutralization. These differences in function correlate with an increase in the affinity of PGT145 binding to Env on the surface of virus-infected cells and to soluble Env trimers. To our knowledge, this represents the first instance of an HIV-1 Env-specific antibody that cross-reacts with SIVsmm/mac Env and illustrates how differences in antibody binding affinity for Env can differentiate sensitivity to ADCC from neutralization

W3_LLC_R1.fastq

V781-PG9 engineered Ramos cells after HIV C108 SOSIP enrichment followed by three rounds of selection using HIV WITO SOSIP. Immunoglobulin lambda light chain variable sequences from RT-PCR and amplification of cell mRNA performed using gene specific primers. TruSeq LT amplicon sequenced using 300x300 paired end reads on the Illumina MiSeq. This is read 1 (R1

D1_nonengineered_R2.fastq

Donor 1 control primary B cells 13 days post nucleofection. Immunoglobulin heavy chain variable regions amplified from cDNA made by RACE with barcoded 5' template switch adapters. NEBnext amplicon sequenced using 400x100 paired end reads on the Illumina MiSeq.This is read 2

D1_nonengineered_R1.fastq

Donor 1 control primary B cells 13 days post nucleofection. Immunoglobulin heavy chain variable regions amplified from cDNA made by RACE with barcoded 5' template switch adapters. NEBnext amplicon sequenced using 400x100 paired end reads on the Illumina MiSeq. This is read 1

WT_LLC_R2.fastq

Ramos (RA 1) ATCC CRL-1596 Homo sapiens Burkitt's cell lymphoma immunoglobulin lambda light chain variable sequences. RT-PCR and amplification from cell mRNA performed using gene specific primers. TruSeq LT amplicon sequenced using 300x300 paired end reads on the Illumina MiSeq. This is read 2 (R2

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 (Ig) 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 (bnAb), 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 (LCs) 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 HIV neutralizing activity. Thus, BCRs engineered in this way retain the genetic flexibility normally required for affinity maturation during adaptive immune responses. Peripheral blood derived primary B cells from three different donors were edited using this strategy. Engineered cells could bind the PG9 epitope and sequenced mRNA showed PG9 HC transcribed as several different isotypes after culture with CD40 ligand and IL-4.This work was supported by the National Institutes of Health, R01-5R01DE025167-04, by The Bill and Melinda Gates Foundation OPP1183956, by CHAVI-ID grant UM1 AI100663, by the Ramon y Cajal Merit Award from Ministerio de Ciencia, Innovacion y Universidades, Spain (RYC-2016–21155 to AG-M), and by a Marie-Curie Fellowship (FP7-PEOPLE-2013-IOF to LEM).Peer reviewe

W3_IgM__R1.fastq

V781-PG9 engineered Ramos cells after HIV C108 SOSIP enrichment followed by three rounds of selection using HIV WITO SOSIP. Immunoglobulin heavy chain variable sequences from RT-PCR and amplification of cell mRNA performed using gene specific primers. TruSeq LT amplicon sequenced using 300x300 paired end reads on the Illumina MiSeq. This is read 1 (R1